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Sample records for fluorescence subtraction imaging

  1. Enhanced image reconstruction of three-dimensional fluorescent assays by subtractive structured-light illumination microscopy.

    Science.gov (United States)

    Choi, Jong-ryul; Kim, Donghyun

    2012-10-01

    We investigate improved image reconstruction of structured light illumination for high-resolution imaging of three-dimensional (3D) cell-based assays. For proof of concept, an in situ fluorescence optical detection system was built with a digital micromirror device as a spatial light modulator, for which phase and tilting angle in a grid pattern were varied to implement specific image reconstruction schemes. Subtractive reconstruction algorithms based on structured light illumination were used to acquire images of fluorescent microbeads deposited as a two-dimensional monolayer or in 3D alginate matrix. We have confirmed that an optical subtraction algorithm improves axial and lateral resolution by effectively removing out-of-focus fluorescence. The results suggest that subtractive image reconstruction can be useful for structured illumination microscopy of broad types of cell-based assays with high image resolution.

  2. Estimating background-subtracted fluorescence transients in calcium imaging experiments: a quantitative approach.

    Science.gov (United States)

    Joucla, Sébastien; Franconville, Romain; Pippow, Andreas; Kloppenburg, Peter; Pouzat, Christophe

    2013-08-01

    Calcium imaging has become a routine technique in neuroscience for subcellular to network level investigations. The fast progresses in the development of new indicators and imaging techniques call for dedicated reliable analysis methods. In particular, efficient and quantitative background fluorescence subtraction routines would be beneficial to most of the calcium imaging research field. A background-subtracted fluorescence transients estimation method that does not require any independent background measurement is therefore developed. This method is based on a fluorescence model fitted to single-trial data using a classical nonlinear regression approach. The model includes an appropriate probabilistic description of the acquisition system's noise leading to accurate confidence intervals on all quantities of interest (background fluorescence, normalized background-subtracted fluorescence time course) when background fluorescence is homogeneous. An automatic procedure detecting background inhomogeneities inside the region of interest is also developed and is shown to be efficient on simulated data. The implementation and performances of the proposed method on experimental recordings from the mouse hypothalamus are presented in details. This method, which applies to both single-cell and bulk-stained tissues recordings, should help improving the statistical comparison of fluorescence calcium signals between experiments and studies.

  3. Optimising Optimal Image Subtraction

    CERN Document Server

    Israel, H; Schuh, S; Israel, Holger; Hessman, Frederic V.; Schuh, Sonja

    2006-01-01

    Difference imaging is a technique for obtaining precise relative photometry of variable sources in crowded stellar fields and, as such, constitutes a crucial part of the data reduction pipeline in surveys for microlensing events or transiting extrasolar planets. The Optimal Image Subtraction (OIS) algorithm permits the accurate differencing of images by determining convolution kernels which, when applied to reference images of particularly good quality, provide excellent matches to the point-spread functions (PSF) in other images of the time series to be analysed. The convolution kernels are built as linear combinations of a set of basis functions, conventionally bivariate Gaussians modulated by polynomials. The kernel parameters must be supplied by the user and should ideally be matched to the PSF, pixel-sampling, and S/N of the data to be analysed. We have studied the outcome of the reduction as a function of the kernel parameters using our implementation of OIS within the TRIPP package. From the analysis o...

  4. Image subtraction by solid state electroradiography

    Energy Technology Data Exchange (ETDEWEB)

    Fallone, B.G.; Podgorsak, E.B. (McGill Univ., Montreal, Quebec (Canada). Dept. of Radiation Oncology; McGill Univ., Montreal, Quebec (Canada). Dept. of Diagnostic Radiology)

    1984-06-01

    An analog radiographic image subtraction technique is presented. A selenium photoconductor is used as the radiation sensitive medium and image subtraction is achieved by electrostatically adding latent images of opposite polarities to get a latent image of the region of interest. In principle, the technique is simple and a comparison with commercial digital techniques shows a similar image quality, suggesting that the electrostatic subtraction technique could be developed into a clinical tool as an inexpensive alternative to digital radiography.

  5. A Wide-Field Fluorescence Microscope Extension for Ultrafast Screening of One-Bead One-Compound Libraries Using a Spectral Image Subtraction Approach.

    Science.gov (United States)

    Heusermann, Wolf; Ludin, Beat; Pham, Nhan T; Auer, Manfred; Weidemann, Thomas; Hintersteiner, Martin

    2016-05-09

    The increasing involvement of academic institutions and biotech companies in drug discovery calls for cost-effective methods to identify new bioactive molecules. Affinity-based on-bead screening of combinatorial one-bead one-compound libraries combines a split-mix synthesis design with a simple protein binding assay operating directly at the bead matrix. However, one bottleneck for academic scale on-bead screening is the unavailability of a cheap, automated, and robust screening platform that still provides a quantitative signal related to the amount of target protein binding to individual beads for hit bead ranking. Wide-field fluorescence microscopy has long been considered unsuitable due to significant broad spectrum autofluorescence of the library beads in conjunction with low detection sensitivity. Herein, we demonstrate how such a standard microscope equipped with LED-based excitation and a modern CMOS camera can be successfully used for selecting hit beads. We show that the autofluorescence issue can be overcome by an optical image subtraction approach that yields excellent signal-to-noise ratios for the detection of bead-associated target proteins. A polymer capillary attached to a semiautomated bead-picking device allows the operator to efficiently isolate individual hit beads in less than 20 s. The system can be used for ultrafast screening of >200,000 bead-bound compounds in 1.5 h, thereby making high-throughput screening accessible to a wider group within the scientific community.

  6. Astronomical Image Subtraction by Cross-Convolution

    Science.gov (United States)

    Yuan, Fang; Akerlof, Carl W.

    2008-04-01

    In recent years, there has been a proliferation of wide-field sky surveys to search for a variety of transient objects. Using relatively short focal lengths, the optics of these systems produce undersampled stellar images often marred by a variety of aberrations. As participants in such activities, we have developed a new algorithm for image subtraction that no longer requires high-quality reference images for comparison. The computational efficiency is comparable with similar procedures currently in use. The general technique is cross-convolution: two convolution kernels are generated to make a test image and a reference image separately transform to match as closely as possible. In analogy to the optimization technique for generating smoothing splines, the inclusion of an rms width penalty term constrains the diffusion of stellar images. In addition, by evaluating the convolution kernels on uniformly spaced subimages across the total area, these routines can accommodate point-spread functions that vary considerably across the focal plane.

  7. Intensity Weighted Subtraction Microscopy Approach for Image Contrast and Resolution Enhancement

    Science.gov (United States)

    Korobchevskaya, Kseniya; Peres, Chiara; Li, Zhibin; Antipov, Alexei; Sheppard, Colin J. R.; Diaspro, Alberto; Bianchini, Paolo

    2016-05-01

    We propose and demonstrate a novel subtraction microscopy algorithm, exploiting fluorescence emission difference or switching laser mode and their derivatives for image enhancement. The key novelty of the proposed approach lies in the weighted subtraction coefficient, adjusted pixel-by-pixel with respect to the intensity distributions of initial images. This method produces significant resolution enhancement and minimizes image distortions. Our theoretical and experimental studies demonstrate that this approach can be applied to any optical microscopy techniques, including label free and non-linear methods, where common super-resolution techniques cannot be used.

  8. Appearance of the canine meninges in subtraction magnetic resonance images.

    Science.gov (United States)

    Lamb, Christopher R; Lam, Richard; Keenihan, Erin K; Frean, Stephen

    2014-01-01

    The canine meninges are not visible as discrete structures in noncontrast magnetic resonance (MR) images, and are incompletely visualized in T1-weighted, postgadolinium images, reportedly appearing as short, thin curvilinear segments with minimal enhancement. Subtraction imaging facilitates detection of enhancement of tissues, hence may increase the conspicuity of meninges. The aim of the present study was to describe qualitatively the appearance of canine meninges in subtraction MR images obtained using a dynamic technique. Images were reviewed of 10 consecutive dogs that had dynamic pre- and postgadolinium T1W imaging of the brain that was interpreted as normal, and had normal cerebrospinal fluid. Image-anatomic correlation was facilitated by dissection and histologic examination of two canine cadavers. Meningeal enhancement was relatively inconspicuous in postgadolinium T1-weighted images, but was clearly visible in subtraction images of all dogs. Enhancement was visible as faint, small-rounded foci compatible with vessels seen end on within the sulci, a series of larger rounded foci compatible with vessels of variable caliber on the dorsal aspect of the cerebral cortex, and a continuous thin zone of moderate enhancement around the brain. Superimposition of color-encoded subtraction images on pregadolinium T1- and T2-weighted images facilitated localization of the origin of enhancement, which appeared to be predominantly dural, with relatively few leptomeningeal structures visible. Dynamic subtraction MR imaging should be considered for inclusion in clinical brain MR protocols because of the possibility that its use may increase sensitivity for lesions affecting the meninges.

  9. Blind Source Separation Algorithms for PSF Subtraction from Direct Imaging

    Science.gov (United States)

    Shapiro, Jacob; Ranganathan, Nikhil; Savransky, Dmitry; Ruffio, Jean-Baptise; Macintosh, Bruce; GPIES Team

    2017-01-01

    The principal difficulty with detecting planets via direct imaging is that the target signal is similar in magnitude, or fainter, than the noise sources in the image. To compensate for this, several methods exist to subtract the PSF of the host star and other confounding noise sources. One of the most effective methods is Karhunen-Loève Image Processing (KLIP). The core algorithm within KLIP is Principal Component Analysis, which is a member of a class of algorithms called Blind Source Separation (BSS).We examine three other BSS algorithms that may potentially also be used for PSF subtraction: Independent Component Analysis, Stationary Subspace Analysis, and Common Spatial Pattern Filtering. The underlying principles of each of the algorithms is discussed, as well as the processing steps needed to achieve PSF subtraction. The algorithms are examined both as primary PSF subtraction techniques, as well as additional postprocessing steps used with KLIP.These algorithms have been used on data from the Gemini Planet Imager, analyzing images of β Pic b. To build a reference library, both Angular Differential Imaging and Spectral Differential Imaging were used. To compare to KLIP, three major metrics are examined: computation time, signal-to-noise ratio, and astrometric and photometric biases in different image regimes (e.g., speckle-dominated compared to Poisson-noise dominated). Preliminary results indicate that these BSS algorithms improve performance when used as an enhancement for KLIP, and that they can achieve similar SNR when used as the primary method of PSF subtraction.

  10. Subtraction gadolinium enhanced magnetic resonance for head and neck imaging

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    Lloyd, G.A.S.; Barker, P.G.; Phelps, P.D. (Royal National Throat, Nose and Ear Hospital, London (United Kingdom))

    1993-01-01

    The subtraction method of Des Plantes was applied to gadolinium enhanced magnetic resonance imaging (GdMR). Using short acquisition times, T[sub 1] weighted spin echo pulse sequences are made immediately before and after intravenous administration of gadolinium DTPA. The needle is placed in the vein prior to putting the patient into the scanner and is irrigated with saline while the control series is obtained. 42 patients with naso-sinus or skull base tumours have been successfully investigated by this technique and satisfactory subtraction studies are now obtained on all patients other than the claustrophobic. Subtraction GdMR provides the best demonstration of the effects of gadolinium DTPA on the magnetic resonance signal for both normal and abnormal tissues. Tbe signal recorded on the subtraction image is dependent on tissue blood supply and provides a more accurate record of tumour extent than that shown by unsubtracted GdMR scans. (author).

  11. Subtraction and dynamic MR images of breast cancer

    Energy Technology Data Exchange (ETDEWEB)

    Murakami, Yoshitaka; Aoki, Manabu; Harada, Junta (Jikei Univ., Tokyo (Japan). School of Medicine)

    1993-04-01

    The purpose of this study was to evaluate the diagnostic effectiveness of subtraction and dynamic MR imaging in patients with breast masses. In 23 breast cancers and six fibroadenomas, spin echo T1 images were obtained at 0.2 Tesla before and every minute after intravenous injection of Gd-DTPA (0.1 or 0.2 mmol/kg). Subtraction images were obtained sequentially on the CRT monitor. All breast masses were enhanced after gadolinium and stood out as bright lesions on subtraction images. The tumor margin and its extension were more precisely evaluated on subtraction MR images than on conventional postcontrast MR images. Breast cancer showed a characteristic time-intensity curve with an early peak, in contrast to fibroadenoma, which showed a gradual increase in signal intensity. Subtraction MR imaging is a simple method for the evaluation of breast masses, and further, the time-intensity curve obtained by dynamic study is helpful in the differential diagnosis of lesions. (author).

  12. Proper image subtraction - optimal transient detection, photometry and hypothesis testing

    CERN Document Server

    Zackay, Barak; Gal-Yam, Avishay

    2016-01-01

    Transient detection and flux measurement via image subtraction stand at the base of time domain astronomy. Due to the varying seeing conditions, the image subtraction process is non-trivial, and existing solutions suffer from a variety of problems. Starting from basic statistical principles, we develop the optimal statistic for transient detection, flux measurement and any image-difference hypothesis testing. We derive a closed-form statistic that: (i) Is mathematically proven to be the optimal transient detection statistic in the limit of background-dominated noise; (ii) Is numerically stable; (iii) For accurately registered, adequately sampled images, does not leave subtraction or deconvolution artifacts; (iv) Allows automatic transient detection to the theoretical sensitivity limit by providing credible detection significance; (v) Has uncorrelated white noise; (vi) Is a sufficient statistic for any further statistical test on the difference image, and in particular, allows to distinguish particle hits and ...

  13. Image Segmentation in Video Sequences Using Modified Background Subtraction

    Directory of Open Access Journals (Sweden)

    D. W. Chinchkhede

    2012-03-01

    Full Text Available In computer vision, "Background subtraction" is a technique for finding moving objects in a video sequences for example vehicle driving on a freeway. For to detect non stationary (dynamic objects, it is necessary to subtracting current image from a time-averaged background image. There are various background subtraction algorithms for detecting moving vehicles or any moving object(s like pedestrians in urban traffic video sequences. A crude approximation to the task of classifying each pixel on the frame of current image, locate slow-moving objects or in poor image qualities of videos and distinguish shadows from moving objects by using modified background subtraction method. While classifying each pixel on the frame of the current image, it is to be detect the moving object at foreground and background conditional environment that we can classify each pixel using a model of how that pixellooks when it is part of video frame classes. A mixture of Gaussians classification model for each pixel using an unsupervised technique is an efficient, incremental version of Expectation Maximization (EM is used for the purpose. Unlike standard image-averaging approach, this method automatically updates the mixture component for each video frame class according to likelihood of membership; hence slowmoving objects and poor image quality of videos are also being handled perfectly. Our approach identifies and eliminates shadows much more effectively than other techniques like thresholding.

  14. Direct Imaging of Exoplanets Without Background Subtraction: Implications for ELTs

    CERN Document Server

    Frazin, Richard A

    2015-01-01

    The ultra-high contrast capability required to form images of other solar systems is arguably the highest-profile challenge in astronomy today. The current high-contrast imaging efforts all require background subtraction to separate the planetary image from the image of the host star. Background estimation is difficult due to the presence of non-common path aberrations (NCPAs) that change with time. The only major source of information that is not being utilized by current efforts is the random encoding of the planetary image and the NCPAs by the atmosphere on millisecond time-scales. Here, a method that utilizes this information in order to avoid background subtraction altogether is proposed. This new paradigm will allow simultaneous estimation of the time-dependent NCPAs and the planetary image via rigorous statistical inference procedures. These procedures are fully compatible with other information sources, such as diurnal field rotation and spectral diversity. Given the open-ended nature of the backgroun...

  15. A method for dynamic subtraction MR imaging of the liver

    Directory of Open Access Journals (Sweden)

    Setti Ernesto

    2006-06-01

    Full Text Available Abstract Background Subtraction of Dynamic Contrast-Enhanced 3D Magnetic Resonance (DCE-MR volumes can result in images that depict and accurately characterize a variety of liver lesions. However, the diagnostic utility of subtraction images depends on the extent of co-registration between non-enhanced and enhanced volumes. Movement of liver structures during acquisition must be corrected prior to subtraction. Currently available methods are computer intensive. We report a new method for the dynamic subtraction of MR liver images that does not require excessive computer time. Methods Nineteen consecutive patients (median age 45 years; range 37–67 were evaluated by VIBE T1-weighted sequences (TR 5.2 ms, TE 2.6 ms, flip angle 20°, slice thickness 1.5 mm acquired before and 45s after contrast injection. Acquisition parameters were optimized for best portal system enhancement. Pre and post-contrast liver volumes were realigned using our 3D registration method which combines: (a rigid 3D translation using maximization of normalized mutual information (NMI, and (b fast 2D non-rigid registration which employs a complex discrete wavelet transform algorithm to maximize pixel phase correlation and perform multiresolution analysis. Registration performance was assessed quantitatively by NMI. Results The new registration procedure was able to realign liver structures in all 19 patients. NMI increased by about 8% after rigid registration (native vs. rigid registration 0.073 ± 0.031 vs. 0.078 ± 0.031, n.s., paired t-test and by a further 23% (0.096 ± 0.035 vs. 0.078 ± 0.031, p t-test after non-rigid realignment. The overall average NMI increase was 31%. Conclusion This new method for realigning dynamic contrast-enhanced 3D MR volumes of liver leads to subtraction images that enhance diagnostic possibilities for liver lesions.

  16. Intermediate Palomar Transient Factory: Realtime Image Subtraction Pipeline

    CERN Document Server

    Cao, Yi; Kasliwal, Mansi M

    2016-01-01

    A fast-turnaround pipeline for realtime data reduction plays an essential role in discovering and permitting follow-up observations to young supernovae and fast-evolving transients in modern time-domain surveys. In this paper, we present the realtime image subtraction pipeline in the intermediate Palomar Transient Factory. By using high-performance computing, efficient database, and machine learning algorithms, this pipeline manages to reliably deliver transient candidates within ten minutes of images being taken. Our experience in using high performance computing resources to process big data in astronomy serves as a trailblazer to dealing with data from large-scale time-domain facilities in near future.

  17. Fluorescence and Spectral Imaging

    Directory of Open Access Journals (Sweden)

    Ralph S. DaCosta

    2007-01-01

    Full Text Available Early identification of dysplasia remains a critical goal for diagnostic endoscopy since early discovery directly improves patient survival because it allows endoscopic or surgical intervention with disease localized without lymph node involvement. Clinical studies have successfully used tissue autofluorescence with conventional white light endoscopy and biopsy for detecting adenomatous colonic polyps, differentiating benign hyperplastic from adenomas with acceptable sensitivity and specificity. In Barrett's esophagus, the detection of dysplasia remains problematic because of background inflammation, whereas in the squamous esophagus, autofluorescence imaging appears to be more dependable. Point fluorescence spectroscopy, although playing a crucial role in the pioneering mechanistic development of fluorescence endoscopic imaging, does not seem to have a current function in endoscopy because of its nontargeted sampling and suboptimal sensitivity and specificity. Other point spectroscopic modalities, such as Raman spectroscopy and elastic light scattering, continue to be evaluated in clinical studies, but still suffer the significant disadvantages of being random and nonimaging. A recent addition to the fluorescence endoscopic imaging arsenal is the use of confocal fluorescence endomicroscopy, which provides real-time optical biopsy for the first time. To improve detection of dysplasia in the gastrointestinal tract, a new and exciting development has been the use of exogenous fluorescence contrast probes that specifically target a variety of disease-related cellular biomarkers using conventional fluorescent dyes and novel potent fluorescent nanocrystals (i.e., quantum dots. This is an area of great promise, but still in its infancy, and preclinical studies are currently under way.

  18. Stroboscopic fluorescence lifetime imaging.

    Science.gov (United States)

    Holton, Mark D; Silvestre, Oscar R; Errington, Rachel J; Smith, Paul J; Matthews, Daniel R; Rees, Paul; Summers, Huw D

    2009-03-30

    We report a fluorescence lifetime imaging technique that uses the time integrated response to a periodic optical excitation, eliminating the need for time resolution in detection. A Dirac pulse train of variable period is used to probe the frequency response of the total fluorescence per pulse leading to a frequency roll-off that is dependent on the relaxation rate of the fluorophores. The technique is validated by demonstrating wide-field, realtime, lifetime imaging of the endocytosis of inorganic quantum dots by a cancer cell line. Surface charging of the dots in the intra-cellular environment produces a switch in the fluorescence lifetime from approximately 40 ns to technique offers lifetime based imaging at video rates with standard CCD cameras and has application in probing millisecond cell dynamics and in high throughput imaging assays.

  19. Digital subtraction peripheral angiography using image stacking: initial clinical results.

    Science.gov (United States)

    Kump, K S; Sachs, P B; Wilson, D L

    2001-07-01

    Using clinically acquired x-ray angiography image sequences, we compared three algorithms for creating a single diagnostic quality image that combined input images containing flowing contrast agent. These image-stacking algorithms were: maximum opacity with the minimum gray-scale value across time recorded at each spatial location, (REC) recursive temporal filtering followed by a maximum opacity operation, and (AMF) an approximate matched filter consisting of a convolution with a kernel approximating the matched filter followed by a maximum opacity operation. Eighteen clinical exams of the peripheral arteries of the legs were evaluated. AMF gave 2.7 times greater contrast to noise ratio than the single best subtraction image and 1.3 times improvement over REC, the second best stacking algorithm. This is consistent with previous simulations showing that AMF performs nearly equal to the optimal result from matched filtering without the well-known limitations. For example, unlike matched filtering, AMF filter coefficients were obtained automatically using an image-processing algorithm. AMF effectively brought out small collateral arteries, otherwise difficult to see, without degrading artery sharpness or stenosis grading. Comparing results using reduced and full contrast agent volumes demonstrated that contrast agent load could be reduced to one-third of the conventional amount with AMF processing. By simulating reduced x-ray exposures on clinical exams, we determined that x-ray exposure could be reduced by 80% with AMF processing. We conclude that AMF is a promising, potential technique for reducing contrast agent load and for improving vessel visibility, both very important characteristics for vascular imaging.

  20. A symmetrical subtraction combined with interpolated values for eliminating scattering from fluorescence EEM data

    Science.gov (United States)

    Xu, Jing; Liu, Xiaofei; Wang, Yutian

    2016-08-01

    Parallel factor analysis is a widely used method to extract qualitative and quantitative information of the analyte of interest from fluorescence emission-excitation matrix containing unknown components. Big amplitude of scattering will influence the results of parallel factor analysis. Many methods of eliminating scattering have been proposed. Each of these methods has its advantages and disadvantages. The combination of symmetrical subtraction and interpolated values has been discussed. The combination refers to both the combination of results and the combination of methods. Nine methods were used for comparison. The results show the combination of results can make a better concentration prediction for all the components.

  1. Dual energy subtraction method for breast calcification imaging

    Science.gov (United States)

    Koukou, Vaia; Martini, Niki; Fountos, George; Michail, Christos; Sotiropoulou, Panagiota; Bakas, Athanasios; Kalyvas, Nektarios; Kandarakis, Ioannis; Speller, Robert; Nikiforidis, George

    2017-03-01

    The aim of this work was to present an experimental dual energy (DE) method for the visualization of microcalcifications (μCs). A modified radiographic X-ray tube combined with a high resolution complementary metal-oxide-semiconductor (CMOS) active pixel sensor (APS) X-ray detector was used. A 40/70 kV spectral combination was filtered with 100 μm cadmium (Cd) and 1000 μm copper (Cu) for the low/high-energy combination. Homogenous and inhomogeneous breast phantoms and two calcification phantoms were constructed with various calcification thicknesses, ranging from 16 to 152 μm . Contrast-to-noise ratio (CNR) was calculated from the DE subtracted images for various entrance surface doses. A calcification thickness of 152 μm was visible, with mean glandular doses (MGD) in the acceptable levels (below 3 mGy). Additional post-processing on the DE images of the inhomogeneous breast phantom resulted in a minimum visible calcification thickness of 93 μm (MGD=1.62 mGy). The proposed DE method could potentially improve calcification visibility in DE breast calcification imaging.

  2. Development of a voxel-matching technique for substantial reduction of subtraction artifacts in temporal subtraction images obtained from thoracic MDCT.

    Science.gov (United States)

    Itai, Yoshinori; Kim, Hyoungseop; Ishikawa, Seiji; Katsuragawa, Shigehiko; Doi, Kunio

    2010-02-01

    A temporal subtraction image, which is obtained by subtraction of a previous image from a current one, can be used for enhancing interval changes (such as formation of new lesions and changes in existing abnormalities) on medical images by removing most of the normal structures. However, subtraction artifacts are commonly included in temporal subtraction images obtained from thoracic computed tomography and thus tend to reduce its effectiveness in the detection of pulmonary nodules. In this study, we developed a new method for substantially removing the artifacts on temporal subtraction images of lungs obtained from multiple-detector computed tomography (MDCT) by using a voxel-matching technique. Our new method was examined on 20 clinical cases with MDCT images. With this technique, the voxel value in a warped (or nonwarped) previous image is replaced by a voxel value within a kernel, such as a small cube centered at a given location, which would be closest (identical or nearly equal) to the voxel value in the corresponding location in the current image. With the voxel-matching technique, the correspondence not only between the structures but also between the voxel values in the current and the previous images is determined. To evaluate the usefulness of the voxel-matching technique for removal of subtraction artifacts, the magnitude of artifacts remaining in the temporal subtraction images was examined by use of the full width at half maximum and the sum of a histogram of voxel values, which may indicate the average contrast and the total amount, respectively, of subtraction artifacts. With our new method, subtraction artifacts due to normal structures such as blood vessels were substantially removed on temporal subtraction images. This computerized method can enhance lung nodules on chest MDCT images without disturbing misregistration artifacts.

  3. [The three-dimension spectral subtraction fluorescence to check adulteration of diesel oil by 120# solvent naphtha].

    Science.gov (United States)

    Yang, Ren-jie; Xu, Xiao-xuan; Shang, Li-ping; Xu, Jia-lin; Zhang, Cun-zhou

    2006-01-01

    The characteristics of a adulterated sample of diesel oil by 120# solvent naphtha were studied with three-dimension spectral subtraction fluorescence technique, and it is shown that the technique could determine quickly whether there is a adulterated sample of 120# solvent naphtha in diesel. At the same time, by measuring the volume of three-dimension spectral subtraction fluorescence graph, the authors found that this total volume is directly related to the concentration of the adulterant present in the sample. The study is important for us to analyse whether there is a adulterated sample of 120# solvent naphtha in diesel, and is of significance for us to maintain the stability of market.

  4. Improved Savitzky-Golay-method-based fluorescence subtraction algorithm for rapid recovery of Raman spectra.

    Science.gov (United States)

    Chen, Kun; Zhang, Hongyuan; Wei, Haoyun; Li, Yan

    2014-08-20

    In this paper, we propose an improved subtraction algorithm for rapid recovery of Raman spectra that can substantially reduce the computation time. This algorithm is based on an improved Savitzky-Golay (SG) iterative smoothing method, which involves two key novel approaches: (a) the use of the Gauss-Seidel method and (b) the introduction of a relaxation factor into the iterative procedure. By applying a novel successive relaxation (SG-SR) iterative method to the relaxation factor, additional improvement in the convergence speed over the standard Savitzky-Golay procedure is realized. The proposed improved algorithm (the RIA-SG-SR algorithm), which uses SG-SR-based iteration instead of Savitzky-Golay iteration, has been optimized and validated with a mathematically simulated Raman spectrum, as well as experimentally measured Raman spectra from non-biological and biological samples. The method results in a significant reduction in computing cost while yielding consistent rejection of fluorescence and noise for spectra with low signal-to-fluorescence ratios and varied baselines. In the simulation, RIA-SG-SR achieved 1 order of magnitude improvement in iteration number and 2 orders of magnitude improvement in computation time compared with the range-independent background-subtraction algorithm (RIA). Furthermore the computation time of the experimentally measured raw Raman spectrum processing from skin tissue decreased from 6.72 to 0.094 s. In general, the processing of the SG-SR method can be conducted within dozens of milliseconds, which can provide a real-time procedure in practical situations.

  5. Value of blood-pool subtraction in cardiac indium-111-labeled platelet imaging

    Energy Technology Data Exchange (ETDEWEB)

    Machac, J.; Vallabhajosula, S.; Goldman, M.E.; Goldsmith, S.J.; Palestro, C.; Strashun, A.; Vaquer, R.; Phillips, R.A.; Fuster, V. (Mt. Sinai Medical Center, New York, NY (USA))

    1989-09-01

    Blood-pool subtraction has been proposed to enhance {sup 111}In-labeled platelet imaging of intracardiac thrombi. We tested the accuracy of labeled platelet imaging, with and without blood-pool subtraction, in ten subjects with cardiac thrombi of varying age, eight with endocarditis being treated with antimicrobial therapy and ten normal controls. Imaging was performed early after labeled platelet injection (24 hr or less) and late (48 hr or more). Blood-pool subtraction was carried out. All images were graded subjectively by four experienced, blinded readers. Detection accuracy was measured by the sensitivity at three fixed levels of specificity estimated from receiver operator characteristic curve analysis and tested by three-way analysis of variance. Detection accuracy was generally improved on delayed images. Blood-pool subtraction did not improve accuracy. Although blood-pool subtraction increased detection sensitivity, this was offset by decreased specificity. For this population studied, blood-pool subtraction did not improve subjective detection of abnormal platelet deposition by 111In platelet imaging.

  6. Tumor scintigraphy by the method for subtracting the initial image with technetium-99m labeled antibody

    Energy Technology Data Exchange (ETDEWEB)

    Karube, Yoshiharu; Katsuno, Kentaro; Ito, Sanae; Matsunaga, Kazuhisa; Takata, Jiro [Fukuoka Univ. (Japan). Faculty of Pharmaceutical Sciences; Kuroki, Masahide; Murakami, Masaaki; Matsuoka, Yuji

    1999-12-01

    The method for subtracting the initial image from the localization image was evaluated for radioimmunoscintigraphy of tumors with technetium-99m (Tc-99m) labeled antibodies. Monoclonal antibodies were parental mouse and mouse-human chimeric antibodies to carcinoembryonic antigen (CEA), designated F11-39 and ChF11-39, respectively, both of which have been found to discriminate CEA in tumor tissues from the CEA-related antigens. After reduction of the intrinsic disulfide bonds, these antibodies were labeled with Tc-99m. In vivo studies were performed on athymic nude mice bearing the human CEA-producing gastric carcinoma xenografts. Though biodistribution results showed selective and progressive accumulation of Tc-99m labeled antibodies at the tumor site, high radioactivity in blood was inappropriate for scintigraphic visualization of the tumors within a few hours. We examined the subtraction of the initial Tc-99m image from the Tc-99m localization image after a few hours. Subtracted images of the same count reflected the in vivo behavior of the Tc-99m radioactivity. The subtracted scintigrams revealed excellent tumor images with no significant extrarenal background. Visualization of the tumor site was dependent on antigen-specific binding and nonspecific exudation. These results demonstrate that a method of subtraction of the initial image may serve as a potentially useful diagnostic method for an abnormal site for agents with a low pharmacokinetic value. (author)

  7. Assessing Photosynthesis by Fluorescence Imaging

    Science.gov (United States)

    Saura, Pedro; Quiles, Maria Jose

    2011-01-01

    This practical paper describes a novel fluorescence imaging experiment to study the three processes of photochemistry, fluorescence and thermal energy dissipation, which compete during the dissipation of excitation energy in photosynthesis. The technique represents a non-invasive tool for revealing and understanding the spatial heterogeneity in…

  8. Procedures for imaging of hemodialysis fistulas with particular reference to digital subtraction angiography (DSA)

    Energy Technology Data Exchange (ETDEWEB)

    Neufang, K.F.R.; Erasmi-Koerber, H.; Wimmer, G.

    1983-05-01

    All angiographic procedures established for imaging of hemodialysis fistulas, such as direct venous angiography, intravenous subtraction angiography and arteriography by direct puncture of the brachial artery by Seldinger's transfemoral technique, can also be effected with digital image processing. Depending on the angiographic technique, the use of digital subtraction angiography has several advantages: lower doses and concentrations of the contrast agent, lower risk of complications (thrombosis of the fistula, vasospasm) and freedom from pain. In addition, there is a marked reduction of examination time and film cost.

  9. Subtraction of point sources from interferometric radio images through an algebraic forward modeling scheme

    CERN Document Server

    Bernardi, G; Ord, S M; Greenhill, L J; Pindor, B; Wayth, R B; Wyithe, J S B

    2010-01-01

    We present a method for subtracting point sources from interferometric radio images via forward modeling of the instrument response and involving an algebraic nonlinear minimization. The method is applied to simulated maps of the Murchison Wide-field Array but is generally useful in cases where only image data are available. After source subtraction, the residual maps have no statistical difference to the expected thermal noise distribution at all angular scales, indicating high effectiveness in the subtraction. Simulations indicate that the errors in recovering the source parameters decrease with increasing signal-to-noise ratio, which is consistent with the theoretical measurement errors. In applying the technique to simulated snapshot observations with the Murchison Wide-field Array, we found that all 101 sources present in the simulation were recovered with an average position error of 10 arcsec and an average flux density error of 0.15%. This led to a dynamic range increase of approximately 3 orders of m...

  10. Bone images from dual-energy subtraction chest radiography in the detection of rib fractures

    Energy Technology Data Exchange (ETDEWEB)

    Szucs-Farkas, Zsolt, E-mail: zsolt.szuecs@insel.ch [Department of Diagnostic, Interventional and Pediatric Radiology, University Hospital Bern, Freiburgstrasse 4, Bern CH-3010 (Switzerland); Lautenschlager, Katrin, E-mail: katrin@students.unibe.ch [Department of Diagnostic, Interventional and Pediatric Radiology, University Hospital Bern, Freiburgstrasse 4, Bern CH-3010 (Switzerland); Flach, Patricia M., E-mail: patricia.flach@irm.unibe.ch [Institute of Forensic Medicine, University of Bern, Freiburgstrasse 4, Bern CH-3010 (Switzerland); Ott, Daniel, E-mail: daniel.ott@insel.ch [Department of Diagnostic, Interventional and Pediatric Radiology, University Hospital Bern, Freiburgstrasse 4, Bern CH-3010 (Switzerland); Strautz, Tamara, E-mail: tamara.strautz@insel.ch [Department of Diagnostic, Interventional and Pediatric Radiology, University Hospital Bern, Freiburgstrasse 4, Bern CH-3010 (Switzerland); Vock, Peter, E-mail: peter.vock@insel.ch [Department of Diagnostic, Interventional and Pediatric Radiology, University Hospital Bern, Freiburgstrasse 4, Bern CH-3010 (Switzerland); Ruder, Thomas D., E-mail: thomas.ruder@irm.unibe.ch [Institute of Forensic Medicine, University of Bern, Freiburgstrasse 4, Bern CH-3010 (Switzerland)

    2011-08-15

    Objective: To assess the sensitivity and image quality of chest radiography (CXR) with or without dual-energy subtracted (ES) bone images in the detection of rib fractures. Materials and methods: In this retrospective study, 39 patients with 204 rib fractures and 24 subjects with no fractures were examined with a single exposure dual-energy subtraction digital radiography system. Three blinded readers first evaluated the non-subtracted posteroanterior and lateral chest radiographs alone, and 3 months later they evaluated the non-subtracted images together with the subtracted posteroanterior bone images. The locations of rib fractures were registered with confidence levels on a 3-grade scale. Image quality was rated on a 5-point scale. Marks by readers were compared with fracture localizations in CT as a standard of reference. Results: The sensivity for fracture detection using both methods was very similar (34.3% with standard CXR and 33.5% with ES-CXR, p = 0.92). At the patient level, both sensitivity (71.8%) and specificity (92.9%) with or without ES were identical. Diagnostic confidence was not significantly different (2.61 with CXR and 2.75 with ES-CXR, p = 0.063). Image quality with ES was rated higher than that on standard CXR (4.08 vs. 3.74, p < 0.001). Conclusions: Despite a better image quality, adding ES bone images to standard radiographs of the chest does not provide better sensitivity or improved diagnostic confidence in the detection of rib fractures.

  11. Real-time optical image subtraction by a holographic shear lens

    Science.gov (United States)

    Rao, V. Venkateswara; Joenathan, C.; Sirohi, R. S.

    1985-08-01

    A new optical method of image subtraction by employing a holographic shear lens is proposed. The principle underlying this technique is that of optical interference between two sheared fields produced by the holographic shear lens (HSL). Two dissimilar inputs with some common characters are subtracted in real time while keeping the HSL at the Fourier plane of a well corrected lens. The difference is detectable only when zero fringe is obtained in the interferogram. Experimental verification is presented with the results. The basic advantages of this technique are the simplicity in aligning the input transparencies and the real time operation.

  12. A Feature Subtraction Method for Image Based Kinship Verification under Uncontrolled Environments

    DEFF Research Database (Denmark)

    Duan, Xiaodong; Tan, Zheng-Hua

    2015-01-01

    The most fundamental problem of local feature based kinship verification methods is that a local feature can capture the variations of environmental conditions and the differences between two persons having a kin relation, which can significantly decrease the performance. To address this problem...... the feature distance between face image pairs with kinship and maximize the distance between non-kinship pairs. Based on the subtracted feature, the verification is realized through a simple Gaussian based distance comparison method. Experiments on two public databases show that the feature subtraction method...

  13. A practical method for randoms subtraction in volume imaging PET from detector singles countrate measurements

    Energy Technology Data Exchange (ETDEWEB)

    Smith, R.J.; Karp, J.S. [Univ. of Pennsylvania, Philadelphia, PA (United States). Dept. of Radiology

    1996-06-01

    Randoms subtraction in a volume imaging PET scanner is a significant problem due to the high singles countrates experienced. The delayed coincidence method requires double counting of randoms events and results in a lowered countrate capability. Calculations based on detector singles countrates require complex corrections for countrate dependent livetime and event acceptance due to the camera coincidence processing between the detector and rebinned randoms countrates. The profile distribution method has been used to estimate and subtract both scatter and randoms background but this method is a compromise and couples these 2 sources of background together. In order to avoid these problems and provide accurate subtraction of both the distribution and magnitude of randoms contamination in the scan data the authors have developed an alternative singles based method. The singles distributions are measured across the detectors and are used to construct a randoms distribution sinogram. This distribution is scaled to the appropriate rebinned randoms countrate by means of a lookup table of randoms countrate vs detector singles countrate, generated from phantom calibrations. The advantages of performing randoms subtraction by this method are: (1) there is no increase in camera deadtime, (2) the method compensates for nonuniformities in randoms distributions due to both the activity distribution and nonuniform geometric response of the camera for on and off bankpairs, and (3) it deals with randoms subtraction independently of scatter so that different scatter correction routines may then be applied to the data.

  14. Evaluation of carotid vessel wall enhancement with image subtraction after gadobenate dimeglumine-enhanced MR angiography

    Energy Technology Data Exchange (ETDEWEB)

    Sardanelli, Francesco [Department of Medical and Surgical Sciences, University of Milan School of Medicine, via Morandi 30, 20097 San Donato Milanese, Milan (Italy); Unit of Radiology, IRCCS Policlinico San Donato, via Morandi 30, 20097 San Donato Milanese, Milan (Italy)], E-mail: f.sardanelli@grupposandonato.it; Di Leo, Giovanni [Unit of Radiology, IRCCS Policlinico San Donato, via Morandi 30, 20097 San Donato Milanese, Milan (Italy)], E-mail: gianni.dileo77@virgilio.it; Aliprandi, Alberto [Unit of Radiology, IRCCS Policlinico San Donato, via Morandi 30, 20097 San Donato Milanese, Milan (Italy)], E-mail: a.aliprandi@grupposandonato.it; Flor, Nicola [Department of Diagnostic and Interventional Radiology, University of Milan School of Medicine, San Paolo Hospital, via di Rudini 8, 20142 Milan (Italy)], E-mail: flornic@hotmail.com; Papini, Giacomo D.E. [Unit of Radiology, IRCCS Policlinico San Donato, via Morandi 30, 20097 San Donato Milanese, Milan (Italy)], E-mail: giacomo.papini@fastwebnet.it; Roccatagliata, Luca [Unit of Radiology, IRCCS Policlinico San Donato, via Morandi 30, 20097 San Donato Milanese, Milan (Italy); Department of Neuroscience, Ophthalmology and Genetics, University of Genoa, via De Toni 5, 16132 Genoa (Italy)], E-mail: lroccatagliata@neurologia.unige.it; Cotticelli, Biagio [Unit of Radiology, IRCCS Policlinico San Donato, via Morandi 30, 20097 San Donato Milanese, Milan (Italy)], E-mail: neurobiagio@tiscali.it; Nano, Giovanni [Department of Medical and Surgical Sciences, University of Milan School of Medicine, via Morandi 30, 20097 San Donato Milanese, Milan (Italy); Unit of Vascular Surgery, IRCCS Policlinico San Donato, via Morandi 30, 20097 Milan (Italy)], E-mail: gianni.nano@libero.it; Cornalba, Gianpaolo [Department of Diagnostic and Interventional Radiology, University of Milan School of Medicine, San Paolo Hospital, via di Rudini 8, 20142 Milan (Italy)], E-mail: gianpaolo.cornalba@unimi.it

    2009-06-15

    Objectives: This study was aimed at testing the value of image subtraction for evaluating carotid vessel wall enhancement in contrast-enhanced MR angiography (MRA). Materials and methods: IRB approval was obtained. The scans of 81 consecutive patients who underwent carotid MRA with 0.1 mmol/kg of gadobenate dimeglumine were reviewed. Axial carotid 3D T1-weighted fast low-angle shot sequence before and 3 min after contrast injection were acquired and subtracted (enhanced minus unenhanced). Vessel wall enhancement was assigned a four-point score using native or subtracted images from 0 (no enhancement) to 3 (strong enhancement). Stenosis degree was graded according to NASCET. Results: With native images, vessel wall enhancement was detected in 20/81 patients (25%) and in 20/161 carotids (12%), and scored 2.0 {+-} 0.6 (mean {+-} standard deviation); with subtracted images, in 21/81 (26%) and 22/161 (14%), and scored 2.5 {+-} 0.6, respectively (P < 0.001, Sign test). The overall stenosis degree distribution was: mild, 41/161 (25%); moderate, 77/161 (48%); severe, 43/161 (27%). Carotids with moderate stenosis showed vessel wall enhancement with a frequency (17/77, 22%) significantly higher than that observed in carotids with mild stenosis (1/41, 2%) (P = 0.005, Fisher exact test) and higher, even though with borderline significance (P = 0.078, Fisher exact test), than that observed in carotids with severe stenosis (4/43, 9%). Conclusion: Roughly a quarter of patients undergoing carotid MRA showed vessel wall enhancement. Image subtraction improved vessel wall enhancement conspicuity. Vessel wall enhancement seems to be an event relatively independent from the degree of stenosis. Further studies are warranted to define the relation between vessel wall enhancement and histopathology, inflammatory status, and instability.

  15. The Characterization, Subtraction, and Addition of Astronomical Images

    Science.gov (United States)

    Lupton, R.

    2007-11-01

    Astronomical images are characterized by a spatially-varying Point Spread Function (``blur''; PSF) which is usually not known ab initio. Knowledge of the PSF is crucial for the correct analysis of images (e.g. separating stars from galaxies; estimating the deconvolved shapes of objects), and we have ways of modelling the PSF, but are not currently solving the problem especially well --- the fields may be (very) crowded; the data may be undersampled; and the spatial variation of the PSF may be considerable (in the case of X-ray data, varying in width by an order of magnitude). A related issue is that of finding what has changed between a pair of images taken under different conditions. Rather than solve for the PSF of both images, we generally solve for the convolution kernel that transforms one into the other, but the question of how to represent this spatial structure of this kernel is similar to the (unsolved) problem of the previous paragraph. Finally, we are often faced with the question of how to handle a set of several to many images of the same part of the sky taken under different conditions, and which are, in general, marginally sampled. A variety of ad-hoc solutions exist to recover the properties of the objects in the field, but I do not believe that we currently solve the problem optimally.

  16. The Reduction Of Motion Artifacts In Digital Subtraction Angiography By Geometrical Image Transformation

    Science.gov (United States)

    Fitzpatrick, J. Michael; Pickens, David R.; Mandava, Venkateswara R.; Grefenstette, John J.

    1988-06-01

    In the diagnosis of arteriosclerosis, radio-opaque dye is injected into the interior of the arteries to make them visible. Because of its increased contrast sensitivity, digital subtraction angiography has the potential for providing diagnostic images of arteries with reduced dye volumes. In the conventional technique, a mask image, acquired before the introduction of the dye, is subtracted from the contrast image, acquired after the dye is introduced, to produce a difference image in which only the dye in the arteries is visible. The usefulness of this technique has been severely limited by the image degradation caused by patient motion during image acquisition. This motion produces artifacts in the difference image that obscure the arteries. One technique for dealing with this problem is to reduce the degradation by means of image registration. The registration is carried out by means of a geometrical transformation of the mask image before subtraction so that it is in registration with the contrast image. This paper describes our technique for determining an optimal transformation. We employ a one-to-one elastic mapping and the Jacobian of that mapping to produce a geometrical image transformation. We choose a parameterized class of such mappings and use a heuristic search algorithm to optimize the parameters to minimize the severity of the motion artifacts. To increase the speed of the optimization process we use a statistical image comparison technique that provides a quick approximate evaluation of each image transformation. We present the experimental results of the application of our registration system to mask-contrast pairs, for images acquired from a specially designed phantom (described in a companion paper), and for clinical images.

  17. Metal artifact reduction method using metal streaks image subtraction

    Energy Technology Data Exchange (ETDEWEB)

    Pua, Rizza D.; Cho, Seung Ryong [Dept. of Nuclear and Quantum Engineering, Korea Advanced Institute of Science and Technology, Daejeon (Korea, Republic of)

    2014-04-15

    Many studies have been dedicated for metal artifact reduction (MAR); however, the methods are successful to varying degrees depending on situations. Sinogram in-painting, filtering, iterative method are some of the major categories of MAR. Each has its own merits and weaknesses. A combination of these methods or hybrid methods have also been developed to make use of the different benefits of two techniques and minimize the unfavorable results. Our method focuses on the in-paitning approach and a hybrid MAR described by Xia et al. Although in-painting scheme is an effective technique in reducing the primary metal artifacts, a major drawback is the reintroduction of new artifacts that can be caused by an inaccurate interpolation process. Furthermore, combining the segmented metal image to the corrected nonmetal image in the final step of a conventional inpainting approach causes an issue of incorrect metal pixel values. Our proposed method begins with a sinogram in-painting approach and ends with an image-based metal artifact reduction scheme. This work provides a simple, yet effective solution for reducing metal artifacts and acquiring the original metal pixel information. The proposed method demonstrated its effectiveness in a simulation setting. The proposed method showed image quality that is comparable to the standard MAR; however, quantitatively more accurate than the standard MAR.

  18. Multi Spectral Fluorescence Imager (MSFI)

    Science.gov (United States)

    Caron, Allison

    2016-01-01

    Genetic transformation with in vivo reporter genes for fluorescent proteins can be performed on a variety of organisms to address fundamental biological questions. Model organisms that may utilize an ISS imager include unicellular organisms (Saccharomyces cerevisiae), plants (Arabidopsis thaliana), and invertebrates (Caenorhabditis elegans). The multispectral fluorescence imager (MSFI) will have the capability to accommodate 10 cm x 10 cm Petri plates, various sized multi-well culture plates, and other custom culture containers. Features will include programmable temperature and light cycles, ethylene scrubbing (less than 25 ppb), CO2 control (between 400 ppm and ISS-ambient levels in units of 100 ppm) and sufficient airflow to prevent condensation that would interfere with imaging.

  19. Nonlinear ultrasonic imaging method for closed cracks using subtraction of responses at different external loads.

    Science.gov (United States)

    Ohara, Yoshikazu; Horinouchi, Satoshi; Hashimoto, Makoto; Shintaku, Yohei; Yamanaka, Kazushi

    2011-08-01

    To improve the selectivity of closed cracks for objects other than cracks in ultrasonic imaging, we propose an extension of a novel imaging method, namely, subharmonic phased array for crack evaluation (SPACE) as well as another approach using the subtraction of responses at different external loads. By applying external static or dynamic loads to closed cracks, the contact state in the cracks varies, resulting in an intensity change of responses at cracks. In contrast, objects other than cracks are independent of external load. Therefore, only cracks can be extracted by subtracting responses at different loads. In this study, we performed fundamental experiments on a closed fatigue crack formed in an aluminum alloy compact tension (CT) specimen using the proposed method. We examined the static load dependence of SPACE images and the dynamic load dependence of linear phased array (PA) images by simulating the external loads with a servohydraulic fatigue testing machine. By subtracting the images at different external loads, we show that this method is useful in extracting only the intensity change of responses related to closed cracks, while canceling the responses of objects other than cracks.

  20. The IPAC Image Subtraction and Discovery Pipeline for the Intermediate Palomar Transient Factory

    Science.gov (United States)

    Masci, Frank J.; Laher, Russ R.; Rebbapragada, Umaa D.; Doran, Gary B.; Miller, Adam A.; Bellm, Eric; Kasliwal, Mansi; Ofek, Eran O.; Surace, Jason; Shupe, David L.; Grillmair, Carl J.; Jackson, Ed; Barlow, Tom; Yan, Lin; Cao, Yi; Cenko, S. Bradley; Storrie-Lombardi, Lisa J.; Helou, George; Prince, Thomas A.; Kulkarni, Shrinivas R.

    2017-01-01

    We describe the near real-time transient-source discovery engine for the intermediate Palomar Transient Factory (iPTF), currently in operations at the Infrared Processing and Analysis Center (IPAC), Caltech. We coin this system the IPAC/iPTF Discovery Engine (or IDE). We review the algorithms used for PSF-matching, image subtraction, detection, photometry, and machine-learned (ML) vetting of extracted transient candidates. We also review the performance of our ML classifier. For a limiting signal-to-noise ratio of 4 in relatively unconfused regions, bogus candidates from processing artifacts and imperfect image subtractions outnumber real transients by ≃10:1. This can be considerably higher for image data with inaccurate astrometric and/or PSF-matching solutions. Despite this occasionally high contamination rate, the ML classifier is able to identify real transients with an efficiency (or completeness) of ≃97% for a maximum tolerable false-positive rate of 1% when classifying raw candidates. All subtraction-image metrics, source features, ML probability-based real-bogus scores, contextual metadata from other surveys, and possible associations with known Solar System objects are stored in a relational database for retrieval by the various science working groups. We review our efforts in mitigating false-positives and our experience in optimizing the overall system in response to the multitude of science projects underway with iPTF.

  1. The IPAC Image Subtraction and Discovery Pipeline for the Intermediate Palomar Transient Factory

    Science.gov (United States)

    Masci, Frank J.; Laher, Russ R.; Rebbapragada, Umaa D.; Doran, Gary B.; Miller, Adam A.; Bellm, Eric; Kasliwal, Mansi; Ofek, Eran O.; Surace, Jason; Shupe, David L.; hide

    2016-01-01

    We describe the near real-time transient-source discovery engine for the intermediate Palomar Transient Factory (iPTF), currently in operations at the Infrared Processing and Analysis Center (IPAC), Caltech. We coin this system the IPAC/iPTF Discovery Engine (or IDE). We review the algorithms used for PSF-matching, image subtraction, detection, photometry, and machine-learned (ML) vetting of extracted transient candidates. We also review the performance of our ML classifier. For a limiting signal-to-noise ratio of 4 in relatively unconfused regions, bogus candidates from processing artifacts and imperfect image subtractions outnumber real transients by approximately equal to 10:1. This can be considerably higher for image data with inaccurate astrometric and/or PSF-matching solutions. Despite this occasionally high contamination rate, the ML classifier is able to identify real transients with an efficiency (or completeness) of approximately equal to 97% for a maximum tolerable false-positive rate of 1% when classifying raw candidates. All subtraction-image metrics, source features, ML probability-based real-bogus scores, contextual metadata from other surveys, and possible associations with known Solar System objects are stored in a relational database for retrieval by the various science working groups. We review our efforts in mitigating false-positives and our experience in optimizing the overall system in response to the multitude of science projects underway with iPTF.

  2. Use of Caval Subtraction 2D Phase-Contrast MR Imaging to Measure Total Liver and Hepatic Arterial Blood Flow: Preclinical Validation and Initial Clinical Translation.

    Science.gov (United States)

    Chouhan, Manil D; Mookerjee, Rajeshwar P; Bainbridge, Alan; Walker-Samuel, Simon; Davies, Nathan; Halligan, Steve; Lythgoe, Mark F; Taylor, Stuart A

    2016-09-01

    Purpose To validate caval subtraction two-dimensional (2D) phase-contrast magnetic resonance (MR) imaging measurements of total liver blood flow (TLBF) and hepatic arterial fraction in an animal model and evaluate consistency and reproducibility in humans. Materials and Methods Approval from the institutional ethical committee for animal care and research ethics was obtained. Fifteen Sprague-Dawley rats underwent 2D phase-contrast MR imaging of the portal vein (PV) and infrahepatic and suprahepatic inferior vena cava (IVC). TLBF and hepatic arterial flow were estimated by subtracting infrahepatic from suprahepatic IVC flow and PV flow from estimated TLBF, respectively. Direct PV transit-time ultrasonography (US) and fluorescent microsphere measurements of hepatic arterial fraction were the standards of reference. Thereafter, consistency of caval subtraction phase-contrast MR imaging-derived TLBF and hepatic arterial flow was assessed in 13 volunteers (mean age, 28.3 years ± 1.4) against directly measured phase-contrast MR imaging PV and proper hepatic arterial inflow; reproducibility was measured after 7 days. Bland-Altman analysis of agreement and coefficient of variation comparisons were undertaken. Results There was good agreement between PV flow measured with phase-contrast MR imaging and that measured with transit-time US (mean difference, -3.5 mL/min/100 g; 95% limits of agreement [LOA], ±61.3 mL/min/100 g). Hepatic arterial fraction obtained with caval subtraction agreed well with those with fluorescent microspheres (mean difference, 4.2%; 95% LOA, ±20.5%). Good consistency was demonstrated between TLBF in humans measured with caval subtraction and direct inflow phase-contrast MR imaging (mean difference, -1.3 mL/min/100 g; 95% LOA, ±23.1 mL/min/100 g). TLBF reproducibility at 7 days was similar between the two methods (95% LOA, ±31.6 mL/min/100 g vs ±29.6 mL/min/100 g). Conclusion Caval subtraction phase-contrast MR imaging is a simple and clinically

  3. Feasibility of coronary calcium and stent image subtraction using 320-detector row CT angiography

    DEFF Research Database (Denmark)

    Fuchs, Andreas; Kühl, J Tobias; Chen, Marcus Y;

    2015-01-01

    . We defined target segments on CCTAconv as motion-free coronary segments with calcification or stent and low reader confidence. The effect of CCTAsub was assessed. No approval from the ethics committee was required according to Danish law. RESULTS: A total of 76 target segments were identified....... The use of coronary calcium image subtraction improved the reader confidence in 66% of these segments. In target segments, specificity (86% vs 65%; P

  4. Three-dimensional labeling of newly formed bone using synchrotron radiation barium K-edge subtraction imaging

    Science.gov (United States)

    Panahifar, Arash; Swanston, Treena M.; Pushie, M. Jake; Belev, George; Chapman, Dean; Weber, Lynn; Cooper, David M. L.

    2016-07-01

    Bone is a dynamic tissue which exhibits complex patterns of growth as well as continuous internal turnover (i.e. remodeling). Tracking such changes can be challenging and thus a high resolution imaging-based tracer would provide a powerful new perspective on bone tissue dynamics. This is, particularly so if such a tracer can be detected in 3D. Previously, strontium has been demonstrated to be an effective tracer which can be detected by synchrotron-based dual energy K-edge subtraction (KES) imaging in either 2D or 3D. The use of strontium is, however, limited to very small sample thicknesses due to its low K-edge energy (16.105 keV) and thus is not suitable for in vivo application. Here we establish proof-of-principle for the use of barium as an alternative tracer with a higher K-edge energy (37.441 keV), albeit for ex vivo imaging at the moment, which enables application in larger specimens and has the potential to be developed for in vivo imaging of preclinical animal models. New bone formation within growing rats in 2D and 3D was demonstrated at the Biomedical Imaging and Therapy bending magnet (BMIT-BM) beamline of the Canadian Light Source synchrotron. Comparative x-ray fluorescence imaging confirmed those patterns of uptake detected by KES. This initial work provides a platform for the further development of this tracer and its exploration of applications for in vivo development.

  5. Ability of subtraction and dynamic MR imaging to detect breast tumors. Comparison with ultrasonography and mammography

    Energy Technology Data Exchange (ETDEWEB)

    Terao, Eri; Takeuchi, Hiroaki; Iwamura, Akira; Murakami, Yoshitaka; Harada, Junta; Tada, Shinpei (Jikei Univ., Tokyo (Japan). School of Medicine)

    1994-09-01

    We evaluated the ability of subtraction and dynamic MR imaging to accurately detect breast tumors. Sixty-five breast carcinomas and 24 fibroadenomas were examined by an SE pulse sequence using a 0.2 Tesla unit. Subtraction MR images were obtained every minute during dynamic study with Gd-DTPA. Almost all breast tumors were seen as very bright masses, and the margin of the mass was clearly demonstrated on subtraction MR images. Breast carcinomas and fibroadenomas showed characteristic time-intensity curves on dynamic study. Time-intensity curves of the early peak type and plateau type were seen in 97% of breast carcinomas, while the gradually increasing type was seen in 92% of fibroadenomas. The detectability of breast carcinoma was 98% by MRI, 98% by ultrasonography, and 87% by mammography. That of fibroadenoma was 95% by MRI, 91% by ultrasonography and 60% by mammography. Sensitivity and specificity for breast carcinoma were 98% and 92% for MRI and 97% and 71% for ultrasonography. For fibroadenoma, they were 96% and 98% for MRI and 89% and 92% for ultrasonography. (author).

  6. Cooperative Moving Object Segmentation using Two Cameras based on Background Subtraction and Image Registration

    Directory of Open Access Journals (Sweden)

    Zhigao Cui

    2014-03-01

    Full Text Available Moving camera, such as PTZ (pan-tilt-zoom camera, has been widely applied in visual surveillance system. However, it’s difficult to extract moving objects because of the dynamic background caused by the camera motion. In this paper, a novel framework for moving object segmentation exploiting two cameras collaboration is presented by combining background subtraction and image registration method. The proposed method uses one static camera to capture large-view images at low resolution, and one moving camera (i.e. PTZ camera to capture local-view images at high resolution. Different with methods using a single moving camera, the moving objects can be effectively segmented in the static camera image by background subtraction method. Then image registration method can be applied to extract moving region in the moving camera image. To deal with the resolution and intensity discrepancy between two synchronized images, we design a practical three-step image registration method, which has higher registration accuracy than traditional feature based method. Experimental results on outdoor scene demonstrate the effectiveness and robustness of proposed approach.

  7. Comparison of the diagnostic accuracy of direct digital radiography system, filtered images, and subtraction radiography

    Directory of Open Access Journals (Sweden)

    Wilton Mitsunari Takeshita

    2013-01-01

    Full Text Available Background: To compare the diagnostic accuracy of three different imaging systems: Direct digital radiography system (DDR-CMOS, four types of filtered images, and a priori and a posteriori registration of digital subtraction radiography (DSR in the diagnosis of proximal defects. Materials and Methods: The teeth were arranged in pairs in 10 blocks of vinyl polysiloxane, and proximal defects were performed with drills of 0.25, 0.5, and 1 mm diameter. Kodak RVG 6100 sensor was used to capture the images. A posteriori DSR registrations were done with Regeemy 0.2.43 and subtraction with Image Tool 3.0. Filtered images were obtained with Kodak Dental Imaging 6.1 software. Images (n = 360 were evaluated by three raters, all experts in dental radiology. Results: Sensitivity and specificity of the area under the receiver operator characteristic (ROC curve (Az were higher for DSR images with all three drills (Az = 0.896, 0.979, and 1.000 for drills 0.25, 0.5, and 1 mm, respectively. The highest values were found for 1-mm drills and the lowest for 0.25-mm drills, with negative filter having the lowest values of all (Az = 0.631. Conclusion: The best method of diagnosis was by using a DSR. The negative filter obtained the worst results. Larger drills showed the highest sensitivity and specificity values of the area under the ROC curve.

  8. Computed tomography lung iodine contrast mapping by image registration and subtraction

    Science.gov (United States)

    Goatman, Keith; Plakas, Costas; Schuijf, Joanne; Beveridge, Erin; Prokop, Mathias

    2014-03-01

    Pulmonary embolism (PE) is a relatively common and potentially life threatening disease, affecting around 600,000 people annually in the United States alone. Prompt treatment using anticoagulants is effective and saves lives, but unnecessary treatment risks life threatening haemorrhage. The specificity of any diagnostic test for PE is therefore as important as its sensitivity. Computed tomography (CT) angiography is routinely used to diagnose PE. However, there are concerns it may over-report the condition. Additional information about the severity of an occlusion can be obtained from an iodine contrast map that represents tissue perfusion. Such maps tend to be derived from dual-energy CT acquisitions. However, they may also be calculated by subtracting pre- and post-contrast CT scans. Indeed, there are technical advantages to such a subtraction approach, including better contrast-to-noise ratio for the same radiation dose, and bone suppression. However, subtraction relies on accurate image registration. This paper presents a framework for the automatic alignment of pre- and post-contrast lung volumes prior to subtraction. The registration accuracy is evaluated for seven subjects for whom pre- and post-contrast helical CT scans were acquired using a Toshiba Aquilion ONE scanner. One hundred corresponding points were annotated on the pre- and post-contrast scans, distributed throughout the lung volume. Surface-to-surface error distances were also calculated from lung segmentations. Prior to registration the mean Euclidean landmark alignment error was 2.57mm (range 1.43-4.34 mm), and following registration the mean error was 0.54mm (range 0.44-0.64 mm). The mean surface error distance was 1.89mm before registration and 0.47mm after registration. There was a commensurate reduction in visual artefacts following registration. In conclusion, a framework for pre- and post-contrast lung registration has been developed that is sufficiently accurate for lung subtraction

  9. Temporal subtraction system on torso FDG-PET scans based on statistical image analysis

    Science.gov (United States)

    Shimizu, Yusuke; Hara, Takeshi; Fukuoka, Daisuke; Zhou, Xiangrong; Muramatsu, Chisako; Ito, Satoshi; Hakozaki, Kenta; Kumita, Shin-ichiro; Ishihara, Kei-ichi; Katafuchi, Tetsuro; Fujita, Hiroshi

    2013-02-01

    Diagnostic imaging on FDG-PET scans was often used to evaluate chemotherapy results of cancer patients. Radiologists compare the changes of lesions' activities between previous and current examinations for the evaluation. The purpose of this study was to develop a new computer-aided detection (CAD) system with temporal subtraction technique for FDGPET scans and to show the fundamental usefulness based on an observer performance study. Z-score mapping based on statistical image analysis was newly applied to the temporal subtraction technique. The subtraction images can be obtained based on the anatomical standardization results because all of the patients' scans were deformed into standard body shape. An observer study was performed without and with computer outputs to evaluate the usefulness of the scheme by ROC (receiver operating characteristics) analysis. Readers responded as confidence levels on a continuous scale from absolutely no change to definitely change between two examinations. The recognition performance of the computer outputs for the 43 pairs was 96% sensitivity with 31.1 false-positive marks per scan. The average of area-under-the-ROC-curve (AUC) from 4 readers in the observer performance study was increased from 0.85 without computer outputs to 0.90 with computer outputs (p=0.0389, DBM-MRMC). The average of interpretation time was slightly decreased from 42.11 to 40.04 seconds per case (p=0.625, Wilcoxon test). We concluded that the CAD system for torso FDG-PET scans with temporal subtraction technique might improve the diagnostic accuracy of radiologist in cancer therapy evaluation.

  10. Multi-face detection based on downsampling and modified subtractive clustering for color images

    Institute of Scientific and Technical Information of China (English)

    KONG Wan-zeng; ZHU Shan-an

    2007-01-01

    This paper presents a multi-face detection method for color images. The method is based on the assumption that faces are well separated from the background by skin color detection. These faces can be located by the proposed method which modifies the subtractive clustering. The modified clustering algorithm proposes a new definition of distance for multi-face detection, and its key parameters can be predetermined adaptively by statistical information of face objects in the image. Downsampling is employed to reduce the computation of clustering and speed up the process of the proposed method. The effectiveness of the proposed method is illustrated by three experiments.

  11. Fluorescence imaging spectrometer optical design

    Science.gov (United States)

    Taiti, A.; Coppo, P.; Battistelli, E.

    2015-09-01

    The optical design of the FLuORescence Imaging Spectrometer (FLORIS) studied for the Fluorescence Explorer (FLEX) mission is discussed. FLEX is a candidate for the ESA's 8th Earth Explorer opportunity mission. FLORIS is a pushbroom hyperspectral imager foreseen to be embarked on board of a medium size satellite, flying in tandem with Sentinel-3 in a Sun synchronous orbit at a height of about 815 km. FLORIS will observe the vegetation fluorescence and reflectance within a spectral range between 500 and 780 nm. Multi-frames acquisitions on matrix detectors during the satellite movement will allow the production of 2D Earth scene images in two different spectral channels, called HR and LR with spectral resolution of 0.3 and 2 nm respectively. A common fore optics is foreseen to enhance by design the spatial co-registration between the two spectral channels, which have the same ground spatial sampling (300 m) and swath (150 km). An overlapped spectral range between the two channels is also introduced to simplify the spectral coregistration. A compact opto-mechanical solution with all spherical and plane optical elements is proposed, and the most significant design rationales are described. The instrument optical architecture foresees a dual Babinet scrambler, a dioptric telescope and two grating spectrometers (HR and LR), each consisting of a modified Offner configuration. The developed design is robust, stable vs temperature, easy to align, showing very high optical quality along the whole field of view. The system gives also excellent correction for transverse chromatic aberration and distortions (keystone and smile).

  12. Quantitative imaging with fluorescent biosensors.

    Science.gov (United States)

    Okumoto, Sakiko; Jones, Alexander; Frommer, Wolf B

    2012-01-01

    Molecular activities are highly dynamic and can occur locally in subcellular domains or compartments. Neighboring cells in the same tissue can exist in different states. Therefore, quantitative information on the cellular and subcellular dynamics of ions, signaling molecules, and metabolites is critical for functional understanding of organisms. Mass spectrometry is generally used for monitoring ions and metabolites; however, its temporal and spatial resolution are limited. Fluorescent proteins have revolutionized many areas of biology-e.g., fluorescent proteins can report on gene expression or protein localization in real time-yet promoter-based reporters are often slow to report physiologically relevant changes such as calcium oscillations. Therefore, novel tools are required that can be deployed in specific cells and targeted to subcellular compartments in order to quantify target molecule dynamics directly. We require tools that can measure enzyme activities, protein dynamics, and biophysical processes (e.g., membrane potential or molecular tension) with subcellular resolution. Today, we have an extensive suite of tools at our disposal to address these challenges, including translocation sensors, fluorescence-intensity sensors, and Förster resonance energy transfer sensors. This review summarizes sensor design principles, provides a database of sensors for more than 70 different analytes/processes, and gives examples of applications in quantitative live cell imaging.

  13. Scatter-to-primary based scatter fractions for transmission-dependent convolution subtraction of SPECT images.

    Science.gov (United States)

    Larsson, Anne; Johansson, Lennart

    2003-11-21

    In single photon emission computed tomography (SPECT), transmission-dependent convolution subtraction has been shown to be useful when correcting for scattered events. The method is based on convolution subtraction, but includes a matrix of scatter fractions instead of a global scatter fraction. The method can be extended to iteratively improve the scatter estimate, but in this note we show that this requires a modification of the theory to use scatter-to-total scatter fractions for the first iteration only and scatter-to-primary fractions thereafter. To demonstrate this, scatter correction is performed on a Monte Carlo simulated image of a point source of activity in water. The modification of the theory is compared to corrections where the scatter fractions are based on the scatter-to-total ratio, using one and ten iterations. The resulting ratios of subtracted to original counts are compared to the true scatter-to-total ratio of the simulation and the most accurate result is found for our modification of the theory.

  14. Digital subtraction laryngography

    Energy Technology Data Exchange (ETDEWEB)

    Brown, B.W.; Enzmann, D.R.; Hopp, M.L.; Castellino, R.A.

    1983-06-01

    Digital subtraction laryngography was used to evaluate laryngeal function in 8 patients: 4 with normal larynxes and 4 with laryngeal disease. Subtracted digital images provided a dynamic display of the extent and symmetry of vocal cord excursions and pyriform sinus inflation, and the vocal cord resting position was also clearly depicted. The technical details of digital subtraction laryngography and its application are described.

  15. The IPAC Image Subtraction and Discovery Pipeline for the intermediate Palomar Transient Factory

    CERN Document Server

    Masci, Frank; Rebbapragada, Umaa; Doran, Gary; Miller, Adam; Bell, Eric; Kasliwal, Mansi; Ofek, Eran; Surace, Jason; Shupe, David; Grillmair, Carl; Jackson, Ed; Barlow, Tom; Yan, Lin; Cao, Yi; Cenko, S Bradley; Storrie-Lombardi, Lisa; Helou, George; Prince, Thomas; Kulkarni, Shrinivas

    2016-01-01

    We describe the near real-time transient-source discovery engine for the intermediate Palomar Transient Factory (iPTF), currently in operations at the Infrared Processing and Analysis Center (IPAC), Caltech. We coin this system the IPAC/iPTF Discovery Engine (or IDE). We review the algorithms used for PSF-matching, image subtraction, detection, photometry, and machine-learned (ML) vetting of extracted transient candidates. We also review the performance of our ML classifier. For a limiting signal-to-noise ratio of 4 in relatively unconfused regions, "bogus" candidates from processing artifacts and imperfect image subtractions outnumber real transients by ~ 10:1. This can be considerably higher for image data with inaccurate astrometric and/or PSF-matching solutions. Despite this occasionally high contamination rate, the ML classifier is able to identify real transients with an efficiency (or completeness) of ~ 97% for a maximum tolerable false-positive rate of 1% when classifying raw candidates. All subtracti...

  16. Automated Transient Recovery Algorithm using Discrete Zernike Polynomials on Image-Subtracted Data

    Science.gov (United States)

    Ackley, Kendall; Eikenberry, Stephen S.; Klimenko, Sergey

    2016-01-01

    We present an unsupervised algorithm for the automated identification of astrophysical transients recovered through image subtraction techniques. We use a set of discrete Zernike polynomials to decompose and characterize residual energy discovered in the final subtracted image, identifying candidate sources which appear point-like in nature. This work is motivated for use in collaboration with Advanced gravitational wave (GW) interferometers, such as Advanced LIGO and Virgo, where multiwavelength electromagnetic (EM) emission is expected in parallel with gravitational radiation from compact binary object mergers of neutron stars (NS-NS) and stellar-mass black holes (NS-BH). Imaging an EM counterpart coincident with a GW trigger will help to constrain the multi-dimensional GW parameter space as well as aid in the resolution of long-standing astrophysical mysteries, such as the true nature of the progenitor relationship between short-duration GRBs and massive compact binary mergers. We are working on making our method an open-source package optimized for low-latency response for community use during the upcoming era of GW astronomy.

  17. Quantitative Method for the Optimal Subtraction of Continuum Emission from Narrow-band Images : Skewness Transition Analysis

    CERN Document Server

    Hong, Sungryong; Dickinson, Mark

    2013-01-01

    We present an objective method to remove the stellar continuum emission from narrow-band images to derive emission-line images. The method is based on the skewness of the pixel histogram of the residual images. Specifically, we exploit a transition in the skewness of the signal in the continuum-subtracted image, which appears when the image changes from being under-subtracted to over-subtracted. Tests on one-dimensional artificial images demonstrate that the transition identifies the optimal scaling factor {\\mu} to be used on the broad-band image IB in order to produce the optimal line-emission image IE, i.e., IE =IN - {\\mu} IB, with IN the original (un-subtracted) narrow-band image. The advantage of this method is that it uses all information-bearing pixels in the final image, and not just a sub-set of those pixels (the latter being common in many traditional approaches to stellar continuum removal from narrow-band images). We apply our method to actual images, both from ground-based and space facilities, in...

  18. Dual-Energy Subtraction Imaging for Diagnosing Vocal Cord Paralysis with Flat Panel Detector Radiography

    Energy Technology Data Exchange (ETDEWEB)

    Machida, Haruhiko; Yoda, Keiko; Arai, Yasuko [Tokyo Women' s Medical University Medical Center East, Tokyo (Japan)] (and others)

    2010-06-15

    To investigate the clinical feasibility of dual energy subtraction (DES) imaging to improve the delineation of the vocal cord and diagnostic accuracy of vocal cord paralysis as compared with the anterior-posterior view of flat panel detector (FPD) neck radiography. For 122 consecutive patients who underwent both a flexible laryngoscopy and conventional/DES FPD radiography, three blinded readers retrospectively graded the radiographs during phonation and inspiration on a scale of 1 (poor) to 5 (excellent) for the delineation of the vocal cord, and in consensus, reviewed the diagnostic accuracy of vocal cord paralysis employing the laryngoscopy as the reference. We compared vocal cord delineation scores and accuracy of vocal cord paralysis diagnosis by both conventional and DES techniques using ({kappa}statistics and assessing the area under the receiver operating characteristic curve (AUC). Vocal cord delineation scores by DES (mean, 4.2 {+-} 0.4) were significantly higher than those by conventional imaging (mean, 3.3 {+-} 0.5) (p < 0.0001). Sensitivity for diagnosing vocal cord paralysis by the conventional technique was 25%, whereas the specificity was 94%. Sensitivity by DES was 75%, whereas the specificity was 96%. The diagnostic accuracy by DES was significantly superior (({kappa}= 0.60, AUC = 0.909) to that by conventional technique ({kappa}= 0.18, AUC = 0.852) (p = 0.038). Dual energy subtraction is a superior method compared to the conventional FPD radiography for delineating the vocal cord and accurately diagnosing vocal cord paralysis.

  19. A LabVIEW Platform for Preclinical Imaging Using Digital Subtraction Angiography and Micro-CT.

    Science.gov (United States)

    Badea, Cristian T; Hedlund, Laurence W; Johnson, G Allan

    2013-01-01

    CT and digital subtraction angiography (DSA) are ubiquitous in the clinic. Their preclinical equivalents are valuable imaging methods for studying disease models and treatment. We have developed a dual source/detector X-ray imaging system that we have used for both micro-CT and DSA studies in rodents. The control of such a complex imaging system requires substantial software development for which we use the graphical language LabVIEW (National Instruments, Austin, TX, USA). This paper focuses on a LabVIEW platform that we have developed to enable anatomical and functional imaging with micro-CT and DSA. Our LabVIEW applications integrate and control all the elements of our system including a dual source/detector X-ray system, a mechanical ventilator, a physiological monitor, and a power microinjector for the vascular delivery of X-ray contrast agents. Various applications allow cardiac- and respiratory-gated acquisitions for both DSA and micro-CT studies. Our results illustrate the application of DSA for cardiopulmonary studies and vascular imaging of the liver and coronary arteries. We also show how DSA can be used for functional imaging of the kidney. Finally, the power of 4D micro-CT imaging using both prospective and retrospective gating is shown for cardiac imaging.

  20. In vivo small-animal imaging using micro-CT and digital subtraction angiography

    Energy Technology Data Exchange (ETDEWEB)

    Badea, C T; Johnson, G A [Center for In Vivo Microscopy, Department of Radiology, Duke University, Durham, NC 27710 (United States); Drangova, M; Holdsworth, D W [Imaging Research Laboratories, Robarts Research Institute, University of Western Ontario, London, ON N6A 5K8 (United States)], E-mail: Cristian.Badea@duke.edu

    2008-10-07

    Small-animal imaging has a critical role in phenotyping, drug discovery and in providing a basic understanding of mechanisms of disease. Translating imaging methods from humans to small animals is not an easy task. The purpose of this work is to review in vivo x-ray based small-animal imaging, with a focus on in vivo micro-computed tomography (micro-CT) and digital subtraction angiography (DSA). We present the principles, technologies, image quality parameters and types of applications. We show that both methods can be used not only to provide morphological, but also functional information, such as cardiac function estimation or perfusion. Compared to other modalities, x-ray based imaging is usually regarded as being able to provide higher throughput at lower cost and adequate resolution. The limitations are usually associated with the relatively poor contrast mechanisms and potential radiation damage due to ionizing radiation, although the use of contrast agents and careful design of studies can address these limitations. We hope that the information will effectively address how x-ray based imaging can be exploited for successful in vivo preclinical imaging. (topical reviews)

  1. K-edge digital subtraction imaging based on a dichromatic and compact x-ray source

    Science.gov (United States)

    Sarnelli, A.; Taibi, A.; Tuffanelli, A.; Baldazzi, G.; Bollini, D.; Cabal Rodriguez, A. E.; Gombia, M.; Prino, F.; Ramello, L.; Tomassi, E.; Gambaccini, M.

    2004-07-01

    This work proposes a compact dichromatic imaging system for the application of the K-edge digital subtraction technique based on a conventional x-ray tube and a monochromator system. A quasi-monochromatic x-ray beam at the energy of iodine K-edge is produced by Bragg diffraction on a mosaic crystal. Two thin adjacent beams with energies that bracket the K-edge discontinuity are obtained from the diffracted beam by means of a proper collimation system. They are then detected using an array of Si detectors. A home-made phantom is used to study the image quality as a function of iodine concentration. Signal and signal-to-noise ratio analysis has also been performed. The results are compared with theoretical expectations.

  2. K-edge digital subtraction imaging based on a dichromatic and compact x-ray source

    Energy Technology Data Exchange (ETDEWEB)

    Sarnelli, A [Dipartimento di Fisica dell' Universita di Ferrara and INFN Sezione di Ferrara, Via Paradiso 12, I-44100 Ferrara (Italy); Taibi, A [Dipartimento di Fisica dell' Universita di Ferrara and INFN Sezione di Ferrara, Via Paradiso 12, I-44100 Ferrara (Italy); Tuffanelli, A [Dipartimento di Fisica dell' Universita di Ferrara and INFN Sezione di Ferrara, Via Paradiso 12, I-44100 Ferrara (Italy); Baldazzi, G [Dipartimento di Fisica dell' Universita di Bologna and INFN Sezione di Bologna, Viale Berti Pichat 64/2, 40127 Bologna (Italy); Bollini, D [Dipartimento di Fisica dell' Universita di Bologna and INFN Sezione di Bologna, Viale Berti Pichat 64/2, 40127 Bologna (Italy); Rodriguez, A E Cabal [CAEDAN, Havana (Cuba); Gombia, M [Dipartimento di Fisica dell' Universita di Bologna and INFN Sezione di Bologna, Viale Berti Pichat 64/2, 40127 Bologna (Italy); Prino, F [Dipartimento di Scienze e Tecnologie Avanzate, Universita del Piemonte Orientale and INFN Sezione di Alessandria, C.so, Borsalino 54, I-15100 Alessandria (Italy); Ramello, L [Dipartimento di Scienze e Tecnologie Avanzate, Universita del Piemonte Orientale and INFN Sezione di Alessandria, C.so, Borsalino 54, I-15100 Alessandria (Italy); Tomassi, E [Dipartimento di Scienze e Tecnologie Avanzate, Universita del Piemonte Orientale and INFN Sezione di Alessandria, C.so, Borsalino 54, I-15100 Alessandria (Italy); Gambaccini, M [Dipartimento di Fisica dell' Universita di Ferrara and INFN Sezione di Ferrara, Via Paradiso 12, I-44100 Ferrara (Italy)

    2004-07-21

    This work proposes a compact dichromatic imaging system for the application of the K-edge digital subtraction technique based on a conventional x-ray tube and a monochromator system. A quasi-monochromatic x-ray beam at the energy of iodine K-edge is produced by Bragg diffraction on a mosaic crystal. Two thin adjacent beams with energies that bracket the K-edge discontinuity are obtained from the diffracted beam by means of a proper collimation system. They are then detected using an array of Si detectors. A home-made phantom is used to study the image quality as a function of iodine concentration. Signal and signal-to-noise ratio analysis has also been performed. The results are compared with theoretical expectations.

  3. A comparison of different energy window subtraction methods to correct for scatter and downscatter in I-123 SPECT imaging

    DEFF Research Database (Denmark)

    Lagerburg, Vera; de Nijs, Robin; Holm, Søren

    2012-01-01

    One of the main problems in quantification of single photon emission computer tomography imaging is scatter. In iodine-123 (I-123) imaging, both the primary 159 keV photons and photons of higher energies are scattered. In this experimental study, different scatter correction methods, based on ene...... on energy window subtraction, have been compared with each other....

  4. Deep UV Native Fluorescence Imaging of Antarctic Cryptoendolithic Communities

    Science.gov (United States)

    Storrie-Lombardi, M. C.; Douglas, S.; Sun, H.; McDonald, G. D.; Bhartia, R.; Nealson, K. H.; Hug, W. F.

    2001-01-01

    An interdisciplinary team at the Jet Propulsion Laboratory Center for Life Detection has embarked on a project to provide in situ chemical and morphological characterization of Antarctic cryptoendolithic microbial communities. We present here in situ deep ultraviolet (UV) native fluorescence and environmental scanning electron microscopy images transiting 8.5 mm into a sandstone sample from the Antarctic Dry Valleys. The deep ultraviolet imaging system employs 224.3, 248.6, and 325 nm lasers to elicit differential fluorescence and resonance Raman responses from biomolecules and minerals. The 224.3 and 248.6 nm lasers elicit a fluorescence response from the aromatic amino and nucleic acids. Excitation at 325 nm may elicit activity from a variety of biomolecules, but is more likely to elicit mineral fluorescence. The resultant fluorescence images provide in situ chemical and morphological maps of microorganisms and the associated organic matrix. Visible broadband reflectance images provide orientation against the mineral background. Environmental scanning electron micrographs provided detailed morphological information. The technique has made possible the construction of detailed fluorescent maps extending from the surface of an Antarctic sandstone sample to a depth of 8.5 mm. The images detect no evidence of microbial life in the superficial 0.2 mm crustal layer. The black lichen component between 0.3 and 0.5 mm deep absorbs all wavelengths of both laser and broadband illumination. Filamentous deep ultraviolet native fluorescent activity dominates in the white layer between 0.6 mm and 5.0 mm from the surface. These filamentous forms are fungi that continue into the red (iron-rich) region of the sample extending from 5.0 to 8.5 mm. Using differential image subtraction techniques it is possible to identify fungal nuclei. The ultraviolet response is markedly attenuated in this region, apparently from the absorption of ultraviolet light by iron-rich particles coating

  5. Statistical image segmentation for the detection of skin lesion borders in UV fluorescence excitation

    Science.gov (United States)

    Ortega-Martinez, Antonio; Padilla-Martinez, Juan Pablo; Franco, Walfre

    2016-04-01

    The skin contains several fluorescent molecules or fluorophores that serve as markers of structure, function and composition. UV fluorescence excitation photography is a simple and effective way to image specific intrinsic fluorophores, such as the one ascribed to tryptophan which emits at a wavelength of 345 nm upon excitation at 295 nm, and is a marker of cellular proliferation. Earlier, we built a clinical UV photography system to image cellular proliferation. In some samples, the naturally low intensity of the fluorescence can make it difficult to separate the fluorescence of cells in higher proliferation states from background fluorescence and other imaging artifacts -- like electronic noise. In this work, we describe a statistical image segmentation method to separate the fluorescence of interest. Statistical image segmentation is based on image averaging, background subtraction and pixel statistics. This method allows to better quantify the intensity and surface distributions of fluorescence, which in turn simplify the detection of borders. Using this method we delineated the borders of highly-proliferative skin conditions and diseases, in particular, allergic contact dermatitis, psoriatic lesions and basal cell carcinoma. Segmented images clearly define lesion borders. UV fluorescence excitation photography along with statistical image segmentation may serve as a quick and simple diagnostic tool for clinicians.

  6. Classification-based summation of cerebral digital subtraction angiography series for image post-processing algorithms

    Energy Technology Data Exchange (ETDEWEB)

    Schuldhaus, D; Spiegel, M; Polyanskaya, M; Hornegger, J [Pattern Recognition Lab, University Erlangen-Nuremberg (Germany); Redel, T [Siemens AG Healthcare Sector, Forchheim (Germany); Struffert, T; Doerfler, A, E-mail: martin.spiegel@informatik.uni-erlangen.de [Department of Neuroradiology, University Erlangen-Nuremberg (Germany)

    2011-03-21

    X-ray-based 2D digital subtraction angiography (DSA) plays a major role in the diagnosis, treatment planning and assessment of cerebrovascular disease, i.e. aneurysms, arteriovenous malformations and intracranial stenosis. DSA information is increasingly used for secondary image post-processing such as vessel segmentation, registration and comparison to hemodynamic calculation using computational fluid dynamics. Depending on the amount of injected contrast agent and the duration of injection, these DSA series may not exhibit one single DSA image showing the entire vessel tree. The interesting information for these algorithms, however, is usually depicted within a few images. If these images would be combined into one image the complexity of segmentation or registration methods using DSA series would drastically decrease. In this paper, we propose a novel method automatically splitting a DSA series into three parts, i.e. mask, arterial and parenchymal phase, to provide one final image showing all important vessels with less noise and moving artifacts. This final image covers all arterial phase images, either by image summation or by taking the minimum intensities. The phase classification is done by a two-step approach. The mask/arterial phase border is determined by a Perceptron-based method trained from a set of DSA series. The arterial/parenchymal phase border is specified by a threshold-based method. The evaluation of the proposed method is two-sided: (1) comparison between automatic and medical expert-based phase selection and (2) the quality of the final image is measured by gradient magnitudes inside the vessels and signal-to-noise (SNR) outside. Experimental results show a match between expert and automatic phase separation of 93%/50% and an average SNR increase of up to 182% compared to summing up the entire series.

  7. Classification-based summation of cerebral digital subtraction angiography series for image post-processing algorithms

    Science.gov (United States)

    Schuldhaus, D.; Spiegel, M.; Redel, T.; Polyanskaya, M.; Struffert, T.; Hornegger, J.; Doerfler, A.

    2011-03-01

    X-ray-based 2D digital subtraction angiography (DSA) plays a major role in the diagnosis, treatment planning and assessment of cerebrovascular disease, i.e. aneurysms, arteriovenous malformations and intracranial stenosis. DSA information is increasingly used for secondary image post-processing such as vessel segmentation, registration and comparison to hemodynamic calculation using computational fluid dynamics. Depending on the amount of injected contrast agent and the duration of injection, these DSA series may not exhibit one single DSA image showing the entire vessel tree. The interesting information for these algorithms, however, is usually depicted within a few images. If these images would be combined into one image the complexity of segmentation or registration methods using DSA series would drastically decrease. In this paper, we propose a novel method automatically splitting a DSA series into three parts, i.e. mask, arterial and parenchymal phase, to provide one final image showing all important vessels with less noise and moving artifacts. This final image covers all arterial phase images, either by image summation or by taking the minimum intensities. The phase classification is done by a two-step approach. The mask/arterial phase border is determined by a Perceptron-based method trained from a set of DSA series. The arterial/parenchymal phase border is specified by a threshold-based method. The evaluation of the proposed method is two-sided: (1) comparison between automatic and medical expert-based phase selection and (2) the quality of the final image is measured by gradient magnitudes inside the vessels and signal-to-noise (SNR) outside. Experimental results show a match between expert and automatic phase separation of 93%/50% and an average SNR increase of up to 182% compared to summing up the entire series.

  8. Classification-based summation of cerebral digital subtraction angiography series for image post-processing algorithms.

    Science.gov (United States)

    Schuldhaus, D; Spiegel, M; Redel, T; Polyanskaya, M; Struffert, T; Hornegger, J; Doerfler, A

    2011-03-21

    X-ray-based 2D digital subtraction angiography (DSA) plays a major role in the diagnosis, treatment planning and assessment of cerebrovascular disease, i.e. aneurysms, arteriovenous malformations and intracranial stenosis. DSA information is increasingly used for secondary image post-processing such as vessel segmentation, registration and comparison to hemodynamic calculation using computational fluid dynamics. Depending on the amount of injected contrast agent and the duration of injection, these DSA series may not exhibit one single DSA image showing the entire vessel tree. The interesting information for these algorithms, however, is usually depicted within a few images. If these images would be combined into one image the complexity of segmentation or registration methods using DSA series would drastically decrease. In this paper, we propose a novel method automatically splitting a DSA series into three parts, i.e. mask, arterial and parenchymal phase, to provide one final image showing all important vessels with less noise and moving artifacts. This final image covers all arterial phase images, either by image summation or by taking the minimum intensities. The phase classification is done by a two-step approach. The mask/arterial phase border is determined by a Perceptron-based method trained from a set of DSA series. The arterial/parenchymal phase border is specified by a threshold-based method. The evaluation of the proposed method is two-sided: (1) comparison between automatic and medical expert-based phase selection and (2) the quality of the final image is measured by gradient magnitudes inside the vessels and signal-to-noise (SNR) outside. Experimental results show a match between expert and automatic phase separation of 93%/50% and an average SNR increase of up to 182% compared to summing up the entire series.

  9. Cryo-imaging of fluorescently labeled single cells in a mouse

    Science.gov (United States)

    Steyer, Grant J.; Roy, Debashish; Salvado, Olivier; Stone, Meredith E.; Wilson, David L.

    2009-02-01

    We developed a cryo-imaging system to provide single-cell detection of fluorescently labeled cells in mouse, with particular applicability to stem cells and metastatic cancer. The Case cryoimaging system consists of a fluorescence microscope, robotic imaging positioner, customized cryostat, PC-based control system, and visualization/analysis software. The system alternates between sectioning (10-40 μm) and imaging, collecting color brightfield and fluorescent blockface image volumes >60GB. In mouse experiments, we imaged quantum-dot labeled stem cells, GFP-labeled cancer and stem cells, and cell-size fluorescent microspheres. To remove subsurface fluorescence, we used a simplified model of light-tissue interaction whereby the next image was scaled, blurred, and subtracted from the current image. We estimated scaling and blurring parameters by minimizing entropy of subtracted images. Tissue specific attenuation parameters were found [uT : heart (267 +/- 47.6 μm), liver (218 +/- 27.1 μm), brain (161 +/- 27.4 μm)] to be within the range of estimates in the literature. "Next image" processing removed subsurface fluorescence equally well across multiple tissues (brain, kidney, liver, adipose tissue, etc.), and analysis of 200 microsphere images in the brain gave 97+/-2% reduction of subsurface fluorescence. Fluorescent signals were determined to arise from single cells based upon geometric and integrated intensity measurements. Next image processing greatly improved axial resolution, enabled high quality 3D volume renderings, and improved enumeration of single cells with connected component analysis by up to 24%. Analysis of image volumes identified metastatic cancer sites, found homing of stem cells to injury sites, and showed microsphere distribution correlated with blood flow patterns. We developed and evaluated cryo-imaging to provide single-cell detection of fluorescently labeled cells in mouse. Our cryo-imaging system provides extreme (>60GB), micron

  10. Boronic acids for fluorescence imaging of carbohydrates.

    Science.gov (United States)

    Sun, Xiaolong; Zhai, Wenlei; Fossey, John S; James, Tony D

    2016-02-28

    "Fluorescence imaging" is a particularly exciting and rapidly developing area of research; the annual number of publications in the area has increased ten-fold over the last decade. The rapid increase of interest in fluorescence imaging will necessitate the development of an increasing number of molecular receptors and binding agents in order to meet the demand in this rapidly expanding area. Carbohydrate biomarkers are particularly important targets for fluorescence imaging given their pivotal role in numerous important biological events, including the development and progression of many diseases. Therefore, the development of new fluorescent receptors and binding agents for carbohydrates is and will be increasing in demand. This review highlights the development of fluorescence imaging agents based on boronic acids a particularly promising class of receptors given their strong and selective binding with carbohydrates in aqueous media.

  11. Averaged subtracted polarization imaging for endoscopic diagnostics of surface microstructures on translucent mucosae

    Science.gov (United States)

    Kanamori, Katsuhiro

    2016-07-01

    An endoscopic image processing technique for enhancing the appearance of microstructures on translucent mucosae is described. This technique employs two pairs of co- and cross-polarization images under two different linearly polarized lights, from which the averaged subtracted polarization image (AVSPI) is calculated. Experiments were then conducted using an acrylic phantom and excised porcine stomach tissue using a manual experimental setup with ring-type lighting, two rotating polarizers, and a color camera; better results were achieved with the proposed method than with conventional color intensity image processing. An objective evaluation method that uses texture analysis was developed and used to evaluate the enhanced microstructure images. This paper introduces two types of online, rigid-type, polarimetric endoscopic implementations using a polarized ring-shaped LED and a polarimetric camera. The first type uses a beam-splitter-type color polarimetric camera, and the second uses a single-chip monochrome polarimetric camera. Microstructures on the mucosa surface were enhanced robustly with these online endoscopes regardless of the difference in the extinction ratio of each device. These results show that polarimetric endoscopy using AVSPI is both effective and practical for hardware implementation.

  12. Quality Index for Stereoscopic Images by Separately Evaluating Adding and Subtracting.

    Directory of Open Access Journals (Sweden)

    Jiachen Yang

    Full Text Available The human visual system (HVS plays an important role in stereo image quality perception. Therefore, it has aroused many people's interest in how to take advantage of the knowledge of the visual perception in image quality assessment models. This paper proposes a full-reference metric for quality assessment of stereoscopic images based on the binocular difference channel and binocular summation channel. For a stereo pair, the binocular summation map and binocular difference map are computed first by adding and subtracting the left image and right image. Then the binocular summation is decoupled into two parts, namely additive impairments and detail losses. The quality of binocular summation is obtained as the adaptive combination of the quality of detail losses and additive impairments. The quality of binocular summation is computed by using the Contrast Sensitivity Function (CSF and weighted multi-scale (MS-SSIM. Finally, the quality of binocular summation and binocular difference is integrated into an overall quality index. The experimental results indicate that compared with existing metrics, the proposed metric is highly consistent with the subjective quality assessment and is a robust measure. The result have also indirectly proved hypothesis of the existence of binocular summation and binocular difference channels.

  13. Fluorescence goggle for intraoperative breast cancer imaging

    Science.gov (United States)

    Liu, Yang; Bauer, Adam Q.; Akers, Walter; Sudlow, Gail; Liang, Kexian; Charanya, Tauseef; Mondal, Suman; Culver, Joseph P.; Achilefu, Samuel

    2012-03-01

    We have developed a fluorescence goggle device for intraoperative oncologic imaging. With our system design, the surgeon can directly visualize the fluorescence information from the eyepieces in real time without any additional monitor, which can improve one's coordination and surgical accuracy. In conjunction with targeting fluorescent dyes, the goggle device can successfully detect tumor margins and small nodules that are not obvious to naked eye. This can potentially decrease the incidence of incomplete resection.

  14. High speed multispectral fluorescence lifetime imaging

    NARCIS (Netherlands)

    Fereidouni, F.; Reitsma, K.; Gerritsen, H.C.

    2013-01-01

    We report a spectrally resolved fluorescence lifetime imaging system based on time gated single photon detection with a fixed gate width of 200 ps and 7 spectral channels. Time gated systems can operate at high count rates but usually have large gate widths and sample only part of the fluorescence d

  15. Enhancing in vivo tumor boundary delineation with structured illumination fluorescence molecular imaging and spatial gradient mapping

    Science.gov (United States)

    Sun, Jessica; Miller, Jessica P.; Hathi, Deep; Zhou, Haiying; Achilefu, Samuel; Shokeen, Monica; Akers, Walter J.

    2016-08-01

    Fluorescence imaging, in combination with tumor-avid near-infrared (NIR) fluorescent molecular probes, provides high specificity and sensitivity for cancer detection in preclinical animal models, and more recently, assistance during oncologic surgery. However, conventional camera-based fluorescence imaging techniques are heavily surface-weighted such that surface reflection from skin or other nontumor tissue and nonspecific fluorescence signals dominate, obscuring true cancer-specific signals and blurring tumor boundaries. To address this challenge, we applied structured illumination fluorescence molecular imaging (SIFMI) in live animals for automated subtraction of nonspecific surface signals to better delineate accumulation of an NIR fluorescent probe targeting α4β1 integrin in mice bearing subcutaneous plasma cell xenografts. SIFMI demonstrated a fivefold improvement in tumor-to-background contrast when compared with other full-field fluorescence imaging methods and required significantly reduced scanning time compared with diffuse optical spectroscopy imaging. Furthermore, the spatial gradient mapping enhanced highlighting of tumor boundaries. Through the relatively simple hardware and software modifications described, SIFMI can be integrated with clinical fluorescence imaging systems, enhancing intraoperative tumor boundary delineation from the uninvolved tissue.

  16. A computer-aided diagnosis system to detect pathologies in temporal subtraction images of chest radiographs

    Science.gov (United States)

    Looper, Jared; Harrison, Melanie; Armato, Samuel G.

    2016-03-01

    Radiologists often compare sequential radiographs to identify areas of pathologic change; however, this process is prone to error, as human anatomy can obscure the regions of change, causing the radiologists to overlook pathology. Temporal subtraction (TS) images can provide enhanced visualization of regions of change in sequential radiographs and allow radiologists to better detect areas of change in radiographs. Not all areas of change shown in TS images, however, are actual pathology. The purpose of this study was to create a computer-aided diagnostic (CAD) system that identifies which regions of change are caused by pathology and which are caused by misregistration of the radiographs used to create the TS image. The dataset used in this study contained 120 images with 74 pathologic regions on 54 images outlined by an experienced radiologist. High and low ("light" and "dark") gray-level candidate regions were extracted from the images using gray-level thresholding. Then, sampling techniques were used to address the class imbalance problem between "true" and "false" candidate regions. Next, the datasets of light candidate regions, dark candidate regions, and the combined set of light and dark candidate regions were used as training and testing data for classifiers by using five-fold cross validation. Of the classifiers tested (support vector machines, discriminant analyses, logistic regression, and k-nearest neighbors), the support vector machine on the combined candidates using synthetic minority oversampling technique (SMOTE) performed best with an area under the receiver operating characteristic curve value of 0.85, a sensitivity of 85%, and a specificity of 84%.

  17. Lung iodine mapping by subtraction with image registration allowing for tissue sliding

    Science.gov (United States)

    Mohr, Brian; Brink, Monique; Oostveen, Luuk J.; Schuijf, Joanne D.; Prokop, Mathias

    2016-03-01

    Pulmonary embolism is a fairly common and serious entity, so rapid diagnosis and treatment has a significant impact on morbidity and mortality rates. Iodine maps representing tissue perfusion enhancement are commonly generated by dual-energy CT acquisitions to provide information about the effect of the embolism on pulmonary perfusion. Alternatively, the iodine map can be generated by subtracting pre- from post-contrast CT scans as previously reported. Although accurate image registration is essential, subtraction has the advantage of a higher signal-to-noise ratio and suppression of bone. This paper presents an improvement over the previously reported registration algorithm. Significantly, allowance for sliding motion at tissue boundaries is included in the regularization. Pre- and post-contrast helical CT scans were acquired for thirty subjects using a Toshiba Aquilion ONE scanner. Ten of these subjects were designated for algorithm development, while the remaining twenty were reserved for qualitative clinical evaluation. Quantitative evaluation was performed against the previously reported method and using publicly available data for comparison against other methods. Comparison of 100 landmarks in seven datasets shows no change in the mean Euclidean error of 0.48 mm, compared to the previous method. Evaluation in the publicly available DIR-Lab data with 300 annotations results in a mean Euclidean error of 1.17 mm in the ten 4DCT cases and 3.37 mm in the ten COPDGene cases. Clinical evaluation on a sliding scale from 1 (excellent) to 5 (non-diagnostic) indicates a slight, but non-significant, improvement in registration adequacy from 3.1 to 2.9.

  18. Subtraction MRI versus diffusion weighted imaging: Which is more accurate in assessment of hepatocellular carcinoma after Trans Arterial Chemoembolization (TACE?

    Directory of Open Access Journals (Sweden)

    Noha Abd ElShafy ElSaid

    2016-12-01

    Full Text Available This study aims to compare the role of both subtraction MRI and Diffusion weighted images in assessment of HCC after trans-arterial chemo embolization (TACE. Patients and methods: 32 patients having 54 HCC lesions underwent TACE. Ages ranged between 59 and 73 years (mean age 59.6; there were 26 males and 6 females. All examinations were performed using Philips 1.5 T MRI (Achieva. Pre contrast: T1, T2W and In phase and out phase gradient echo sequence, dynamic: eTHRIVE technique performed. Subtraction of an unenhanced T1-weighted was from the identical post enhancement sequences in early angiographic and late arterial phases. Diffusion imaging: it was performed using single-shot spin-echo echo-planar imaging with b = 0, 300, 600 mm/s2. DWI was used to create ADC maps. Two readers blinded to each other assessed subtraction of dynamic study and DWI technique to evaluate post treatment response. Results: Reader 1 subtraction dynamic MRI sensitivity = 97%, specificity = 100% PPV = 100% and NPV = 95% compared to 70.59%, 75%, 82.76% and 60% respectively in DWI. Reader 2 subtraction dynamic MRI sensitivity = 97%, specificity = 100% PPV = 100% and NPV = 95% compared to 76.5%, 90%, 92.8% and 69% respectively in DWI. Conclusion: DW-MRI had lower accuracy measures compared to subtraction MRI with increased false negative. DWI may act as a supplementary sequence to compensate for the dynamic MRI in patients who couldn't hold their breath adequately.

  19. Imaging an atomic beam using fluorescence

    Institute of Scientific and Technical Information of China (English)

    Ming He(何明); Jin Wang(王谨); Mingsheng Zhan(詹明生)

    2003-01-01

    A fluorescence detection scheme is applied to image an atomic beam. Using two laser diodes as the sources of detection light and pumping light respectively, the fluorescence image of the atomic beam is then observed by a commercial CCD-camera, which is corresponding to the atomic state and velocity distribution. The detection scheme has a great utilization in the experiments of cold atoms and atomic optics.

  20. Cancer detection by quantitative fluorescence image analysis.

    Science.gov (United States)

    Parry, W L; Hemstreet, G P

    1988-02-01

    Quantitative fluorescence image analysis is a rapidly evolving biophysical cytochemical technology with the potential for multiple clinical and basic research applications. We report the application of this technique for bladder cancer detection and discuss its potential usefulness as an adjunct to methods used currently by urologists for the diagnosis and management of bladder cancer. Quantitative fluorescence image analysis is a cytological method that incorporates 2 diagnostic techniques, quantitation of nuclear deoxyribonucleic acid and morphometric analysis, in a single semiautomated system to facilitate the identification of rare events, that is individual cancer cells. When compared to routine cytopathology for detection of bladder cancer in symptomatic patients, quantitative fluorescence image analysis demonstrated greater sensitivity (76 versus 33 per cent) for the detection of low grade transitional cell carcinoma. The specificity of quantitative fluorescence image analysis in a small control group was 94 per cent and with the manual method for quantitation of absolute nuclear fluorescence intensity in the screening of high risk asymptomatic subjects the specificity was 96.7 per cent. The more familiar flow cytometry is another fluorescence technique for measurement of nuclear deoxyribonucleic acid. However, rather than identifying individual cancer cells, flow cytometry identifies cellular pattern distributions, that is the ratio of normal to abnormal cells. Numerous studies by others have shown that flow cytometry is a sensitive method to monitor patients with diagnosed urological disease. Based upon results in separate quantitative fluorescence image analysis and flow cytometry studies, it appears that these 2 fluorescence techniques may be complementary tools for urological screening, diagnosis and management, and that they also may be useful separately or in combination to elucidate the oncogenic process, determine the biological potential of tumors

  1. Cubosomes for in vivo fluorescence lifetime imaging

    Science.gov (United States)

    Biffi, Stefania; Andolfi, Laura; Caltagirone, Claudia; Garrovo, Chiara; Falchi, Angela M.; Lippolis, Vito; Lorenzon, Andrea; Macor, Paolo; Meli, Valeria; Monduzzi, Maura; Obiols-Rabasa, Marc; Petrizza, Luca; Prodi, Luca; Rosa, Antonella; Schmidt, Judith; Talmon, Yeshayahu; Murgia, Sergio

    2017-02-01

    Herein we provided the first proof of principle for in vivo fluorescence optical imaging application using monoolein-based cubosomes in a healthy mouse animal model. This formulation, administered at a non-cytotoxic concentration, was capable of providing both exogenous contrast for NIR fluorescence imaging with very high efficiency and chemospecific information upon lifetime analysis. Time-resolved measurements of fluorescence after the intravenous injection of cubosomes revealed that the dye rapidly accumulated mainly in the liver, while lifetimes profiles obtained in vivo allowed for discriminating between free dye or dye embedded within the cubosome nanostructure after injection.

  2. Cubosomes for in vivo fluorescence lifetime imaging.

    Science.gov (United States)

    Biffi, Stefania; Andolfi, Laura; Caltagirone, Claudia; Garrovo, Chiara; Falchi, Angela M; Lippolis, Vito; Lorenzon, Andrea; Macor, Paolo; Meli, Valeria; Monduzzi, Maura; Obiols-Rabasa, Marc; Petrizza, Luca; Prodi, Luca; Rosa, Antonella; Schmidt, Judith; Talmon, Yeshayahu; Murgia, Sergio

    2017-02-03

    Herein we provided the first proof of principle for in vivo fluorescence optical imaging application using monoolein-based cubosomes in a healthy mouse animal model. This formulation, administered at a non-cytotoxic concentration, was capable of providing both exogenous contrast for NIR fluorescence imaging with very high efficiency and chemospecific information upon lifetime analysis. Time-resolved measurements of fluorescence after the intravenous injection of cubosomes revealed that the dye rapidly accumulated mainly in the liver, while lifetimes profiles obtained in vivo allowed for discriminating between free dye or dye embedded within the cubosome nanostructure after injection.

  3. K-edge digital subtraction imaging with dichromatic x-ray sources: SNR and dose studies

    Science.gov (United States)

    Sarnelli, A.; Elleaume, H.; Taibi, A.; Gambaccini, M.; Bravin, A.

    2006-09-01

    The aim of the present work is to analytically evaluate the signal to noise ratio (SNR) and the delivered dose in K-edge digital subtraction imaging (KES) using two types of x-ray sources: a monochromatic x-ray source (available at synchrotron radiation facilities and considered as gold standard) and a quasi-monochromatic compact source. The energy separation ΔE between the two monochromatic beams is 1 keV and 4 keV for the two sources, respectively. The evaluation has been performed for both radiography and computed tomography. Different geometries have been studied to mimic clinical situations. In mammography, a pathology perfused by a contrast agent has been modelled; in angiography, a vessel superimposed to a ventricle or a stand-alone artery stenosis has been studied. The SNR and the skin dose have been calculated as a function of the detail diameter, the contrast agent (iodine and gadolinium), and its concentration in the tissues. Results show that for ΔE = 4 keV a slightly higher delivered dose is required to obtain the same SNR with respect to ΔE < 1 keV. A similar study has been performed for KES-CT. Computer simulations of CT images performed with Snark software are shown to validate the analytical calculations.

  4. Multiparameter fluorescence spectroscopic imaging of cell function

    Science.gov (United States)

    Bright, Gary R.

    1994-08-01

    The ability to quantitate physiological parameters in single living cells using fluorescence spectroscopic imaging has expanded our understanding of many cell regulatory processes. Previous studies have focussed on the measurement of single parameters, such as the concentration of calcium, and more recently two parameters, such as calcium and pH using fluorescence ratio imaging. The complexity of the interrelationships among cell biochemical reactions suggests a need to extend the measurement scheme to several parameters. Expansion of the number of parameters involves several complexities associated with fluorescent probe selection and instrumentation design as well as the processing and management of the data. A system has been assembled which provides maximum flexibility in multiparameter fluorescence imaging measurements. The system provides multiple combinations of excitation, dichroic mirror, and emission wavelengths. It has automatic acquisition of any number of parameters. The number of parameters is primarily limited by the selection of fluorescent probes with nonoverlapping spectra. We demonstrate the utility of the system by the coordinated monitoring of stimulated changes in the concentrations of calcium, magnesium, and pH using fluorescence ratio imaging coupled with a conventional transmitted light image of single smooth muscle cells. The results demonstrate coordinated changes in some instances but uncoordinated changes in others.

  5. Subtractive Leadership

    Science.gov (United States)

    Larwin, K. H.; Thomas, Eugene M.; Larwin, David A.

    2015-01-01

    This paper introduces a new term and concept to the leadership discourse: Subtractive Leadership. As an extension of the distributive leadership model, the notion of subtractive leadership refers to a leadership style that detracts from organizational culture and productivity. Subtractive leadership fails to embrace and balance the characteristics…

  6. Hyperspectral Fluorescence and Reflectance Imaging Instrument

    Science.gov (United States)

    Ryan, Robert E.; O'Neal, S. Duane; Lanoue, Mark; Russell, Jeffrey

    2008-01-01

    The system is a single hyperspectral imaging instrument that has the unique capability to acquire both fluorescence and reflectance high-spatial-resolution data that is inherently spatially and spectrally registered. Potential uses of this instrument include plant stress monitoring, counterfeit document detection, biomedical imaging, forensic imaging, and general materials identification. Until now, reflectance and fluorescence spectral imaging have been performed by separate instruments. Neither a reflectance spectral image nor a fluorescence spectral image alone yields as much information about a target surface as does a combination of the two modalities. Before this system was developed, to benefit from this combination, analysts needed to perform time-consuming post-processing efforts to co-register the reflective and fluorescence information. With this instrument, the inherent spatial and spectral registration of the reflectance and fluorescence images minimizes the need for this post-processing step. The main challenge for this technology is to detect the fluorescence signal in the presence of a much stronger reflectance signal. To meet this challenge, the instrument modulates artificial light sources from ultraviolet through the visible to the near-infrared part of the spectrum; in this way, both the reflective and fluorescence signals can be measured through differencing processes to optimize fluorescence and reflectance spectra as needed. The main functional components of the instrument are a hyperspectral imager, an illumination system, and an image-plane scanner. The hyperspectral imager is a one-dimensional (line) imaging spectrometer that includes a spectrally dispersive element and a two-dimensional focal plane detector array. The spectral range of the current imaging spectrometer is between 400 to 1,000 nm, and the wavelength resolution is approximately 3 nm. The illumination system consists of narrowband blue, ultraviolet, and other discrete

  7. Multiphoton fluorescence lifetime imaging of human hair.

    Science.gov (United States)

    Ehlers, Alexander; Riemann, Iris; Stark, Martin; König, Karsten

    2007-02-01

    In vivo and in vitro multiphoton imaging was used to perform high resolution optical sectioning of human hair by nonlinear excitation of endogenous as well as exogenous fluorophores. Multiphoton fluorescence lifetime imaging (FLIM) based on time-resolved single photon counting and near-infrared femtosecond laser pulse excitation was employed to analyze the various fluorescent hair components. Time-resolved multiphoton imaging of intratissue pigments has the potential (i) to identify endogenous keratin and melanin, (ii) to obtain information on intrahair dye accumulation, (iii) to study bleaching effects, and (iv) to monitor the intratissue diffusion of pharmaceutical and cosmetical components along hair shafts.

  8. Fluorescence imaging of soybean flavonol isolines

    Science.gov (United States)

    Kim, Moon S.; Lee, Edward H.; Mulchi, Charles L.; McMurtrey, James E., III; Chappelle, Emmett W.; Rowland, Randy A.

    1998-07-01

    Experiments were conducted to characterize the fluorescence emission of leaves from four soybean ('Harosoy') plants containing different concentrations of flavonols (kaempferol glycosides). The investigation utilized genetically mutated soybean flavonol isolines grown in a constant environment, thus limiting factors known to affect fluorescence emission characteristics other than different kaempferol glycosides concentrations. Flavonol isolines included OX922, OX941, OX942, OX944. The first two isolines contain kaempferol (K) glycosides; K3, K6, and K9, and the latter two did not have K3, K6, and K9. A fluorescence imaging system (FIS) was used to characterize steady state florescence images of the sample leaves measured at wavelengths centered at 450, 550, 680, and 740 nm with an excitation at 360 nm. Images taken with FIS greatly complement non-imaging fluorescence measurements by characterizing the spatial variation of fluorescence within leaves. We also acquired fluorescence emission spectra to characterize spectral features of the soybean flavonol isolines. The emission spectral shape of the fluorescence emission characteristics were not significantly different between the soybeans that contain kaempferol glycosides and the ones that do not contain kaempferol glycosides. Typical emission maxima of green vegetation in the blue, green, red, and far-red bands were noticed in all four soybean isolines. However, plants containing kaempferol glycosides, OX922 and OX941 had significantly lower intensities throughout the wavelength regions. These results imply that fluorescence emission intensities in the fluorescence emission bands studied are significantly affected by the presence and absence of kaempferol glycosides concentrations (UV radiation screening compounds). Pure kaempferol glycoside dissolved in solution show minimal fluorescence emission when excited with the absorption maximum radiation at 365 nm. However, a broad band emission can be seen in the green

  9. Value of subtraction MRI in assessing treatment response following image-guided loco-regional therapies for hepatocellular carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Winters, S.D. [Department of Radiology and Diagnostic Imaging, Royal Alexandra Hospital, Edmonton (Canada); Department of Radiology and Diagnostic Imaging, University of Alberta Hospital, Edmonton (Canada); 1 Canadian Field Hospital, Department of National Defence, Government of Canada (Canada); Jackson, S. [Department of Radiology and Diagnostic Imaging, University of Alberta Hospital, Edmonton (Canada); Armstrong, G.A. [Department of Radiology and Diagnostic Imaging, Royal Alexandra Hospital, Edmonton (Canada); Department of Radiology and Diagnostic Imaging, University of Alberta Hospital, Edmonton (Canada); Birchall, I.W. [Department of Radiology and Diagnostic Imaging, University of Alberta Hospital, Edmonton (Canada); Lee, K.H.Y. [Department of Radiology and Diagnostic Imaging, Royal Alexandra Hospital, Edmonton (Canada); Low, G., E-mail: timgy@yahoo.com [Department of Radiology and Diagnostic Imaging, University of Alberta Hospital, Edmonton (Canada)

    2012-07-15

    Aim: To compare contrast-enhanced subtraction magnetic resonance imaging (MRI) with contrast-enhanced standard MRI in assessing treatment response following loco-regional therapies for hepatocellular carcinoma (HCC). Method and materials: Institutional review board approval was obtained and informed consent was waived for this retrospective study. All patients were analysed from our institution's liver tumour database that had loco-regional HCC therapy and the following: (1) a contrast-enhanced MRI {<=}6 weeks post-treatment, (2) an unenhanced T1-weighted high-signal treatment zone (TZ) {>=}1 cm, (3) follow-up contrast-enhanced MRI performed {>=}6 months post-treatment. Randomized standard and subtraction TZ datasets were independently assessed by three blinded radiology readers for either complete treatment necrosis or residual disease. The standard of reference (SOR) comprised a consensus read by two radiologists with knowledge of the follow-up MRI and all available clinical data. Statistical analyses were performed using receiver operating characteristics (ROC), t-test, and kappa statistic. Results: Twenty-six patients (19 male and seven female patients; mean age 60 years, standard deviation 10.9 years, range 46-88 years) had a total of 45 corresponding HCCs and TZs. For ROC, the area under the curve (AUC) was 0.93 (subtraction protocol) versus 0.90 (standard protocol; p = 0.49). For the t-test, the mean reader confidence level was 4.4, 3.6, and 4.4 (subtraction protocol) versus 3, 3, and 3.7 (standard protocol; p {<=} 0.011). The kappa statistic for reader-to-SOR agreement was 0.83, 0.63, and 0.71 (subtraction protocol) versus 0.51, 0.36, and 0.64 (standard protocol). Conclusion: Subtraction MRI significantly improves the reader confidence level in the assessment of treatment response following loco-regional therapies for HCC.

  10. Optical image encryption based on real-valued coding and subtracting with the help of QR code

    Science.gov (United States)

    Deng, Xiaopeng

    2015-08-01

    A novel optical image encryption based on real-valued coding and subtracting is proposed with the help of quick response (QR) code. In the encryption process, the original image to be encoded is firstly transformed into the corresponding QR code, and then the corresponding QR code is encoded into two phase-only masks (POMs) by using basic vector operations. Finally, the absolute values of the real or imaginary parts of the two POMs are chosen as the ciphertexts. In decryption process, the QR code can be approximately restored by recording the intensity of the subtraction between the ciphertexts, and hence the original image can be retrieved without any quality loss by scanning the restored QR code with a smartphone. Simulation results and actual smartphone collected results show that the method is feasible and has strong tolerance to noise, phase difference and ratio between intensities of the two decryption light beams.

  11. Image-Subtraction Photometry of Variable Stars in the Field of the Globular Cluster NGC 6934

    CERN Document Server

    Kaluzny, J; Stanek, K Z

    2001-01-01

    (Abriged) We present CCD BVI photometry of 85 variable stars from the field of the globular cluster NGC 6934. The photometry was obtained with the image subtraction package ISIS. 35 variables are new identifications: 24 RRab stars, 5 RRc stars, 2 eclipsing binaries of W UMa-type, one SX Phe star, and 3 variables of other types. Both detected contact binaries are foreground stars. The SX Phe variable belongs most likely to the group of cluster blue stragglers. Large number of newly found RR Lyr variables in this cluster, as well as in other clusters recently observed by us, indicates that total RR Lyr population identified up to date in nearby galactic globular clusters is significantly (>30%) incomplete. Fourier decomposition of the light curves of RRc variables was used to estimate the basic properties of these stars: M=0.63 M_odot, log(L/L_odot)=1.72, T_eff=7300 and Y=0.27. Using RRab variables we obtain: M_V=0.81, [Fe/H]=-1.53 and T_eff=6450. From the B-V color at minimum light of the RRab variables we obt...

  12. Laser-induced fluorescence imaging of bacteria

    Science.gov (United States)

    Hilton, Peter J.

    1998-12-01

    This paper outlines a method for optically detecting bacteria on various backgrounds, such as meat, by imaging their laser induced auto-fluorescence response. This method can potentially operate in real-time, which is many times faster than current bacterial detection methods, which require culturing of bacterial samples. This paper describes the imaging technique employed whereby a laser spot is scanned across an object while capturing, filtering, and digitizing the returned light. Preliminary results of the bacterial auto-fluorescence are reported and plans for future research are discussed. The results to date are encouraging with six of the eight bacterial strains investigated exhibiting auto-fluorescence when excited at 488 nm. Discrimination of these bacterial strains against red meat is shown and techniques for reducing background fluorescence discussed.

  13. Technique of Hadamard transform microscope fluorescence image analysis

    Institute of Scientific and Technical Information of China (English)

    梅二文; 顾文芳; 曾晓斌; 陈观铨; 曾云鹗

    1995-01-01

    Hadamard transform spatial multiplexed imaging technique is combined with fluorescence microscope and an instrument of Hadamard transform microscope fluorescence image analysis is developed. Images acquired by this instrument can provide a lot of useful information simultaneously, including three-dimensional Hadamard transform microscope cell fluorescence image, the fluorescence intensity and fluorescence distribution of a cell, the background signal intensity and the signal/noise ratio, etc.

  14. Evaluation of the rockburst potential in longwall coal mining using passive seismic velocity tomography and image subtraction technique

    Science.gov (United States)

    Hosseini, Navid

    2017-09-01

    Rockburst is a typical dynamic disaster in underground coal mines which its occurrences relate to the mechanical quality of coal seam and surrounding rock mass and also the condition of stress distribution. The main aim of this paper is to study the potential of rockburst in a longwall coal mine by using of passive seismic velocity tomography and image subtraction technique. For this purpose, first by mounting an array of receivers on the surface above the active panel, the mining-induced seismic data as a passive source for several continuous days were recorded. Then, the three-dimensional tomograms using simultaneous iteration reconstruction technique (SIRT) for each day are created and by employing the velocity filtering, the overstressed zones are detected. In addition, the two-dimensional seismic velocity tomograms in coal seam level by slicing the three-dimensional tomograms are obtained. Then the state of stress changes in successive days by applying the image subtraction technique on these two-dimensional tomograms is considered. The results show that the compilation of filtered three-dimensional tomograms and subtracted images is an appropriate approach for detecting the overstressed zones around the panel and subsequent evaluation of rockburst potential. The research conclusion proves that the applied approach in this study in combination with field observations of rock mass status can effectively identify the rockburst-prone areas during the mining operation and help to improve the safety condition.

  15. Projection-based energy weighting on photon-counting X-ray images in digital subtraction mammography: a feasibility study

    Science.gov (United States)

    Choi, Sung-Hoon; Lee, Seung-Wan; Choi, Yu-Na; Lee, Young-Jin; Kim, Hee-Joung

    2014-03-01

    In digital subtraction mammography where subtracts the one image (with contrast medium) from the other (anatomical background) for observing the tumor structure, tumors which include more blood vessels than normal tissue could be distinguished through the enhancement of contrast-to-noise ratio (CNR). In order to improve CNR, we adopted projection-based energy weighting for iodine solutions with four different concentrations embedded in a breast phantom (50% adipose and 50% glandular tissues). In this study, a Monte Carlo simulation was used to simulate a 40 mm thickness breast phantom, which has 15 and 30 mg/cm3 iodine solutions with two different thicknesses, and an energy resolving photon-counting system. The input energy spectrum was simulated in a range of 20 to 45 keV in order to reject electronic noise and include k-edge energy of iodine (33.2 keV). The results showed that the projection-based energy weighting improved the CNR by factors of 1.05-1.86 compared to the conventional integrating images. Consequently, the CNR of images from the digital subtraction mammography could be improved by the projection-based energy weighting with photon-counting detectors.

  16. Fluorescein Derivatives in Intravital Fluorescence Imaging

    Directory of Open Access Journals (Sweden)

    Michael S. Roberts

    2013-08-01

    Full Text Available Intravital fluorescence microscopy enables the direct imaging of fluorophores in vivo and advanced techniques such as fluorescence lifetime imaging (FLIM enable the simultaneous detection of multiple fluorophores. Consequently, it is now possible to record distribution and metabolism of a chemical in vivo and to optimise the delivery of fluorophores in vivo. Recent clinical applications with fluorescein and other intravital fluorescent stains have occurred in neurosurgery, dermatology [including photodynamic therapy (PDT] and endomicroscopy. Potential uses have been identified in periodontal disease, skin graft and cancer surgery. Animal studies have demonstrated that diseased tissue can be specifically stained with fluorophore conjugates. This review focuses on the fluorescein derived fluorophores in common clinical use and provides examples of novel applications from studies in tissue samples.

  17. Developing an imaging bi-spectrometer for fluorescent materials

    Science.gov (United States)

    Mohammadi, Mahnaz

    Fluorescent effects have been observed for thousands of years. Stokes, in 1852, began the science of fluorescence culminating in his law of fluorescence, which explained that fluorescence emission occurs at longer wavelengths than the excitation wavelength. This phenomenon is observed extensively in the art world. Daylight fluorescent colors known as Day-GloRTM have become an artistic medium since the 1960s. Modern artists exploit these saturated and brilliant colors to glitter their painting. Multipsectral imaging as a noninvasive technique has been used for archiving by museums and cultural-heritage institutions for about a decade. The complex fluorescence phenomenon has been often ignored in the multispectral projects. The ignored fluorescence results in errors in digital imaging of artwork containing fluorescent colors. The illuminant-dependency of the fluorescence radiance makes the fluorescence colorimetry and consequently spectral imaging more complex. In this dissertation an abridged imaging bi-spectrometer for artwork containing both fluorescent and non-fluorescent colors was developed. The method developed included two stages of reconstruction of the spectral reflected radiance factor and prediction of the fluorescent radiance factor. The estimation of the reflected radiance factor as a light source independent component was achieved by imaging with a series of short-wavelength cutoff filters placed in the illumination path. The fluorescent radiance factor, a light source dependent component, was estimated based on a proposed model, the abridged two-monochromator method. The abridged two-monochromator method was developed for reconstructing the bi-spectral matrix of a fluorescent color based on a calibrated UV-fluorescence imaging. In this way, one could predict the fluorescence radiance factor under any desired illuminant and consequently a better color evaluation and rendering could be obtained. Furthermore, this method easily fitted in a general system

  18. [Examination of reducing misregistration for lower tube voltage of the mask image in CT angiography using subtraction method].

    Science.gov (United States)

    Nakatani, Kasumi; Fukunishi, Yasunobu

    2015-05-01

    Computed tomographic angiography (CTA) has been used recently for the evaluation of intracerebral aneurysms, but it is difficult to use this technique to visualize aneurysms near the base of the skull because of the presence of bone. So, subtracted CTA has been used to separate vessels from bony structures. However, we see some misregistration when using subtraction method because of the patient moving, the disaccord of the X-ray tube orbit between the mask image and the live image, the expanding focus, and the bed bending. So, attentioning the difference of bone CT number in any tube voltages, we examined to make the image containing less misregistration by changing the tube voltage of mask image. Making a sham blood vessel, we examined the bone misregistration, the blood vessel volume, and the smoothness when changing the tube voltages of mask images. Comparing with 120 kV, as the tube voltage of the mask image was 80 kV, the bone misregistration decreased significantly, however the blood vessel volume decreased. As for the tube voltage of 100 kV, the bone misregistration decreased significantly, and the blood vessel volume and the smoothness were not significantly different so we could get coordinative image of 120 kV. When the tube voltage of the mask image becomes lower than that of the live image and the effective energy becomes different, the effect of misregistration is less. This method deals with changing the tube voltage only. So, it may be easy to make volume rendering (VR) image and this method may be used in every facility.

  19. NOTE: Scatter-to-primary based scatter fractions for transmission-dependent convolution subtraction of SPECT images

    Science.gov (United States)

    Larsson, Anne; Johansson, Lennart

    2003-11-01

    In single photon emission computed tomography (SPECT), transmission-dependent convolution subtraction has been shown to be useful when correcting for scattered events. The method is based on convolution subtraction, but includes a matrix of scatter fractions instead of a global scatter fraction. The method can be extended to iteratively improve the scatter estimate, but in this note we show that this requires a modification of the theory to use scatter-to-total scatter fractions for the first iteration only and scatter-to-primary fractions thereafter. To demonstrate this, scatter correction is performed on a Monte Carlo simulated image of a point source of activity in water. The modification of the theory is compared to corrections where the scatter fractions are based on the scatter-to-total ratio, using one and ten iterations. The resulting ratios of subtracted to original counts are compared to the true scatter-to-total ratio of the simulation and the most accurate result is found for our modification of the theory.

  20. Predictors of Morbidity and Cleavage Plane in Surgical Resection of Pure Convexity Meningiomas Using Cerebrospinal Fluid Sensitive Image Subtraction Magnetic Resonance Imaging

    Science.gov (United States)

    THENIER-VILLA, José LUIS; CAMPOVERDE, Raúl ALEJANDRO GALÁRRAGA; DE LA LAMA ZARAGOZA, Adolfo RAMÓN; ALONSO, Cesáreo CONDE

    2017-01-01

    Meningiomas are the most common primary intracranial tumors. Since the adhesions in the plane of dissection are of interest in surgical planning, we suggest that digital image subtraction of FLAIR data from the T2 sequence of MRI may represent better the CSF spaces in the brain–tumor interface and may be a predictor of the intraoperative cleavage plane. From 2006 to 2016, 83 convexity meningiomas were resected in the Department of Neurosurgery of the University Hospital Complex of Vigo, an analysis of preoperative MRI was performed to assess peritumoral edema (PTE), tumor volume, among others; a digital subtraction of T2-FLAIR sequences was performed and analyzed in relationship to the cleavage plane described in the intraoperative report and postoperative neurological deficits. Simpson grade 1 resection was achieved in 85.54%, the overall 5-year PFS was 93.75%. Our rate of permanent new neurological deficit was 4.82% and the overall complication rate of 14.46%. The grade of PTE was proportional to tumor volume, 20 ± 2.8, 30 ± 5.3, and 34 ± 4.3 cm3 for grades 1, 2, and 3, respectively, positive cleft sign on image subtraction was predictive of good intraoperative cleavage plane and low grade cleavage plane (P = 0.04), and was a protective factor for postoperative neurological deficit (P = 0.02). Positive cleft sign in T2-FLAIR digital subtraction image is an independent predictor of good intraoperative cleavage plane, PTE is an independent predictor of the bad cleavage plane. Negative cleft sign in the image subtraction and a bad intraoperative cleavage plane are predictors of postoperative neurological deficit. PMID:27580930

  1. Intra-Arterial MR Perfusion Imaging of Meningiomas: Comparison to Digital Subtraction Angiography and Intravenous MR Perfusion Imaging

    Science.gov (United States)

    Martin, Alastair J.; Alexander, Matthew D.; McCoy, David B.; Cooke, Daniel L.; Lillaney, Prasheel; Moftakhar, Parham; Amans, Matthew R.; Settecase, Fabio; Nicholson, Andrew; Dowd, Christopher F.; Halbach, Van V.; Higashida, Randall T.; McDermott, Michael W.; Saloner, David; Hetts, Steven W.

    2016-01-01

    Background and Purpose To evaluate the ability of IA MR perfusion to characterize meningioma blood supply. Methods Studies were performed in a suite comprised of an x-ray angiography unit and 1.5T MR scanner that permitted intraprocedural patient movement between the imaging modalities. Patients underwent intra-arterial (IA) and intravenous (IV) T2* dynamic susceptibility MR perfusion immediately prior to meningioma embolization. Regional tumor arterial supply was characterized by digital subtraction angiography and classified as external carotid artery (ECA) dural, internal carotid artery (ICA) dural, or pial. MR perfusion data regions of interest (ROIs) were analyzed in regions with different vascular supply to extract peak height, full-width at half-maximum (FWHM), relative cerebral blood flow (rCBF), relative cerebral blood volume (rCBV), and mean transit time (MTT). Linear mixed modeling was used to identify perfusion curve parameter differences for each ROI for IA and IV MR imaging techniques. IA vs. IV perfusion parameters were also directly compared for each ROI using linear mixed modeling. Results 18 ROIs were analyzed in 12 patients. Arterial supply was identified as ECA dural (n = 11), ICA dural (n = 4), or pial (n = 3). FWHM, rCBV, and rCBF showed statistically significant differences between ROIs for IA MR perfusion. Peak Height and FWHM showed statistically significant differences between ROIs for IV MR perfusion. RCBV and MTT were significantly lower for IA perfusion in the Dural ECA compared to IV perfusion. Relative CBF in IA MR was found to be significantly higher in the Dural ICA region and MTT significantly lower compared to IV perfusion. PMID:27802268

  2. Fluorescence confocal endomicroscopy in biological imaging

    Science.gov (United States)

    Delaney, Peter; Thomas, Steven; Allen, John; McLaren, Wendy; Murr, Elise; Harris, Martin

    2007-02-01

    In vivo fluorescence microscopic imaging of biological systems in human disease states and animal models is possible with high optical resolution and mega pixel point-scanning performance using optimised off-the-shelf turn-key devices. There are however various trade-offs between tissue access and instrument performance when miniaturising in vivo microscopy systems. A miniature confocal scanning technology that was developed for clinical human endoscopy has been configured into a portable device for direct hand-held interrogation of living tissue in whole animal models (Optiscan FIVE-1 system). Scanning probes of 6.3mm diameter with a distal tip diameter of 5.0mm were constructed either in a 150mm length for accessible tissue, or a 300mm probe for laparoscopic interrogation of internal tissues in larger animal models. Both devices collect fluorescence confocal images (excitation 488 nm; emission >505 or >550 nm) comprised of 1024 x 1204 sampling points/image frame, with lateral resolution 0.7um; axial resolution 7um; FOV 475 x 475um. The operator can dynamically control imaging depth from the tissue surface to approx 250um in 4um steps via an internally integrated zaxis actuator. Further miniaturisation is achieved using an imaging contact probe based on scanning the proximal end of a high-density optical fibre bundle (~30,000 fibres) of sheep and pigs was fluorescently stained with calcein-AM or fluorescein. Surface and sub-surface cellular and sub-cellular details could be readily visualised in vivo at high resolution. In rodent disease models, in vivo endomicroscopy with appropriate fluorescent agents allowed examination of thrombosis formation, tumour microvasculature and liver metastases, diagnosis and staging of ulcerative colitis, liver necrosis and glomerulonephritis. Miniaturised confocal endomicroscopy allows rapid in vivo molecular and subsurface microscopy of normal and pathologic tissue at high resolution in small and large whole animal models

  3. Suppression of high-density artefacts in x-ray CT images using temporal digital subtraction with application to cryotherapy

    Energy Technology Data Exchange (ETDEWEB)

    Baissalov, R.; Sandison, G.A.; Rewcastle, J.C. [Department of Medical Physics, Tom Baker Cancer Center, Calgary, Canada, T2N 4N2 2 Department of Physics and Astronomy, University of Calgary, Calgary T2N 2N4 (Canada); Donnelly, B.J. [Department of Surgery, Tom Baker Cancer Center, Calgary, Canada, T2N 4N2 4 Department of Surgery, Foothills Hospital, Calgary T2N 2T7 (Canada); Saliken, J.C. [Department of Surgery, Tom Baker Cancer Center, Calgary T2N 4N2 (Canada); Department of Diagnostic Imaging, Foothills Hospital, Calgary T2N 2T7 (Canada); McKinnon, J.G. [Department of Surgery, Foothills Hospital, Calgary T2N 2T7 (Canada); Muldrew, K. [Department of Surgery, Faculty of Medicine, University of Calgary, Calgary T2N 2T7 (Canada)

    2000-05-01

    Image guidance in cryotherapy is usually performed using ultrasound. Although not currently in routine clinical use, x-ray CT imaging is an alternative means of guidance that can display the full 3D structure of the iceball, including frozen and unfrozen regions. However, the quality of x-ray CT images is compromised by the presence of high-density streak artefacts. To suppress these artefacts we applied temporal digital subtraction (TDS). This TDS method has the added advantage of improving the grey-scale contrast between frozen and unfrozen tissue in the CT images. Two sets of CT images were taken of a phantom material, cryoprobes and a urethral warmer (UW) before and during the cryoprobe freeze cycle. The high-density artefacts persisted in both image sets. TDS was performed on these two image sets using the corresponding mask image of unfrozen material and the same geometrical configuration of the cryoprobes and the UW. The resultant difference image had a significantly reduced artefact content. Thus TDS can be used to significantly suppress or eliminate high-density CT streak artefacts without reducing the metallic content of the cryoprobes. In vivo study needs to be conducted to establish the utility of this TDS procedure for CT assisted prostate or liver cryotherapy. Applying TDS in x-ray CT guided cryotherapy will facilitate estimation of the number and location of all frozen and unfrozen regions, potentially making cryotherapy safer and less operator dependent. (author)

  4. Suppression of high-density artifacts in x-ray CT images using temporal digital subtraction with application to cryotherapy

    Science.gov (United States)

    Baissalov, Roustem; Sandison, George A.; Donnelly, Bryan J.; Saliken, John C.; Muldrew, Ken; Rewcastle, John C.

    2000-06-01

    Image guidance of cryotherapy is usually performed using ultrasound or x-ray CT. Despite the ability of CT to display the full 3D structure of the iceball, including frozen and unfrozen regions, the quality of the images is compromised by the presence of high density streak artifacts. To suppress these artifacts we applied Temporal Digital Subtraction (TDS). This TDS method has the added advantage of improving the gray scale contrast between frozen and unfrozen tissue in the CT images. Two sets of CT images were taken of a phantom material, cryoprobes and a urethral warmer (UW) before and during the cryoprobe freeze cycle. The high density artifacts persisted in both image sets. TDS was performed on these two image sets using the corresponding mask image of unfrozen material and the same geometrical configuration of the cryoprobes and the UW. The resultant difference image had a significantly reduced content of the artifacts. This TDS can be used in x-ray CT assisted cryotherapy to significantly suppress or eliminate high density x-ray CT streak artifacts by digitally processing x-ray CT images. Applying TDS in cryotherapy will facilitate estimation of the amount and location of all frozen and unfrozen regions, potentially making cryotherapy safer and less operator dependent.

  5. Subtracted geometry

    Science.gov (United States)

    Saleem, Zain Hamid

    In this thesis we study a special class of black hole geometries called subtracted geometries. Subtracted geometry black holes are obtained when one omits certain terms from the warp factor of the metric of general charged rotating black holes. The omission of these terms allows one to write the wave equation of the black hole in a completely separable way and one can explicitly see that the wave equation of a massless scalar field in this slightly altered background of a general multi-charged rotating black hole acquires an SL(2, R) x SL(2, R) x SO(3) symmetry. The "subtracted limit" is considered an appropriate limit for studying the internal structure of the non-subtracted black holes because new 'subtracted' black holes have the same horizon area and periodicity of the angular and time coordinates in the near horizon regions as the original black hole geometry it was constructed from. The new geometry is asymptotically conical and is physically similar to that of a black hole in an asymptotically confining box. We use the different nice properties of these geometries to understand various classically and quantum mechanically important features of general charged rotating black holes.

  6. Assessing the use of an infrared spectrum hyperpixel array imager to measure temperature during additive and subtractive manufacturing

    Science.gov (United States)

    Whitenton, Eric; Heigel, Jarred; Lane, Brandon; Moylan, Shawn

    2016-05-01

    Accurate non-contact temperature measurement is important to optimize manufacturing processes. This applies to both additive (3D printing) and subtractive (material removal by machining) manufacturing. Performing accurate single wavelength thermography suffers numerous challenges. A potential alternative is hyperpixel array hyperspectral imaging. Focusing on metals, this paper discusses issues involved such as unknown or changing emissivity, inaccurate greybody assumptions, motion blur, and size of source effects. The algorithm which converts measured thermal spectra to emissivity and temperature uses a customized multistep non-linear equation solver to determine the best-fit emission curve. Emissivity dependence on wavelength may be assumed uniform or have a relationship typical for metals. The custom software displays residuals for intensity, temperature, and emissivity to gauge the correctness of the greybody assumption. Initial results are shown from a laser powder-bed fusion additive process, as well as a machining process. In addition, the effects of motion blur are analyzed, which occurs in both additive and subtractive manufacturing processes. In a laser powder-bed fusion additive process, the scanning laser causes the melt pool to move rapidly, causing a motion blur-like effect. In machining, measuring temperature of the rapidly moving chip is a desirable goal to develop and validate simulations of the cutting process. A moving slit target is imaged to characterize how the measured temperature values are affected by motion of a measured target.

  7. Continuum subtracting Lyman-alpha images: Low redshift studies using the Solar Blind Channel of HST/ACS

    CERN Document Server

    Hayes, Matthew; Mas-Hesse, J Miguel; Kunth, Daniel

    2008-01-01

    (Abridged) We are undertaking an imaging study of local star-forming galaxies in the Lyman-alpha (Lya) emission line using the Solar Blind Channel (SBC) of the ACS onboard HST. Observations have been obtained in Lya and H-alpha and six line-free continuum filters between ~1500 AA and the I-band. In a previous article Hayes et al. (2005) we demonstrated that the production of Lya line-only images (i.e. continuum subtraction) in the SBC-only data-set is non-trivial and that supporting data is a requirement. We here develop various methods of continuum subtraction and assess their relative performance for given input spectral energy distributions (SED). We show that simple assumptions about the behavior of the UV continuum consistently lead to results that are significantly in error, and determine that a spectral fitting approach is essential. Furthermore, fitting of a single component stellar or stellar+nebular spectrum is not always sufficient for realistic template SEDs and, in order to successfully recover t...

  8. Fluorescence lifetime imaging of oxygen in living cells

    NARCIS (Netherlands)

    Gerritsen, H.C.; Sanders, R.; Draaijer, A.; Ince, C.; Levine, Y.K.

    1997-01-01

    The usefulness of the fluorescent probe ruthenium tris(2,2′-dipyridyl) dichloride hydrate (RTDP) for the quantitative imaging of oxygen in single cells was investigated utilizing fluorescence life-time imaging. The results indicate that the fluorescence behavior of RTDP in the presence of oxygen can

  9. Creating Panoramic Images for Bladder Fluorescence Endoscopy

    Directory of Open Access Journals (Sweden)

    A. Behrens

    2008-01-01

    Full Text Available The medical diagnostic analysis and therapy of urinary bladder cancer based on endoscopes are state of the art in urological medicine. Due to the limited field of view of endoscopes, the physician can examine only a small part of the whole operating field at once. This constraint makes visual control and navigation difficult, especially in hollow organs. A panoramic image, covering a larger field of view, can overcome this difficulty. Directly motivated by a physician we developed an image mosaicing algorithm for endoscopic bladder fluorescence video sequences. In this paper, we present an approach which is capable of stitching single endoscopic video images to a combined panoramic image. Based on SIFT features we estimate a 2-D homography for each image pair, using an affine model and an iterative model-fitting algorithm. We then apply the stitching process and perform a mutual linear interpolation. Our panoramic image results show a correct stitching and lead to a better overview and understanding of the operation field. 

  10. Performance evaluation of spot detection algorithms in fluorescence microscopy images

    CSIR Research Space (South Africa)

    Mabaso, M

    2012-10-01

    Full Text Available Detection of messenger Ribonucleic Acid (mRNA) spots in fluorescence microscopy images is of great importance for biologists seeking better understanding of cell functionality. Fluorescence microscopy and specific staining methods make biological...

  11. In vivo macroscopic HPD fluorescence reflectance imaging on small animals bearing surface ARO/NPA tumor

    Science.gov (United States)

    Autiero, Maddalena; Celentano, Luigi; Laccetti, Paolo; Marotta, Marcello; Mettivier, Giovanni; Montesi, Maria C.; Riccio, Patrizia; Russo, Paolo; Roberti, Giuseppe

    2005-08-01

    Recently multimodal imaging systems have been devised because the combination of different imaging modalities results in the complementarity and integration of the techniques and in a consequent improvement of the diagnostic capabilities of the multimodal system with respect to each separate imaging modality. We developed a simple and reliable HematoPorphyrin (HP) mediated Fluorescence Reflectance Imaging (FRI) system that allows for in vivo real time imaging of surface tumors with a large field of view. The tumor cells are anaplastic human thyroid carcinoma-derived ARO cells, or human papillary thyroid carcinoma-derived NPA cells. Our measurements show that the optical contrast of the tumor region image is increased by a simple digital subtraction of the background fluorescence and that HP fluorescence emissivity of ARO tumors is about 2 times greater than that of NPA tumors, and about 4 times greater than that of healthy tissues. This is also confirmed by spectroscopic measurements on histological sections of tumor and healthy tissues. It was shown also the capability of this system to distinguish the tumor type on the basis of the different intensity of the fluorescence emission, probably related to the malignancy degree. The features of this system are complementary with those ones of a pixel radionuclide detection system, which allows for relatively time expensive, narrow field of view measurements, and applicability to tumors also deeply imbedded in tissues. The fluorescence detection could be used as a large scale and quick analysis tool and could be followed by narrow field, higher resolution radionuclide measurements on previously determined highly fluorescent regions.

  12. A Monte Carlo simulation study of an improved K-edge log-subtraction X-ray imaging using a photon counting CdTe detector

    Science.gov (United States)

    Lee, Youngjin; Lee, Amy Candy; Kim, Hee-Joung

    2016-09-01

    Recently, significant effort has been spent on the development of photons counting detector (PCD) based on a CdTe for applications in X-ray imaging system. The motivation of developing PCDs is higher image quality. Especially, the K-edge subtraction (KES) imaging technique using a PCD is able to improve image quality and useful for increasing the contrast resolution of a target material by utilizing contrast agent. Based on above-mentioned technique, we presented an idea for an improved K-edge log-subtraction (KELS) imaging technique. The KELS imaging technique based on the PCDs can be realized by using different subtraction energy width of the energy window. In this study, the effects of the KELS imaging technique and subtraction energy width of the energy window was investigated with respect to the contrast, standard deviation, and CNR with a Monte Carlo simulation. We simulated the PCD X-ray imaging system based on a CdTe and polymethylmethacrylate (PMMA) phantom which consists of the various iodine contrast agents. To acquired KELS images, images of the phantom using above and below the iodine contrast agent K-edge absorption energy (33.2 keV) have been acquired at different energy range. According to the results, the contrast and standard deviation were decreased, when subtraction energy width of the energy window is increased. Also, the CNR using a KELS imaging technique is higher than that of the images acquired by using whole energy range. Especially, the maximum differences of CNR between whole energy range and KELS images using a 1, 2, and 3 mm diameter iodine contrast agent were acquired 11.33, 8.73, and 8.29 times, respectively. Additionally, the optimum subtraction energy width of the energy window can be acquired at 5, 4, and 3 keV for the 1, 2, and 3 mm diameter iodine contrast agent, respectively. In conclusion, we successfully established an improved KELS imaging technique and optimized subtraction energy width of the energy window, and based on

  13. A Monte Carlo simulation study of an improved K-edge log-subtraction X-ray imaging using a photon counting CdTe detector

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Youngjin, E-mail: radioyoungj@gmail.com [Department of Radiological Science, Eulji University, 553, Sanseong-daero, Sujeong-gu, Seongnam-si, Gyeonggi-do (Korea, Republic of); Lee, Amy Candy [Department of Mathematics and Statistics, McGill University (Canada); Kim, Hee-Joung [Department of Radiological Science and Radiation Convergence Engineering, Yonsei University (Korea, Republic of)

    2016-09-11

    Recently, significant effort has been spent on the development of photons counting detector (PCD) based on a CdTe for applications in X-ray imaging system. The motivation of developing PCDs is higher image quality. Especially, the K-edge subtraction (KES) imaging technique using a PCD is able to improve image quality and useful for increasing the contrast resolution of a target material by utilizing contrast agent. Based on above-mentioned technique, we presented an idea for an improved K-edge log-subtraction (KELS) imaging technique. The KELS imaging technique based on the PCDs can be realized by using different subtraction energy width of the energy window. In this study, the effects of the KELS imaging technique and subtraction energy width of the energy window was investigated with respect to the contrast, standard deviation, and CNR with a Monte Carlo simulation. We simulated the PCD X-ray imaging system based on a CdTe and polymethylmethacrylate (PMMA) phantom which consists of the various iodine contrast agents. To acquired KELS images, images of the phantom using above and below the iodine contrast agent K-edge absorption energy (33.2 keV) have been acquired at different energy range. According to the results, the contrast and standard deviation were decreased, when subtraction energy width of the energy window is increased. Also, the CNR using a KELS imaging technique is higher than that of the images acquired by using whole energy range. Especially, the maximum differences of CNR between whole energy range and KELS images using a 1, 2, and 3 mm diameter iodine contrast agent were acquired 11.33, 8.73, and 8.29 times, respectively. Additionally, the optimum subtraction energy width of the energy window can be acquired at 5, 4, and 3 keV for the 1, 2, and 3 mm diameter iodine contrast agent, respectively. In conclusion, we successfully established an improved KELS imaging technique and optimized subtraction energy width of the energy window, and based on

  14. Smartphone microendoscopy for high resolution fluorescence imaging

    CERN Document Server

    Hong, Xiangqian; Mugler, Dale H; Yu, Bing

    2016-01-01

    High resolution optical endoscopes are increasingly used in diagnosis of various medical conditions of internal organs, such as the gastrointestinal tracts, but they are too expensive for use in resource-poor settings. On the other hand, smartphones with high resolution cameras and Internet access have become more affordable, enabling them to diffuse into most rural areas and developing countries in the past decade. In this letter we describe a smartphone microendoscope that can take fluorescence images with a spatial resolution of 3.1 {\\mu}m. Images collected from ex vivo, in vitro and in vivo samples using the device are also presented. The compact and cost-effective smartphone microendoscope may be envisaged as a powerful tool for detecting pre-cancerous lesions of internal organs in low and middle income countries.

  15. A novel multiwavelength fluorescence image-guided surgery imaging system

    Science.gov (United States)

    Volpi, D.; Tullis, I. D. C.; Laios, A.; Pathiraja, P. N. J.; Haldar, K.; Ahmed, A. A.; Vojnovic, B.

    2014-02-01

    We describe the development and performance analysis of two clinical near-infrared fluorescence image-guided surgery (FIGS) devices that aim to overcome some of the limitations of current FIGS systems. The devices operate in a widefield-imaging mode and can work (1) in conjunction with a laparoscope, during minimally invasive surgery, and (2) as a hand-held, open surgery imaging system. In both cases, narrow-band excitation light, delivered at multiple wavelengths, is efficiently combined with white reflectance light. Light is delivered to ~100 cm2 surgical field at 1-2 mW/cm2 for white light and 3-7 mW/cm2 (depending on wavelength) of red - near infrared excitation, at a typical working distance of 350 mm for the hand-held device and 100 mm for the laparoscope. A single, sensitive, miniaturized color camera collects both fluorescence and white reflectance light. The use of a single imager eliminates image alignment and software overlay complexity. A novel filtering and illumination arrangement allows simultaneous detection of white reflectance and fluorescence emission from multiple dyes in real-time. We will present both fluorescence detection sensitivity modeling and practical performance data. We have demonstrated the efficiency and the advantages of the devices both pre-clinically and during live surgery on humans. Both the hand-held and the laparoscopic systems have proved to be reliable and beneficial in an ongoing clinical trial involving sentinel lymph node detection in gynecological cancers. We will show preliminary results using two clinically approved dyes, Methylene blue and indocyanine green. We anticipate that this technology can be integrated and routinely used in a larger variety of surgical procedures.

  16. Carbon Quantum Dots for Zebrafish Fluorescence Imaging

    Science.gov (United States)

    Kang, Yan-Fei; Li, Yu-Hao; Fang, Yang-Wu; Xu, Yang; Wei, Xiao-Mi; Yin, Xue-Bo

    2015-07-01

    Carbon quantum dots (C-QDs) are becoming a desirable alternative to metal-based QDs and dye probes owing to their high biocompatibility, low toxicity, ease of preparation, and unique photophysical properties. Herein, we describe fluorescence bioimaging of zebrafish using C-QDs as probe in terms of the preparation of C-QDs, zebrafish husbandry, embryo harvesting, and introduction of C-QDs into embryos and larvae by soaking and microinjection. The multicolor of C-QDs was validated with their imaging for zebrafish embryo. The distribution of C-QDs in zebrafish embryos and larvae were successfully observed from their fluorescence emission. the bio-toxicity of C-QDs was tested with zebrafish as model and C-QDs do not interfere to the development of zebrafish embryo. All of the results confirmed the high biocompatibility and low toxicity of C-QDs as imaging probe. The absorption, distribution, metabolism and excretion route (ADME) of C-QDs in zebrafish was revealed by their distribution. Our work provides the useful information for the researchers interested in studying with zebrafish as a model and the applications of C-QDs. The operations related zebrafish are suitable for the study of the toxicity, adverse effects, transport, and biocompatibility of nanomaterials as well as for drug screening with zebrafish as model.

  17. Imaging carious dental tissues with multiphoton fluorescence lifetime imaging microscopy

    Science.gov (United States)

    Lin, Po-Yen; Lyu, Hong-Chou; Hsu, Chin-Ying Stephen; Chang, Chia-Seng; Kao, Fu-Jen

    2011-01-01

    In this study, multiphoton excitation was utilized to image normal and carious dental tissues noninvasively. Unique structures in dental tissues were identified using the available multimodality (second harmonic, autofluorescence, and fluorescence lifetime analysis) without labeling. The collagen in dentin exhibits a strong second harmonic response. Both dentin and enamel emit strong autofluorescence that reveals in detail morphological features (such as dentinal tubules and enamel rods) and, despite their very similar spectral profiles, can be differentiated by lifetime analysis. Specifically, the carious dental tissue exhibits a greatly reduced autofluorescence lifetime, which result is consistent with the degree of demineralization, determined by micro-computed tomography. Our findings suggest that two-photon excited fluorescence lifetime imaging may be a promising tool for diagnosing and monitoring dental caries. PMID:21326645

  18. Behavior Subtraction

    CERN Document Server

    Jodoin, P M; Konrad, J

    2009-01-01

    Background subtraction has been a driving engine for many computer vision and video analytics tasks. Although its many variants exist, they all share the underlying assumption that photometric scene properties are either static or exhibit temporal stationarity. While this works in some applications, the model fails when one is interested in discovering {\\it changes in scene dynamics} rather than those in a static background; detection of unusual pedestrian and motor traffic patterns is but one example. We propose a new model and computational framework that address this failure by considering stationary scene dynamics as a ``background'' with which observed scene dynamics are compared. Central to our approach is the concept of an {\\it event}, that we define as short-term scene dynamics captured over a time window at a specific spatial location in the camera field of view. We compute events by time-aggregating motion labels, obtained by background subtraction, as well as object descriptors (e.g., object size)....

  19. Background and Scattered-Light Subtraction in the High-Resolution Echelle Modes of the Space Telescope Imaging Spectrograph

    Science.gov (United States)

    Howk, J. Christopher; Sembach, Kenneth R.

    2000-05-01

    We present a simple, effective approach for estimating the on-order backgrounds of spectra taken with the highest resolution modes of the Space Telescope Imaging Spectrograph (STIS) on board the Hubble Space Telescope. Our scheme for determining the on-order background spectrum for STIS E140H and E230H observations uses moderate-order polynomial fits to the interorder scattered light visible in the two-dimensional STIS MAMA images. We present a suite of high-resolution STIS spectra to demonstrate that our background-subtraction routine produces the correct overall zero point as judged by the small residual flux levels in the centers of strongly saturated interstellar absorption lines. Although there are multiple sources of background light in STIS echelle mode data, this simple approach works very well for wavelengths longward of Lyα (λ>~1215 Å). At shorter wavelengths, the smaller order separation and generally lower signal-to-noise ratios of the data can reduce the effectiveness of our background estimation procedure. Slight artifacts in the background-subtracted spectrum can be seen in some cases, particularly at wavelengths of B2B and the GHRS first-order G160M observations of the early-type star HD 218915. We find no significant differences between the GHRS data and the STIS data reduced with our method in either case. Based on observations made with the NASA/ESA Hubble Space Telescope, obtained from the data archive at the Space Telescope Science Institute, which is operated by the Association of Universities for Research in Astronomy, Inc., under NASA contract NAS 5-26555.

  20. Data acquisition and analysis for the energy-subtraction Compton scatter camera for medical imaging

    Science.gov (United States)

    Khamzin, Murat Kamilevich

    In response to the shortcomings of the Anger camera currently being used in conventional SPECT, particularly the trade-off between sensitivity and spatial resolution, a novel energy-subtraction Compton scatter camera, or the ESCSC, has been proposed. A successful clinical implementation of the ESCSC could revolutionize the field of SPECT. Features of this camera include utilization of silicon and CdZnTe detectors in primary and secondary detector systems, list-mode time stamping data acquisition, modular architecture, and post-acquisition data analysis. Previous ESCSC studies were based on Monte Carlo modeling. The objective of this work is to test the theoretical framework developed in previous studies by developing the data acquisition and analysis techniques necessary to implement the ESCSC. The camera model working in list-mode with time stamping was successfully built and tested thus confirming potential of the ESCSC that was predicted in previous simulation studies. The obtained data were processed during the post-acquisition data analysis based on preferred event selection criteria. Along with the construction of a camera model and proving the approach, the post-acquisition data analysis was further extended to include preferred event weighting based on the likelihood of a preferred event to be a true preferred event. While formulated to show ESCSC capabilities, the results of this study are important for any Compton scatter camera implementation as well as for coincidence data acquisition systems in general.

  1. Nanoprobes for super-resolution fluorescence imaging at the nanoscale

    Institute of Scientific and Technical Information of China (English)

    HOU ShangGuo; LIANG Le; DENG SuHui; CHEN JianFang; HUANG Qing; CHENG Ya; FAN ChunHai

    2014-01-01

    Compared with other imaging techniques,fluorescence microscopy has become an essential tool to study cell biology due to its high compatibility with living cells.Owing to the resolution limit set by the diffraction of light,fluorescence microscopy could not resolve the nanostructures in the range of〈200 nm.Recently,many techniques have been emerged to overcome the diffraction barrier,providing nanometer spatial resolution.In the course of development,the progress in fluorescent probes has helped to promote the development of the high-resolution fluorescence nanoscopy.Here,we describe the contributions of the fluorescent probes to far-field super resolution imaging,focusing on concepts of the existing super-resolution nanoscopy based on the photophysics of fluorescent nanoprobes,like photoswitching,bleaching and blinking.Fluorescent probe technology is crucial in the design and implementation of super-resolution imaging methods.

  2. Sensor noise in direct digital imaging (the RadioVisioGraphy, Sens-a-Ray, and Visualix/Vixa systems) evaluated by subtraction radiography.

    Science.gov (United States)

    Wenzel, A

    1994-01-01

    The aim of this study was to evaluate sensor noise with the use of the subtraction method in radiographs captured with three direct digital intraoral systems. Ten radiographs were taken of the lower left molar region of a phantom head at each of three exposure times: 0.20 seconds, 0.46 seconds, and 0.60 seconds with the use of the RadioVisioGraphy (Trophy Radiologie, Vincennes, France), Sens-a-Ray (Regam Medical Systems, AB, Sundsvall, Sweden), and Visualix (Gendex, Philips Medical Systems, Inc., Monza, Italy) systems. Neither the x-ray tube nor the phantom were moved between exposures, and the three sensors were identically positioned. The images were stored in the tagged image file format provided by the systems in 8-bit depth and imported by a subtraction program. Subtractions were performed between identical images taken with the three systems. The standard deviation for the distribution of the shades of grey in the subtraction image histogram served as an expression for image noise. Paired t tests evaluated differences between the standard deviations of the subtraction images from the three systems. The standard deviation increased with increasing exposure time for all three systems (p < 0.00001). The standard deviation for the images performed with Visualix were 6.47, 10.34, and 11.16 at exposure times 0.20, 0.46, and 0.60, respectively. For the RadioVisioGraphy, these values were 1.61, 2.03, and 2.18, and for Sens-a-Ray 2.90, 3.98, and 3.96, respectively. The differences between the systems were highly statistically significant (p < 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)

  3. SPECT {sup 99m}Tc-sestamibi/{sup 123}I subtraction images merged to the scanner: interest of patients with hyperparathyroidism, candidates to surgery; Images de soustraction SPECT 99mTc-Sestamibi/123 I fusionnees au scanner: interet chez des patients avec hyperparathyroidie, candidats a la chirurgie

    Energy Technology Data Exchange (ETDEWEB)

    Poullias, X.; Hapdey, S.; Salles, A.; Vera, P.; Edet-Sanson, A. [Centre Henri-Becquerel, 76 - Rouen (France); Guernou, M. [Centre cardiologique du Nord, 93 - Saint-Denis (France); Hitzel, A. [CHU de Toulouse, 31 (France)

    2010-07-01

    Purpose: the aim of this study is to evaluate the interest of SPECT subtraction images merged to the scanner (S/CT), compared to planar subtraction (S/PL) and to echography, in the framework of hyperparathyroidism. Conclusions: Although subtraction SPECT images merged on CT have a sensitivity close to planar subtraction images, making this modality often allows to visualize the lesion to define its size and anatomical reports. These elements are a help for surgical management. (N.C.)

  4. Behavior subtraction.

    Science.gov (United States)

    Jodoin, Pierre-Marc; Saligrama, Venkatesh; Konrad, Janusz

    2012-09-01

    Background subtraction has been a driving engine for many computer vision and video analytics tasks. Although its many variants exist, they all share the underlying assumption that photometric scene properties are either static or exhibit temporal stationarity. While this works in many applications, the model fails when one is interested in discovering changes in scene dynamics instead of changes in scene's photometric properties; the detection of unusual pedestrian or motor traffic patterns are but two examples. We propose a new model and computational framework that assume the dynamics of a scene, not its photometry, to be stationary, i.e., a dynamic background serves as the reference for the dynamics of an observed scene. Central to our approach is the concept of an event, which we define as short-term scene dynamics captured over a time window at a specific spatial location in the camera field of view. Unlike in our earlier work, we compute events by time-aggregating vector object descriptors that can combine multiple features, such as object size, direction of movement, speed, etc. We characterize events probabilistically, but use low-memory, low-complexity surrogates in a practical implementation. Using these surrogates amounts to behavior subtraction, a new algorithm for effective and efficient temporal anomaly detection and localization. Behavior subtraction is resilient to spurious background motion, such as due to camera jitter, and is content-blind, i.e., it works equally well on humans, cars, animals, and other objects in both uncluttered and highly cluttered scenes. Clearly, treating video as a collection of events rather than colored pixels opens new possibilities for video analytics.

  5. Fluorescence Imaging Study of Impinging Underexpanded Jets

    Science.gov (United States)

    Inman, Jennifer A.; Danehy, Paul M.; Nowak, Robert J.; Alderfer, David W.

    2008-01-01

    An experiment was designed to create a simplified simulation of the flow through a hole in the surface of a hypersonic aerospace vehicle and the subsequent impingement of the flow on internal structures. In addition to planar laser-induced fluorescence (PLIF) flow visualization, pressure measurements were recorded on the surface of an impingement target. The PLIF images themselves provide quantitative spatial information about structure of the impinging jets. The images also help in the interpretation of impingement surface pressure profiles by highlighting the flow structures corresponding to distinctive features of these pressure profiles. The shape of the pressure distribution along the impingement surface was found to be double-peaked in cases with a sufficiently high jet-exit-to-ambient pressure ratio so as to have a Mach disk, as well as in cases where a flow feature called a recirculation bubble formed at the impingement surface. The formation of a recirculation bubble was in turn found to depend very sensitively upon the jet-exit-to-ambient pressure ratio. The pressure measured at the surface was typically less than half the nozzle plenum pressure at low jet pressure ratios and decreased with increasing jet pressure ratios. Angled impingement cases showed that impingement at a 60deg angle resulted in up to a factor of three increase in maximum pressure at the plate compared to normal incidence.

  6. Phase-sensitive fluorescent imaging with coherent reconstruction

    CERN Document Server

    Field, Jeffrey J; Bartels, Randy A

    2015-01-01

    Optical imaging plays a critical role in advancing our understanding of three dimensional dynamics of biological systems. Coherent imaging (CI) methods exploit spatial phase information, encoded through propagation of coherent signal light emerging from a specimen, to extract a three-dimensional representation of the object from a single high-speed measurement. Until now, CI methods could not be applied to incoherent light, severely limiting their ability to image the most powerful biological probes available - fluorescent molecules - with sufficient speed and volume to observe important processes, such as neural processing in live specimens. We introduce a new imaging technique that transfers the spatial propagation phase of coherent illumination light to incoherent fluorescent light emission. The transfer of propagation phase allows CI techniques to be applied to fluorescent light imaging, and leads to large increases in imaging speed and depth of field. With this advance, biological imaging of fluorescent ...

  7. Subtraction of unidirectionally encoded images for suppression of heavily isotropic objects (SUSHI) for selective visualization of peripheral nerves

    Energy Technology Data Exchange (ETDEWEB)

    Takahara, Taro; Kwee, Thomas C.; Hendrikse, Jeroen; Niwa, Tetsu; Mali, Willem P.T.M.; Luijten, Peter R. [University Medical Center Utrecht, Department of Radiology, Utrecht (Netherlands); Van Cauteren, Marc [Philips Healthcare, Asia Pacific, Tokyo (Japan); Koh, Dow-Mu [Royal Marsden Hospital, Department of Radiology, Sutton (United Kingdom)

    2011-02-15

    The aim of this study was to introduce and assess a new magnetic resonance (MR) technique for selective peripheral nerve imaging, called ''subtraction of unidirectionally encoded images for suppression of heavily isotropic objects'' (SUSHI). Six volunteers underwent diffusion-weighted MR neurography (DW-MRN) of the brachial plexus, and seven volunteers underwent DW-MRN of the sciatic, common peroneal, and tibial nerves at the level of the knee, at 1.5 T. DW-MRN images with SUSHI (DW-MRN{sub SUSHI}) and conventional DW-MRN images (DW-MRN{sub AP}) were displayed using a coronal maximum intensity projection and evaluated by two independent observers regarding signal suppression of lymph nodes, bone marrow, veins, and articular fluids and regarding signal intensity of nerves and ganglia, using five-point grading scales. Scores of DW-MRN{sub SUSHI} were compared to those of DW-MRN{sub AP} using Wilcoxon tests. Suppression of lymph nodes around the brachial plexus and suppression of articular fluids at the level of the knee at DW-MRN{sub SUSHI} was significantly better than that at DW-MRN{sub AP} (P < 0.05). However, overall signal intensity of brachial plexus nerves and ganglia at DW-MRN{sub SUSHI} was significantly lower than that at DW-MRN{sub AP} (P < 0.05). On the other hand, signal intensity of the sciatic, common peroneal, and tibial nerves at the level of the knee at DW-MRN{sub SUSHI} was judged as significantly better than that at DW-MRN{sub AP} (P < 0.05). The SUSHI technique allows more selective visualization of the sciatic, common peroneal, and tibial nerves at the level of the knee but is less useful for brachial plexus imaging because signal intensity of the brachial plexus nerves and ganglia can considerably be decreased. (orig.)

  8. A Study of Stellar Photometric Variability Within the Central 4 pc of the Galactic Center with Infrared Image Subtraction

    CERN Document Server

    Peeples, Molly S; De Poy, D L

    2007-01-01

    We present a catalog of 110 variable stars within ~1' of Sgr A* based on image subtraction of near-infrared (H and K) photometry. Our images were obtained over 133 nights from 2000--2002 in H-band and over 134 nights from 2001--2002 in K-band; the typical FWHM is 1.4''. We match the catalog to other near-infrared, X-ray, and radio (i.e., maser) data, and we discuss some of the more interesting objects. The catalog includes 14 periodic sources, several known long-period variables and three new LPV candidates. We associate IRS 10* with OH, SiO, and H2O masers and a bright X-ray point source; this analysis suggests IRS 10* is an AGB star with an accreting companion. Among the approximately 90 newly discovered sources are a probable cataclysmic variable, a potential edge-on contact 84 day period eclipsing binary, and a possible 41 day period pulsating variable.

  9. Combined dynamic contrast-enhancement and serial 3D-subtraction analysis in magnetic resonance imaging of osteoid osteomas

    Energy Technology Data Exchange (ETDEWEB)

    Kalle, T. von; Winkler, P. [Klinikum Stuttgart Olgahospital, Department of Paediatric Radiology, Stuttgart (Germany); Langendoerfer, M.; Fernandez, F.F. [Klinikum Stuttgart Olgahospital, Department of Paediatric Orthopaedics, Stuttgart (Germany)

    2009-10-15

    The purpose of this study was to retrospectively correlate the results of dynamic contrast-enhanced magnetic resonance imaging (MRI) with histological and clinical diagnoses in patients with osteoid osteomas. Fifty-four patients with the MR diagnosis of osteoid osteoma were studied. MRI (1.5 Tesla) consisted of thin-section STIR sequences, dynamic 3D T1 gradient echo sequences during application of contrast material, and high-resolution postcontrast T1 spin echo sequences with fat saturation (maximum voxel size 0.6 x 0.6 x 3.0 mm). Evaluation was focused on serial image subtraction during the early phase after contrast injection and on time-intensity curves. The surrounding edema was helpful in finding the nidus in each lesion. In 49 of 54 patients (90.7%), the diagnosis of osteoid osteoma was certain or highly probable (sensitivity 1.0, positive predictive value 0.91). A total of 38 of 54 osteoid osteomas were histologically proven. Five MRI diagnoses were regarded as false positives. A similar proportion has been reported for computed tomography. Tailored high-resolution MR examinations with dynamic contrast enhancement can reliably diagnose osteoid osteomas and exactly localize the nidus without radiation exposure. We propose a stepwise approach with STIR sequences, dynamic contrast-enhanced scanning, and high-resolution postcontrast T1 spin echo sequences with fat saturation. (orig.)

  10. Variability-based AGN selection using image subtraction in the SDSS and LSST era

    CERN Document Server

    Choi, Yumi; Becker, Andrew C; Ivezić, \\vZeljko; Connolly, Andrew J; MacLeod, Chelsea L; Ruan, John J; Anderson, Scott F

    2013-01-01

    With upcoming all sky surveys such as LSST poised to generate a deep digital movie of the optical sky, variability-based AGN selection will enable the construction of highly-complete catalogs with minimum contamination. In this study, we generate $g$-band difference images and construct light curves for QSO/AGN candidates listed in SDSS Stripe 82 public catalogs compiled from different methods, including spectroscopy, optical colors, variability, and X-ray detection. Image differencing excels at identifying variable sources embedded in complex or blended emission regions such as Type II AGNs and other low-luminosity AGNs that may be omitted from traditional photometric or spectroscopic catalogs. To separate QSOs/AGNs from other sources using our difference image light curves, we explore several light curve statistics and parameterize optical variability by the characteristic damping timescale ($\\tau$) and variability amplitude. By virtue of distinguishable variability parameters of AGNs, we are able to select...

  11. CT perfusion imaging and CT subtraction angiography in the diagnosis of ischemic cerebrovascular disease within 24 hours

    Institute of Scientific and Technical Information of China (English)

    管小亭; 于学英; 刘翔; 龙洁; 戴建平

    2003-01-01

    Objective To evaluate the value of the clinical use of CT perfusion imaging (CTPI) and CT subtraction angiography (CTSA) for diagnosing acute ischemic cerebrovascular disease (AICVD). Methods Twenty-four patients with AICVD onset within 24 hours were examined with regular CT, CTPI, and CTSA. Some cases received CTPI, magnetic resonance imaging (MRI), magnetic resonance angiography (MRA), digital subtraction angiography (DSA) or single photon emission computer tomography (SPECT) during follow-up examinations.Results Of the 24 cases, 11 had negative results from regular CT scans 3-6 hours after onset of stroke in 6 cases, 6-12 hours in 3 cases, and 12-24 hours in 2 cases. Ten of these cases were then confirmed by CTPI as having ischemic lesions, 2 with middle cerebral artery occlusion (MCAO), and 1 case with transient ischemic attack (TIA) with CTPI negative. Of the 24 cases, 13 had positive results from regular CT, 9 were diagnosed with ischemic lesions larger by using CTPI than regular CT, 1 case had MCAO and 1 had internal carotid artery occlusion (ICAO). There were 4 cases with ischemic lesions observed with regular CT having nearly the same range as that of lacunar infarctions using CTPI. Another 4 cases had more than 2 lesion areas. The peak time (PT), mean transit time (MTT) and relative flow (RF) of 24 cases were markedly different. The sides of ischemic lesions compared to each other and the core of the lesion compared to peripheral zones were also altered significantly (P<0.01).Conclusions Combined CTPI with CTSA can detect acute ischemic lesions at early and hyper-early stages and could distinguish between TIA, lacunar infarction and a larger area of infarction. Using semiquantitative blood perfusion analysis status, CTPI with CTSA could define position, area and range of the ischemic lesion and penumbra. These scans can also analyze the brain blood perfusion status. It is important to early diagnose the occlusion of the entire division of the internal

  12. An automatic fuzzy-based multi-temporal brain digital subtraction angiography image fusion algorithm using curvelet transform and content selection strategy.

    Science.gov (United States)

    Momeni, Saba; Pourghassem, Hossein

    2014-08-01

    Recently image fusion has prominent role in medical image processing and is useful to diagnose and treat many diseases. Digital subtraction angiography is one of the most applicable imaging to diagnose brain vascular diseases and radiosurgery of brain. This paper proposes an automatic fuzzy-based multi-temporal fusion algorithm for 2-D digital subtraction angiography images. In this algorithm, for blood vessel map extraction, the valuable frames of brain angiography video are automatically determined to form the digital subtraction angiography images based on a novel definition of vessel dispersion generated by injected contrast material. Our proposed fusion scheme contains different fusion methods for high and low frequency contents based on the coefficient characteristic of wrapping second generation of curvelet transform and a novel content selection strategy. Our proposed content selection strategy is defined based on sample correlation of the curvelet transform coefficients. In our proposed fuzzy-based fusion scheme, the selection of curvelet coefficients are optimized by applying weighted averaging and maximum selection rules for the high frequency coefficients. For low frequency coefficients, the maximum selection rule based on local energy criterion is applied to better visual perception. Our proposed fusion algorithm is evaluated on a perfect brain angiography image dataset consisting of one hundred 2-D internal carotid rotational angiography videos. The obtained results demonstrate the effectiveness and efficiency of our proposed fusion algorithm in comparison with common and basic fusion algorithms.

  13. Multispectral fluorescence imaging techniques for nondestructive food safety inspection

    Science.gov (United States)

    Kim, Moon S.; Lefcourt, Alan M.; Chen, Yud-Ren

    2004-03-01

    The use of spectral sensing has gained acceptance as a rapid means for nondestructive inspection of postharvest food produce. Current technologies generally use color or a single wavelength camera technology. The applicability and sensitivity of these techniques can be expanded through the use of multiple wavelengths. Reflectance in the Vis/NIR is the prevalent spectral technique. Fluorescence, compared to reflectance, is regarded as a more sensitive technique due to its dynamic responses to subtle changes in biological entities. Our laboratory has been exploring fluorescence as a potential means for detection of quality and wholesomeness of food products. Applications of fluorescence sensing require an understanding of the spectral characteristics emanating from constituents and potential contaminants. A number of factors affecting fluorescence emission characteristics are discussed. Because of relatively low fluorescence quantum yield from biological samples, a system with a powerful pulse light source such as a laser coupled with a gated detection device is used to harvest fluorescence, in the presence of ambient light. Several fluorescence sensor platforms developed in our laboratory, including hyperspectral imaging, and laser-induced fluorescence (LIF) and steady-state fluorescence imaging systems with multispectral capabilities are presented. We demonstrate the potential uses of recently developed fluorescence imaging platforms in food safety inspection of apples contaminated with animal feces.

  14. Temporal Subtraction of Digital Breast Tomosynthesis Images for Improved Mass Detection

    Science.gov (United States)

    2007-10-01

    number of low-dose cone-beam projection images", Medical Physics 30 (3), 365 (2003). 9 E.A. Sickles, W.N. Weber, H.B. Galvin, S.H. Ominsky, and R.A...temporal pairs of mammograms for interval change analysis--local affine transformation for improved localization", Medical Physics 28 (6), 1070 (2001...aided classification of malignant and benign breast masses", Medical Physics 28 (11), 2309 (2001). 25 K. Marias, C. Behrenbruch, S. Parbhoo, A

  15. Variability-based Active Galactic Nucleus Selection Using Image Subtraction in the SDSS and LSST Era

    Science.gov (United States)

    Choi, Yumi; Gibson, Robert R.; Becker, Andrew C.; Ivezić, Željko; Connolly, Andrew J.; MacLeod, Chelsea L.; Ruan, John J.; Anderson, Scott F.

    2014-02-01

    With upcoming all-sky surveys such as LSST poised to generate a deep digital movie of the optical sky, variability-based active galactic nucleus (AGN) selection will enable the construction of highly complete catalogs with minimum contamination. In this study, we generate g-band difference images and construct light curves (LCs) for QSO/AGN candidates listed in Sloan Digital Sky Survey Stripe 82 public catalogs compiled from different methods, including spectroscopy, optical colors, variability, and X-ray detection. Image differencing excels at identifying variable sources embedded in complex or blended emission regions such as Type II AGNs and other low-luminosity AGNs that may be omitted from traditional photometric or spectroscopic catalogs. To separate QSOs/AGNs from other sources using our difference image LCs, we explore several LC statistics and parameterize optical variability by the characteristic damping timescale (τ) and variability amplitude. By virtue of distinguishable variability parameters of AGNs, we are able to select them with high completeness of 93.4% and efficiency (i.e., purity) of 71.3%. Based on optical variability, we also select highly variable blazar candidates, whose infrared colors are consistent with known blazars. One-third of them are also radio detected. With the X-ray selected AGN candidates, we probe the optical variability of X-ray detected optically extended sources using their difference image LCs for the first time. A combination of optical variability and X-ray detection enables us to select various types of host-dominated AGNs. Contrary to the AGN unification model prediction, two Type II AGN candidates (out of six) show detectable variability on long-term timescales like typical Type I AGNs. This study will provide a baseline for future optical variability studies of extended sources.

  16. Effect of injection technique on temporal parametric imaging derived from digital subtraction angiography in patient specific phantoms

    Science.gov (United States)

    Ionita, Ciprian N.; Garcia, Victor L.; Bednarek, Daniel R.; Snyder, Kenneth V.; Siddiqui, Adnan H.; Levy, Elad I.; Rudin, Stephen

    2014-03-01

    Parametric imaging maps (PIM's) derived from digital subtraction angiography (DSA) for the cerebral arterial flow assessment in clinical settings have been proposed, but experiments have yet to determine the reliability of such studies. For this study, we have observed the effects of different injection techniques on PIM's. A flow circuit set to physiologic conditions was created using an internal carotid artery phantom. PIM's were derived for two catheter positions, two different contrast bolus injection volumes (5ml and 10 ml), and four injection rates (5, 10, 15 and 20 ml/s). Using a gamma variate fitting approach, we derived PIM's for mean-transit-time (MTT), time-to-peak (TTP) and bolus-arrivaltime (BAT). For the same injection rates, a larger bolus resulted in an increased MTT and TTP, while a faster injection rate resulted in a shorter MTT, TTP, and BAT. In addition, the position of the catheter tip within the vasculature directly affected the PIM. The experiment showed that the PIM is strongly correlated with the injection conditions, and, therefore, they have to be interpreted with caution. PIM images must be taken from the same patient to be able to be meaningfully compared. These comparisons can include pre- and post-treatment images taken immediately before and after an interventional procedure or simultaneous arterial flow comparisons through the left and right cerebral hemispheres. Due to the strong correlation between PIM and injection conditions, this study indicates that this assessment method should be used only to compare flow changes before and after treatment within the same patient using the same injection conditions.

  17. Refractive Index Sensing of Green Fluorescent Proteins in Living Cells Using Fluorescence Lifetime Imaging Microscopy

    NARCIS (Netherlands)

    Manen, van Henk-Jan; Verkuijlen, Paul; Wittendorp, Paul; Subramaniam, Vinod; Berg, van den Timo K.; Roos, Dirk; Otto, Cees

    2008-01-01

    We show that fluorescence lifetime imaging microscopy (FLIM) of green fluorescent protein (GFP) molecules in cells can be used to report on the local refractive index of intracellular GFP. We expressed GFP fusion constructs of Rac2 and gp91phox, which are both subunits of the phagocyte NADPH oxidase

  18. Bilateral image subtraction features for multivariate automated classification of breast cancer risk

    Science.gov (United States)

    Celaya-Padilla, Jose M.; Rodriguez-Rojas, Juan; Galván-Tejada, Jorge I.; Martínez-Torteya, Antonio; Treviño, Victor; Tamez-Peña, José G.

    2014-03-01

    Early tumor detection is key in reducing breast cancer deaths and screening mammography is the most widely available method for early detection. However, mammogram interpretation is based on human radiologist, whose radiological skills, experience and workload makes radiological interpretation inconsistent. In an attempt to make mammographic interpretation more consistent, computer aided diagnosis (CADx) systems has been introduced. This paper presents an CADx system aimed to automatically triage normal mammograms form suspicious mammograms. The CADx system co-reregister the left and breast images, then extracts image features from the co-registered mammographic bilateral sets. Finally, an optimal logistic multivariate model is generated by means of an evolutionary search engine. In this study, 440 subjects form the DDSM public data sets were used: 44 normal mammograms, 201 malignant mass mammograms, and 195 mammograms with malignant calci cations. The results showed a cross validation accuracy of 0.88 and an area under receiver operating characteristic (AUC) of 0.89 for the calci cations vs. normal mammograms. The optimal mass vs. normal mammograms model obtained an accuracy of 0.85 and an AUC of 0.88.

  19. Gadofosveset-enhanced MR angiography of carotid arteries: does steady-state imaging improve accuracy of first-pass imaging? Comparison with selective digital subtraction angiography.

    Science.gov (United States)

    Anzidei, Michele; Napoli, Alessandro; Marincola, Beatrice Cavallo; Nofroni, Italo; Geiger, Daniel; Zaccagna, Fulvio; Catalano, Carlo; Passariello, Roberto

    2009-05-01

    To evaluate the diagnostic accuracy of gadofosveset-enhanced magnetic resonance (MR) angiography in the assessment of carotid artery stenosis, with digital subtraction angiography (DSA) as the reference standard, and to determine the value of reading first-pass, steady-state, and "combined" (first-pass plus steady-state) MR angiograms. This study was approved by the local ethics committee, and all subjects gave written informed consent. MR angiography and DSA were performed in 84 patients (56 men, 28 women; age range, 61-76 years) with carotid artery stenosis at Doppler ultrasonography. Three readers reviewed the first-pass, steady-state, and combined MR data sets, and one independent observer evaluated the DSA images to assess stenosis degree, plaque morphology and ulceration, stenosis length, and tandem lesions. Interobserver agreement regarding MR angiographic findings was analyzed by using intraclass correlation and Cohen kappa coefficients. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated by using the McNemar test to determine possible significant differences (P < .05). Interobserver agreement regarding all MR angiogram readings was substantial. For grading stenosis, sensitivity, specificity, PPV, and NPV were, respectively, 90%, 92%, 91%, and 91% for first-pass imaging; 95% each for steady-state imaging; and 96%, 99%, 99%, and 97% for combined imaging. For evaluation of plaque morphology, respective values were 84%, 86%, 88%, and 82% for first-pass imaging; 98%, 97%, 98%, and 97% for steady-state imaging; and 98%, 100%, 100%, and 97% for combined imaging. Differences between the first-pass, steady-state, and combined image readings for assessment of stenosis degree and plaque morphology were significant (P < .001). Gadofosveset-enhanced MR angiography is a promising technique for imaging carotid artery stenosis. Steady-state image reading is superior to first-pass image reading, but the combined

  20. Detection of rheumatoid arthritis in humans by fluorescence imaging

    Science.gov (United States)

    Ebert, Bernd; Dziekan, Thomas; Weissbach, Carmen; Mahler, Marianne; Schirner, Michael; Berliner, Birgitt; Bauer, Daniel; Voigt, Jan; Berliner, Michael; Bahner, Malte L.; Macdonald, Rainer

    2010-02-01

    The blood pool agent indo-cyanine green (ICG) has been investigated in a prospective clinical study for detection of rheumatoid arthritis using fluorescence imaging. Temporal behavior as well as spatial distribution of fluorescence intensity are suited to differentiate healthy and inflamed finger joints after i.v. injection of an ICG bolus.

  1. Hyperspectral imaging fluorescence excitation scanning for colon cancer detection

    Science.gov (United States)

    Leavesley, Silas J.; Walters, Mikayla; Lopez, Carmen; Baker, Thomas; Favreau, Peter F.; Rich, Thomas C.; Rider, Paul F.; Boudreaux, Carole W.

    2016-10-01

    Optical spectroscopy and hyperspectral imaging have shown the potential to discriminate between cancerous and noncancerous tissue with high sensitivity and specificity. However, to date, these techniques have not been effectively translated to real-time endoscope platforms. Hyperspectral imaging of the fluorescence excitation spectrum represents new technology that may be well suited for endoscopic implementation. However, the feasibility of detecting differences between normal and cancerous mucosa using fluorescence excitation-scanning hyperspectral imaging has not been evaluated. The goal of this study was to evaluate the initial feasibility of using fluorescence excitation-scanning hyperspectral imaging for measuring changes in fluorescence excitation spectrum concurrent with colonic adenocarcinoma using a small pre-pilot-scale sample size. Ex vivo analysis was performed using resected pairs of colorectal adenocarcinoma and normal mucosa. Adenocarcinoma was confirmed by histologic evaluation of hematoxylin and eosin (H&E) permanent sections. Specimens were imaged using a custom hyperspectral imaging fluorescence excitation-scanning microscope system. Results demonstrated consistent spectral differences between normal and cancerous tissues over the fluorescence excitation range of 390 to 450 nm that could be the basis for wavelength-dependent detection of colorectal cancers. Hence, excitation-scanning hyperspectral imaging may offer an alternative approach for discriminating adenocarcinoma from surrounding normal colonic mucosa, but further studies will be required to evaluate the accuracy of this approach using a larger patient cohort.

  2. Bilateral Image Subtraction and Multivariate Models for the Automated Triaging of Screening Mammograms

    Directory of Open Access Journals (Sweden)

    José Celaya-Padilla

    2015-01-01

    Full Text Available Mammography is the most common and effective breast cancer screening test. However, the rate of positive findings is very low, making the radiologic interpretation monotonous and biased toward errors. This work presents a computer-aided diagnosis (CADx method aimed to automatically triage mammogram sets. The method coregisters the left and right mammograms, extracts image features, and classifies the subjects into risk of having malignant calcifications (CS, malignant masses (MS, and healthy subject (HS. In this study, 449 subjects (197 CS, 207 MS, and 45 HS from a public database were used to train and evaluate the CADx. Percentile-rank (p-rank and z-normalizations were used. For the p-rank, the CS versus HS model achieved a cross-validation accuracy of 0.797 with an area under the receiver operating characteristic curve (AUC of 0.882; the MS versus HS model obtained an accuracy of 0.772 and an AUC of 0.842. For the z-normalization, the CS versus HS model achieved an accuracy of 0.825 with an AUC of 0.882 and the MS versus HS model obtained an accuracy of 0.698 and an AUC of 0.807. The proposed method has the potential to rank cases with high probability of malignant findings aiding in the prioritization of radiologists work list.

  3. A framework for creating realistic synthetic fluorescence microscopy image sequences

    CSIR Research Space (South Africa)

    Mabaso, M

    2016-02-01

    Full Text Available of the 9th International Joint Conference on Biomedical Engineering Systems and Technologies, Rome, Italy. 21-23 February, 2016 A Framework for Creating Realistic Synthetic Fluorescence Microscopy Image Sequences Matsilele Mabaso1, Daniel Withey1...

  4. Timing and Operating Mode Design for Time-Gated Fluorescence Lifetime Imaging Microscopy

    OpenAIRE

    Chao Liu; Xinwei Wang; Yan Zhou; Yuliang Liu

    2013-01-01

    Steady-state fluorence imaging and time-resolved fluorescence imaging are two important areas in fluorescence imaging research. Fluorescence lifetime imaging is an absolute measurement method which is independent of excitation laser intensity, fluorophore concentration, and photobleaching compared to fluorescence intensity imaging techniques. Time-gated fluorescence lifetime imaging microscopy (FLIM) can provide high resolution and high imaging frame during mature FLIM methods. An abstract ti...

  5. Image processing for drift compensation in fluorescence microscopy

    DEFF Research Database (Denmark)

    Petersen, Steffen; Thiagarajan, Viruthachalam; Coutinho, Isabel

    2013-01-01

    Fluorescence microscopy is characterized by low background noise, thus a fluorescent object appears as an area of high signal/noise. Thermal gradients may result in apparent motion of the object, leading to a blurred image. Here, we have developed an image processing methodology that may remove....../reduce blur significantly for any type of microscopy. A total of ~100 images were acquired with a pixel size of 30 nm. The acquisition time for each image was approximately 1second. We can quantity the drift in X and Y using the sub pixel accuracy computed centroid location of an image object in each frame....... We can measure drifts down to approximately 10 nm in size and a drift-compensated image can therefore be reconstructed on a grid of the same size using the “Shift and Add” approach leading to an image of identical size asthe individual image. We have also reconstructed the image using a 3 fold larger...

  6. Fluorogen-based reporters for fluorescence imaging: a review

    Science.gov (United States)

    Jullien, Ludovic; Gautier, Arnaud

    2015-12-01

    Fluorescence bioimaging has recently jumped into a new area of spatiotemporal resolution and sensitivity thanks to synergistic advances in both optical physics and probe/biosensor design. This review focuses on the recent development of genetically encodable fluorescent reporters that bind endogenously present or exogenously applied fluorogenic chromophores (so-called fluorogens) and activate their fluorescence. We highlight the innovative engineering and design that gave rise to these new natural and synthetic fluorescent reporters, and describe some of the emerging applications in imaging and biosensing.

  7. Fluorescence Image Analyzer - FLIMA: software for quantitative analysis of fluorescence in situ hybridization.

    Science.gov (United States)

    Silva, H C M; Martins-Júnior, M M C; Ribeiro, L B; Matoso, D A

    2017-03-30

    The Fluorescence Image Analyzer (FLIMA) software was developed for the quantitative analysis of images generated by fluorescence in situ hybridization (FISH). Currently, the images of FISH are examined without a coefficient that enables a comparison between them. Through GD Graphics Library, the FLIMA software calculates the amount of pixels on image and recognizes each present color. The coefficient generated by the algorithm shows the percentage of marks (probes) hybridized on the chromosomes. This software can be used for any type of image generated by a fluorescence microscope and is able to quantify digoxigenin probes exhibiting a red color, biotin probes exhibiting a green color, and double-FISH probes (digoxigenin and biotin used together), where the white color is displayed.

  8. Use of fluorescence lifetime imaging (FLIM) for latent fingerprints detection

    Science.gov (United States)

    Wang, Peng; Chao, Zhi Xia; Seah, Leong K.; Murukeshan, Vadakke M.

    2005-04-01

    Fluorescence lifetime imaging (FLIM) in frequency domain enables the mapping of the spatial distribution of fluorescence lifetimes of a specimen. FLIM can provide unique information about fluorophores and hence is widely used in biology and for medical diagnostics. In this paper, a theoretical analysis for the fluorescence lifetime determination of latent fingerprint samples is described, which is followed by the feasibility study of using FLIM in frequency domain for latent fingerprints detection. Experiments are carried out with fingerprint on green paper substrate and postcard substrate treated with certain fluorescent powder. The total phase lag and demodulation factor are calculated to determine the lifetimes pixel by pixel. The resulting fluorescence lifetime image of fingerprint revealed an improvement in the contrast, and was able to detect the latent fingerprint clearly.

  9. Fluorescence polarization imaging for delineating nonmelanoma skin cancers

    Science.gov (United States)

    Yaroslavsky, A. N.; Neel, V.; Anderson, R. R.

    2004-09-01

    We present a method for detecting nonmelanoma skin cancers using exogenous fluorescence polarization. We built an automated system that permits exogenous fluorescence polarization imaging. It includes a tunable linearly polarized monochromatic light source and a CCD camera equipped with a rotating linear polarizer and a filter to reject excitation light. Two fluorophores that are retained in tumors, toluidine blue and methylene blue, are employed. We demonstrate that fluorescence polarization imaging can be used for accurate delineation of nonmelanoma cancers. The results suggest that this optical technique may be suitable for real-time noninvasive demarcation of epithelial cancers.

  10. Chlorophyll Fluorescence Imaging Uncovers Photosynthetic Fingerprint of Citrus Huanglongbing

    Directory of Open Access Journals (Sweden)

    Haiyan Cen

    2017-08-01

    Full Text Available Huanglongbing (HLB is one of the most destructive diseases of citrus, which has posed a serious threat to the global citrus production. This research was aimed to explore the use of chlorophyll fluorescence imaging combined with feature selection to characterize and detect the HLB disease. Chlorophyll fluorescence images of citrus leaf samples were measured by an in-house chlorophyll fluorescence imaging system. The commonly used chlorophyll fluorescence parameters provided the first screening of HLB disease. To further explore the photosynthetic fingerprint of HLB infected leaves, three feature selection methods combined with the supervised classifiers were employed to identify the unique fluorescence signature of HLB and perform the three-class classification (i.e., healthy, HLB infected, and nutrient deficient leaves. Unlike the commonly used fluorescence parameters, this novel data-driven approach by using the combination of the mean fluorescence parameters and image features gave the best classification performance with the accuracy of 97%, and presented a better interpretation for the spatial heterogeneity of photochemical and non-photochemical components in HLB infected citrus leaves. These results imply the potential of the proposed approach for the citrus HLB disease diagnosis, and also provide a valuable insight for the photosynthetic response to the HLB disease.

  11. Chlorophyll Fluorescence Imaging Uncovers Photosynthetic Fingerprint of Citrus Huanglongbing.

    Science.gov (United States)

    Cen, Haiyan; Weng, Haiyong; Yao, Jieni; He, Mubin; Lv, Jingwen; Hua, Shijia; Li, Hongye; He, Yong

    2017-01-01

    Huanglongbing (HLB) is one of the most destructive diseases of citrus, which has posed a serious threat to the global citrus production. This research was aimed to explore the use of chlorophyll fluorescence imaging combined with feature selection to characterize and detect the HLB disease. Chlorophyll fluorescence images of citrus leaf samples were measured by an in-house chlorophyll fluorescence imaging system. The commonly used chlorophyll fluorescence parameters provided the first screening of HLB disease. To further explore the photosynthetic fingerprint of HLB infected leaves, three feature selection methods combined with the supervised classifiers were employed to identify the unique fluorescence signature of HLB and perform the three-class classification (i.e., healthy, HLB infected, and nutrient deficient leaves). Unlike the commonly used fluorescence parameters, this novel data-driven approach by using the combination of the mean fluorescence parameters and image features gave the best classification performance with the accuracy of 97%, and presented a better interpretation for the spatial heterogeneity of photochemical and non-photochemical components in HLB infected citrus leaves. These results imply the potential of the proposed approach for the citrus HLB disease diagnosis, and also provide a valuable insight for the photosynthetic response to the HLB disease.

  12. Multiplexed Spectral Imaging of 120 Different Fluorescent Labels.

    Directory of Open Access Journals (Sweden)

    Alex M Valm

    Full Text Available The number of fluorescent labels that can unambiguously be distinguished in a single image when acquired through band pass filters is severely limited by the spectral overlap of available fluorophores. The recent development of spectral microscopy and the application of linear unmixing algorithms to spectrally recorded image data have allowed simultaneous imaging of fluorophores with highly overlapping spectra. However, the number of distinguishable fluorophores is still limited by the unavoidable decrease in signal to noise ratio when fluorescence signals are fractionated over multiple wavelength bins. Here we present a spectral image analysis algorithm to greatly expand the number of distinguishable objects labeled with binary combinations of fluorophores. Our algorithm utilizes a priori knowledge about labeled specimens and imposes a binary label constraint on the unmixing solution. We have applied our labeling and analysis strategy to identify microbes labeled by fluorescence in situ hybridization and here demonstrate the ability to distinguish 120 differently labeled microbes in a single image.

  13. Photobleaching correction in fluorescence microscopy images

    Energy Technology Data Exchange (ETDEWEB)

    Vicente, Nathalie B; Diaz Zamboni, Javier E; Adur, Javier F; Paravani, Enrique V; Casco, Victor H [Microscopy Laboratory, School of Engineering - Bioengineering, National University of Entre Rios (UNER), Ruta 11, Km 10 (3101), Oro Verde, Entre Rios (Argentina)

    2007-11-15

    Fluorophores are used to detect molecular expression by highly specific antigen-antibody reactions in fluorescence microscopy techniques. A portion of the fluorophore emits fluorescence when irradiated with electromagnetic waves of particular wavelengths, enabling its detection. Photobleaching irreversibly destroys fluorophores stimulated by radiation within the excitation spectrum, thus eliminating potentially useful information. Since this process may not be completely prevented, techniques have been developed to slow it down or to correct resulting alterations (mainly, the decrease in fluorescent signal). In the present work, the correction by photobleaching curve was studied using E-cadherin (a cell-cell adhesion molecule) expression in Bufo arenarum embryos. Significant improvements were observed when applying this simple, inexpensive and fast technique.

  14. Wide field-of-view fluorescence imaging of coral reefs.

    Science.gov (United States)

    Treibitz, Tali; Neal, Benjamin P; Kline, David I; Beijbom, Oscar; Roberts, Paul L D; Mitchell, B Greg; Kriegman, David

    2015-01-13

    Coral reefs globally are declining rapidly because of both local and global stressors. Improved monitoring tools are urgently needed to understand the changes that are occurring at appropriate temporal and spatial scales. Coral fluorescence imaging tools have the potential to improve both ecological and physiological assessments. Although fluorescence imaging is regularly used for laboratory studies of corals, it has not yet been used for large-scale in situ assessments. Current obstacles to effective underwater fluorescence surveying include limited field-of-view due to low camera sensitivity, the need for nighttime deployment because of ambient light contamination, and the need for custom multispectral narrow band imaging systems to separate the signal into meaningful fluorescence bands. Here we describe the Fluorescence Imaging System (FluorIS), based on a consumer camera modified for greatly increased sensitivity to chlorophyll-a fluorescence, and we show high spectral correlation between acquired images and in situ spectrometer measurements. This system greatly facilitates underwater wide field-of-view fluorophore surveying during both night and day, and potentially enables improvements in semi-automated segmentation of live corals in coral reef photographs and juvenile coral surveys.

  15. A Method of Time-Intensity Curve Calculation for Vascular Perfusion of Uterine Fibroids Based on Subtraction Imaging with Motion Correction

    Science.gov (United States)

    Zhu, Xinjian; Wu, Ruoyu; Li, Tao; Zhao, Dawei; Shan, Xin; Wang, Puling; Peng, Song; Li, Faqi; Wu, Baoming

    2016-12-01

    The time-intensity curve (TIC) from contrast-enhanced ultrasound (CEUS) image sequence of uterine fibroids provides important parameter information for qualitative and quantitative evaluation of efficacy of treatment such as high-intensity focused ultrasound surgery. However, respiration and other physiological movements inevitably affect the process of CEUS imaging, and this reduces the accuracy of TIC calculation. In this study, a method of TIC calculation for vascular perfusion of uterine fibroids based on subtraction imaging with motion correction is proposed. First, the fibroid CEUS recording video was decoded into frame images based on the record frame rate. Next, the Brox optical flow algorithm was used to estimate the displacement field and correct the motion between two frames based on warp technique. Then, subtraction imaging was performed to extract the positional distribution of vascular perfusion (PDOVP). Finally, the average gray of all pixels in the PDOVP from each image was determined, and this was considered the TIC of CEUS image sequence. Both the correlation coefficient and mutual information of the results with proposed method were larger than those determined using the original method. PDOVP extraction results have been improved significantly after motion correction. The variance reduction rates were all positive, indicating that the fluctuations of TIC had become less pronounced, and the calculation accuracy has been improved after motion correction. This proposed method can effectively overcome the influence of motion mainly caused by respiration and allows precise calculation of TIC.

  16. Fluorescence and Bioluminescence Imaging of Orthotopic Brain Tumors in Mice.

    Science.gov (United States)

    McKinnon, Emilie; Moore, Alfred; Dixit, Suraj; Zhu, Yun; Broome, Ann-Marie

    2017-01-01

    Optical imaging strategies, such as fluorescence and bioluminescence imaging, are non-invasive, in vivo whole body imaging techniques utilized to study cancer. Optical imaging is widely used in preclinical work because of its ease of use and cost-friendliness. It also provides the opportunity to study animals and biological responses longitudinally over time. Important considerations include depth of tissue penetration, photon scattering, absorption and the choice of light emitting probe, all of which affect the resolution (image quality and data information) and the signal to noise ratio of the image. We describe how to use bioluminescence and fluorescence imaging to track a chemotherapeutic delivery nanocarrier conjugated with a fluorophore to determine its localization in vivo.

  17. Hyperspectral fluorescence imaging with multi wavelength LED excitation

    Science.gov (United States)

    Luthman, A. Siri; Dumitru, Sebastian; Quirós-Gonzalez, Isabel; Bohndiek, Sarah E.

    2016-04-01

    Hyperspectral imaging (HSI) can combine morphological and molecular information, yielding potential for real-time and high throughput multiplexed fluorescent contrast agent imaging. Multiplexed readout from targets, such as cell surface receptors overexpressed in cancer cells, could improve both sensitivity and specificity of tumor identification. There remains, however, a need for compact and cost effective implementations of the technology. We have implemented a low-cost wide-field multiplexed fluorescence imaging system, which combines LED excitation at 590, 655 and 740 nm with a compact commercial solid state HSI system operating in the range 600 - 1000 nm. A key challenge for using reflectance-based HSI is the separation of contrast agent fluorescence from the reflectance of the excitation light. Here, we illustrate how it is possible to address this challenge in software, using two offline reflectance removal methods, prior to least-squares spectral unmixing. We made a quantitative comparison of the methods using data acquired from dilutions of contrast agents prepared in well-plates. We then established the capability of our HSI system for non-invasive in vivo fluorescence imaging in small animals using the optimal reflectance removal method. The HSI presented here enables quantitative unmixing of at least four fluorescent contrast agents (Alexa Fluor 610, 647, 700 and 750) simultaneously in living mice. A successful unmixing of the four fluorescent contrast agents was possible both using the pure contrast agents and with mixtures. The system could in principle also be applied to imaging of ex vivo tissue or intraoperative imaging in a clinical setting. These data suggest a promising approach for developing clinical applications of HSI based on multiplexed fluorescence contrast agent imaging.

  18. A dual oxygenation and fluorescence imaging platform for reconstructive surgery

    Science.gov (United States)

    Ashitate, Yoshitomo; Nguyen, John N.; Venugopal, Vivek; Stockdale, Alan; Neacsu, Florin; Kettenring, Frank; Lee, Bernard T.; Frangioni, John V.; Gioux, Sylvain

    2013-03-01

    There is a pressing clinical need to provide image guidance during surgery. Currently, assessment of tissue that needs to be resected or avoided is performed subjectively, leading to a large number of failures, patient morbidity, and increased healthcare costs. Because near-infrared (NIR) optical imaging is safe, noncontact, inexpensive, and can provide relatively deep information (several mm), it offers unparalleled capabilities for providing image guidance during surgery. These capabilities are well illustrated through the clinical translation of fluorescence imaging during oncologic surgery. In this work, we introduce a novel imaging platform that combines two complementary NIR optical modalities: oxygenation imaging and fluorescence imaging. We validated this platform during facial reconstructive surgery on large animals approaching the size of humans. We demonstrate that NIR fluorescence imaging provides identification of perforator arteries, assesses arterial perfusion, and can detect thrombosis, while oxygenation imaging permits the passive monitoring of tissue vital status, as well as the detection and origin of vascular compromise simultaneously. Together, the two methods provide a comprehensive approach to identifying problems and intervening in real time during surgery before irreparable damage occurs. Taken together, this novel platform provides fully integrated and clinically friendly endogenous and exogenous NIR optical imaging for improved image-guided intervention during surgery.

  19. Fluorescence lifetime imaging of oxygen in dental biofilm

    Science.gov (United States)

    Gerritsen, Hans C.; de Grauw, Cees J.

    2000-12-01

    Dental biofilm consists of micro-colonies of bacteria embedded in a matrix of polysaccharides and salivary proteins. pH and oxygen concentration are of great importance in dental biofilm. Both can be measured using fluorescence techniques. The imaging of dental biofilm is complicated by the thickness of the biofilms that can be up to several hundred micrometers thick. Here, we employed a combination of two-photon excitation microscopy with fluorescence lifetime imaging to quantify the oxygen concentration in dental biofilm. Collisional quenching of fluorescent probes by molecular oxygen leads to a reduction of the fluorescence lifetime of the probe. We employed this mechanism to measure the oxygen concentration distribution in dental biofilm by means of fluorescence lifetime imaging. Here, TRIS Ruthenium chloride hydrate was used as an oxygen probe. A calibration procedure on buffers was use to measure the lifetime response of this Ruthenium probe. The results are in agreement with the Stern-Volmer equation. A linear relation was found between the ratio of the unquenched and the quenched lifetime and the oxygen concentration. The biofilm fluorescence lifetime imaging results show a strong oxygen gradient at the buffer - biofilm interface and the average oxygen concentration in the biofilm amounted to 50 μM.

  20. In vivo cellular imaging using fluorescent proteins - Methods and Protocols

    Directory of Open Access Journals (Sweden)

    M. Monti

    2012-12-01

    Full Text Available The discovery and genetic engineering of fluorescent proteins has revolutionized cell biology. What was previously invisible to the cell often can be made visible with the use of fluorescent proteins. With this words, Robert M. Hoffman introduces In vivo Cellular Imaging Using Fluorescent proteins, the eighteen chapters book dedicated to the description of how fluorescence proteins have changed the way to analyze cellular processes in vivo. Modern researches aim to study new and less invasive methods able to follow the behavior of different cell types in different biological contexts: for example, how cancer cells migrate or how they respond to different therapies. Also, in vivo systems can help researchers to better understand animal embryonic development so as how fluorescence proteins may be used to monitor different processes in living organisms at the molecular and cellular level.

  1. Fluorescent Dendrimer Nanoconjugates as Advanced Probes for Biological Imaging

    Science.gov (United States)

    Reilly, Daniel; Kim, Sung Hoon; Katzenellenbogen, John A.; Schroeder, Charles M.

    2014-03-01

    Recent advances in fluorescence microscopy have enabled improvements in spatial resolution for biological imaging. However, there is a strong need for development of advanced fluorescent probes to enable a molecular-scale understanding of biological events. In this work, we report the development of a new class of probes for fluorescence imaging based on dye-conjugated dendrimer nanoconjugates. We utilize molecular-scale dendritic scaffolds as fluorescent probes, thereby enabling conjugation of multiple dyes and linkers to the scaffold periphery. In particular, we use polyamidoamine dendrimers as molecular scaffolds, wherein dye conjugation can be varied over a wide range. Single molecule fluorescence imaging shows that dendrimer nanoconjugates are far brighter than single fluorophores, resulting in increased localization precision. In addition, we further developed a new set of remarkably photostable probes by conjugating photoprotective triplet state quenchers directly onto the dendritic scaffold. We observe large increases in the photobleaching times compared to single dyes and reduced transient dark states (blinking). Overall, we believe that these new probes will allow for single molecule imaging over long time scales, enabling new vistas in biological imaging.

  2. Modulated electron-multiplied fluorescence lifetime imaging microscope: all-solid-state camera for fluorescence lifetime imaging

    NARCIS (Netherlands)

    Zhao, Q.; Schelen, B.; Schouten, R., et al.

    2012-01-01

    We have built an all-solid-state camera that is directly modulated at the pixel level for frequency-domain fluorescence lifetime imaging microscopy (FLIM) measurements. This novel camera eliminates the need for an image intensifier through the use of an application-specific charge coupled device des

  3. Enhanced speed in fluorescence imaging using beat frequency multiplexing

    Science.gov (United States)

    Mikami, Hideharu; Kobayashi, Hirofumi; Wang, Yisen; Hamad, Syed; Ozeki, Yasuyuki; Goda, Keisuke

    2016-03-01

    Fluorescence imaging using radiofrequency-tagged emission (FIRE) is an emerging technique that enables higher imaging speed (namely, temporal resolution) in fluorescence microscopy compared to conventional fluorescence imaging techniques such as confocal microscopy and wide-field microscopy. It works based on the principle that it uses multiple intensity-modulated fields in an interferometric setup as excitation fields and applies frequency-division multiplexing to fluorescence signals. Unfortunately, despite its high potential, FIRE has limited imaging speed due to two practical limitations: signal bandwidth and signal detection efficiency. The signal bandwidth is limited by that of an acousto-optic deflector (AOD) employed in the setup, which is typically 100-200 MHz for the spectral range of fluorescence excitation (400-600 nm). The signal detection efficiency is limited by poor spatial mode-matching between two interfering fields to produce a modulated excitation field. Here we present a method to overcome these limitations and thus to achieve higher imaging speed than the prior version of FIRE. Our method achieves an increase in signal bandwidth by a factor of two and nearly optimal mode matching, which enables the imaging speed limited by the lifetime of the target fluorophore rather than the imaging system itself. The higher bandwidth and better signal detection efficiency work synergistically because higher bandwidth requires higher signal levels to avoid the contribution of shot noise and amplifier noise to the fluorescence signal. Due to its unprecedentedly high-speed performance, our method has a wide variety of applications in cancer detection, drug discovery, and regenerative medicine.

  4. Photoacoustic imaging of a near-infrared fluorescent marker based on dual wavelength pump-probe excitation

    Science.gov (United States)

    Märk, Julia; Theiss, Christoph; Schmitt, Franz-Josef; Laufer, Jan

    2014-03-01

    Photoacoustic imaging has been used to determine the spatial distribution of fluorophores, such as exogenous dyes and genetically expressed proteins, from images acquired in phantoms and in vivo. Most methods involve the acquisition of multiwavelength images and rely on differences in the absorption spectra of the tissue chromophores to estimate the spatial distribution and abundance of the latter using spectral decomposition techniques, such as model based inversion schemes. However, the inversion of 3-D images can be computationally expensive. Experimental approaches to localising contrast agents may therefore be useful, especially if quantification is not essential. This work aims to develop a method for determining the spatial distribution of a near-infrared fluorescent cell marker from images acquired using dual wavelength excitation. The excitation wavelengths coincided with the absorption and emission spectrum of the fluorophore. The contrast mechanism relies on reducing the excited state lifetime of the fluorophore by inducing stimulated emission. This changes the amount of energy thermalized by the fluorophore, and hence the photoacoustic signal amplitude. Since this is not observed in endogenous chromophores, the background may be removed by subtracting two images acquired with and without pulse delay between the pump and probe pulses. To characterise the fluorophore, the signal amplitude is measured in a cuvette as a function of pulse delay, concentration, and fluence. The spatial distribution of the fluorophore is determined from images acquired in realistic tissue phantoms. This method may be suitable for in vivo applications, such as imaging of exogenous or genetically expressed fluorescent cell markers.

  5. Imaging of a targeted PDT drug with fluorescence tomography

    Science.gov (United States)

    Muffoletto, Dan; Gupta, Anurag; Xu, Zhiqiang; Mahrer, Chris; Bauer, Gretchen; Galas, Scott; Pandey, Ravindra K.; Sunar, Ulas

    2009-02-01

    We constructed a whole-body fluorescence tomography instrument to monitor novel bifunctional phototherapeutic drugs (e.g., HPPH-Cyanine dye conjugate) in small animals. The instrument allows dense source and detector sampling with a fast galvo scanner and a CCD detector for improved resolution and sensitivity (Patwardhan et al., 2005). Here we report tissue phantom measurements to evaluate the imaging performance with a newly constructed tomography instrument. Phantom measurements showed that strong fluorescence generated by HPPH-Cyanine dye (HPPH-CD), having high fluorescence quantum yield and long wavelength fluorescence emission, allowed deep tissue imaging. We also report in vivo fluorescence measurements of the conjugate in Nude mice bearing A549 human non-small cell lung carcinoma (NSCLC) tumors at 24 hr post injection to evaluate tumor detection ability of the conjugate. Our results indicate that the HPPH-CD shows preferential uptake in tumors compared to surrounding normal tissue at 24 hr post injection. This study demonstrates a potential use of HPPH-CD in detection (fluorescence imaging) and treatment (PDT) of deeply seated tumors.

  6. A portable fluorescence microscopic imaging system for cholecystectomy

    Science.gov (United States)

    Ye, Jian; Yang, Chaoyu; Gan, Qi; Ma, Rong; Zhang, Zeshu; Chang, Shufang; Shao, Pengfei; Zhang, Shiwu; Liu, Chenhai; Xu, Ronald

    2016-03-01

    In this paper we proposed a portable fluorescence microscopic imaging system to prevent iatrogenic biliary injuries from occurring during cholecystectomy due to misidentification of the cystic structures. The system consisted of a light source module, a CMOS camera, a Raspberry Pi computer and a 5 inch HDMI LCD. Specifically, the light source module was composed of 690 nm and 850 nm LEDs, allowing the CMOS camera to simultaneously acquire both fluorescence and background images. The system was controlled by Raspberry Pi using Python programming with the OpenCV library under Linux. We chose Indocyanine green(ICG) as a fluorescent contrast agent and then tested fluorescence intensities of the ICG aqueous solution at different concentration levels by our fluorescence microscopic system compared with the commercial Xenogen IVIS system. The spatial resolution of the proposed fluorescence microscopic imaging system was measured by a 1951 USAF resolution target and the dynamic response was evaluated quantitatively with an automatic displacement platform. Finally, we verified the technical feasibility of the proposed system in mouse models of bile duct, performing both correct and incorrect gallbladder resection. Our experiments showed that the proposed system can provide clear visualization of the confluence between the cystic duct and common bile duct or common hepatic duct, suggesting that this is a potential method for guiding cholecystectomy. The proposed portable system only cost a total of $300, potentially promoting its use in resource-limited settings.

  7. Rapid global fitting of large fluorescence lifetime imaging microscopy datasets.

    Directory of Open Access Journals (Sweden)

    Sean C Warren

    Full Text Available Fluorescence lifetime imaging (FLIM is widely applied to obtain quantitative information from fluorescence signals, particularly using Förster Resonant Energy Transfer (FRET measurements to map, for example, protein-protein interactions. Extracting FRET efficiencies or population fractions typically entails fitting data to complex fluorescence decay models but such experiments are frequently photon constrained, particularly for live cell or in vivo imaging, and this leads to unacceptable errors when analysing data on a pixel-wise basis. Lifetimes and population fractions may, however, be more robustly extracted using global analysis to simultaneously fit the fluorescence decay data of all pixels in an image or dataset to a multi-exponential model under the assumption that the lifetime components are invariant across the image (dataset. This approach is often considered to be prohibitively slow and/or computationally expensive but we present here a computationally efficient global analysis algorithm for the analysis of time-correlated single photon counting (TCSPC or time-gated FLIM data based on variable projection. It makes efficient use of both computer processor and memory resources, requiring less than a minute to analyse time series and multiwell plate datasets with hundreds of FLIM images on standard personal computers. This lifetime analysis takes account of repetitive excitation, including fluorescence photons excited by earlier pulses contributing to the fit, and is able to accommodate time-varying backgrounds and instrument response functions. We demonstrate that this global approach allows us to readily fit time-resolved fluorescence data to complex models including a four-exponential model of a FRET system, for which the FRET efficiencies of the two species of a bi-exponential donor are linked, and polarisation-resolved lifetime data, where a fluorescence intensity and bi-exponential anisotropy decay model is applied to the analysis

  8. Compressive Fluorescence Microscopy for Biological and Hyperspectral Imaging

    CERN Document Server

    Studer, Vincent; Chahid, Makhlad; Moussavi, Hamed; Candes, Emmanuel; Dahan, Maxime

    2012-01-01

    The mathematical theory of compressed sensing (CS) asserts that one can acquire signals from measurements whose rate is much lower than the total bandwidth. Whereas the CS theory is now well developed, challenges concerning hardware implementations of CS-based acquisition devices---especially in optics---have only started being addressed. This paper presents an implementation of compressive sensing in fluorescence microscopy and its applications to biomedical imaging. Our CS microscope combines a dynamic structured wide-field illumination and a fast and sensitive single-point fluorescence detection to enable reconstructions of images of fluorescent beads, cells and tissues with undersampling ratios (between the number of pixels and number of measurements) up to 32. We further demonstrate a hyperspectral mode and record images with 128 spectral channels and undersampling ratios up to 64, illustrating the potential benefits of CS acquisition for higher dimensional signals which typically exhibits extreme redund...

  9. Fluorescence Imaging of Fast Retrograde Axonal Transport in Living Animals

    Directory of Open Access Journals (Sweden)

    Dawid Schellingerhout

    2009-11-01

    Full Text Available Our purpose was to enable an in vivo imaging technology that can assess the anatomy and function of peripheral nerve tissue (neurography. To do this, we designed and tested a fluorescently labeled molecular probe based on the nontoxic C fragment of tetanus toxin (TTc. TTc was purified, labeled, and subjected to immunoassays and cell uptake assays. The compound was then injected into C57BL/6 mice (N = 60 for in vivo imaging and histologic studies. Image analysis and immunohistochemistry were performed. We found that TTc could be labeled with fluorescent moieties without loss of immunoreactivity or biologic potency in cell uptake assays. In vivo fluorescent imaging experiments demonstrated uptake and retrograde transport of the compound along the course of the sciatic nerve and in the spinal cord. Ex vivo imaging and immunohistochemical studies confirmed the presence of TTc in the sciatic nerve and spinal cord, whereas control animals injected with human serum albumin did not exhibit these features. We have demonstrated neurography with a fluorescently labeled molecular imaging contrast agent based on the TTc.

  10. 3-D Image Analysis of Fluorescent Drug Binding

    Directory of Open Access Journals (Sweden)

    M. Raquel Miquel

    2005-01-01

    Full Text Available Fluorescent ligands provide the means of studying receptors in whole tissues using confocal laser scanning microscopy and have advantages over antibody- or non-fluorescence-based method. Confocal microscopy provides large volumes of images to be measured. Histogram analysis of 3-D image volumes is proposed as a method of graphically displaying large amounts of volumetric image data to be quickly analyzed and compared. The fluorescent ligand BODIPY FL-prazosin (QAPB was used in mouse aorta. Histogram analysis reports the amount of ligand-receptor binding under different conditions and the technique is sensitive enough to detect changes in receptor availability after antagonist incubation or genetic manipulations. QAPB binding was concentration dependent, causing concentration-related rightward shifts in the histogram. In the presence of 10 μM phenoxybenzamine (blocking agent, the QAPB (50 nM histogram overlaps the autofluorescence curve. The histogram obtained for the 1D knockout aorta lay to the left of that of control and 1B knockout aorta, indicating a reduction in 1D receptors. We have shown, for the first time, that it is possible to graphically display binding of a fluorescent drug to a biological tissue. Although our application is specific to adrenergic receptors, the general method could be applied to any volumetric, fluorescence-image-based assay.

  11. Snapshot imaging Fraunhofer line discriminator for detection of plant fluorescence

    Science.gov (United States)

    Gupta Roy, S.; Kudenov, M. W.

    2015-05-01

    Non-invasive quantification of plant health is traditionally accomplished using reflectance based metrics, such as the normalized difference vegetative index (NDVI). However, measuring plant fluorescence (both active and passive) to determine photochemistry of plants has gained importance. Due to better cost efficiency, lower power requirements, and simpler scanning synchronization, detecting passive fluorescence is preferred over active fluorescence. In this paper, we propose a high speed imaging approach for measuring passive plant fluorescence, within the hydrogen alpha Fraunhofer line at ~656 nm, using a Snapshot Imaging Fraunhofer Line Discriminator (SIFOLD). For the first time, the advantage of snapshot imaging for high throughput Fraunhofer Line Discrimination (FLD) is cultivated by our system, which is based on a multiple-image Fourier transform spectrometer and a spatial heterodyne interferometer (SHI). The SHI is a Sagnac interferometer, which is dispersion compensated using blazed diffraction gratings. We present data and techniques for calibrating the SIFOLD to any particular wavelength. This technique can be applied to quantify plant fluorescence at low cost and reduced complexity of data collection.

  12. Tumor-stem cells interactions by fluorescence imaging

    Science.gov (United States)

    Meleshina, Aleksandra V.; Cherkasova, Elena I.; Sergeeva, Ekaterina; Turchin, Ilya V.; Kiseleva, Ekaterina V.; Dashinimaev, Erdem B.; Shirmanova, Marina V.; Zagaynova, Elena V.

    2013-02-01

    Recently, great deal of interest is investigation the function of the stem cells (SC) in tumors. In this study, we studied «recipient-tumor- fluorescent stem cells » system using the methods of in vivo imaging and laser scanning microscopy (LSM). We used adipose-derived adult stem (ADAS) cells of human lentiviral transfected with the gene of fluorescent protein Turbo FP635. ADAS cells were administrated into nude mice with transplanted tumor HeLa Kyoto (human cervical carcinoma) at different stages of tumor growth (0-8 days) intravenously or into tumor. In vivo imaging was performed on the experimental setup for epi - luminescence bioimaging (IAP RAS, Nizhny Novgorod). The results of the imaging showed localization of fluorophore tagged stem cells in the spleen on day 5-9 after injection. The sensitivity of the technique may be improved by spectral separation autofluorescence and fluorescence of stem cells. We compared the results of in vivo imaging and confocal laser scanning microscopy (LSM 510 META, Carl Zeiss, Germany). Internal organs of the animals and tumor tissue were investigated. It was shown that with i.v. injection of ADAS, bright fluorescent structures with spectral characteristics corresponding to TurboFP635 protein are locally accumulated in the marrow, lungs and tumors of animals. These findings indicate that ADAS cells integrate in the animal body with transplanted tumor and can be identified by fluorescence bioimaging techniques in vivo and ex vivo.

  13. Coded Aperture Imaging for Fluorescent X-rays-Biomedical Applications

    Energy Technology Data Exchange (ETDEWEB)

    Haboub, Abdel; MacDowell, Alastair; Marchesini, Stefano; Parkinson, Dilworth

    2013-06-01

    Employing a coded aperture pattern in front of a charge couple device pixilated detector (CCD) allows for imaging of fluorescent x-rays (6-25KeV) being emitted from samples irradiated with x-rays. Coded apertures encode the angular direction of x-rays and allow for a large Numerical Aperture x- ray imaging system. The algorithm to develop the self-supported coded aperture pattern of the Non Two Holes Touching (NTHT) pattern was developed. The algorithms to reconstruct the x-ray image from the encoded pattern recorded were developed by means of modeling and confirmed by experiments. Samples were irradiated by monochromatic synchrotron x-ray radiation, and fluorescent x-rays from several different test metal samples were imaged through the newly developed coded aperture imaging system. By choice of the exciting energy the different metals were speciated.

  14. Whole mount nuclear fluorescent imaging: convenient documentation of embryo morphology.

    Science.gov (United States)

    Sandell, Lisa L; Kurosaka, Hiroshi; Trainor, Paul A

    2012-11-01

    Here, we describe a relatively inexpensive and easy method to produce high quality images that reveal fine topological details of vertebrate embryonic structures. The method relies on nuclear staining of whole mount embryos in combination with confocal microscopy or conventional wide field fluorescent microscopy. In cases where confocal microscopy is used in combination with whole mount nuclear staining, the resulting embryo images can rival the clarity and resolution of images produced by scanning electron microscopy (SEM). The fluorescent nuclear staining may be performed with a variety of cell permeable nuclear dyes, enabling the technique to be performed with multiple standard microscope/illumination or confocal/laser systems. The method may be used to document morphology of embryos of a variety of organisms, as well as individual organs and tissues. Nuclear stain imaging imposes minimal impact on embryonic specimens, enabling imaged specimens to be utilized for additional assays.

  15. APPLICATION OF MODULATED CHLOROPHYLL FLUORESCENCE AND MODULATED CHLOROPHYLL FLUORESCENCE IMAGING IN STUDYING ENVIRONMENTAL STRESSES EFFECT

    Directory of Open Access Journals (Sweden)

    L. Guidi

    2016-03-01

    Full Text Available Chlorophyll (Chl a fluorescence is a widely used tool to monitor the photosynthetic process in plants subjected to environmental stresses.this review reports the theoretical bases of Chl fluorescence, and the significance of the most important Chl fluorescence parameters. it also reportshow these parameters can be utilised to estimate changes in photosystem ii (PSII photochemistry, linear electron flux and energy dissipationmechanisms. the relation between actual PSII photochemistry and CO2 assimilation is discussed, as is the role of photochemical andnon-photochemical quenching in inducing changes in PSII activity. the application of Chl fluorescence imaging to study heterogeneity on leaflamina is also considered. this review summarises only some of the results obtained by this methodology to study the effects of differentenvironmental stresses, namely water and nutrients availability, pollutants, temperature and salinity.

  16. Breast MRI at Very Short TE (minTE): Image Analysis of minTE Sequences on Non-Fat-Saturated, Subtracted T1-Weighted Images.

    Science.gov (United States)

    Wenkel, Evelyn; Janka, Rolf; Geppert, Christian; Kaemmerer, Nadine; Hartmann, Arndt; Uder, Michael; Hammon, Matthias; Brand, Michael

    2017-02-01

    Purpose The aim was to evaluate a minimum echo time (minTE) protocol for breast magnetic resonance imaging (MRI) in patients with breast lesions compared to a standard TE (nTE) time protocol. Methods Breasts of 144 women were examined with a 1.5 Tesla MRI scanner. Additionally to the standard gradient-echo sequence with nTE (4.8 ms), a variant with minimum TE (1.2 ms) was used in an interleaved fashion which leads to a better temporal resolution and should reduce the scan time by approximately 50 %. Lesion sizes were measured and the signal-to-noise ratio (SNR) as well as the contrast-to-noise ratio (CNR) were calculated. Subjective confidence was evaluated using a 3-point scale before looking at the nTE sequences (1 = very sure that I can identify a lesion and classify it, 2 = quite sure that I can identify a lesion and classify it, 3 = definitely want to see nTE for final assessment) and the subjective image quality of all examinations was evaluated using a four-grade scale (1 = sharp, 2 = slight blur, 3 = moderate blur and 4 = severe blur/not evaluable) for lesion and skin sharpness. Lesion morphology and contrast enhancement were also evaluated. Results With minTE sequences, no lesion was rated with "definitely want to see nTE sequences for final assessment". The difference of the longitudinal and transverse diameter did not differ significantly (p > 0.05). With minTE, lesions and skin were rated to be significantly more blurry (p Image Analysis of minTE Sequences on Non-Fat-Saturated, Subtracted T1-Weighted Images. Fortschr Röntgenstr 2017; 189: 137 - 145.

  17. Color-matched esophagus phantom for fluorescent imaging

    Science.gov (United States)

    Yang, Chenying; Hou, Vivian; Nelson, Leonard Y.; Seibel, Eric J.

    2013-02-01

    We developed a stable, reproducible three-dimensional optical phantom for the evaluation of a wide-field endoscopic molecular imaging system. This phantom mimicked a human esophagus structure with flexibility to demonstrate body movements. At the same time, realistic visual appearance and diffuse spectral reflectance properties of the tissue were simulated by a color matching methodology. A photostable dye-in-polymer technology was applied to represent biomarker probed "hot-spot" locations. Furthermore, fluorescent target quantification of the phantom was demonstrated using a 1.2mm ultrathin scanning fiber endoscope with concurrent fluorescence-reflectance imaging.

  18. Fluorescence Lifetime Imaging of Quantum Dot Labeled DNA Microarrays

    Directory of Open Access Journals (Sweden)

    Jonathan G. Terry

    2009-04-01

    Full Text Available Quantum dot (QD labeling combined with fluorescence lifetime imaging microscopy is proposed as a powerful transduction technique for the detection of DNA hybridization events. Fluorescence lifetime analysis of DNA microarray spots of hybridized QD labeled target indicated a characteristic lifetime value of 18.8 ns, compared to 13.3 ns obtained for spots of free QD solution, revealing that QD labels are sensitive to the spot microenvironment. Additionally, time gated detection was shown to improve the microarray image contrast ratio by 1.8, achieving femtomolar target sensitivity. Finally, lifetime multiplexing based on Qdot525 and Alexa430 was demonstrated using a single excitation-detection readout channel.

  19. Using digital subtraction in computer simulated images as a tool to aid the visual detection of masked lesions in dense breasts

    Science.gov (United States)

    Schiabel, Homero; Guimarães, Luciana T.; Sousa, Maria A. Z.

    2015-03-01

    This work proposes a simulation model involving subtraction of digital mammography images obtained at different X-ray beam levels of energy to aid the detection of breast malignant lesions. Absorption coefficients behavior of 3 main structures of clinical interest - adipose tissue, fiber glandular tissue and the typical carcinoma - as a function of the beam energy from a Mo X-ray tube was the basis to develop a computer simulation of the possible acquired images. The simulation has considered a typical compressed breast with 4.5cm in thickness, and variations of the carcinoma and glandular tissues thicknesses - 0.4 up to 2.0cm and 4.1 to 2.5cm, respectively - were evaluated as a function of the photons mean energy - 14 up to 25 keV, in the typical mammography energy range. Results have shown that: (a) if the carcinoma thickness is over 0.4cm, its detection may be feasible even masked by fiber tissue with exposures in the range of 19 to 25 keV; (b) for masked carcinoma with thickness in the range of 0.4-2.0cm, the proposed procedure can enhance it in the image resulting from the digital subtraction between images obtained at 14 and at 22 keV. Therefore such results indicate that this simulation procedure can be a useful tool in aiding the identification of possible missed malignant lesions which could not be detected in the typical exam, mainly considering dense breasts.

  20. A Practical Approach to Quantitative Processing and Analysis of Small Biological Structures by Fluorescent Imaging

    Science.gov (United States)

    Noller, Crystal M.; Boulina, Maria; McNamara, George; Szeto, Angela; McCabe, Philip M.

    2016-01-01

    Standards in quantitative fluorescent imaging are vaguely recognized and receive insufficient discussion. A common best practice is to acquire images at Nyquist rate, where highest signal frequency is assumed to be the highest obtainable resolution of the imaging system. However, this particular standard is set to insure that all obtainable information is being collected. The objective of the current study was to demonstrate that for quantification purposes, these correctly set acquisition rates can be redundant; instead, linear size of the objects of interest can be used to calculate sufficient information density in the image. We describe optimized image acquisition parameters and unbiased methods for processing and quantification of medium-size cellular structures. Sections of rabbit aortas were immunohistochemically stained to identify and quantify sympathetic varicosities, >2 μm in diameter. Images were processed to reduce background noise and segment objects using free, open-access software. Calculations of the optimal sampling rate for the experiment were based on the size of the objects of interest. The effect of differing sampling rates and processing techniques on object quantification was demonstrated. Oversampling led to a substantial increase in file size, whereas undersampling hindered reliable quantification. Quantification of raw and incorrectly processed images generated false structures, misrepresenting the underlying data. The current study emphasizes the importance of defining image-acquisition parameters based on the structure(s) of interest. The proposed postacquisition processing steps effectively removed background and noise, allowed for reliable quantification, and eliminated user bias. This customizable, reliable method for background subtraction and structure quantification provides a reproducible tool for researchers across biologic disciplines. PMID:27182204

  1. 3D Beam Reconstruction by Fluorescence Imaging

    CERN Document Server

    Radwell, Neal; Franke-Arnold, Sonja

    2013-01-01

    We present a technique for mapping the complete 3D spatial intensity profile of a laser beam from its fluorescence in an atomic vapour. We propagate shaped light through a rubidium vapour cell and record the resonant scattering from the side. From a single measurement we obtain a camera limited resolution of 200 x 200 transverse points and 659 longitudinal points. In constrast to invasive methods in which the camera is placed in the beam path, our method is capable of measuring patterns formed by counterpropagating laser beams. It has high resolution in all 3 dimensions, is fast and can be completely automated. The technique has applications in areas which require complex beam shapes, such as optical tweezers, atom trapping and pattern formation.

  2. Quasi-real-time fluorescence imaging with lifetime dependent contrast

    Science.gov (United States)

    Jiang, Pei-Chi; Grundfest, Warren S.; Stafsudd, Oscar M.

    2011-08-01

    Conventional fluorescence lifetime imaging requires complicated algorithms to extract lifetimes of fluorophores and acquisition of multiple data points at progressively longer delay times to characterize tissues. To address diminishing signal-to-noise ratios at these progressively longer time delays, we report a time-resolved fluorescence imaging method, normalized fluorescence yield imaging that does not require the extraction of lifetimes. The concept is to extract the ``contrast'' instead of the lifetime value of the fluorophores by using simple mathematical algorithms. This process converts differences in decay times directly to different intensities. The technique was verified experimentally using a gated iCCD camera and an ultraviolet light-emitting diode light source. It was shown that this method can distinguish between chemical dyes (Fluorescein and Rhodamine-B) and biomedical samples, such as powders of elastin and collagen. Good contrast was obtained between fluorophores that varied by less than 6% in lifetime. Additionally, it was shown that long gate times up to 16 ns achieve good contrast depending upon the samples to be studied. These results support the feasibility of time-resolved fluorescence imaging without lifetime extraction, which has a potential clinical role in noninvasive real-time imaging.

  3. Inherently fluorescent polystyrene microspheres for coating, sensing and cellular imaging.

    Science.gov (United States)

    Qu, Jian-Bo; Xu, Yu-Liang; Liu, Yu; Wang, Yanan; Sui, Yuanhong; Liu, Jian-Guo; Wang, Xiaojuan

    2017-04-01

    Commercially available polystyrene (PS) fluorescent microspheres are widely used in biological field for tracing, in vivo imaging and calibration of flow cytometry, among other applications. However, these particles do suffer from some drawbacks such as the leakage and photobleaching of organic dyes within them. In the present study, inherently fluorescent properties of PS based microspheres have been explored for the first time. Here we find that a simple chloromethylation reaction endows the polystyrene particles with inherent fluorescence without any subsequent conjugation of an external fluorophore. A possible mechanism for fluorescence is elucidated by synthesizing and investigating p-ethylbenzyl chloride, a compound with similar structure. Significantly, no photobleaching or leaking issues were observed owing to the stable structure of the microspheres. Chloromethylated PS (CMPS) microspheres can keep their perpetual blue fluorescence even in dry powder state making them attractive as a potential coating material. Furthermore, the chloromethyl groups on CMPS microspheres make them very convenient for further functionalization. Poly(ethylene glycol) (PEG) grafted microspheres showed good biocompatibility and negligible cytotoxicity, and could be used to image intracellular Fe(3+) due to the selective fluorescence quenching effect of aqueous Fe(3+) in cytoplasm.

  4. Enhanced 3D fluorescence live cell imaging on nanoplasmonic substrate

    Energy Technology Data Exchange (ETDEWEB)

    Gartia, Manas Ranjan [Department of Nuclear, Plasma and Radiological Engineering, University of Illinois, Urbana, IL 61801 (United States); Hsiao, Austin; Logan Liu, G [Department of Bioengineering, University of Illinois, Urbana, IL 61801 (United States); Sivaguru, Mayandi [Institute for Genomic Biology, University of Illinois, Urbana, IL 61801 (United States); Chen Yi, E-mail: loganliu@illinois.edu [Department of Electrical and Computer Engineering, University of Illinois, Urbana, IL 61801 (United States)

    2011-09-07

    We have created a randomly distributed nanocone substrate on silicon coated with silver for surface-plasmon-enhanced fluorescence detection and 3D cell imaging. Optical characterization of the nanocone substrate showed it can support several plasmonic modes (in the 300-800 nm wavelength range) that can be coupled to a fluorophore on the surface of the substrate, which gives rise to the enhanced fluorescence. Spectral analysis suggests that a nanocone substrate can create more excitons and shorter lifetime in the model fluorophore Rhodamine 6G (R6G) due to plasmon resonance energy transfer from the nanocone substrate to the nearby fluorophore. We observed three-dimensional fluorescence enhancement on our substrate shown from the confocal fluorescence imaging of chinese hamster ovary (CHO) cells grown on the substrate. The fluorescence intensity from the fluorophores bound on the cell membrane was amplified more than 100-fold as compared to that on a glass substrate. We believe that strong scattering within the nanostructured area coupled with random scattering inside the cell resulted in the observed three-dimensional enhancement in fluorescence with higher photostability on the substrate surface.

  5. Doped semiconductor nanocrystal based fluorescent cellular imaging probes.

    Science.gov (United States)

    Maity, Amit Ranjan; Palmal, Sharbari; Basiruddin, S K; Karan, Niladri Sekhar; Sarkar, Suresh; Pradhan, Narayan; Jana, Nikhil R

    2013-06-21

    Doped semiconductor nanocrystals such as Mn doped ZnS, Mn doped ZnSe and Cu doped InZnS, are considered as new classes of fluorescent biological probes with low toxicity. Although the synthesis in high quality of such nanomaterials is now well established, transforming them into functional fluorescent probes remains a challenge. Here we report a fluorescent cellular imaging probe made of high quality doped semiconductor nanocrystals. We have identified two different coating approaches suitable for transforming the as synthesized hydrophobic doped semiconductor nanocrystals into water-soluble functional nanoparticles. Following these approaches we have synthesized TAT-peptide- and folate-functionalized nanoparticles of 10-80 nm hydrodynamic diameter and used them as a fluorescent cell label. The results shows that doped semiconductor nanocrystals can be an attractive alternative for conventional cadmium based quantum dots with low toxicity.

  6. Optimization of microsatellite DNA Gelred fluorescence imaging ...

    African Journals Online (AJOL)

    user1

    2012-10-11

    Oct 11, 2012 ... In order to explore the best microsatellite DNA Gelred imaging technology, this study .... The cycling parameters were: 4 min at 94°C, followed by 35 cycles of ..... double-strand breaks in mammalian cells. Nucleic Acids Res.

  7. Polyacrylamide based ICG nanocarriers for enhanced fluorescence and photoacoustic imaging

    Science.gov (United States)

    Ray, Aniruddha; Yoon, Hyung Ki; Ryu, HeeJu; Koo Lee, Yong-Eun; Kim, Gwangseong; Wang, Xueding; Kopelman, Raoul

    2013-02-01

    Indocyanine green (ICG) is an FDA approved tricarbocyanine dye. This dye, with a strong absorbance in the near infrared (NIR) region, has been extensively used for fluorescence and photoacoustic imaging in vivo. ICG in its free form, however, has a few drawbacks that limit its in vivo applications, such as non-targetability, tendency to form aggregates which changes its optical properties, fast degradation, short plasma lifetime and reduced fluorescence at body temperature. In order to bypass these inherent drawbacks, we demonstrate a polyacrylamide based nanocarrier that was particularly designed to carry the negatively charged ICG molecules. These nanocarriers are biodegradable, biocompatible and can be specifically targeted to any cell or tissue. Using these nanocarriers we avoid all the problems associated with free ICG, such as degradation, aggregation and short plasma lifetime, and also enhance demonstrate its ability towards photoacoustics and fluorescence imaging.

  8. The Cyan Fluorescent Protein (CFP) Transgenic Mouse as a Model for Imaging Pancreatic Exocrine Cells

    Science.gov (United States)

    Cao, Hop S Tran; Kimura, Hiroaki; Kaushal, Sharmeela; Snyder, Cynthia S; Reynoso, Jose; Hoffman, Robert M; Bouvet, Michael

    2015-01-01

    Context The use of fluorescent proteins for in vivo imaging has opened many new areas of research. Among the important advances in the field have been the development of transgenic mice expressing various fluorescent proteins. Objective To report whole-body and organ-specific fluorescence imaging to characterize the transgenic cyan fluorescent protein mouse. Design Mice were imaged using two devices. Brightfield images were obtained with the OV100 Small Animal Imaging System (Olympus Corp., Tokyo, Japan). Fluorescence imaging was performed under the cyan fluorescent protein filter using the iBox Small Animal Imaging System (UVP, Upland, CA, USA). Intervention All animals were sacrificed immediately before imaging. They were imaged before and throughout multiple steps of a complete necropsy. Harvested organs were also imaged with both devices. Selected organs were then frozen and processed for histology, fluorescence microscopy, and H&E staining. Fluorescence microscopy was performed with an Olympus IMT-2 inverted fluorescence microscope. Main outcome measure Determination of fluorescence intensity of different organs. Results Surprisingly, we found that there is differential enhancement of fluorescence among organs; most notably, the pancreas stands out from the rest of the gastrointestinal tract, displaying the strongest fluorescence of all organs in the mouse. Fluorescence microscopy demonstrated that the cyan fluorescent protein fluorescence resided in the acinar cells of the pancreas and not the islet cells. Conclusions The cyan fluorescent protein mouse should lead to a deeper understanding of pancreatic function and pathology, including cancer. PMID:19287108

  9. The Cyan Fluorescent Protein (CFP Transgenic Mouse as a Model for Imaging Pancreatic Exocrine Cells

    Directory of Open Access Journals (Sweden)

    Hop S Tran Cao

    2009-03-01

    Full Text Available The use of fluorescent proteins for in vivo imaging has opened many new areas of research. Among the important advances in the field have been the development of transgenic mice expressing various fluorescent proteins. Objective To report whole-body and organ-specific fluorescence imaging to characterize the transgenic cyan fluorescent protein mouse. Design Mice were imaged using two devices. Brightfield images were obtained with the OV100 Small Animal Imaging System (Olympus Corp., Tokyo, Japan. Fluorescence imaging was performed under the cyan fluorescent protein filter using the iBox Small Animal Imaging System (UVP, Upland, CA, USA. Intervention All animals were sacrificed immediately before imaging. They were imaged before and throughout multiple steps of a complete necropsy. Harvested organs were also imaged with both devices. Selected organs were then frozen and processed for histology, fluorescence microscopy, and H&E staining. Fluorescence microscopy was performed with an Olympus IMT-2 inverted fluorescence microscope. Main outcome measure Determination of fluorescence intensity of different organs. Results Surprisingly, we found that there is differential enhancement of fluorescence among organs; most notably, the pancreas stands out from the rest of the gastrointestinal tract, displaying the strongest fluorescence of all organs in the mouse. Fluorescence microscopy demonstrated that the cyan fluorescent protein fluorescence resided in the acinar cells of the pancreas and not the islet cells. Conclusions The cyan fluorescent protein mouse should lead to a deeper understanding of pancreatic function and pathology, including cancer.

  10. Fluorescence Lifetime Imaging of Free and Protein-Bound NADH

    Science.gov (United States)

    Lakowicz, Joseph R.; Szmacinski, Henryk; Nowaczyk, Kazimierz; Johnson, Michael L.

    1992-02-01

    We introduce a methodology, fluorescence lifetime imaging (FLIM), in which the contrast depends on the fluorescence lifetime at each point in a two-dimensional image and not on the local concentration and/or intensity of the fluorophore. We used FLIM to create lifetime images of NADH when free in solution and when bound to malate dehydrogenase. This represents a challenging case for lifetime imaging because the NADH decay times are just 0.4 and 1.0 ns in the free and bound states, respectively. In the present apparatus, lifetime images are created from a series of phase-sensitive images obtained with a gain-modulated image intensifier and recorded with a charge-coupled device (CCD) camera. The intensifier gain is modulated at the light-modulation frequency or a harmonic thereof. A series of stationary phase-sensitive images, each obtained with various phase shifts of the gain-modulation signal, is used to determine the phase angle or modulation of the emission at each pixel, which is in essence the lifetime image. We also describe an imaging procedure that allows specific decay times to be suppressed, allowing in this case suppression of the emission from either free or bound NADH. Since the fluorescence lifetimes of probes are known to be sensitive to numerous chemical and physical factors such as pH, oxygen, temperature, cations, polarity, and binding to macromolecules, this method allows imaging of the chemical or property of interest in macroscopic and microscopic samples. The concept of FLIM appears to have numerous potential applications in the biosciences.

  11. Polyester Fabric's Fluorescent Dyeing in Supercritical Carbon Dioxide and its Fluorescence Imaging.

    Science.gov (United States)

    Xiong, Xiaoqing; Xu, Yanyan; Zheng, Laijiu; Yan, Jun; Zhao, Hongjuan; Zhang, Juan; Sun, Yanfeng

    2017-03-01

    As one of the most important coumarin-like dyes, disperse fluorescent Yellow 82 exhibits exceptionally large two-photon effects. Here, it was firstly introduced into the supercritical CO2 dyeing polyester fabrics in this work. Results of the present work showed that the dyeing parameters such as the dyeing time, pressure and temperature had remarkable influences on the color strength of fabrics. The optimized dyeing condition in supercritical CO2 dyeing has been proposed that the dyeing time was 60 min; the pressure was 25 MPa and the temperature was 120 °C. As a result, acceptable products were obtained with the wash and rub fastness rating at 5 or 4-5. The polyester fabrics dyed with fluorescent dyes can be satisfied for the requirement of manufacturing warning clothing. Importantly, the confocal microscopy imaging technology was successfully introduced into textile fields to observe the distribution and fluorescence intensity of disperse fluorescent Yellow 82 on polyester fabrics. As far as we know, this is the first report about supercritical CO2 dyeing polyester fabrics based on disperse fluorescent dyes. It will be very helpful for the further design of new fluorescent functional dyes suitable for supercritical CO2 dyeing technique.

  12. Two-photon fluorescence and fluorescence imaging of two styryl heterocyclic dyes combined with DNA.

    Science.gov (United States)

    Gao, Chao; Liu, Shu-yao; Zhang, Xian; Liu, Ying-kai; Qiao, Cong-de; Liu, Zhao-e

    2016-03-01

    Two new styryl heterocyclic two-photon (TP) materials, 4-[4-(N-methyl)styrene]-imidazo [4,5-f][1,10] phenanthroline-benzene iodated salt (probe-1) and 4,4-[4-(N-methyl)styrene]-benzene iodated salt (probe-2) were successfully synthesized and studied as potential fluorescent probes of DNA detection. The linear and nonlinear photophysical properties of two compounds in different solvents were investigated. The absorption, one- and two-photon fluorescent spectra of the free dye and dye-DNA complex were also examined to evaluate their photophysical properties. The binding constants of dye-DNA were obtained according to Scatchard equation with good values. The results showed that two probes could be used as fluorescent DNA probes by two-photon excitation, and TP fluorescent properties of probe-1 are superior to that of probe-2. The fluorescent method date indicated that the mechanisms of dye-DNA complex interaction may be groove binding for probe-1 and electrostatic interaction for probe-2, respectively. The MTT assay experiments showed two probes are low toxicity. Moreover, the TP fluorescence imaging of DNA detection in living cells at 800 nm indicated that the ability to locate in cell nuclei of probe-1 is better than that of probe-2.

  13. Theoretical and experimental comparison of image signal and noise for dual-energy subtraction angiography and conventional x-ray angiography

    Science.gov (United States)

    Burton, Christiane S.; Mayo, John R.; Cunningham, I. A.

    2015-03-01

    Cardiovascular disease is currently the leading cause of mortality worldwide. Digital subtraction angiography (DSA) is widely used to enhance the visibility of small vessels and vasculature obscurred by overlying bone and lung fields by subtracting a mask and contrast image. However, motion between these mask and contrast images can introduce artifacts that can render a study non-diagnostic. This makes DSA particularly unsuccessful for cardiac imaging. A method called dual-energy, or energy subtraction angiography (ESA), was proposed in the past as an alternative for vascular imaging, however it was not pursued because experimental results suggested that image quality was deemed as poor and inferior to DSA. Image quality for angiography comes down to iodine signal and noise. In this paper we investigate the fundamental iodine signal and noise analysis of ESA and compare it to DSA. Method: We developed a polyenergetic and monoenergetic theoretical model for iodine signal and noise for both ESA and DSA. We validated our polyenergetic model by experiment where ESA and DSA images of a vascular phantom were acquired using an x-ray system with a flat panel CsI Xmaru1215CF-MPTM (Rayence Co., Ltd., Republic of Korea) detector. For ESA low and high applied tube voltages of 50 kV and 120 kV (2.5 mmCu), respectively, and for DSA the applied tube voltage was 80 kV. Iodine signal-to-noise ratio (SNR) per entrance exposure was calculated for each iodine concentration for both ESA and DSA. Results: Our measured iodine SNR agreed well with theoretical calculations. Iodine SNR for ESA was relatively higher than DSA for low iodine mass loadings, and as iodine mass loading increases iodine SNR decreases. Conclusions: We have developed a model for iodine SNR for both DSA and ESA. Our model was validated with experiment and showed excellent agreement. We have shown that there is potential for obtaining iodine-specific images using ESA that are similar to DSA.

  14. Photon budget analysis for fluorescence lifetime imaging microscopy

    NARCIS (Netherlands)

    Zhao, Q.; Young, I.T.; De Jong, J.G.S.

    2011-01-01

    We have constructed a mathematical model to analyze the photon efficiency of frequency-domain fluorescence lifetime imaging microscopy (FLIM). The power of the light source needed for illumination in a FLIM system and the signal-to-noise ratio of the detector have led us to a photon “budget.” These

  15. Fluorescence lifetime imaging in biosciences: technologies and applications

    Institute of Scientific and Technical Information of China (English)

    Raluca NIESNER; Karl-Heinz GERICKE

    2008-01-01

    The biosciences require the development of methods that allow a non-invasive and rapid investigation of biological systems. In this aspect, high-end imaging tech-niques allow intravital microscopy in real-time, providing information on a molecular basis. Far-field fluorescence imaging techniques are some of the most adequate methods for such investigations. However, there are great differences between the common fluorescence imaging techniques, i.e., wide-field, confocal one-photon and two-photon microscopy, as far as their applicability in diverse bioscientific research areas is concerned. In the first part of this work, we briefly compare these techniques. Standard methods used in the biosciences, i.e., steady-state techniques based on the analy-sis of the total fluorescence signal originating from the sam-ple, can successfully be employed in the study of cell, tissue and organ morphology as well as in monitoring the macro-scopic tissue function. However, they are mostly inadequate for the quantitative investigation of the cellular function at the molecular level. The intrinsic disadvantages of steady-state techniques are countered by using time-resolved tech-niques. Among these fluorescence lifetime imaging (FLIM) is currently the most common. Different FLIM principles as well as applications of particular relevance for the biosci-ences, especially for fast intravital studies are discussed in this work.

  16. Colorful protein-based fluorescent probes for collagen imaging.

    Directory of Open Access Journals (Sweden)

    Stijn J A Aper

    Full Text Available Real-time visualization of collagen is important in studies on tissue formation and remodeling in the research fields of developmental biology and tissue engineering. Our group has previously reported on a fluorescent probe for the specific imaging of collagen in live tissue in situ, consisting of the native collagen binding protein CNA35 labeled with fluorescent dye Oregon Green 488 (CNA35-OG488. The CNA35-OG488 probe has become widely used for collagen imaging. To allow for the use of CNA35-based probes in a broader range of applications, we here present a toolbox of six genetically-encoded collagen probes which are fusions of CNA35 to fluorescent proteins that span the visible spectrum: mTurquoise2, EGFP, mAmetrine, LSSmOrange, tdTomato and mCherry. While CNA35-OG488 requires a chemical conjugation step for labeling with the fluorescent dye, these protein-based probes can be easily produced in high yields by expression in E. coli and purified in one step using Ni2+-affinity chromatography. The probes all bind specifically to collagen, both in vitro and in porcine pericardial tissue. Some first applications of the probes are shown in multicolor imaging of engineered tissue and two-photon imaging of collagen in human skin. The fully-genetic encoding of the new probes makes them easily accessible to all scientists interested in collagen formation and remodeling.

  17. A Review of Indocyanine Green Fluorescent Imaging in Surgery

    Directory of Open Access Journals (Sweden)

    Jarmo T. Alander

    2012-01-01

    Full Text Available The purpose of this paper is to give an overview of the recent surgical intraoperational applications of indocyanine green fluorescence imaging methods, the basics of the technology, and instrumentation used. Well over 200 papers describing this technique in clinical setting are reviewed. In addition to the surgical applications, other recent medical applications of ICG are briefly examined.

  18. Behavior Subtraction applied to radar

    NARCIS (Netherlands)

    Rossum, W.L. van; Caro Cuenca, M.

    2014-01-01

    An algorithm developed for optical images has been applied to radar data. The algorithm, Behavior Subtraction, is based on capturing the dynamics of a scene and detecting anomalous behavior. The radar application is the detection of small surface targets at sea. The sea surface yields the expected s

  19. Fluorescence lifetime to image epidermal ionic concentrations

    Science.gov (United States)

    Behne, Martin J.; Barry, Nicholas P.; Moll, Ingrid; Gratton, Enrico; Mauro, Theodora M.

    2004-09-01

    Measurements of ionic concentrations in skin have traditionally been performed with an array of methods which either did not reveal detailed localization information, or only provided qualitative, not quantitative information. FLIM combines a number of advantages into a method ideally suited to visualize concentrations of ions such as H+ in intact, unperturbed epidermis and stratum corneum (SC). Fluorescence lifetime is dye concentration-independent, the method requires only low light intensities and is therefore not prone to photobleaching or phototoxic artifacts, and because multiphoton lasers of IR wavelength are used, light penetrates deep into intact tissue. The standard method to measure SC pH is the flat pH electrode, which provides reliable information only about surface pH changes, without further vertical or subcellular spatial resolution; i.e., specific microdomains such as the corneocyte interstices are not resolved, and the deeper SC is inaccessible without resorting to inherently disruptive stripping methods. Furthermore, the concept of a gradient of pH through the SC stems from such stripping experiments, but other confirmation for this concept is lacking. Our investigations into the SC pH distribution so far have revealed the crucial role of the Sodium/Hydrogen Antiporter NHE1 in generation of SC acidity, the colocalization of enzymatic lipid processing activity in the SC with acidic domains of the SC, and the timing and localization of emerging acidity in the SC of newborns. Together, these results have led to an improved understanding of the SC pH, its distribution, origin, and regulation. Future uses for this method include measurements of other ions important for epidermal processes, such as Ca2+, and a quantitative approach to topical drug penetration.

  20. A simple protocol for attenuating the auto-fluorescence of cyanobacteria for optimized fluorescence in situ hybridization (FISH) imaging.

    Science.gov (United States)

    Zeller, Perrine; Ploux, Olivier; Méjean, Annick

    2016-03-01

    Cyanobacteria contain pigments, which generate auto-fluorescence that interferes with fluorescence in situ hybridization (FISH) imaging of cyanobacteria. We describe simple chemical treatments using CuSO4 or H2O2 that significantly reduce the auto-fluorescence of Microcystis strains. These protocols were successfully applied in FISH experiments using 16S rRNA specific probes and filamentous cyanobacteria.

  1. NOTE: Suppression of high-density artefacts in x-ray CT images using temporal digital subtraction with application to cryotherapy

    Science.gov (United States)

    Baissalov, R.; Sandison, G. A.; Donnelly, B. J.; Saliken, J. C.; McKinnon, J. G.; Muldrew, K.; Rewcastle, J. C.

    2000-05-01

    Image guidance in cryotherapy is usually performed using ultrasound. Although not currently in routine clinical use, x-ray CT imaging is an alternative means of guidance that can display the full 3D structure of the iceball, including frozen and unfrozen regions. However, the quality of x-ray CT images is compromised by the presence of high-density streak artefacts. To suppress these artefacts we applied temporal digital subtraction (TDS). This TDS method has the added advantage of improving the grey-scale contrast between frozen and unfrozen tissue in the CT images. Two sets of CT images were taken of a phantom material, cryoprobes and a urethral warmer (UW) before and during the cryoprobe freeze cycle. The high-density artefacts persisted in both image sets. TDS was performed on these two image sets using the corresponding mask image of unfrozen material and the same geometrical configuration of the cryoprobes and the UW. The resultant difference image had a significantly reduced artefact content. Thus TDS can be used to significantly suppress or eliminate high-density CT streak artefacts without reducing the metallic content of the cryoprobes. In vivo study needs to be conducted to establish the utility of this TDS procedure for CT assisted prostate or liver cryotherapy. Applying TDS in x-ray CT guided cryotherapy will facilitate estimation of the number and location of all frozen and unfrozen regions, potentially making cryotherapy safer and less operator dependent.

  2. Fluorescence Lifetime Imaging System for in Vivo Studies

    Directory of Open Access Journals (Sweden)

    Moinuddin Hassan

    2007-07-01

    Full Text Available In this article, a fluorescence lifetime imaging system for small animals is presented. Data were collected by scanning a region of interest with a measurement head, a linear fiber array with fixed separations between a single source fiber and several detection fibers. The goal was to localize tumors and monitor their progression using specific fluorescent markers. We chose a near-infrared contrast agent, Alexa Fluor 750 (Invitrogen Corp., Carlsbad, CA. Preliminary results show that the fluorescence lifetime for this dye was sensitive to the immediate environment of the fluorophore (in particular, pH, making it a promising candidate for reporting physiologic changes around a fluorophore. To quantify the intrinsic lifetime of deeply embedded fluorophores, we performed phantom experiments to investigate the contribution of photon migration effects on observed lifetime by calculating the fluorescence intensity decay time. A previously proposed theoretical model of migration, based on random walk theory, is also substantiated by new experimental data. The developed experimental system has been used for in vivo mouse imaging with Alexa Fluor 750 contrast agent conjugated to tumor-specific antibodies (trastuzumab [Herceptin]. Three-dimensional mapping of the fluorescence lifetime indicates lower lifetime values in superficial breast cancer tumors in mice.

  3. Novel fluorescence molecular imaging of chemotherapy-induced intestinal apoptosis

    Science.gov (United States)

    Levin, Galit; Shirvan, Anat; Grimberg, Hagit; Reshef, Ayelet; Yogev-Falach, Merav; Cohen, Avi; Ziv, Ilan

    2009-09-01

    Chemotherapy-induced enteropathy (CIE) is one of the most serious complications of anticancer therapy, and tools for its early detection and monitoring are highly needed. We report on a novel fluorescence method for detection of CIE, based on molecular imaging of the related apoptotic process. The method comprises systemic intravenous administration of the ApoSense fluorescent biomarker (N,N'-didansyl-L-cystine DDC) in vivo and subsequent fluorescence imaging of the intestinal mucosa. In the reported proof-of-concept studies, mice were treated with either taxol+cyclophosphamide or doxil. DDC was administered in vivo at various time points after drug administration, and tracer uptake by ileum tissue was subsequently evaluated by ex vivo fluorescent microscopy. Chemotherapy caused marked and selective uptake of DDC in ileal epithelial cells, in correlation with other hallmarks of apoptosis (i.e., DNA fragmentation and Annexin-V binding). Induction of DDC uptake occurred early after chemotherapy, and its temporal profile was parallel to that of the apoptotic process, as assessed histologically. DDC may therefore serve as a useful tool for detection of CIE. Future potential integration of this method with fluorescent endoscopic techniques, or development of radio-labeled derivatives of DDC for emission tomography, may advance early diagnosis and monitoring of this severe adverse effect of chemotherapy.

  4. A new fluorescent imaging of renal inflammation with RCP.

    Science.gov (United States)

    Nakamura, Kentaro; Tabata, Yasuhiko

    2010-12-20

    The objective of this study is to design a fluorescent imaging agent with R-Gel, one of the recombinant polymers (RCP), for renal inflammation. The R-Gel based on human type I collagen has multiple Arg-Gly-Asp (RGD) motifs which are ligands for some types of integrin receptors on the cell surface. After intravenous administration of R-Gel labeled by Cy7 of a fluorescent dye to three animal models of nephritis mousse, interstitial nephritis (by using UUO model mice), glomerulonephritis (HIGA mice), and ischemia-reperfusion injured kidney (I/R mice), the extent of fluorescent imaging at the renal inflammation was assessed. The Cy7-labeled R-Gel was accumulated in the inflammation site to a significantly greater extent than in the normal one at 24h after administration. The renal pattern of fluorescent imaging was similar to that of administration anti-Mac1 antibody. Taken together, it is conceivable that the R-Gel was targeted to macrophages infiltrated into the inflammation site of kidney. Copyright © 2010 Elsevier B.V. All rights reserved.

  5. Infrared imaging of LED lighting tubes and fluorescent tubes

    Science.gov (United States)

    Siikanen, Sami; Kivi, Sini; Kauppinen, Timo; Juuti, Mikko

    2011-05-01

    The low energy efficiency of conventional light sources is mainly caused by generation of waste heat. We used infrared (IR) imaging in order to monitor the heating of both LED tube luminaires and ordinary T8 fluorescent tubes. The IR images showed clearly how the surface temperatures of the fluorescent tube ends quickly rose up to about +50...+70°C, whereas the highest surface temperatures seen on the LED tubes were only about +30...+40°C. The IR images demonstrated how the heat produced by the individual LED chips can be efficiently guided to the supporting structure in order to keep the LED emitters cool and hence maintain efficient operation. The consumed electrical power and produced illuminance were also recorded during 24 hour measurements. In order to assess the total luminous efficacy of the luminaires, separate luminous flux measurements were made in a large integrating sphere. The currently available LED tubes showed efficacies of up to 88 lm/W, whereas a standard "cool white" T8 fluorescent tube produced ca. 75 lm/W. Both lamp types gave ca. 110 - 130 lx right below the ceiling-mounted luminaire, but the LED tubes consume only 40 - 55% of the electric power compared to fluorescent tubes.

  6. Mueller matrix signature in advanced fluorescence microscopy imaging

    Science.gov (United States)

    Mazumder, Nirmal; Qiu, Jianjun; Kao, Fu-Jen; Diaspro, Alberto

    2017-02-01

    We have demonstrated the measurement and characterization of the polarization properties of a fluorescence signal using four-channel photon counting based Stokes-Mueller polarization microscopy. Thus, Lu-Chipman decomposition was applied to extract the critical polarization properties such as depolarization, linear retardance and the optical rotation of collagen type I fiber. We observed the spatial distribution of anisotropic and helical molecules of collagen from the reconstructed 2D Mueller images based on the fluorescence signal in a pixel-by-pixel manner.

  7. Dynamic fluorescence lifetime imaging based on acousto-optic deflectors

    Science.gov (United States)

    Yan, Wei; Peng, Xiao; Qi, Jing; Gao, Jian; Fan, Shunping; Wang, Qi; Qu, Junle; Niu, Hanben

    2014-11-01

    We report a dynamic fluorescence lifetime imaging (D-FLIM) system that is based on a pair of acousto-optic deflectors for the random regions of interest (ROI) study in the sample. The two-dimensional acousto-optic deflector devices are used to rapidly scan the femtosecond excitation laser beam across the sample, providing specific random access to the ROI. Our experimental results using standard fluorescent dyes in live cancer cells demonstrate that the D-FLIM system can dynamically monitor the changing process of the microenvironment in the ROI in live biological samples.

  8. Highly stable organic fluorescent nanorods for living- cell imaging

    Institute of Scientific and Technical Information of China (English)

    Minhuan Lan[1,3; Jinfeng Zhang[1,3; Xiaoyue Zhu[1; Pengfei Wang[2; Xianfeng Chen[1; Chun-Sing Lee[1; Wenjun Zhang[1

    2015-01-01

    Metal-free, organic-dye-based fluorescent nanorods were fabricated through a simple solvent-exchange procedure. The as-prepared nanorods exhibit low toxicity to living cells and excellent photostability. Furthermore, they are stable in solutions of various pHs and high ionic strength and in solutions with interfering metal ions. Compared with the free DPP-Br molecules in THF, these nanorods exhibit larger Stokes shift, broader absorption spectra, and greatly improved photostability. We successfully demonstrated the application of the nanorods, including their aforementioned beneficial characteristics, as a good fluorescence probe for bio-imaging.

  9. Correction for scatter and septal penetration using convolution subtraction methods and model-based compensation in {sup 123}I brain SPECT imaging-a Monte Carlo study

    Energy Technology Data Exchange (ETDEWEB)

    Larsson, Anne [Department of Radiation Sciences, Radiation Physics, Umeaa University, SE-901 87 Umeaa (Sweden); Ljungberg, Michael [Medical Radiation Physics, Department of Clinical Sciences, Lund, Lund University, SE-221 85 Lund (Sweden); Mo, Susanna Jakobson [Department of Radiation Sciences, Diagnostic Radiology, Umeaa University, SE-901 87 Umeaa (Sweden); Riklund, Katrine [Department of Radiation Sciences, Diagnostic Radiology, Umeaa University, SE-901 87 Umeaa (Sweden); Johansson, Lennart [Department of Radiation Sciences, Radiation Physics, Umeaa University, SE-901 87 Umeaa (Sweden)

    2006-11-21

    Scatter and septal penetration deteriorate contrast and quantitative accuracy in single photon emission computed tomography (SPECT). In this study four different correction techniques for scatter and septal penetration are evaluated for {sup 123}I brain SPECT. One of the methods is a form of model-based compensation which uses the effective source scatter estimation (ESSE) for modelling scatter, and collimator-detector response (CDR) including both geometric and penetration components. The other methods, which operate on the 2D projection images, are convolution scatter subtraction (CSS) and two versions of transmission dependent convolution subtraction (TDCS), one of them proposed by us. This method uses CSS for correction for septal penetration, with a separate kernel, and TDCS for scatter correction. The corrections are evaluated for a dopamine transporter (DAT) study and a study of the regional cerebral blood flow (rCBF), performed with {sup 123}I. The images are produced using a recently developed Monte Carlo collimator routine added to the program SIMIND which can include interactions in the collimator. The results show that the method included in the iterative reconstruction is preferable to the other methods and that the new TDCS version gives better results compared with the other 2D methods.

  10. Correction for scatter and septal penetration using convolution subtraction methods and model-based compensation in 123I brain SPECT imaging-a Monte Carlo study.

    Science.gov (United States)

    Larsson, Anne; Ljungberg, Michael; Mo, Susanna Jakobson; Riklund, Katrine; Johansson, Lennart

    2006-11-21

    Scatter and septal penetration deteriorate contrast and quantitative accuracy in single photon emission computed tomography (SPECT). In this study four different correction techniques for scatter and septal penetration are evaluated for 123I brain SPECT. One of the methods is a form of model-based compensation which uses the effective source scatter estimation (ESSE) for modelling scatter, and collimator-detector response (CDR) including both geometric and penetration components. The other methods, which operate on the 2D projection images, are convolution scatter subtraction (CSS) and two versions of transmission dependent convolution subtraction (TDCS), one of them proposed by us. This method uses CSS for correction for septal penetration, with a separate kernel, and TDCS for scatter correction. The corrections are evaluated for a dopamine transporter (DAT) study and a study of the regional cerebral blood flow (rCBF), performed with 123I. The images are produced using a recently developed Monte Carlo collimator routine added to the program SIMIND which can include interactions in the collimator. The results show that the method included in the iterative reconstruction is preferable to the other methods and that the new TDCS version gives better results compared with the other 2D methods.

  11. The study of blue LED to induce fluorescence spectroscopy and fluorescence imaging for oral carcinoma detection

    Science.gov (United States)

    Zheng, Longjiang; Hu, Yuanting

    2009-07-01

    Fluorescence spectroscopy and fluorescence imaging diagnosis of malignant lesions provides us with a new method to diagnose diseases in precancerous stage. Early diagnosis of disease has significant importance in cancer treatment, because most cancers can be cured well in precancerous, especially when the diffusion of cancer is limited in a restricted region. In this study, Golden hamster models were applied to 5% 9, 10 dimethyl-1, 2-benzanthracene (DMBA) to induce hamster buccal cheek pouch carcinoma three times a week. Rose Bengal, which has been used in clinican for years and avoids visible side-effect to human was chosen as photosensitizer. 405 nm blue LED was used to induce the fluorescence of photosensitizer. After topical application of photosensitizer, characteristic red emission fluorescence peak was observed around 600nm. Similar, normal oral cavity has special luminescence around 480nm. Fluorescence spectroscopy technology is based on analysing emission peaks of photosensitizer in the areas of oral carcinoma, moreover, red-to-green (IR/IG) intensity ratio is also applied as a diagnostic algorithm. A CCD which is connected with a computer is used to take pictures at carcinoma areas through different filters. Fluorescence images from normal hamster buccal cheek pouch are compared with those from carcinogen-induced models of carcinoma, and morphological differences between normal and lesion tissue can be distinguished. The pictures are analyzed by Matlab and shown on the screen of computer. This paper demonstrates that Rose Bengal could be used as photosensitizer to detect oral carcinoma, and blue LED as excitation source could not only have a good effect to diagnose oral carcinoma, but also decrease cost greatly.

  12. Nanoscale fluorescence lifetime imaging with a single diamond NV center

    CERN Document Server

    Beams, Ryan; Johnson, Timothy W; Oh, Sang-Hyun; Novotny, Lukas; Vamivakas, Nick

    2013-01-01

    Solid-state quantum emitters, such as artificially engineered quantum dots or naturally occurring defects in solids, are being investigated for applications ranging from quantum information science and optoelectronics to biomedical imaging. Recently, these same systems have also been studied from the perspective of nanoscale metrology. In this letter we study the near-field optical properties of a diamond nanocrystal hosting a single nitrogen vacancy center. We find that the nitrogen vacancy center is a sensitive probe of the surrounding electromagnetic mode structure. We exploit this sensitivity to demonstrate nanoscale fluorescence lifetime imaging microscopy (FLIM) with a single nitrogen vacancy center by imaging the local density of states of an optical antenna.

  13. A Pico Projector Source for Confocal Fluorescence and Ophthalmic Imaging.

    Science.gov (United States)

    Muller, Matthew S

    2012-09-02

    A Pico digital light projector has been implemented as an integrated illumination source and spatial light modulator for confocal imaging. The target is illuminated with a series of rapidly projected lines or points to simulate scanning. Light returning from the target is imaged onto a 2D rolling shutter CMOS sensor. By controlling the spatio-temporal relationship between the rolling shutter and illumination pattern, light returning from the target is spatially filtered. Confocal retinal, fluorescence, and Fourier-domain optical coherence tomography implementations of this novel imaging technique are presented.

  14. In Vivo Metal Ion Imaging Using Fluorescent Sensors.

    Science.gov (United States)

    Van de Bittner, Genevieve C; Hirayama, Tasuku

    2016-01-01

    In vivo imaging in living animals provides the ability to monitor alterations of signaling molecules, ions, and other biological components during various life stages and in disease. The data gained from in vivo imaging can be used for biological discovery or to determine elements of disease progression and can inform the development and translation of therapeutics. Herein, we present theories behind small-molecule, fluorescent, metal ion sensors as well as the methods for their successful application to in vivo metal ion imaging, including ex vivo validation.

  15. Chlorophyll fluorescence imaging of cadmium-treated white cabbage plants

    Directory of Open Access Journals (Sweden)

    Borek M.

    2013-04-01

    Full Text Available The chlorophyll fluorescence imaging technique is a valuable tool to study the impact of heavy metal stress in plants. The aim of this paper was to investigate the influence of Cd on photosynthetic apparatus of white cabbage (Brassica oleracea subsp. capitata f. alba plants. Two cabbage cultivars ‘Ditmarska Najwcześniejsza’ (‘DN’; early and ‘Amager Polana’ (‘AP’; late were used. Cd was applied before planting seedlings (10 mg Cd kg−1 DM of soil.. Measurements were performed at the 3rd leaf after 2 weeks of planting. The level of Cd-induced stress to plants was estimated by chlorophyll (Chl content (photometrically and analyses of images and numeric values of the major fluorescence parameters of Chl (Chl fluorescence imaging system FluorCam. Cd negatively affected the chlorophyll content and Fv/Fm, Fv’/Fm’, Φ PSII and qP in leaves of early cultivar of white cabbage. However, in the case of late cv. we did not observe such distinct changes. It suggests that late cultivars. are more resistant to Cd than the early ones. Considering methodological aspect of the study, Chl fluorescence imaging can better reveal some alterations within the leaf, because numeric values of specific parameters, which are the averaged data collected from the whole leaf, cannot reflect the tissue specificity. Abbreviations: HM – heavy metal, Cd – cadmium, Chl – chlorophyll, Fv/Fm – photochemical efficiency of PSII in the dark-adapted state, F‘v’/F‘m’ – PSII maximum efficiency, Φ PSII – quantum efficiency of PSII electron transport, NPQ – nonphotochemical quenching of maximal Chl fluorescence, qP – photochemical quenching coefficient.

  16. Fluorescent ligand for human progesterone receptor imaging in live cells.

    Science.gov (United States)

    Weinstain, Roy; Kanter, Joan; Friedman, Beth; Ellies, Lesley G; Baker, Michael E; Tsien, Roger Y

    2013-05-15

    We employed molecular modeling to design and then synthesize fluorescent ligands for the human progesterone receptor. Boron dipyrromethene (BODIPY) or tetramethylrhodamine were conjugated to the progesterone receptor antagonist RU486 (Mifepristone) through an extended hydrophilic linker. The fluorescent ligands demonstrated comparable bioactivity to the parent antagonist in live cells and triggered nuclear translocation of the receptor in a specific manner. The BODIPY labeled ligand was applied to investigate the dependency of progesterone receptor nuclear translocation on partner proteins and to show that functional heat shock protein 90 but not immunophilin FKBP52 activity is essential. A tissue distribution study indicated that the fluorescent ligand preferentially accumulates in tissues that express high levels of the receptor in vivo. The design and properties of the BODIPY-labeled RU486 make it a potential candidate for in vivo imaging of PR by positron emission tomography through incorporation of (18)F into the BODIPY core.

  17. Fluorescence imaging to study cancer burden on lymph nodes

    Science.gov (United States)

    D'Souza, Alisha V.; Elliott, Jonathan T.; Gunn, Jason R.; Samkoe, Kimberley S.; Tichauer, Kenneth M.; Pogue, Brian W.

    2015-03-01

    Morbidity and complexity involved in lymph node staging via surgical resection and biopsy calls for staging techniques that are less invasive. While visible blue dyes are commonly used in locating sentinel lymph nodes, since they follow tumor-draining lymphatic vessels, they do not provide a metric to evaluate presence of cancer. An area of active research is to use fluorescent dyes to assess tumor burden of sentinel and secondary lymph nodes. The goal of this work was to successfully deploy and test an intra-nodal cancer-cell injection model to enable planar fluorescence imaging of a clinically relevant blue dye, specifically methylene blue along with a cancer targeting tracer, Affibody labeled with IRDYE800CW and subsequently segregate tumor-bearing from normal lymph nodes. This direct-injection based tumor model was employed in athymic rats (6 normal, 4 controls, 6 cancer-bearing), where luciferase-expressing breast cancer cells were injected into axillary lymph nodes. Tumor presence in nodes was confirmed by bioluminescence imaging before and after fluorescence imaging. Lymphatic uptake from the injection site (intradermal on forepaw) to lymph node was imaged at approximately 2 frames/minute. Large variability was observed within each cohort.

  18. Pulse subtraction Doppler

    Science.gov (United States)

    Mahue, Veronique; Mari, Jean Martial; Eckersley, Robert J.; Caro, Colin G.; Tang, Meng-Xing

    2010-01-01

    Recent advances have demonstrated the feasibility of molecular imaging using targeted microbubbles and ultrasound. One technical challenge is to selectively detect attached bubbles from those freely flowing bubbles and surrounding tissue. Pulse Inversion Doppler is an imaging technique enabling the selective detection of both static and moving ultrasound contrast agents: linear scatterers generate a single band Doppler spectrum, while non-linear scatterers generate a double band spectrum, one being uniquely correlated with the presence of contrast agents and non-linear tissue signals. We demonstrate that similar spectrums, and thus the same discrimination, can be obtained through a Doppler implementation of Pulse Subtraction. This is achieved by reconstructing a virtual echo using the echo generated from a short pulse transmission. Moreover by subtracting from this virtual echo the one generated from a longer pulse transmission, it is possible to fully suppress the echo from linear scatterers, while for non-linear scatterers, a signal will remain, allowing classical agent detection. Simulations of a single moving microbubble and a moving linear scatterer subject to these pulses show that when the virtual echo and the long pulse echo are used to perform pulsed Doppler, the power Doppler spectrum allows separation of linear and non-linear moving scattering. Similar results are obtained on experimental data acquired on a flow containing either microbubble contrast agents or linear blood mimicking fluid. This new Doppler method constitutes an alternative to Pulse Inversion Doppler and preliminary results suggest that similar dual band spectrums could be obtained by the combination of any non-linear detection technique with Doppler demodulation.

  19. [Intraoperative graft assessment using fluorescent imaging system (SPY)].

    Science.gov (United States)

    Kawashima, T; Naraoka, S; Kakizaki, T

    2009-07-01

    We investigated the efficacy of intraoperative fluorescent imaging system for the assessment of coronary artery bypass grafting (CABG). We used SPY imaging system in 100 CABG (57 off-pump and 43 on-pump CABG), totalling 287 distal anastomoses. The total graft patency rate on postoperative angiography in this series was 96.2% (276/287). Graft revision was done in 10 cases (10.0%) and 13 anastomoses (4.5%) by SPY imaging, which all resulted in good patency at postoperative angiography. On the other hand, 7 distal anastomoses and 1 mammary graft (2.8%) appeared to be successful on intraoperative SPY imaging, but were revealed to be occluded by postoperative angiography. SPY imaging system is useful for graft validation, and may contribute to improvement of coronary bypass graft patency.

  20. Proton-induced x-ray fluorescence CT imaging.

    Science.gov (United States)

    Bazalova-Carter, Magdalena; Ahmad, Moiz; Matsuura, Taeko; Takao, Seishin; Matsuo, Yuto; Fahrig, Rebecca; Shirato, Hiroki; Umegaki, Kikuo; Xing, Lei

    2015-02-01

    To demonstrate the feasibility of proton-induced x-ray fluorescence CT (pXFCT) imaging of gold in a small animal sized object by means of experiments and Monte Carlo (MC) simulations. First, proton-induced gold x-ray fluorescence (pXRF) was measured as a function of gold concentration. Vials of 2.2 cm in diameter filled with 0%-5% Au solutions were irradiated with a 220 MeV proton beam and x-ray fluorescence induced by the interaction of protons, and Au was detected with a 3 × 3 mm(2) CdTe detector placed at 90° with respect to the incident proton beam at a distance of 45 cm from the vials. Second, a 7-cm diameter water phantom containing three 2.2-diameter vials with 3%-5% Au solutions was imaged with a 7-mm FWHM 220 MeV proton beam in a first generation CT scanning geometry. X-rays scattered perpendicular to the incident proton beam were acquired with the CdTe detector placed at 45 cm from the phantom positioned on a translation/rotation stage. Twenty one translational steps spaced by 3 mm at each of 36 projection angles spaced by 10° were acquired, and pXFCT images of the phantom were reconstructed with filtered back projection. A simplified geometry of the experimental data acquisition setup was modeled with the MC TOPAS code, and simulation results were compared to the experimental data. A linear relationship between gold pXRF and gold concentration was observed in both experimental and MC simulation data (R(2) > 0.99). All Au vials were apparent in the experimental and simulated pXFCT images. Specifically, the 3% Au vial was detectable in the experimental [contrast-to-noise ratio (CNR) = 5.8] and simulated (CNR = 11.5) pXFCT image. Due to fluorescence x-ray attenuation in the higher concentration vials, the 4% and 5% Au contrast were underestimated by 10% and 15%, respectively, in both the experimental and simulated pXFCT images. Proton-induced x-ray fluorescence CT imaging of 3%-5% gold solutions in a small animal sized water phantom has been demonstrated

  1. Proton-induced x-ray fluorescence CT imaging

    Energy Technology Data Exchange (ETDEWEB)

    Bazalova-Carter, Magdalena, E-mail: bazalova@stanford.edu; Xing, Lei [Department of Radiation Oncology, Stanford University, Stanford, California 94305-5847 and Global Station for Quantum Medical Science and Engineering, Global Institution for Collaborative Research and Education (GI-CoRE), Hokkaido University, Sapporo 060-8648 (Japan); Ahmad, Moiz [Department of Radiation Oncology, Stanford University, Stanford, California 94305-5847 (United States); Matsuura, Taeko; Takao, Seishin; Shirato, Hiroki; Umegaki, Kikuo [Department of Medical Physics, Proton Beam Therapy Center, Hokkaido University Hospital, Sapporo 060-8648, Japan and Global Station for Quantum Medical Science and Engineering, Global Institution for Collaborative Research and Education (GI-CoRE), Hokkaido University, Sapporo 060-8648 (Japan); Matsuo, Yuto [Department of Medical Physics, Proton Beam Therapy Center, Hokkaido University Hospital, Sapporo 060-8648 (Japan); Fahrig, Rebecca [Department of Radiology, Stanford University, Stanford, California 94305 (United States)

    2015-02-15

    Purpose: To demonstrate the feasibility of proton-induced x-ray fluorescence CT (pXFCT) imaging of gold in a small animal sized object by means of experiments and Monte Carlo (MC) simulations. Methods: First, proton-induced gold x-ray fluorescence (pXRF) was measured as a function of gold concentration. Vials of 2.2 cm in diameter filled with 0%–5% Au solutions were irradiated with a 220 MeV proton beam and x-ray fluorescence induced by the interaction of protons, and Au was detected with a 3 × 3 mm{sup 2} CdTe detector placed at 90° with respect to the incident proton beam at a distance of 45 cm from the vials. Second, a 7-cm diameter water phantom containing three 2.2-diameter vials with 3%–5% Au solutions was imaged with a 7-mm FWHM 220 MeV proton beam in a first generation CT scanning geometry. X-rays scattered perpendicular to the incident proton beam were acquired with the CdTe detector placed at 45 cm from the phantom positioned on a translation/rotation stage. Twenty one translational steps spaced by 3 mm at each of 36 projection angles spaced by 10° were acquired, and pXFCT images of the phantom were reconstructed with filtered back projection. A simplified geometry of the experimental data acquisition setup was modeled with the MC TOPAS code, and simulation results were compared to the experimental data. Results: A linear relationship between gold pXRF and gold concentration was observed in both experimental and MC simulation data (R{sup 2} > 0.99). All Au vials were apparent in the experimental and simulated pXFCT images. Specifically, the 3% Au vial was detectable in the experimental [contrast-to-noise ratio (CNR) = 5.8] and simulated (CNR = 11.5) pXFCT image. Due to fluorescence x-ray attenuation in the higher concentration vials, the 4% and 5% Au contrast were underestimated by 10% and 15%, respectively, in both the experimental and simulated pXFCT images. Conclusions: Proton-induced x-ray fluorescence CT imaging of 3%–5% gold solutions in a

  2. Directional bilateral filters for smoothing fluorescence microscopy images

    Directory of Open Access Journals (Sweden)

    Manasij Venkatesh

    2015-08-01

    Full Text Available Images obtained through fluorescence microscopy at low numerical aperture (NA are noisy and have poor resolution. Images of specimens such as F-actin filaments obtained using confocal or widefield fluorescence microscopes contain directional information and it is important that an image smoothing or filtering technique preserve the directionality. F-actin filaments are widely studied in pathology because the abnormalities in actin dynamics play a key role in diagnosis of cancer, cardiac diseases, vascular diseases, myofibrillar myopathies, neurological disorders, etc. We develop the directional bilateral filter as a means of filtering out the noise in the image without significantly altering the directionality of the F-actin filaments. The bilateral filter is anisotropic to start with, but we add an additional degree of anisotropy by employing an oriented domain kernel for smoothing. The orientation is locally adapted using a structure tensor and the parameters of the bilateral filter are optimized for within the framework of statistical risk minimization. We show that the directional bilateral filter has better denoising performance than the traditional Gaussian bilateral filter and other denoising techniques such as SURE-LET, non-local means, and guided image filtering at various noise levels in terms of peak signal-to-noise ratio (PSNR. We also show quantitative improvements in low NA images of F-actin filaments.

  3. Increasing precision of lifetime determination in fluorescence lifetime imaging

    Science.gov (United States)

    Chang, Ching-Wei; Mycek, Mary-Ann

    2010-02-01

    The interest in fluorescence lifetime imaging microscopy (FLIM) is increasing, as commercial FLIM modules become available for confocal and multi-photon microscopy. In biological FLIM applications, low fluorescence signals from samples can be a challenge, and this causes poor precision in lifetime. In this study, for the first time, we applied wavelet-based denoising methods in time-domain FLIM, and compared them with our previously developed total variation (TV) denoising methods. They were first tested using artificial FLIM images. We then applied them to lowlight live-cell images. The results demonstrated that our TV methods could improve lifetime precision multi-fold in FLIM images and preserve the overall lifetime and pre-exponential term values when improving local lifetime fitting, while wavelet-based methods were faster. The results here can enhance the precision of FLIM, especially for low-light and / or fast video-rate imaging, to improve current and rapidly emerging new applications of FLIM such as live-cell, in vivo whole-animal, or endoscopic imaging.

  4. Fluorescence imaging of dendritic spines of Golgi-Cox-stained neurons using brightening background

    Science.gov (United States)

    Ai, Min; Xiong, Hanqing; Yang, Tao; Shang, Zhenhua; Chen, Muqing; Liu, Xiuli; Zeng, Shaoqun

    2015-01-01

    We report a novel fluorescence imaging approach to imaging nonfluorescence-labeled biological tissue samples. The method was demonstrated by imaging neurons in Golgi-Cox-stained and epoxy-resin-embedded samples through the excitation of the background fluorescence of the specimens. The dark neurons stood out clearly against background fluorescence in the images, enabling the tracing of a single dendritic spine using both confocal and wide-field fluorescence microscopy. The results suggest that the reported fluorescence imaging method would provide an effective alternative solution to image nonfluorescence-labeled samples, and it allows tracing the dendritic spine structure of neurons.

  5. Fluorescence lifetime imaging microscopy of nanodiamonds in vivo

    Science.gov (United States)

    Kuo, Yung; Hsu, Tsung-Yuan; Wu, Yi-Chun; Hsu, Jui-Hung; Chang, Huan-Cheng

    2013-03-01

    The negatively charged nitrogen-vacancy (NV-) center in bulk diamond is a photostable fluorophore with a radiative lifetime of 11.6 ns at room temperature. The lifetime substantially increases to ~20 ns for diamond nanoparticles (size ~ 100 nm) suspended in water due to the change in refractive index of the surrounding medium of the NV- centers. This fluorescence decay time is much longer than that (typically 1 - 4 ns) of endogenous and exogenous fluorophores commonly used in biological imaging, making it possible to detect NV--containing nanodiamonds in vivo at the single particle level by fluorescence lifetime imaging microscopy (FLIM). We demonstrate the feasibility of this approach using Caenorhabditis elegans (C. elegans) as a model organism.

  6. Novel fluorescent carbonic nanomaterials for sensing and imaging

    Science.gov (United States)

    Demchenko, Alexander P.; Dekaliuk, Mariia O.

    2013-12-01

    Small brightly fluorescent carbon nanoparticles have emerged as a new class of materials important for sensing and imaging applications. We analyze comparatively the properties of nanodiamonds, graphene and graphene oxide ‘dots’, of modified carbon nanotubes and of diverse carbon nanoparticles known as ‘C-dots’ obtained by different methods. The mechanisms of their light absorption and luminescence emission are still unresolved and the arguments are presented for their common origin. Regarding present and potential applications, we provide critical comparison with the other types of fluorescence reporters, such as organic dyes and semiconductor quantum dots. Their most prospective applications in sensing (based on the changes of intensity, FRET and lifetime) and in imaging technologies on the level of living cells and whole bodies are overviewed. The possibilities for design on their basis of multifunctional nanocomposites on a broader scale of theranostics are outlined.

  7. Modulated electron-multiplied fluorescence lifetime imaging microscope: all-solid-state camera for fluorescence lifetime imaging.

    Science.gov (United States)

    Zhao, Qiaole; Schelen, Ben; Schouten, Raymond; van den Oever, Rein; Leenen, René; van Kuijk, Harry; Peters, Inge; Polderdijk, Frank; Bosiers, Jan; Raspe, Marcel; Jalink, Kees; Geert Sander de Jong, Jan; van Geest, Bert; Stoop, Karel; Young, Ian Ted

    2012-12-01

    We have built an all-solid-state camera that is directly modulated at the pixel level for frequency-domain fluorescence lifetime imaging microscopy (FLIM) measurements. This novel camera eliminates the need for an image intensifier through the use of an application-specific charge coupled device design in a frequency-domain FLIM system. The first stage of evaluation for the camera has been carried out. Camera characteristics such as noise distribution, dark current influence, camera gain, sampling density, sensitivity, linearity of photometric response, and optical transfer function have been studied through experiments. We are able to do lifetime measurement using our modulated, electron-multiplied fluorescence lifetime imaging microscope (MEM-FLIM) camera for various objects, e.g., fluorescein solution, fixed green fluorescent protein (GFP) cells, and GFP-actin stained live cells. A detailed comparison of a conventional microchannel plate (MCP)-based FLIM system and the MEM-FLIM system is presented. The MEM-FLIM camera shows higher resolution and a better image quality. The MEM-FLIM camera provides a new opportunity for performing frequency-domain FLIM.

  8. Neural imaging in songbirds using fiber optic fluorescence microscopy

    Science.gov (United States)

    Nooshabadi, Fatemeh; Hearn, Gentry; Lints, Thierry; Maitland, Kristen C.

    2012-02-01

    The song control system of juvenile songbirds is an important model for studying the developmental acquisition and generation of complex learned vocal motor sequences, two processes that are fundamental to human speech and language. To understand the neural mechanisms underlying song production, it is critical to characterize the activity of identified neurons in the song control system when the bird is singing. Neural imaging in unrestrained singing birds, although technically challenging, will advance our understanding of neural ensemble coding mechanisms in this system. We are exploring the use of a fiber optic microscope for functional imaging in the brain of behaving and singing birds in order to better understand the contribution of a key brain nucleus (high vocal center nucleus; HVC) to temporal aspects of song motor control. We have constructed a fluorescence microscope with LED illumination, a fiber bundle for transmission of fluorescence excitation and emission light, a ~2x GRIN lens, and a CCD for image acquisition. The system has 2 μm resolution, 375 μm field of view, 200 μm working distance, and 1 mm outer diameter. As an initial characterization of this setup, neurons in HVC were imaged using the fiber optic microscope after injection of quantum dots or fluorescent retrograde tracers into different song nuclei. A Lucid Vivascope confocal microscope was used to confirm the imaging results. Long-term imaging of the activity of these neurons in juvenile birds during singing may lead us to a better understanding of the central motor codes for song and the central mechanism by which auditory experience modifies song motor commands to enable vocal learning and imitation.

  9. Non-invasive imaging of skin cancer with fluorescence lifetime imaging using two photon tomography

    Science.gov (United States)

    Patalay, Rakesh; Talbot, Clifford; Alexandrov, Yuriy; Munro, Ian; Breunig, Hans Georg; König, Karsten; Warren, Sean; Neil, Mark A. A.; French, Paul M. W.; Chu, Anthony; Stamp, Gordon W.; Dunsby, Christopher

    2011-07-01

    Multispectral fluorescence lifetime imaging (FLIM) using two photon microscopy as a non-invasive technique for the diagnosis of skin lesions is described. Skin contains fluorophores including elastin, keratin, collagen, FAD and NADH. This endogenous contrast allows tissue to be imaged without the addition of exogenous agents and allows the in vivo state of cells and tissues to be studied. A modified DermaInspect® multiphoton tomography system was used to excite autofluorescence at 760 nm in vivo and on freshly excised ex vivo tissue. This instrument simultaneously acquires fluorescence lifetime images in four spectral channels between 360-655 nm using time-correlated single photon counting and can also provide hyperspectral images. The multispectral fluorescence lifetime images were spatially segmented and binned to determine lifetimes for each cell by fitting to a double exponential lifetime model. A comparative analysis between the cellular lifetimes from different diagnoses demonstrates significant diagnostic potential.

  10. In Vivo Dual Fluorescence Imaging to Detect Joint Destruction.

    Science.gov (United States)

    Cho, Hongsik; Bhatti, Fazal-Ur-Rehman; Lee, Sangmin; Brand, David D; Yi, Ae-Kyung; Hasty, Karen A

    2016-10-01

    Diagnosis of cartilage damage in early stages of arthritis is vital to impede the progression of disease. In this regard, considerable progress has been made in near-infrared fluorescence (NIRF) optical imaging technique. Arthritis can develop due to various mechanisms but one of the main contributors is the production of matrix metalloproteinases (MMPs), enzymes that can degrade components of the extracellular matrix. Especially, MMP-1 and MMP-13 have main roles in rheumatoid arthritis and osteoarthritis because they enhance collagen degradation in the process of arthritis. We present here a novel NIRF imaging strategy that can be used to determine the activity of MMPs and cartilage damage simultaneously by detection of exposed type II collagen in cartilage tissue. In this study, retro-orbital injection of mixed fluorescent dyes, MMPSense 750 FAST (MMP750) dye and Alexa Fluor 680 conjugated monoclonal mouse antibody immune-reactive to type II collagen, was administered in the arthritic mice. Both dyes were detected with different intensity according to degree of joint destruction in the animal. Thus, our dual fluorescence imaging method can be used to detect cartilage damage as well as MMP activity simultaneously in early stage arthritis.

  11. Fluorescent quantum dots: synthesis, biomedical optical imaging, and biosafety assessment.

    Science.gov (United States)

    Ji, Xiaoyuan; Peng, Fei; Zhong, Yiling; Su, Yuanyuan; He, Yao

    2014-12-01

    The marriage of nanomaterials with biology has significantly promoted advancement of biological techniques, profoundly facilitating basic research and practical applications in biological and biomedical fields. Taking advantages of unique optical properties (e.g., strong fluorescence, robust photostability, size-tunable emission wavelengths, etc.), fluorescent quantum dots (QDs), appearing as high-performance biological fluorescent nanoprobes, have been extensively explored for a variety of biomedical optical imaging applications. In this review, we present representative synthetic strategies for preparation of QDs and their applications in biomedical optical imaging, as well as risk assessments in vitro and in vivo. Briefly, we first summarize recent progress in fabrication of QDs via two rudimentary approaches, i.e., organometallic route and aqueous synthesis. Next we present representative achievement in QDs-based in vitro and in vivo biomedical optical imaging applications. We further discuss the toxicity assessment of QDs, ranging from cell studies to animal models. In the final section, we discuss challenges and perspectives for the QDs-relative bioapplications in the future.

  12. All-optically integrated multimodality imaging system: combined photoacoustic microscopy, optical coherence tomography, and fluorescence imaging

    Science.gov (United States)

    Chen, Zhongjiang; Yang, Sihua; Xing, Da

    2016-10-01

    We have developed a multimodality imaging system by optically integrating all-optical photoacoustic microscopy (AOPAM), optical coherence tomography (OCT) and fluorescence microscopy (FLM) to provide complementary information including optical absorption, optical back-scattering and fluorescence contrast of biological tissue. By sharing the same low-coherence Michelson interferometer, AOPAM and OCT could be organically optically combined to obtain the absorption and scattering information of the biological tissues. Also, owing to using the same laser source and objective lens, intrinsically registered photoacoustic and fluorescence signals are obtained to present the radiative and nonradiative transition process of absorption. Simultaneously photoacoustic angiography, tissue structure and fluorescence molecular in vivo images of mouse ear were acquired to demonstrate the capabilities of the optically integrated trimodality imaging system, which can present more information to study tumor angiogenesis, vasculature, anatomical structure and microenvironments in vivo.

  13. Refractive index sensing of green fluorescent proteins in living cells using fluorescence lifetime imaging microscopy.

    Science.gov (United States)

    van Manen, Henk-Jan; Verkuijlen, Paul; Wittendorp, Paul; Subramaniam, Vinod; van den Berg, Timo K; Roos, Dirk; Otto, Cees

    2008-04-15

    We show that fluorescence lifetime imaging microscopy (FLIM) of green fluorescent protein (GFP) molecules in cells can be used to report on the local refractive index of intracellular GFP. We expressed GFP fusion constructs of Rac2 and gp91(phox), which are both subunits of the phagocyte NADPH oxidase enzyme, in human myeloid PLB-985 cells and showed by high-resolution confocal fluorescence microscopy that GFP-Rac2 and GFP-gp91(phox) are targeted to the cytosol and to membranes, respectively. Frequency-domain FLIM experiments on these PLB-985 cells resulted in average fluorescence lifetimes of 2.70 ns for cytosolic GFP-Rac2 and 2.31 ns for membrane-bound GFP-gp91(phox). By comparing these lifetimes with a calibration curve obtained by measuring GFP lifetimes in PBS/glycerol mixtures of known refractive index, we found that the local refractive indices of cytosolic GFP-Rac2 and membrane-targeted GFP-gp91(phox) are approximately 1.38 and approximately 1.46, respectively, which is in good correspondence with reported values for the cytosol and plasma membrane measured by other techniques. The ability to measure the local refractive index of proteins in living cells by FLIM may be important in revealing intracellular spatial heterogeneities within organelles such as the plasma and phagosomal membrane.

  14. Polymer-encapsulated organic nanoparticles for fluorescence and photoacoustic imaging.

    Science.gov (United States)

    Li, Kai; Liu, Bin

    2014-09-21

    Polymer encapsulated organic nanoparticles have recently attracted increasing attention in the biomedical field because of their unique optical properties, easy fabrication and outstanding performance as imaging and therapeutic agents. Of particular importance is the polymer encapsulated nanoparticles containing conjugated polymers (CP) or fluorogens with aggregation induced emission (AIE) characteristics as the core, which have shown significant advantages in terms of tunable brightness, superb photo- and physical stability, good biocompatibility, potential biodegradability and facile surface functionalization. In this review, we summarize the latest advances in the development of polymer encapsulated CP and AIE fluorogen nanoparticles, including preparation methods, material design and matrix selection, nanoparticle fabrication and surface functionalization for fluorescence and photoacoustic imaging. We also discuss their specific applications in cell labeling, targeted in vitro and in vivo imaging, blood vessel imaging, cell tracing, inflammation monitoring and molecular imaging. We specially focus on strategies to fine-tune the nanoparticle property (e.g. size and fluorescence quantum yield) through precise engineering of the organic cores and careful selection of polymer matrices. The review also highlights the merits and limitations of these nanoparticles as well as strategies used to overcome the limitations. The challenges and perspectives for the future development of polymer encapsulated organic nanoparticles are also discussed.

  15. Two photon fluorescence imaging of lipid membrane domains and potentials using advanced fluorescent probes

    Science.gov (United States)

    Kilin, Vasyl; Darwich, Zeinab; Richert, Ludovic; Didier, Pascal; Klymchenko, Andrey; Mély, Yves

    2013-02-01

    Biomembranes are ordered and dynamic nanoscale structures critical for cell functions. The biological functions of the membranes strongly depend on their physicochemical properties, such as electrostatics, phase state, viscosity, polarity and hydration. These properties are essential for the membrane structure and the proper folding and function of membrane proteins. To monitor these properties, fluorescence techniques and notably, two-photon microscopy appear highly suited due to their exquisite sensitivity and their capability to operate in complex biological systems, such as living cells and tissues. In this context, we have developed multiparametric environment-sensitive fluorescent probes tailored for precise location in the membrane bilayer. We notably developed probes of the 3-hydroxychromone family, characterized by an excited state intramolecular proton transfer reaction, which generates two tautomeric emissive species with well-separated emission bands. As a consequence, the response of these probes to changes in their environment could be monitored through changes in the ratios of the two bands, as well as through changes in the fluorescence lifetimes. Using two-photon ratiometric imaging and FLIM, these probes were used to monitor the surface membrane potential, and were applied to detect apoptotic cells and image membrane domains.

  16. Normalized fluorescence lifetime imaging for tumor identification and margin delineation

    Science.gov (United States)

    Sherman, Adria J.; Papour, Asael; Bhargava, Siddharth; Taylor, Zach; Grundfest, Warren S.; Stafsudd, Oscar M.

    2013-03-01

    Fluorescence lifetime imaging microscopy (FLIM) is a technique that has been proven to produce quantitative and qualitative differentiation and identification of substances with good specificity and sensitivity based on lifetime extracted information. This technique has shown the ability to also differentiate between a wide range of tissue types to identify malignant from benign tissue in vivo and ex vivo. However, the complexity, long duration and effort required to generate this information has limited the adoption of these techniques in a clinical setting. Our group has developed a time-resolved imaging system (patent pending) that does not require the extraction of lifetimes or use of complex curve fitting algorithms to display the needed information. The technique, entitled Lifetime Fluorescence Imaging (LFI, or NoFYI), converts fluorescence lifetime decay information directly into visual contrast. Initial studies using Fluorescein and Rhodamine-B demonstrated the feasibility of this approach. Subsequent studies demonstrated the ability to separate collagen and elastin powders. The technique uses nanosecond pulsed UV LEDs at 375 nm for average illumination intensities of ~4.5 μW on the tissue surface with detection by a gated CCD camera. To date, we have imaged 11 surgical head and neck squamous cell carcinoma and brain cancer biopsy specimens including 5 normal and 6 malignant samples. Images at multiple wavelengths clearly demonstrate differentiation between benign and malignant tissue, which was later confirmed by histology. Contrast was obtained between fluorophores with 35 μm spatial resolution and an SNR of ~30 dB allowing us to clearly define tumor margins in these highly invasive cancers. This method is capable of providing both anatomical and chemical information for the pathologist and the surgeon. These results suggest that this technology has a possible role in identifying tumors in tissue specimens and detecting tumor margins during procedures.

  17. Intracellular temperature mapping with a fluorescent polymeric thermometer and fluorescence lifetime imaging microscopy.

    Science.gov (United States)

    Okabe, Kohki; Inada, Noriko; Gota, Chie; Harada, Yoshie; Funatsu, Takashi; Uchiyama, Seiichi

    2012-02-28

    Cellular functions are fundamentally regulated by intracellular temperature, which influences biochemical reactions inside a cell. Despite the important contributions to biological and medical applications that it would offer, intracellular temperature mapping has not been achieved. Here we demonstrate the first intracellular temperature mapping based on a fluorescent polymeric thermometer and fluorescence lifetime imaging microscopy. The spatial and temperature resolutions of our thermometry were at the diffraction limited level (200 nm) and 0.18-0.58 °C. The intracellular temperature distribution we observed indicated that the nucleus and centrosome of a COS7 cell, both showed a significantly higher temperature than the cytoplasm and that the temperature gap between the nucleus and the cytoplasm differed depending on the cell cycle. The heat production from mitochondria was also observed as a proximal local temperature increase. These results showed that our new intracellular thermometry could determine an intrinsic relationship between the temperature and organelle function.

  18. Simultaneous Fluorescence and Phosphorescence Lifetime Imaging Microscopy in Living Cells.

    Science.gov (United States)

    Jahn, Karolina; Buschmann, Volker; Hille, Carsten

    2015-09-22

    In living cells, there are always a plethora of processes taking place at the same time. Their precise regulation is the basis of cellular functions, since small failures can lead to severe dysfunctions. For a comprehensive understanding of intracellular homeostasis, simultaneous multiparameter detection is a versatile tool for revealing the spatial and temporal interactions of intracellular parameters. Here, a recently developed time-correlated single-photon counting (TCSPC) board was evaluated for simultaneous fluorescence and phosphorescence lifetime imaging microscopy (FLIM/PLIM). Therefore, the metabolic activity in insect salivary glands was investigated by recording ns-decaying intrinsic cellular fluorescence, mainly related to oxidized flavin adenine dinucleotide (FAD) and the μs-decaying phosphorescence of the oxygen-sensitive ruthenium-complex Kr341. Due to dopamine stimulation, the metabolic activity of salivary glands increased, causing a higher pericellular oxygen consumption and a resulting increase in Kr341 phosphorescence decay time. Furthermore, FAD fluorescence decay time decreased, presumably due to protein binding, thus inducing a quenching of FAD fluorescence decay time. Through application of the metabolic drugs antimycin and FCCP, the recorded signals could be assigned to a mitochondrial origin. The dopamine-induced changes could be observed in sequential FLIM and PLIM recordings, as well as in simultaneous FLIM/PLIM recordings using an intermediate TCSPC timing resolution.

  19. Red Fluorescent Carbon Nanoparticle-Based Cell Imaging Probe.

    Science.gov (United States)

    Ali, Haydar; Bhunia, Susanta Kumar; Dalal, Chumki; Jana, Nikhil R

    2016-04-13

    Fluorescent carbon nanoparticle-based probes with tunable visible emission are biocompatible, environment friendly and most suitable for various biomedical applications. However, synthesis of red fluorescent carbon nanoparticles and their transformation into functional nanoparticles are very challenging. Here we report red fluorescent carbon nanoparticle-based nanobioconjugates of nanoparticles are synthesized via high temperature colloid-chemical approach and transformed into water-soluble functional nanoparticles via coating with amphiphilic polymer followed by covalent linking with desired biomolecules. Following this approach, carbon nanoparticles are functionalized with polyethylene glycol, primary amine, glucose, arginine, histidine, biotin and folic acid. These functional nanoparticles can be excited with blue/green light (i.e., 400-550 nm) to capture their emission spanning from 550 to 750 nm. Arginine and folic acid functionalized nanoparticles have been demonstrated as fluorescent cell labels where blue and green excitation has been used for imaging of labeled cells. The presented method can be extended for the development of carbon nanoparticle-based other bioimaging probes.

  20. Development of Fluorescence Imaging Lidar for Boat-Based Coral Observation

    Directory of Open Access Journals (Sweden)

    Sasano Masahiko

    2016-01-01

    Full Text Available A fluorescence imaging lidar system installed in a boat-towable buoy has been developed for the observation of reef-building corals. Long-range fluorescent images of the sea bed can be recorded in the daytime with this system. The viability of corals is clear in these fluorescent images because of the innate fluorescent proteins. In this study, the specifications and performance of the system are shown.

  1. FISHji: New ImageJ macros for the quantification of fluorescence in epifluorescence images

    DEFF Research Database (Denmark)

    Fontenete, Sílvia; Carvalho, Daniel R; Lourenço, Anália

    2016-01-01

    and tools has been trying to overcome this problem, however, the determination of fluorescent intensity in microscopy images still has issues due to the lack of precision in the results and the complexity of existing software. This work presents FISHji, a set of new ImageJ methods for automated...

  2. Single camera imaging system for color and near-infrared fluorescence image guided surgery.

    Science.gov (United States)

    Chen, Zhenyue; Zhu, Nan; Pacheco, Shaun; Wang, Xia; Liang, Rongguang

    2014-08-01

    Near-infrared (NIR) fluorescence imaging systems have been developed for image guided surgery in recent years. However, current systems are typically bulky and work only when surgical light in the operating room (OR) is off. We propose a single camera imaging system that is capable of capturing NIR fluorescence and color images under normal surgical lighting illumination. Using a new RGB-NIR sensor and synchronized NIR excitation illumination, we have demonstrated that the system can acquire both color information and fluorescence signal with high sensitivity under normal surgical lighting illumination. The experimental results show that ICG sample with concentration of 0.13 μM can be detected when the excitation irradiance is 3.92 mW/cm(2) at an exposure time of 10 ms.

  3. Ultrafast superresolution fluorescence imaging with spinning disk confocal microscope optics.

    Science.gov (United States)

    Hayashi, Shinichi; Okada, Yasushi

    2015-05-01

    Most current superresolution (SR) microscope techniques surpass the diffraction limit at the expense of temporal resolution, compromising their applications to live-cell imaging. Here we describe a new SR fluorescence microscope based on confocal microscope optics, which we name the spinning disk superresolution microscope (SDSRM). Theoretically, the SDSRM is equivalent to a structured illumination microscope (SIM) and achieves a spatial resolution of 120 nm, double that of the diffraction limit of wide-field fluorescence microscopy. However, the SDSRM is 10 times faster than a conventional SIM because SR signals are recovered by optical demodulation through the stripe pattern of the disk. Therefore a single SR image requires only a single averaged image through the rotating disk. On the basis of this theory, we modified a commercial spinning disk confocal microscope. The improved resolution around 120 nm was confirmed with biological samples. The rapid dynamics of micro-tubules, mitochondria, lysosomes, and endosomes were observed with temporal resolutions of 30-100 frames/s. Because our method requires only small optical modifications, it will enable an easy upgrade from an existing spinning disk confocal to a SR microscope for live-cell imaging. © 2015 Hayashi and Okada. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  4. Selective Detection of Neurotransmitters by Fluorescence and Chemiluminescence Imaging

    Energy Technology Data Exchange (ETDEWEB)

    Ziqiang Wang; Edward S. Yeung

    2001-08-06

    In recent years, luminescence imaging has been widely employed in neurochemical analysis. It has a number of advantages for the study of neuronal and other biological cells: (1) a particular molecular species or cellular constituent can be selectively visualized in the presence of a large excess of other species in a heterogeneous environment; (2) low concentration detection limits can be achieved because of the inherent sensitivity associated with fluorescence and chemiluminescence; (3) low excitation intensities can be used so that long-term observation can be realized while the viability of the specimen is preserved; and (4) excellent spatial resolution can be obtained with the light microscope so subcellular compartments can be identified. With good sensitivity, temporal and spatial resolution, the flux of ions and molecules and the distribution and dynamics of intracellular species can be measured in real time with specific luminescence probes, substrates, or with native fluorescence. A noninvasive detection scheme based on glutamate dehydrogenase (GDH) enzymatic assay combined with microscopy was developed to measure the glutamate release in cultured cells from the central nervous system (CNS). The enzyme reaction is very specific and sensitive. The detection limit with CCD imaging is down to {micro}M levels of glutamate with reasonable response time. They also found that chemiluminescence associated with the ATP-dependent reaction between luciferase and luciferin can be used to image ATP at levels down to 10 nM in the millisecond time scale. Similar imaging experiments should be feasible in a broad spectrum of biological systems.

  5. Nanoparticles and nanocomposites for fluorescence sensing and imaging

    Science.gov (United States)

    Demchenko, Alexander P.

    2013-06-01

    The assortment of fluorescence reporters is changing dramatically. Traditionally explored intrinsic fluorescence of biological macromolecules and cellular pigments and of externally introduced organic dyes are presently in strong competition with new nanomaterials. Among them are conjugated polymers, semiconductor nanocrystals (quantum dots), up-converting nanocrystals, magic-size clusters of silver and gold, nanodiamonds and carbon dots. They demonstrate diverse photophysical behavior and allow one to obtain diverse information when used in analytical tools or when they form images in biological systems. Based on them, functional nanocomposites displaying a variety of useful features, thus extending dramatically the information content of output data, can be constructed. We describe their properties and compare them with those of small-molecular emitters, such as organic dyes. With their aid, one can modulate over a wide range the wavelengths of excitation and emission, the lifetimes and anisotropies and design the systems with ‘superenhancement’ and ‘superquenching’. Such unlimited possibilities are offered by combining different types of luminophores based on electronic conjugation, plasmonic effects or excited-state resonance energy transfer. This tutorial review provides a comparative analysis of the properties of new nanoscale materials and of their hybrid nanocomposites for applications in fluorescence sensing and imaging.

  6. Photostable and photoswitching fluorescent dyes for super-resolution imaging.

    Science.gov (United States)

    Minoshima, Masafumi; Kikuchi, Kazuya

    2017-01-12

    Super-resolution fluorescence microscopy is a recently developed imaging tool for biological researches. Several methods have been developed for detection of fluorescence signals from molecules in a subdiffraction-limited area, breaking the diffraction limit of the conventional optical microscopies and allowing visualization of detailed macromolecular structures in cells. As objectives are exposed to intense laser in the optical systems, fluorophores for super-resolution microscopy must be tolerated even under severe light irradiation conditions. The fluorophores must also be photoactivatable and photoswitchable for single-molecule localization-based super-resolution microscopy, because the number of active fluorophores must be controlled by light irradiation. This has led to growing interest in these properties in the development of fluorophores. In this mini-review, we focus on the development of photostable and photoswitching fluorescent dyes for super-resolution microscopy. We introduce recent efforts, including improvement of fluorophore photostability and control of photoswitching behaviors of fluorophores based on photochemical and photophysical processes. Understanding and manipulation of chemical reactions in excited fluorophores can develop highly photostable and efficiently photoswitchable fluorophores that are suitable for super-resolution imaging applications.

  7. Portable multispectral fluorescence imaging system for food safety applications

    Science.gov (United States)

    Lefcourt, Alan M.; Kim, Moon S.; Chen, Yud-Ren

    2004-03-01

    Fluorescence can be a sensitive method for detecting food contaminants. Of particular interest is detection of fecal contamination as feces is the source of many pathogenic organisms. Feces generally contain chlorophyll a and related compounds due to ingestion of plant materials, and these compounds can readily be detected using fluorescence techniques. Described is a fluorescence-imaging system consisting primarily of a UV light source, an intensified camera with a six-position filter wheel, and software for controlling the system and automatically analyzing the resulting images. To validate the system, orchard apples artificially contaminated with dairy feces were used in a "hands-on" public demonstration. The contamination sites were easily identified using automated edge detection and threshold detection algorithms. In addition, by applying feces to apples and then washing sets of apples at hourly intervals, it was determined that five h was the minimum contact time that allowed identification of the contamination site after the apples were washed. There are many potential uses for this system, including studying the efficacy of apple washing systems.

  8. RNA Imaging with Multiplexed Error Robust Fluorescence in situ Hybridization

    Science.gov (United States)

    Moffitt, Jeffrey R.; Zhuang, Xiaowei

    2016-01-01

    Quantitative measurements of both the copy number and spatial distribution of large fractions of the transcriptome in single-cells could revolutionize our understanding of a variety of cellular and tissue behaviors in both healthy and diseased states. Single-molecule Fluorescence In Situ Hybridization (smFISH)—an approach where individual RNAs are labeled with fluorescent probes and imaged in their native cellular and tissue context—provides both the copy number and spatial context of RNAs but has been limited in the number of RNA species that can be measured simultaneously. Here we describe Multiplexed Error Robust Fluorescence In Situ Hybridization (MERFISH), a massively parallelized form of smFISH that can image and identify hundreds to thousands of different RNA species simultaneously with high accuracy in individual cells in their native spatial context. We provide detailed protocols on all aspects of MERFISH, including probe design, data collection, and data analysis to allow interested laboratories to perform MERFISH measurements themselves. PMID:27241748

  9. Fluorescence and image guided resection in high grade glioma.

    Science.gov (United States)

    Panciani, Pier Paolo; Fontanella, Marco; Schatlo, Bawarjan; Garbossa, Diego; Agnoletti, Alessandro; Ducati, Alessandro; Lanotte, Michele

    2012-01-01

    The extent of resection in high grade glioma is increasingly been shown to positively effect survival. Nevertheless, heterogeneity and migratory behavior of glioma cells make gross total resection very challenging. Several techniques were used in order to improve the detection of residual tumor. Aim of this study was to analyze advantages and limitations of fluorescence and image guided resection. A multicentric prospective study was designed to evaluate the accuracy of each method. Furthermore, the role of 5-aminolevulinc acid and neuronavigation were reviewed. Twenty-three patients harboring suspected high grade glioma, amenable to complete resection, were enrolled. Fluorescence and image guides were used to perform surgery. Multiple samples were obtained from the resection cavity of each lesion according to 5-ALA staining positivity and boundaries as delineated by neuronavigation. All samples were analyzed by a pathologist blinded to the intra-operative labeling. Decision-making based on fluorescence showed a sensitivity of 91.1% and a specificity of 89.4% (pimage-guided resection accuracy was low (sensitivity: 57.8%; specificity: 57.4%; p=0.346). We observed that the sensitivity of 5-ALA can be improved by the combined use of neuronavigation, but this leads to a significant reduction in specificity. Thus, the use of auxiliary techniques should always be subject to critical skills of the surgeon. We advocate a large-scale study to further improve the assessment of multimodal approaches.

  10. Usefulness of magnetic resonance imaging (MRI) in the detection of the lesions of gestational trophoblastic disease; Comparison with computed tomography and digital subtraction angiography

    Energy Technology Data Exchange (ETDEWEB)

    Takeuchi, Satoshi; Akahori, Taiichiro; Mochizuki, Matsuto; Kono, Michio (Kobe Univ. (Japan). School of Medicine)

    1992-02-01

    Twenty patients with gestational trophoblastic disease (GTN) were examined by magnetic resonance imaging (MRI), computed tomography (CT) and digital subtraction angiography (DSA), to evaluate their usefulness in the diagnosis of the disease. The lesions of hydatidiform mole were mainly composed of molar vesicles, dilated vessels and hemorrhage which were depicted as small round high intensity lesions on the T2-weighted images and as tree-like low intensity lesions and high or low intensity lesions of various shapes in the T1-, T2-weighted images. These MRI findings closely corresponded to the histopathological findings. On the other hand, CT findings obtained with hydatidiform mole were characterized by filling defects or a small round low density area on contrast enhanced images. The detection ratio for intramural lesions of invasive mole and choriocarcinoma by MRI was 83% (5/6), while that by CT was 50% (3/6). The obliteration of the junctional zone and interruption of the myometrium observed in MRI were significant signs suggesting intramural invasion of the disease. In fact, these signs in MRI were observed in all of the six cases of invasive mole or choriocarcinoma examined. In conclusion, MRI is a powerful means for the determining the intramural invasive mole and choriocarcinoma. Thus more accurate diagnosis of GTN will be obtained with the combined use of MRI and DSA. (author).

  11. Image registration and averaging of low laser power two-photon fluorescence images of mouse retina.

    Science.gov (United States)

    Alexander, Nathan S; Palczewska, Grazyna; Stremplewski, Patrycjusz; Wojtkowski, Maciej; Kern, Timothy S; Palczewski, Krzysztof

    2016-07-01

    Two-photon fluorescence microscopy (TPM) is now being used routinely to image live cells for extended periods deep within tissues, including the retina and other structures within the eye . However, very low laser power is a requirement to obtain TPM images of the retina safely. Unfortunately, a reduction in laser power also reduces the signal-to-noise ratio of collected images, making it difficult to visualize structural details. Here, image registration and averaging methods applied to TPM images of the eye in living animals (without the need for auxiliary hardware) demonstrate the structural information obtained with laser power down to 1 mW. Image registration provided between 1.4% and 13.0% improvement in image quality compared to averaging images without registrations when using a high-fluorescence template, and between 0.2% and 12.0% when employing the average of collected images as the template. Also, a diminishing return on image quality when more images were used to obtain the averaged image is shown. This work provides a foundation for obtaining informative TPM images with laser powers of 1 mW, compared to previous levels for imaging mice ranging between 6.3 mW [Palczewska G., Nat Med.20, 785 (2014) Sharma R., Biomed. Opt. Express4, 1285 (2013)].

  12. Fluorescence Imaging In Vivo up to 1700 nm

    CERN Document Server

    Diao, Shuo; Hong, Guosong; Antaris, Alexander L; Chang, Junlei; Wu, Justin Z; Zhang, Bo; Kuo, Calvin J; Dai, Hongjie

    2015-01-01

    Compared to visible and near-infrared regions below ~ 900 nm, imaging in the second near-infrared window beyond 1000 nm (NIR-II, 1000-1700 nm) is promising for deep-tissue high-resolution optical imaging in vivo owing to reduced scattering of photons traversing through tissues. Here, we succeeded fluorescence imaging in vivo in the long 1500-1700 nm (NIR-IIb) region using a novel, chemical separation enriched large-diameter semiconducting single-walled carbon nanotube material. Imaging in the 1500-1700 nm window resolved 3-4 um wide capillary blood vessels at ~ 3 millimeters depth through the intact body and brain of mice with the ability of blood-flow speed mapping in individual capillary vessels. Further, non-invasive single fluorophore imaging inside the tumor of a live mouse was achieved in the 1500-1700 nm window. NIR-IIb imaging can be generalized to a wide range of fluorophores emitting up to 1700 nm for a new paradigm of high performance in vivo optical imaging.

  13. Near-Infrared Fluorescent NanoGUMBOS for Biomedical Imaging

    Energy Technology Data Exchange (ETDEWEB)

    Bwambok, David [Louisiana State University; El-Zahab, Bilal [Lousianna State University; Challa, Santhosh [Louisiana State University; Li, Min [Lousianna State University; Chandler, Lin [Horiba Jobin Yvon Inc.; Baker, Gary A [ORNL; Warner, Isiah M [ORNL

    2009-01-01

    Herein, we report on near-infrared (NIR) fluorescent nanoparticles generated from an emergent class of materials we refer to as a Group of Uniform Materials Based on Organic Salts (GUMBOS). GUMBOS are largely frozen ionic liquids, although the concept is more general and is also easily applied to solid ionic materials with melting points in excess of 100 C. Nanoparticles based on GUMBOS (nanoGUMBOS) derived from a NIR fluorophore are prepared using a reprecipitation method and evaluated for in vivo fluorescence imaging. Due to their uniformity, single-step preparation, and composite nature, nanoGUMBOS help to resolve issues with dye leakage problems innate to alternate cellular stains and unlock a myriad of applications for these materials, highlighting exciting possibilities for multifunctional nanoGUMBOS.

  14. Development of temperature imaging using two-line atomic fluorescence.

    Science.gov (United States)

    Medwell, Paul R; Chan, Qing N; Kalt, Peter A M; Alwahabi, Zeyad T; Dally, Bassam B; Nathan, Graham J

    2009-02-20

    This work aims to advance understanding of the coupling between temperature and soot. The ability to image temperature using the two-line atomic fluorescence (TLAF) technique is demonstrated. Previous TLAF theory is extended from linear excitation into the nonlinear fluence regime. Nonlinear regime two-line atomic fluorescence (NTLAF) provides superior signal and reduces single-shot uncertainty from 250 K for conventional TLAF down to 100 K. NTLAF is shown to resolve the temperature profile across the stoichiometric envelope for hydrogen, ethylene, and natural gas flames, with deviation from thermocouple measurements not exceeding 100 K, and typically ≲30 K. Measurements in flames containing soot demonstrate good capacity of NTLAF to exclude interferences that hamper most two-dimensional thermometry techniques.

  15. Defining a superlens operating regime for imaging fluorescent molecules.

    Directory of Open Access Journals (Sweden)

    Kareem Elsayad

    Full Text Available It has been shown that thin metal-based films can at certain frequencies act as planar near-field lenses for certain polarization components. A desirable property of such "lenses" is that they can also enhance and focus some large transverse spatial frequency components which contain sub-diffraction limit details. Over the last decade there has been much work in optimizing designs to reduce effects (such as material losses and surface roughness that are detrimental to image reconstruction. One design that can reduce some of these undesirable effects, and which has received a fair amount of attention recently, is the stacked metal-dielectric superlens. Here we theoretically explore the imaging ability of such a design for the specific purpose of imaging a fluorescent dye (the common bio-marker GFP in the vicinity of the superlens surface. Our calculations take into consideration the interaction (damping of an oscillating electric dipole with the metallic layers in the superlens. We also assume a Gaussian frequency distribution spectrum for the dipole. We treat the metallic-alloy and dielectric-alloy layers separately using an appropriate effective medium theory. The transmission properties are evaluated via Transfer matrix (-matrix calculations that were performed in the MatLab and MathCad environments. Our study shows that it is in principle possible to image fluorescent molecules using a simple bilayer planar superlens. We find that optimal parameters for such a superlens occur when the peak dipole emission-frequency is slightly offset from the Surface Plasmon resonance frequency of the metal-dielectric interfaces. The best resolution is obtained when the fluorescent molecules are not too close (>/ approximately 10 nm or too far (>/approximately 30 nm from the superlens surface. The realization and application of a superlens with the specified design is possible using current nanofabrication techniques. When combined with e.g. a sub

  16. Materials characterization using micro-x-ray fluorescence elemental imaging.

    Energy Technology Data Exchange (ETDEWEB)

    Havrilla, G. J. (George J.); Miller, T. C. (Thomasin C.); Joseph, M. R. (Martha R.)

    2002-01-01

    Materials characterization continues to be a key challenge in a variety of programs. Although bulk elemental composition provides overall concentration of both major and trace elements, the distribution of these elements both on micro and macro scales can determine the performance and ultimately the physical properties of the materials. Hence elemental imaging can provide a new level of information for major and in some cases bulk trace concentrations of elements. Micro X-ray fluorescence (MXRF) offers unique capabilities in terms of elemental imaging. This approach is based on a meso scale level of resolution around 50 micrometer X-ray spot size. When coupled with a moveable stage, specimens several inches on a side can be imaged with surprising detail. In most instances, qualitative images are sufficient to illustrate the elemental heterogeneity. This information can then be used to determine if the material meets the desired physical characteristics and whether this is due to the observed heterogeneity or in spite of it. Several examples of elemental imaging will be presented. These will include the aging of polymers and the effects of residual organotin catalyst. The tin can be imaged using MXRF and has been show to be mobile within the polymeric material over time. Corrosion is a serious issue throughout the industrial world. A specific example of chloride attack on a metal, which creates problems in waste storage. Finally, MXRF used in high throughput screening in the development of novel peptide receptors will be shown. The advantage of MXRF is that no fluorescent tags need be added to the target molecules. This insures the unhindered interaction of the target molecules and allows for additional characterization using molecular spectroscopic techniques.

  17. TU-CD-207-03: Time Evolution of Texture Parameters of Subtracted Images Obtained by Contrast-Enhanced Digital Mammography (CEDM)

    Energy Technology Data Exchange (ETDEWEB)

    Mateos, M-J; Brandan, M-E [Instituto de Fisica, Universidad Nacional Autonom de Mexico, Mexico, Distrito Federal (Mexico); Gastelum, A; Marquez, J [Centro de Ciencias Aplicadas y Desarrollo Tecnologico Universidad Nacional Autonoma de Mexico, Mexico, Distrito Federal (Mexico)

    2015-06-15

    Purpose: To evaluate the time evolution of texture parameters, based on the gray level co-occurrence matrix (GLCM), in subtracted images of 17 patients (10 malignant and 7 benign) subjected to contrast-enhanced digital mammography (CEDM). The goal is to determine the sensitivity of texture to iodine uptake at the lesion, and its correlation (or lack of) with mean-pixel-value (MPV). Methods: Acquisition of clinical images followed a single-energy CEDM protocol using Rh/Rh/48 kV plus external 0.5 cm Al from a Senographe DS unit. Prior to the iodine-based contrast medium (CM) administration a mask image was acquired; four CM images were obtained 1, 2, 3, and 5 minutes after CM injection. Temporal series were obtained by logarithmic subtraction of registered CM minus mask images.Regions of interest (ROI) for the lesion were drawn by a radiologist and the texture was analyzed. GLCM was evaluated at a 3 pixel distance, 0° angle, and 64 gray-levels. Pixels identified as registration errors were excluded from the computation. 17 texture parameters were chosen, classified according to similarity into 7 groups, and analyzed. Results: In all cases the texture parameters within a group have similar dynamic behavior. Two texture groups (associated to cluster and sum mean) show a strong correlation with MPV; their average correlation coefficient (ACC) is r{sup 2}=0.90. Other two groups (contrast, homogeneity) remain constant with time, that is, a low-sensitivity to CM uptake. Three groups (regularity, lacunarity and diagonal moment) are sensitive to CM uptake but less correlated with MPV; their ACC is r{sup 2}=0.78. Conclusion: This analysis has shown that, at least groups associated to regularity, lacunarity and diagonal moment offer dynamical information additional to the mean pixel value due to the presence of CM at the lesion. The next step will be the analysis in terms of the lesion pathology. Authors thank PAPIIT-IN105813 for support. Consejo Nacional de Ciencia Y

  18. Multispectral fluorescence imaging technique for discrimination of cucumber (Cucumis Sativus) seed viability

    Science.gov (United States)

    In this study, we developed a nondestructive method for discriminating viable cucumber (Cucumis sativus) seeds based on hyperspectral fluorescence imaging. The fluorescence spectra of cucumber seeds in the 420–700 nm range were extracted from hyperspectral fluorescence images obtained using 365 nm u...

  19. Fluorescence and fluorescence-lifetime imaging microscopy (FLIM) to characterize yeast strains by autofluorescence

    Science.gov (United States)

    Bhatta, H.; Goldys, E. M.; Ma, J.

    2006-02-01

    We characterised populations of wild type baking and brewing yeast cells using intrinsic fluorescence and fluorescence lifetime microscopy, in order to obtain quantitative identifiers of different strains. The cell autofluorescence was excited at 405 nm and observed within 440-540 nm range where strong cell to cell variability was observed. The images were analyzed using customised public domain software, which provided information on cell size, intensity and texture-related features. In light of significant diversity of the data, statistical methods were utilized to assess the validity of the proposed quantitative identifiers for strain differentiation. The Kolmogorov-Smirnov test was applied to confirm that empirical distribution functions for size, intensity and entropy for different strains were statistically different. These characteristics were followed with culture age of 24, 48 and 72 h, (the latter corresponding to a stationary growth phase) and size, and to some extent entropy, were found to be independent of age. The fluorescence intensity presented a distinctive evolution with age, different for each of the examined strains. The lifetime analysis revealed a short decay time component of 1.4 ns and a second, longer one with the average value of 3.5 ns and a broad distribution. High variability of lifetime values within cells was observed however a lifetime texture feature in the studied strains was statistically different.

  20. SIMA: Python software for analysis of dynamic fluorescence imaging data

    Directory of Open Access Journals (Sweden)

    Patrick eKaifosh

    2014-09-01

    Full Text Available Fluorescence imaging is a powerful method for monitoring dynamic signals in the nervous system. However, analysis of dynamic fluorescence imaging data remains burdensome, in part due to the shortage of available software tools. To address this need, we have developed SIMA, an open source Python package that facilitates common analysis tasks related to fluorescence imaging. Functionality of this package includes correction of motion artifacts occurring during in vivo imaging with laser-scanning microscopy, segmentation of imaged fields into regions of interest (ROIs, and extraction of signals from the segmented ROIs. We have also developed a graphical user interface (GUI for manual editing of the automatically segmented ROIs and automated registration of ROIs across multiple imaging datasets. This software has been designed with flexibility in mind to allow for future extension with different analysis methods and potential integration with other packages. Software, documentation, and source code for the SIMA package and ROI Buddy GUI are freely available at http://www.losonczylab.org/sima/.

  1. Breast cancer margin delineation with fluorescence lifetime imaging (Conference Presentation)

    Science.gov (United States)

    Phipps, Jennifer E.; Gorpas, Dimitris; Darrow, Morgan; Unger, Jakob; Bold, Richard; Marcu, Laura

    2017-02-01

    The current standard of care for early stages of breast cancer is breast-conserving surgery (BCS). BCS involves a lumpectomy procedure, during which the tumor is removed with a rim of normal tissue-if cancer cells found in that rim of tissue, it is called a positive margin and means part of the tumor remains in the breast. Currently there is no method to determine if cancer cells exist at the margins of lumpectomy specimens aside from time-intensive histology methods that result in reoperations in up to 38% of cases. We used fluorescence lifetime imaging (FLIm) to measure time-resolved autofluorescence from N=13 ex vivo human breast cancer specimens (N=10 patients undergoing lumpectomy or mastectomy) and compared our results to histology. Tumor (both invasive and ductal carcinoma in situ), fibrous tissue, fat and fat necrosis have unique fluorescence signatures. For instance, between 500-580 nm, fluorescence lifetime of tumor was shortest (4.7 +/- 0.4 ns) compared to fibrous tissue (5.5 +/- 0.7 ns) and fat (7.0 +/- 0.1 ns), P<0.05 (ANOVA). These differences are due to the biochemical properties of lipid, nicotineamide adenine dinucleotide (NADH) and collagen fibers in the fat, tumor and fibrous tissue, respectively. Additionally, the FLIm data is augmented to video of the breast tissue with image processing algorithms that track a blue (450 nm) aiming beam used in parallel with the 355 nm excitation beam. This allows for accurate histologic co-registration and in the future will allow for three-dimensional lumpectomy surfaces to be imaged for cancer margin delineation.

  2. FIZICS: fluorescent imaging zone identification system, a novel macro imaging system.

    Science.gov (United States)

    Skwish, Stephen; Asensio, Francisco; King, Greg; Clarke, Glenn; Kath, Gary; Salvatore, Michael J; Dufresne, Claude

    2004-12-01

    Constantly improving biological assay development continues to drive technological requirements. Recently, a specification was defined for capturing white light and fluorescent images of agar plates ranging in size from the NUNC Omni tray (96-well footprint, 128 x 85 mm) to the NUNC Bio Assay Dish (245 x 245 mm). An evaluation of commercially available products failed to identify any system capable of fluorescent macroimaging with discrete wavelength selection. To address the lack of a commercially available system, a custom imaging system was designed and constructed. This system provides the same capabilities of many commercially available systems with the added ability to fluorescently image up to a 245 x 245 mm area using wavelengths in the visible light spectrum.

  3. Cyanine-based probe\\tag-peptide pair fluorescence protein imaging and fluorescence protein imaging methods

    Science.gov (United States)

    Mayer-Cumblidge, M. Uljana; Cao, Haishi

    2013-01-15

    A molecular probe comprises two arsenic atoms and at least one cyanine based moiety. A method of producing a molecular probe includes providing a molecule having a first formula, treating the molecule with HgOAc, and subsequently transmetallizing with AsCl.sub.3. The As is liganded to ethanedithiol to produce a probe having a second formula. A method of labeling a peptide includes providing a peptide comprising a tag sequence and contacting the peptide with a biarsenical molecular probe. A complex is formed comprising the tag sequence and the molecular probe. A method of studying a peptide includes providing a mixture containing a peptide comprising a peptide tag sequence, adding a biarsenical probe to the mixture, and monitoring the fluorescence of the mixture.

  4. Modelling of microcracks image treated with fluorescent dye

    Science.gov (United States)

    Glebov, Victor; Lashmanov, Oleg U.

    2015-06-01

    The main reasons of catastrophes and accidents are high level of wear of equipment and violation of the production technology. The methods of nondestructive testing are designed to find out defects timely and to prevent break down of aggregates. These methods allow determining compliance of object parameters with technical requirements without destroying it. This work will discuss dye penetrant inspection or liquid penetrant inspection (DPI or LPI) methods and computer model of microcracks image treated with fluorescent dye. Usually cracks on image look like broken extended lines with small width (about 1 to 10 pixels) and ragged edges. The used method of inspection allows to detect microcracks with depth about 10 or more micrometers. During the work the mathematical model of image of randomly located microcracks treated with fluorescent dye was created in MATLAB environment. Background noises and distortions introduced by the optical systems are considered in the model. The factors that have influence on the image are listed below: 1. Background noise. Background noise is caused by the bright light from external sources and it reduces contrast on the objects edges. 2. Noises on the image sensor. Digital noise manifests itself in the form of randomly located points that are differing in their brightness and color. 3. Distortions caused by aberrations of optical system. After passing through the real optical system the homocentricity of the bundle of rays is violated or homocentricity remains but rays intersect at the point that doesn't coincide with the point of the ideal image. The stronger the influence of the above-listed factors, the worse the image quality and therefore the analysis of the image for control of the item finds difficulty. The mathematical model is created using the following algorithm: at the beginning the number of cracks that will be modeled is entered from keyboard. Then the point with random position is choosing on the matrix whose size is

  5. Fluorescence-Doped Particles for Simultaneous Temperature and Velocity Imaging

    Science.gov (United States)

    Danehy, Paul M.; Tiemsin, Pacita I.; Wohl, Chrostopher J.; Verkamp, Max; Lowe, T.; Maisto, P.; Byun, G.; Simpson, R.

    2012-01-01

    Polystyrene latex microspheres (PSLs) have been used for particle image velocimetry (PIV) and laser Doppler velocimetry (LDV) measurements for several decades. With advances in laser technologies, instrumentation, and data processing, the capability to collect more information about fluid flow beyond velocity is possible using new seed materials. To provide additional measurement capability, PSLs were synthesized with temperature-sensitive fluorescent dyes incorporated within the particle. These multifunctional PSLs would have the greatest impact if they could be used in large scale facilities with minimal modification to the facilities or the existing instrumentation. Consequently, several potential dyes were identified that were amenable to existing laser systems currently utilized in wind tunnels at NASA Langley Research Center as well as other wind and fluid (water) tunnels. PSLs incorporated with Rhodamine B, dichlorofluorescein (DCF, also known as fluorescein 548 or fluorescein 27) and other dyes were synthesized and characterized for morphology and spectral properties. The resulting particles were demonstrated to exhibit fluorescent emission, which would enable determination of both fluid velocity and temperature. They also would allow near-wall velocity measurements whereas laser scatter from surfaces currently prevents near-wall measurements using undoped seed materials. Preliminary results in a wind tunnel facility located at Virginia Polytechnic Institute and State University (Virginia Tech) have verified fluorescent signal detection and temperature sensitivity of fluorophore-doped PSLs.

  6. Red emitting neutral fluorescent glycoconjugates for membrane optical imaging.

    Science.gov (United States)

    Redon, Sébastien; Massin, Julien; Pouvreau, Sandrine; De Meulenaere, Evelien; Clays, Koen; Queneau, Yves; Andraud, Chantal; Girard-Egrot, Agnès; Bretonnière, Yann; Chambert, Stéphane

    2014-04-16

    A family of neutral fluorescent probes was developed, mimicking the overall structure of natural glycolipids in order to optimize their membrane affinity. Nonreducing commercially available di- or trisaccharidic structures were connected to a push-pull chromophore based on dicyanoisophorone electron-accepting group, which proved to fluoresce in the red region with a very large Stokes shift. This straightforward synthetic strategy brought structural variations to a series of probes, which were studied for their optical, biophysical, and biological properties. The insertion properties of the different probes into membranes were evaluated on a model system using the Langmuir monolayer balance technique. Confocal fluorescence microscopy performed on muscle cells showed completely different localizations and loading efficiencies depending on the structure of the probes. When compared to the commercially available ANEPPS, a family of commonly used membrane imaging dyes, the most efficient probes showed a similar brightness, but a sharper pattern was observed. According to this study, compounds bearing one chromophore, a limited size of the carbohydrate moiety, and an overall rod-like shape gave the best results.

  7. Fluorescence imaging of chromosomal DNA using click chemistry

    Science.gov (United States)

    Ishizuka, Takumi; Liu, Hong Shan; Ito, Kenichiro; Xu, Yan

    2016-09-01

    Chromosome visualization is essential for chromosome analysis and genetic diagnostics. Here, we developed a click chemistry approach for multicolor imaging of chromosomal DNA instead of the traditional dye method. We first demonstrated that the commercially available reagents allow for the multicolor staining of chromosomes. We then prepared two pro-fluorophore moieties that served as light-up reporters to stain chromosomal DNA based on click reaction and visualized the clear chromosomes in multicolor. We applied this strategy in fluorescence in situ hybridization (FISH) and identified, with high sensitivity and specificity, telomere DNA at the end of the chromosome. We further extended this approach to observe several basic stages of cell division. We found that the click reaction enables direct visualization of the chromosome behavior in cell division. These results suggest that the technique can be broadly used for imaging chromosomes and may serve as a new approach for chromosome analysis and genetic diagnostics.

  8. Fluorescent cyanine probe for DNA detection and cellular imaging

    Science.gov (United States)

    Zheng, Yong-Chao; Zheng, Mei-Ling; Zhao, Zhen-Sheng; Duan, Xuan-Ming

    2014-03-01

    In our study, two carbazole-based cyanines, 3,6-bis[2-(1-methylpyridinium)vinyl]-9-methyl carbazole diiodide (A) and 6,6'-bis[2-(1-methylpyridinium)vinyl]-bis(9-methyl-carbazol-3yl)methane diiodide (B) were synthesized and employed as light-up probes for DNA and cell imaging. Both of the cyanine probes possess a symmetric structure and bis-cationic center. The obvious induced circular dichroism signals in circular dichroism spectra reveal that the molecules can specifically interact with DNA. Strong fluorescence enhancement is observed when these two cyanines are bound to DNA. These cyanine probes show high binding affinity to oligonucleotides but different binding preferences to various secondary structures. Confocal microscopy images of fixed cell stained by the probes exhibit strong brightness and high contrast in nucleus with a very low cytoplasmic background.

  9. Fluorescence imaging of chromosomal DNA using click chemistry

    Science.gov (United States)

    Ishizuka, Takumi; Liu, Hong Shan; Ito, Kenichiro; Xu, Yan

    2016-01-01

    Chromosome visualization is essential for chromosome analysis and genetic diagnostics. Here, we developed a click chemistry approach for multicolor imaging of chromosomal DNA instead of the traditional dye method. We first demonstrated that the commercially available reagents allow for the multicolor staining of chromosomes. We then prepared two pro-fluorophore moieties that served as light-up reporters to stain chromosomal DNA based on click reaction and visualized the clear chromosomes in multicolor. We applied this strategy in fluorescence in situ hybridization (FISH) and identified, with high sensitivity and specificity, telomere DNA at the end of the chromosome. We further extended this approach to observe several basic stages of cell division. We found that the click reaction enables direct visualization of the chromosome behavior in cell division. These results suggest that the technique can be broadly used for imaging chromosomes and may serve as a new approach for chromosome analysis and genetic diagnostics. PMID:27620982

  10. Dynamic fluorescence imaging for multiparametric measurement of tumor vasculature

    Science.gov (United States)

    Choi, Myunghwan; Choi, Kyungsun; Ryu, Seung-Wook; Lee, Jungwhoi; Choi, Chulhee

    2011-04-01

    Angiogenesis is essential for tumor growth and a promising target for cancer therapy. Blood vessel monitoring is an indispensable tool for evaluation and development of anti-angiogenic drugs. Here, we report a new noninvasive in vivo imaging tool, named dynamic fluorescence imaging (DyFI), for the simultaneous measurement of multiple vascular parameters including vascular density, perfusion rate, and permeability using spatiotemporal profiles of indocyanine green. Using DyFI in a tumor xenograft model, we quantitatively measured multiple vascular parameters in tumors and normal tissues with high spatial resolution. The multimodality of this method allowed us to find negative spatial correlations between perfusion and permeability. Moreover, DyFI was effective for revealing the early effects of an anti-angiogenic drug. We suggest that DyFI could be a useful tool for the preclinical development of anti-angiogenic drugs.

  11. Sentinel lymph node imaging by a fluorescently labeled DNA tetrahedron.

    Science.gov (United States)

    Kim, Kyoung-Ran; Lee, Yong-Deok; Lee, Taemin; Kim, Byeong-Su; Kim, Sehoon; Ahn, Dae-Ro

    2013-07-01

    Sentinel lymph nodes (SLNs) are the first lymph nodes which cancer cells reach after traveling through lymphatic vessels from the primary tumor. Evaluating the nodal status is crucial in accurate staging of human cancers and accordingly determines prognosis and the most appropriate treatment. The commonly used methods for SLN identification in clinics are based on employment of a colloid of radionuclide or injection of a small dye. Although these methods have certainly contributed to improve surgical practice, new imaging materials are still required to overcome drawbacks of the techniques such as inconvenience of handling radioactive materials and short retention time of small dyes in SLNs. Here, we prepare a fluorescence-labeled DNA tetrahedron and perform SLN imaging by using the DNA nanoconstruct. With a successful identification of SLNs by the DNA nanoconstruct, we suggest that DNA tetrahedron hold great promises for clinical applications.

  12. Non-enhanced magnetic resonance imaging of unruptured intracranial aneurysms at 7 Tesla: Comparison with digital subtraction angiography

    Energy Technology Data Exchange (ETDEWEB)

    Wrede, Karsten H.; Chen, Bixia [University Duisburg-Essen, Erwin L. Hahn Institute for Magnetic Resonance Imaging, Essen (Germany); University Hospital Essen, Department of Neurosurgery, Essen (Germany); Matsushige, Toshinori [University Duisburg-Essen, Erwin L. Hahn Institute for Magnetic Resonance Imaging, Essen (Germany); University Hospital Essen, Department of Neurosurgery, Essen (Germany); Hiroshima University, Department of Neurosurgery, Graduate School of Biomedical and Health Sciences, Hiroshima (Japan); Goericke, Sophia L.; Umutlu, Lale; Forsting, Michael [University Hospital Essen, Department of Diagnostic and Interventional Radiology and Neuroradiology, Essen (Germany); Quick, Harald H. [University Duisburg-Essen, Erwin L. Hahn Institute for Magnetic Resonance Imaging, Essen (Germany); University Hospital Essen, High Field and Hybrid MR Imaging, Essen (Germany); Ladd, Mark E. [University Duisburg-Essen, Erwin L. Hahn Institute for Magnetic Resonance Imaging, Essen (Germany); University Hospital Essen, Department of Diagnostic and Interventional Radiology and Neuroradiology, Essen (Germany); German Cancer Research Center (DKFZ), Division of Medical Physics in Radiology (E020), Heidelberg (Germany); Johst, Soeren [University Duisburg-Essen, Erwin L. Hahn Institute for Magnetic Resonance Imaging, Essen (Germany); Sure, Ulrich [University Hospital Essen, Department of Neurosurgery, Essen (Germany); Schlamann, Marc [University Hospital Essen, Department of Diagnostic and Interventional Radiology and Neuroradiology, Essen (Germany); University Hospital Giessen, Department of Neuroradiology, Giessen (Germany)

    2017-01-15

    To prospectively evaluate non-contrast-enhanced 7-Tesla (T) MRA for delineation of unruptured intracranial aneurysms (UIAs) in comparison with DSA. Forty patients with single or multiple UIAs were enrolled in this IRB-approved trial. Sequences acquired at 7 T were TOF MRA and non-contrast-enhanced MPRAGE. All patients additionally underwent 3D rotational DSA. Two neuroradiologists individually analysed the following aneurysm and image features on a five-point scale in 2D and 3D image reconstructions: delineation of parent vessel, dome and neck; overall image quality; presence of artefacts. Interobserver accordance was assessed by the kappa coefficient. A total of 64 UIAs were detected in DSA and in all 2D and 3D MRA image reconstructions. Ratings showed comparable results for DSA and 7-T MRA when considering all image reconstructions. Highest ratings for individual image reconstructions were given for 2D MPRAGE and 3D TOF MRA. Interobserver accordance was almost perfect for the majority of ratings. This study demonstrates excellent delineation of UIAs using 7-T MRA within a clinical setting comparable to the gold standard, DSA. The combination of 7-T non-enhanced MPRAGE and TOF MRA for assessment of untreated UIAs is a promising clinical application of ultra-high-field MRA. (orig.)

  13. IRDye78 Conjugates for Near-Infrared Fluorescence Imaging

    Directory of Open Access Journals (Sweden)

    Atif Zaheer

    2002-10-01

    Full Text Available The detection of human malignancies by near-infrared (NIR fluorescence will require the conjugation of cancer-specific ligands to NIR fluorophores that have optimal photoproperties and pharmacokinetics. IRDye78, a tetra-sulfonated heptamethine indocyanine NIR fluorophore, meets most of the criteria for an in vivo imaging agent, and is available as an N-hydroxysuccinimide ester for conjugation to low-molecular-weight ligands. However, IRDye78 has a high charge-to-mass ratio, complicating purification of conjugates. It also has a potentially labile linkage between fluorophore and ligand. We have developed an ion-pairing purification strategy for IRDye78 that can be performed with a standard C18 column under neutral conditions, thus preserving the stability of fluorophore, ligand, and conjugate. By employing parallel evaporative light scatter and absorbance detectors, all reactants and products are identified, and conjugate purity is maximized. We describe reversible and irreversible conversions of IRDye78 that can occur during sample purification, and describe methods for preserving conjugate stability. Using seven ligands, spanning several classes of small molecules and peptides (neutral, charged, and/or hydrophobic, we illustrate the robustness of these methods, and confirm that IRDye78 conjugates so purified retain bioactivity and permit NIR fluorescence imaging of specific targets.

  14. A Quantitative Method for Microtubule Analysis in Fluorescence Images.

    Science.gov (United States)

    Lan, Xiaodong; Li, Lingfei; Hu, Jiongyu; Zhang, Qiong; Dang, Yongming; Huang, Yuesheng

    2015-12-01

    Microtubule analysis is of significant value for a better understanding of normal and pathological cellular processes. Although immunofluorescence microscopic techniques have proven useful in the study of microtubules, comparative results commonly rely on a descriptive and subjective visual analysis. We developed an objective and quantitative method based on image processing and analysis of fluorescently labeled microtubular patterns in cultured cells. We used a multi-parameter approach by analyzing four quantifiable characteristics to compose our quantitative feature set. Then we interpreted specific changes in the parameters and revealed the contribution of each feature set using principal component analysis. In addition, we verified that different treatment groups could be clearly discriminated using principal components of the multi-parameter model. High predictive accuracy of four commonly used multi-classification methods confirmed our method. These results demonstrated the effectiveness and efficiency of our method in the analysis of microtubules in fluorescence images. Application of the analytical methods presented here provides information concerning the organization and modification of microtubules, and could aid in the further understanding of structural and functional aspects of microtubules under normal and pathological conditions.

  15. Fluorescence lifetime imaging of membrane lipid order with a ratiometric fluorescent probe.

    Science.gov (United States)

    Kilin, Vasyl; Glushonkov, Oleksandr; Herdly, Lucas; Klymchenko, Andrey; Richert, Ludovic; Mely, Yves

    2015-05-19

    To monitor the lateral segregation of lipids into liquid-ordered (Lo) and -disordered (Ld) phases in lipid membranes, environment-sensitive dyes that partition in both phases but stain them differently have been developed. Of particular interest is the dual-color F2N12S probe, which can discriminate the two phases through the ratio of its two emission bands. These bands are associated with the normal (N(∗)) and tautomer (T(∗)) excited-state species that result from an excited-state intramolecular proton transfer. In this work, we investigated the potency of the time-resolved fluorescence parameters of F2N12S to discriminate lipid phases in model and cell membranes. Both the long and mean lifetime values of the T(∗) form of F2N12S were found to differ by twofold between Ld and Lo phases as a result of the restriction in the relative motions of the two aromatic moieties of F2N12S imposed by the highly packed Lo phase. This differed from the changes in the ratio of the two emission bands between the two phases, which mainly resulted from the decreased hydration of the N(∗) form in the Lo phase. Importantly, the strong difference in lifetimes between the two phases was preserved when cholesterol was added to the Ld phase. The two phases could be imaged with high contrast by fluorescence lifetime imaging microscopy (FLIM) on giant unilamellar vesicles. FLIM images of F2N12S-labeled live HeLa cells confirmed that the plasma membrane was mainly in the Lo-like phase. Furthermore, the two phases were found to be homogeneously distributed all over the plasma membrane, indicating that they are highly mixed at the spatiotemporal resolution of the FLIM setup. Finally, FLIM could also be used to sensitively monitor the change in lipid phase upon cholesterol depletion and apoptosis.

  16. The value of magnetic resonance imaging for the detection of the bleeding source in non-traumatic intracerebral haemorrhages: a comparison with conventional digital subtraction angiography

    Energy Technology Data Exchange (ETDEWEB)

    Lummel, Nina; Lutz, Juergen; Brueckmann, Hartmut; Linn, Jennifer [University of Munich, Department of Neuroradiology, Munich (Germany)

    2012-07-15

    Conventional digital subtraction angiography (DSA) is currently regarded as the gold standard in detecting underlying vascular pathologies in patients with intracerebral haemorrhages (ICH). However, the use of magnetic resonance imaging (MRI) in the diagnostic workup of ICHs has considerably increased in recent years. Our aim was to evaluate the diagnostic accuracy and yield of MRI for the detection of the underlying aetiology in ICH patients. Sixty-seven consecutive patients with an acute ICH who underwent MRI (including magnetic resonance angiography (MRA) and DSA during their diagnostic workup) were included in the study. Magnetic resonance images were retrospectively analysed by two independent neuroradiologists to determine the localisation and cause of the ICH. DSA was used as a reference standard. In seven patients (10.4%), a DSA-positive vascular aetiology was present (one aneurysm, four arteriovenous malformations, one dural arteriovenous fistula and one vasculitis). All of these cases were correctly diagnosed by both readers on MRI. In addition, MRI revealed the following probable bleeding causes in 39 of the 60 DSA-negative patients: cerebral amyloid angiopathy (17), cavernoma (9), arterial hypertension (8), haemorrhagic transformation of an ischaemic infarction (3) and malignant brain tumour with secondary ICH (2). Performing MRI with MRA proved to be an accurate diagnostic tool in detecting vascular malformations in patients with ICH. In addition, MRI provided valuable information regarding DSA-negative ICH causes, and thus had a high diagnostic yield in ICH patients. (orig.)

  17. The background rate of false positives: Combining simulations of gravitational wave events with an unsupervised algorithm for transient identification in crowded image-subtracted data

    Science.gov (United States)

    Ackley, Kendall; Eikenberry, Stephen; Klimenko, Sergey; LIGO Collaboration

    2016-03-01

    We are now entering the era of multimessenger gravitational wave (GW) astronomy with the completion of the first observing run of Advanced LIGO. Multiwavelength electromagnetic (EM) emission is expected to accompany gravitational radiation from compact object binary mergers, such as those between neutron stars and stellar-mass black holes, where Advanced LIGO is most sensitive to their detection. Attempting to perform EM follow-up over the 10-100s deg2 error regions will be faced with many challenges, including the identification and removal of O (105) false positive transients that appear as a commotion of background events and as image artifacts in crowded image-subtracted fields. We present an update to our automated unsupervised algorithm including how our pipeline uses the existing coherent WaveBurst pipeline in an attempt to develop optimized EM follow-up schema. Our end-to-end pipeline combines simulated GW events with actual observational data from a number of ground-based optical observatories, including PTF, ROTSE, and DECam. Our performance is reported both in terms of the number of coincident false positives as well as the efficiency of recovery.

  18. Improved detection of fluorescently labeled microspheres and vessel architecture with an imaging cryomicrotome

    NARCIS (Netherlands)

    P. van Horssen; M. Siebes; I. Hoefer; J.A.E. Spaan; J.P.H.M. van den Wijngaard

    2010-01-01

    Due to spectral overlap, the number of fluorescent labels for imaging cryomicrotome detection was limited to 4. The aim of this study was to increase the separation of fluorescent labels. In the new imaging cryomicrotome, the sample is cut in slices of 40 mu m. Six images are taken for each cutting

  19. A modified phasor approach for analyzing time-gated fluorescence lifetime images

    NARCIS (Netherlands)

    Fereidouni, F.; Esposito, A.; Blab, G.; Gerritsen, H.C.

    2011-01-01

    Fluorescence lifetime imaging is a versatile tool that permits mapping the biochemical environment in the cell. Among various fluorescence lifetime imaging techniques, timecorrelated single photon counting and time-gating methods have been demonstrated to be very efficient and robust for the imaging

  20. Fluorescence imaging and chlorophyll fluorescence to evaluate the role of EDU in UV-B protection in cucumber

    Science.gov (United States)

    Sandhu, Ravinder K.; Kim, Moon S.; Krizek, Donald T.; Middleton, Elizabeth M.

    1997-07-01

    A fluorescence imaging system and chlorophyll fluorescence emissions were used to evaluate whether EDU, N-[2-(2-oxo-1- imidazolidinyl) ethyl]-N'-phenylurea, provided protection against ultraviolet-B (UV-B) irradiation (290 - 320 nm) in cucumber (Cucumis sativus L.) leaves. Plants were grown in growth chambers illuminated for 14 h per day with 400 W high pressure sodium and metal halide lamps. Photosynthetically active radiation (PAR) for 1 hr at the beginning and end of each cycle was provided at 270 micrometers ol m-2 s-1 PAR; during the other 12 hr of the photoperiod, the plants received 840 micrometers ol m-2 s-1 PAR. Beginning on the twelfth day, the plants were exposed to UV-B radiation (0.2 & 18.0 kJ m-2d-1) for 2 days at 8 h per day centered in the photoperiod. Rapidly acquired (less than 1 s), high spatial resolution (less than 1 mm2) images were obtained for whole adaxial leaf surfaces using a fluorescence imaging system. The steady-state fluorescence images were acquired in four spectral regions: blue (F450 nm), green (F550 nm), red (F680 nm), and far-red (F740 nm). Fluorescence emission spectra for leaf pigments extracted in dimethyl sulfoxide (DMSO) were obtained by excitation at 280 and 380 nm (280EX 300 - 530 nm; 380EX 400 - 800 nm). Both UV-B and EDU induced stress responses in cucumber leaves that altered the fluorescence emissions obtained from extracts. In the fluorescence images only UV-B induced stress responses were observed but this damage was detected before it was visually apparent. There was no evidence that EDU afforded protection against UV-B irradiation. Use of fluorescence imaging may provide an early stress detection capability for helping to assess damage to the photosynthetic apparatus of plants.

  1. Fluorescence Imaging of the Cytoskeleton in Plant Roots.

    Science.gov (United States)

    Dyachok, Julia; Paez-Garcia, Ana; Yoo, Cheol-Min; Palanichelvam, Karuppaiah; Blancaflor, Elison B

    2016-01-01

    During the past two decades the use of live cytoskeletal probes has increased dramatically due to the introduction of the green fluorescent protein. However, to make full use of these live cell reporters it is necessary to implement simple methods to maintain plant specimens in optimal growing conditions during imaging. To image the cytoskeleton in living Arabidopsis roots, we rely on a system involving coverslips coated with nutrient supplemented agar where the seeds are directly germinated. This coverslip system can be conveniently transferred to the stage of a confocal microscope with minimal disturbance to the growth of the seedling. For roots with a larger diameter such as Medicago truncatula, seeds are first germinated in moist paper, grown vertically in between plastic trays, and roots mounted on glass slides for confocal imaging. Parallel with our live cell imaging approaches, we routinely process fixed plant material via indirect immunofluorescence. For these methods we typically use non-embedded vibratome-sectioned and whole mount permeabilized root tissue. The clearly defined developmental regions of the root provide us with an elegant system to further understand the cytoskeletal basis of plant development.

  2. Nonnegative matrix factorization: a blind spectra separation method for in vivo fluorescent optical imaging.

    Science.gov (United States)

    Montcuquet, Anne-Sophie; Hervé, Lionel; Navarro, Fabrice; Dinten, Jean-Marc; Mars, Jérôme I

    2010-01-01

    Fluorescence imaging in diffusive media is an emerging imaging modality for medical applications that uses injected fluorescent markers that bind to specific targets, e.g., carcinoma. The region of interest is illuminated with near-IR light and the emitted back fluorescence is analyzed to localize the fluorescence sources. To investigate a thick medium, as the fluorescence signal decreases with the light travel distance, any disturbing signal, such as biological tissues intrinsic fluorescence (called autofluorescence) is a limiting factor. Several specific markers may also be simultaneously injected to bind to different molecules, and one may want to isolate each specific fluorescent signal from the others. To remove the unwanted fluorescence contributions or separate different specific markers, a spectroscopic approach is explored. The nonnegative matrix factorization (NMF) is the blind positive source separation method we chose. We run an original regularized NMF algorithm we developed on experimental data, and successfully obtain separated in vivo fluorescence spectra.

  3. Intraoperative near-infrared fluorescent imaging during robotic operations.

    Science.gov (United States)

    Macedo, Antonio Luiz de Vasconcellos; Schraibman, Vladimir

    2016-01-01

    The intraoperative identification of certain anatomical structures because they are small or visually occult may be challenging. The development of minimally invasive surgery brought additional difficulties to identify these structures due to the lack of complete tactile sensitivity. A number of different forms of intraoperative mapping have been tried. Recently, the near-infrared fluorescence imaging technology with indocyanine green has been added to robotic platforms. In addition, this technology has been tested in several types of operations, and has advantages such as safety, low cost and good results. Disadvantages are linked to contrast distribution in certain clinical scenarios. The intraoperative near-infrared fluorescent imaging is new and promising addition to robotic surgery. Several reports show the utility of this technology in several different procedures. The ideal dose, time and site for dye injection are not well defined. No high quality evidence-based comparative studies and long-term follow-up outcomes have been published so far. Initial results, however, are good and safe. RESUMO A identificação intraoperatória de certas estruturas anatômicas, por seu tamanho ou por elas serem ocultas à visão, pode ser desafiadora. O desenvolvimento da cirurgia minimamente invasiva trouxe dificuldades adicionais, pela falta da sensibilidade tátil completa. Diversas formas de detecção intraoperatória destas estruturas têm sido tentadas. Recentemente, a tecnologia de fluorescência infravermelha com verde de indocianina foi associada às plataformas robóticas. Além disso, essa tecnologia tem sido testada em uma variedade de cirurgias, e suas vantagens parecem estar ligadas a baixo custo, segurança e bons resultados. As desvantagens estão associadas à má distribuição do contraste em determinados cenários. A imagem intraoperatória por fluorescência infravermelha é uma nova e promissora adição à cirurgia robótica. Diversas séries mostram

  4. High resolution fluorescent bio-imaging with electron beam excitation.

    Science.gov (United States)

    Kawata, Yoshimasa; Nawa, Yasunori; Inami, Wataru

    2014-11-01

    We have developed electron beam excitation assisted (EXA) optical microscope[1-3], and demonstrated its resolution higher than 50 nm. In the microscope, a light source in a few nanometers size is excited by focused electron beam in a luminescent film. The microscope makes it possible to observe dynamic behavior of living biological specimens in various surroundings, such as air or liquids. Scan speed of the nanometric light source is faster than that in conventional near-field scanning optical microscopes. The microscope enables to observe optical constants such as absorption, refractive index, polarization, and their dynamic behavior on a nanometric scale. The microscope opens new microscopy applications in nano-technology and nano-science.Figure 1(a) shows schematic diagram of the proposed EXA microscope. An electron beam is focused on a luminescent film. A specimen is put on the luminescent film directly. The inset in Fig. 1(a) shows magnified image of the luminescent film and the specimen. Nanometric light source is excited in the luminescent film by the focused electron beam. The nanometric light source illuminates the specimen, and the scattered or transmitted radiation is detected with a photomultiplier tube (PMT). The light source is scanned by scanning of the focused electron beam in order to construct on image. Figure 1(b) shows a luminescence image of the cells acquired with the EXA microscope, and Fig. 1(c) shows a phase contrast microscope image. Cells were observed in culture solution without any treatments, such as fixation and drying. The shape of each cell was clearly recognized and some bright spots were observed in cells. We believe that the bright spots indicated with arrows were auto-fluorescence of intracellular granules and light- grey regions were auto-fluorescence of cell membranes. It is clearly demonstrated that the EXA microscope is useful tool for observation of living biological cells in physiological conditions.jmicro;63/suppl_1/i

  5. Robust overlay schemes for the fusion of fluorescence and color channels in biological imaging.

    Science.gov (United States)

    Glatz, Jürgen; Symvoulidis, Panagiotis; Garcia-Allende, P Beatriz; Ntziachristos, Vasilis

    2014-04-01

    Molecular fluorescence imaging is a commonly used method in various biomedical fields and is undergoing rapid translation toward clinical applications. Color images are commonly superimposed with fluorescence measurements to provide orientation, anatomical information, and molecular tissue properties in a single image. New adaptive methods that produce a more robust composite image than conventional lime green alpha blending are presented and demonstrated herein. Moreover, visualization through temporal changes is showcased as an alternative for real-time imaging systems.

  6. In Vivo Follow-up of Brain Tumor Growth via Bioluminescence Imaging and Fluorescence Tomography

    Directory of Open Access Journals (Sweden)

    Coralie Genevois

    2016-10-01

    Full Text Available Reporter gene-based strategies are widely used in experimental oncology. Bioluminescence imaging (BLI using the firefly luciferase (Fluc as a reporter gene and d-luciferin as a substrate is currently the most widely employed technique. The present paper compares the performances of BLI imaging with fluorescence imaging using the near infrared fluorescent protein (iRFP to monitor brain tumor growth in mice. Fluorescence imaging includes fluorescence reflectance imaging (FRI, fluorescence diffuse optical tomography (fDOT, and fluorescence molecular Imaging (FMT®. A U87 cell line was genetically modified for constitutive expression of both the encoding Fluc and iRFP reporter genes and assayed for cell, subcutaneous tumor and brain tumor imaging. On cultured cells, BLI was more sensitive than FRI; in vivo, tumors were first detected by BLI. Fluorescence of iRFP provided convenient tools such as flux cytometry, direct detection of the fluorescent protein on histological slices, and fluorescent tomography that allowed for 3D localization and absolute quantification of the fluorescent signal in brain tumors.

  7. Enhanced live cell imaging via photonic crystal enhanced fluorescence microscopy.

    Science.gov (United States)

    Chen, Weili; Long, Kenneth D; Yu, Hojeong; Tan, Yafang; Choi, Ji Sun; Harley, Brendan A; Cunningham, Brian T

    2014-11-21

    We demonstrate photonic crystal enhanced fluorescence (PCEF) microscopy as a surface-specific fluorescence imaging technique to study the adhesion of live cells by visualizing variations in cell-substrate gap distance. This approach utilizes a photonic crystal surface incorporated into a standard microscope slide as the substrate for cell adhesion, and a microscope integrated with a custom illumination source as the detection instrument. When illuminated with a monochromatic light source, angle-specific optical resonances supported by the photonic crystal enable efficient excitation of surface-confined and amplified electromagnetic fields when excited at an on-resonance condition, while no field enhancement occurs when the same photonic crystal is illuminated in an off-resonance state. By mapping the fluorescence enhancement factor for fluorophore-tagged cellular components between on- and off-resonance states and comparing the results to numerical calculations, the vertical distance of labelled cellular components from the photonic crystal substrate can be estimated, providing critical and quantitative information regarding the spatial distribution of the specific components of cells attaching to a surface. As an initial demonstration of the concept, 3T3 fibroblast cells were grown on fibronectin-coated photonic crystals with fluorophore-labelled plasma membrane or nucleus. We demonstrate that PCEF microscopy is capable of providing information about the spatial distribution of cell-surface interactions at the single-cell level that is not available from other existing forms of microscopy, and that the approach is amenable to large fields of view, without the need for coupling prisms, coupling fluids, or special microscope objectives.

  8. Fluorescent Cy5 silica nanoparticles for cancer cell imaging

    Science.gov (United States)

    O'Connell, Claire; Nooney, Robert I.; Glynn, MacDara; Ducree, Jens; McDonagh, Colette

    2015-08-01

    Cancer is a leading cause of death worldwide, with metastasis responsible for the majority of cancer-related deaths. Circulating tumour cells (CTCs) play a central role in metastasis. Fluorescent silica particles (NPs), of diameter ~50 nm which contain a large concentration of Cy5 dye molecules and are extremely bright, have been developed to detect these rare CTCs. Due to this brightness, the particles have superior performance compared to single Cy5 dye molecule labels, for detecting cancer cells. Fluorescence measurements show that the NPs are almost 100 times brighter than the free dye. They do not photo bleach as readily and, due to the biocompatible silica surface, they can be chemically modified, layer-by-layer, in order to bind to cells. The choice of these chemical layers, in particular the NP to antibody linker, along with the incubation period and type of media used in the incubation, has a strong influence on the specific binding abilities of the NPs. In this work, NPs have been shown to selectively bind to the MCF-7 cell line by targeting epithelial cellular adhesion molecule (EpCAM) present on the MCF-7 cell membrane by conjugating anti-EpCAM antibody to the NP surface. Results have shown a high signal to noise ratio for this cell line in comparison to a HeLa control line. NP attachment to cells was verified qualitatively with the use of fluorescence microscopy and quantitatively using image analysis methods. Once the system has been optimised, other dyes will be doped into the silica NPs and their use in multiplexing will be investigated.

  9. Analysis of human aorta using fluorescence lifetime imaging microscopy (FLIM)

    Science.gov (United States)

    Vieira-Damiani, Gislaine; Adur, J.; Ferro, D. P.; Adam, R. L.; Pelegati, V.; Thomáz, A.; Cesar, C. L.; Metze, K.

    2012-03-01

    The use of photonics has improved our understanding of biologic phenomena. For the study of the normal and pathologic architecture of the aorta the use of Two-Photon Excited Fluorescence (TPEF) and Second Harmonic Generation showed interesting details of morphologic changes of the elastin-collagen architecture during aging or development of hypertension in previous studies. In this investigation we tried to apply fluorescence lifetime imaging (FLIM) for the morphologic analysis of human aortas. The aim of our study was to use FLIM in non-stained formalin-fixed and paraffin-embedded samples of the aorta ascendants in hypertensive and normotensive patients of various ages, examining two different topographical regions. The FLIM-spectra of collagen and elastic fibers were clearly distinguishable, thus permitting an exact analysis of unstained material on the microscopic level. Moreover the FLIM spectrum of elastic fibers revealed variations between individual cases, which indicate modifications on a molecular level and might be related to FLIM age or diseases states and reflect modifications on a molecular level.

  10. Application of hyperspectral fluorescence lifetime imaging to tissue autofluorescence: arthritis

    Science.gov (United States)

    Talbot, C. B.; Benninger, R. K. P.; de Beule, P.; Requejo-Isidro, J.; Elson, D. S.; Dunsby, C.; Munro, I.; Neil, M. A.; Sandison, A.; Sofat, N.; Nagase, H.; French, P. M. W.; Lever, M. J.

    2005-08-01

    Tissue contains many natural fluorophores and therefore by exploiting autofluorescence, we can obtain information from tissue with less interference than conventional histological techniques. However, conventional intensity imaging is prone to artifacts since it is an absolute measurement. Fluorescence lifetime and spectral measurements are relative measurements and therefore allow for better measurements. We have applied FLIM and hyperspectral FLIM to the study of articular cartilage and its disease arthritis. We have analyzed normal human articular cartilage and cartilage which was in the early stages of disease. In this case, it was found that FLIM was able to detect changes in the diseased tissue that were not detectable with the conventional diagnosis. Specifically, the fluorescence lifetimes (FL) of the cells were different between the two samples. We have also applied hyperspectral FLIM to degraded cartilage through treatment with interleukin-1. In this case, it was found that there was a shift in the emission spectrum with treatment and that the lifetime had also increased. We also showed that there was greater contrast between the cells and the extracellular matrix (ECM) at longer wavelengths.

  11. Time-of-flight MR angiography at 3 T versus digital subtraction angiography in the imaging follow-up of 51 intracranial aneurysms treated with coils

    Energy Technology Data Exchange (ETDEWEB)

    Ferre, Jean-Christophe [Department of Neuroradiology, Hopital Pontchaillou, University Hospital Rennes, 2 rue Henri Le Guilloux, 35000 Rennes (France)], E-mail: jean-christophe.ferre@chu-rennes.fr; Carsin-Nicol, Beatrice [Department of Neuroradiology, Hopital Pontchaillou, University Hospital Rennes, 2 rue Henri Le Guilloux, 35000 Rennes (France); Morandi, Xavier [Department of Neurosurgery, Hopital Pontchaillou, University Hospital Rennes, 2 rue Henri Le Guilloux, 35000 Rennes (France); Carsin, Michel [Department of Neuroradiology, Hopital Pontchaillou, University Hospital Rennes, 2 rue Henri Le Guilloux, 35000 Rennes (France); Kersaint-Gilly, Axel de [Department of Neuroradiology, Hopital Laennec, University Hospital Nantes, Boulevard Jacques Monod, 44800 Saint-Herblain (France); Gauvrit, Jean-Yves [Department of Neuroradiology, Hopital Pontchaillou, University Hospital Rennes, 2 rue Henri Le Guilloux, 35000 Rennes (France); Desal, Hubert-Armand [Department of Neuroradiology, Hopital Laennec, University Hospital Nantes, Boulevard Jacques Monod, 44800 Saint-Herblain (France)

    2009-12-15

    Objective: To compare 3D time-of-flight MR angiography (TOF-MRA) at 3 Tesla (3 T) with digital subtraction angiography (DSA) for the evaluation of intracranial aneurysm occlusion after endovascular coiling. Methods: In a prospective study, 51 consecutive patients (25 females, 26 males; median age, 51 years) with 51 saccular aneurysms treated with endovascular coiling underwent simultaneous DSA and 3 T TOF-MRA at follow-up. DSA and TOF-MRA images were analyzed independently by two senior neuroradiologists. Findings were assigned to 1 of 3 categories in the Raymond classification: complete obliteration, residual neck or residual aneurysm. Agreement between observers and techniques was evaluated using {kappa} statistics. Results: DSA images were not interpretable for one patient. Interobserver agreement was determined as excellent for DSA ({kappa} = 0.86) and TOF-MRA ({kappa} = 0.80). After reaching a consensus, DSA follow-up showed 26 (51%) complete obliterations, 20 (39%) residual necks and 4 (8%) residual aneurysms. TOF-MRA showed 23 (45%) complete obliterations, 22 (43%) residual necks and 6 (12%) residual aneurysms. Comparison between TOF-MRA and DSA showed excellent agreement between the techniques ({kappa} = 0.86). In the four cases that were misclassified, TOF-MRA findings were assigned to a higher class than for DSA. Conclusion: TOF-MRA at 3 T is at least as efficient as DSA for the evaluation of intracranial aneurysm occlusion after endovascular treatment with detachable coils. We suggest that TOF-MRA at 3 T might be used as the primary method for imaging follow-up of coiled intracranial aneurysms.

  12. Near-infrared (NIR) fluorescence imaging of head and neck squamous cell carcinoma for fluorescence-guided surgery (Conference Presentation)

    Science.gov (United States)

    Moore, Lindsay; Warram, Jason M.; de Boer, Esther; Carroll, William R.; Morlandt, Anthony; Withrow, Kirk P.; Rosenthal, Eben L.

    2016-03-01

    During fluorescence-guided surgery, a cancer-specific optical probe is injected and visualized using a compatible device intraoperatively to provide visual contrast between diseased and normal tissues to maximize resection of cancer and minimize the resection of precious adjacent normal tissues. Six patients with squamous cell carcinomas of the head and neck region (oral cavity (n=4) or cutaneous (n=2)) were injected with an EGFR-targeting antibody (Cetuximab) conjugated to a near-infrared (NIR) fluorescent dye (IRDye800) 3, 4, or 7 days prior to surgical resection of the cancer. Each patient's tumor was then imaged using a commercially available, open-field NIR fluorescence imaging device each day prior to surgery, intraoperatively, and post-operatively. The mean fluorescence intensity (MFI) of the tumor was calculated for each specimen at each imaging time point. Adjacent normal tissue served as an internal anatomic control for each patient to establish a patient-matched "background" fluorescence. Resected tissues were also imaged using a closed-field NIR imaging device. Tumor to background ratios (TBRs) were calculated for each patient using both devices. Fluorescence histology was correlated with traditional pathology assessment to verify the specificity of antibody-dye conjugate binding. Peak TBRs using the open-field device ranged from 2.2 to 11.3, with an average TBR of 4.9. Peak TBRs were achieved between days 1 and 4. This study demonstrated that a commercially available NIR imaging device suited for intraoperative and clinical use can successfully be used with a fluorescently-labeled dye to delineate between diseased and normal tissue in this single cohort human study, illuminated the potential for its use in fluoresence-guided surgery.

  13. Single aflatoxin contaminated corn kernel analysis with fluorescence hyperspectral image

    Science.gov (United States)

    Yao, Haibo; Hruska, Zuzana; Kincaid, Russell; Ononye, Ambrose; Brown, Robert L.; Cleveland, Thomas E.

    2010-04-01

    Aflatoxins are toxic secondary metabolites of the fungi Aspergillus flavus and Aspergillus parasiticus, among others. Aflatoxin contaminated corn is toxic to domestic animals when ingested in feed and is a known carcinogen associated with liver and lung cancer in humans. Consequently, aflatoxin levels in food and feed are regulated by the Food and Drug Administration (FDA) in the US, allowing 20 ppb (parts per billion) limits in food and 100 ppb in feed for interstate commerce. Currently, aflatoxin detection and quantification methods are based on analytical tests including thin-layer chromatography (TCL) and high performance liquid chromatography (HPLC). These analytical tests require the destruction of samples, and are costly and time consuming. Thus, the ability to detect aflatoxin in a rapid, nondestructive way is crucial to the grain industry, particularly to corn industry. Hyperspectral imaging technology offers a non-invasive approach toward screening for food safety inspection and quality control based on its spectral signature. The focus of this paper is to classify aflatoxin contaminated single corn kernels using fluorescence hyperspectral imagery. Field inoculated corn kernels were used in the study. Contaminated and control kernels under long wavelength ultraviolet excitation were imaged using a visible near-infrared (VNIR) hyperspectral camera. The imaged kernels were chemically analyzed to provide reference information for image analysis. This paper describes a procedure to process corn kernels located in different images for statistical training and classification. Two classification algorithms, Maximum Likelihood and Binary Encoding, were used to classify each corn kernel into "control" or "contaminated" through pixel classification. The Binary Encoding approach had a slightly better performance with accuracy equals to 87% or 88% when 20 ppb or 100 ppb was used as classification threshold, respectively.

  14. A 3-D fluorescence imaging system incorporating structured illumination technology

    Science.gov (United States)

    Antos, L.; Emord, P.; Luquette, B.; McGee, B.; Nguyen, D.; Phipps, A.; Phillips, D.; Helguera, M.

    2010-02-01

    A currently available 2-D high-resolution, optical molecular imaging system was modified by the addition of a structured illumination source, OptigridTM, to investigate the feasibility of providing depth resolution along the optical axis. The modification involved the insertion of the OptigridTM and a lens in the path between the light source and the image plane, as well as control and signal processing software. Projection of the OptigridTM onto the imaging surface at an angle, was resolved applying the Scheimpflug principle. The illumination system implements modulation of the light source and provides a framework for capturing depth resolved mages. The system is capable of in-focus projection of the OptigridTM at different spatial frequencies, and supports the use of different lenses. A calibration process was developed for the system to achieve consistent phase shifts of the OptigridTM. Post-processing extracted depth information using depth modulation analysis using a phantom block with fluorescent sheets at different depths. An important aspect of this effort was that it was carried out by a multidisciplinary team of engineering and science students as part of a capstone senior design program. The disciplines represented are mechanical engineering, electrical engineering and imaging science. The project was sponsored by a financial grant from New York State with equipment support from two industrial concerns. The students were provided with a basic imaging concept and charged with developing, implementing, testing and validating a feasible proof-of-concept prototype system that was returned to the originator of the concept for further evaluation and characterization.

  15. Monomeric red fluorescent protein variants used for imaging studies in different species

    NARCIS (Netherlands)

    Mueller-Taubenberger, Annette; Vos, Michel J.; Boettger, Angelika; Lasi, Margherita; Lai, Frank P. L.; Fischer, Markus; Rottner, Klemens

    2006-01-01

    Fluorescent proteins have proven to be excellent tools for live-cell imaging studies. In addition to green fluorescent protein (GFP) and its variants, recent progress was achieved in the development of monomeric red fluorescent proteins (mRFPs) that show improved properties in respect to maturation

  16. Applying two-photon excitation fluorescence lifetime imaging microscopy to study photosynthesis in plant leaves

    NARCIS (Netherlands)

    Broess, K.; Borst, J.W.; Amerongen, van H.

    2009-01-01

    This study investigates to which extent two-photon excitation (TPE) fluorescence lifetime imaging microscopy can be applied to study picosecond fluorescence kinetics of individual chloroplasts in leaves. Using femtosecond 860 nm excitation pulses, fluorescence lifetimes can be measured in leaves of

  17. Inspection of fecal contamination on strawberries using fluorescence imaging

    Science.gov (United States)

    Chuang, Yung-Kun; Yang, Chun-Chieh; Kim, Moon S.; Delwiche, Stephen R.; Lo, Y. Martin; Chen, Suming; Chan, Diane E.

    2013-05-01

    Fecal contamination of produce is a food safety issue associated with pathogens such as Escherichia coli that can easily pollute agricultural products via animal and human fecal matters. Outbreaks of foodborne illnesses associated with consuming raw fruits and vegetables have occurred more frequently in recent years in the United States. Among fruits, strawberry is one high-potential vector of fecal contamination and foodborne illnesses since the fruit is often consumed raw and with minimal processing. In the present study, line-scan LED-induced fluorescence imaging techniques were applied for inspection of fecal material on strawberries, and the spectral characteristics and specific wavebands of strawberries were determined by detection algorithms. The results would improve the safety and quality of produce consumed by the public.

  18. Aptamer-assembled nanomaterials for fluorescent sensing and imaging

    Directory of Open Access Journals (Sweden)

    Lu Danqing

    2017-01-01

    Full Text Available Aptamers, which are selected in vitro by a technology known as the systematic evolution of ligands by exponential enrichment (SELEX, represent a crucial recognition element in molecular sensing. With advantages such as good biocompatibility, facile functionalization, and special optical and physical properties, various nanomaterials can protect aptamers from enzymatic degradation and nonspecific binding in living systems and thus provide a preeminent platform for biochemical applications. Coupling aptamers with various nanomaterials offers many opportunities for developing highly sensitive and selective sensing systems. Here, we focus on the recent applications of aptamer-assembled nanomaterials in fluorescent sensing and imaging. Different types of nanomaterials are examined along with their advantages and disadvantages. Finally, we look toward the future of aptamer-assembled nanomaterials.

  19. Aptamer-assembled nanomaterials for fluorescent sensing and imaging

    Science.gov (United States)

    Lu, Danqing; He, Lei; Zhang, Ge; Lv, Aiping; Wang, Ruowen; Zhang, Xiaobing; Tan, Weihong

    2017-01-01

    Aptamers, which are selected in vitro by a technology known as the systematic evolution of ligands by exponential enrichment (SELEX), represent a crucial recognition element in molecular sensing. With advantages such as good biocompatibility, facile functionalization, and special optical and physical properties, various nanomaterials can protect aptamers from enzymatic degradation and nonspecific binding in living systems and thus provide a preeminent platform for biochemical applications. Coupling aptamers with various nanomaterials offers many opportunities for developing highly sensitive and selective sensing systems. Here, we focus on the recent applications of aptamer-assembled nanomaterials in fluorescent sensing and imaging. Different types of nanomaterials are examined along with their advantages and disadvantages. Finally, we look toward the future of aptamer-assembled nanomaterials.

  20. Flow modification in canine intracranial aneurysm model by an asymmetric stent: studies using digital subtraction angiography (DSA) and image-based computational fluid dynamics (CFD) analyses

    Science.gov (United States)

    Hoi, Yiemeng; Ionita, Ciprian N.; Tranquebar, Rekha V.; Hoffmann, Kenneth R.; Woodward, Scott H.; Taulbee, Dale B.; Meng, Hui; Rudin, Stephen

    2006-03-01

    An asymmetric stent with low porosity patch across the intracranial aneurysm neck and high porosity elsewhere is designed to modify the flow to result in thrombogenesis and occlusion of the aneurysm and yet to reduce the possibility of also occluding adjacent perforator vessels. The purposes of this study are to evaluate the flow field induced by an asymmetric stent using both numerical and digital subtraction angiography (DSA) methods and to quantify the flow dynamics of an asymmetric stent in an in vivo aneurysm model. We created a vein-pouch aneurysm model on the canine carotid artery. An asymmetric stent was implanted at the aneurysm, with 25% porosity across the aneurysm neck and 80% porosity elsewhere. The aneurysm geometry, before and after stent implantation, was acquired using cone beam CT and reconstructed for computational fluid dynamics (CFD) analysis. Both steady-state and pulsatile flow conditions using the measured waveforms from the aneurysm model were studied. To reduce computational costs, we modeled the asymmetric stent effect by specifying a pressure drop over the layer across the aneurysm orifice where the low porosity patch was located. From the CFD results, we found the asymmetric stent reduced the inflow into the aneurysm by 51%, and appeared to create a stasis-like environment which favors thrombus formation. The DSA sequences also showed substantial flow reduction into the aneurysm. Asymmetric stents may be a viable image guided intervention for treating intracranial aneurysms with desired flow modification features.

  1. Dual-modality, fluorescent, PLGA encapsulated bismuth nanoparticles for molecular and cellular fluorescence imaging and computed tomography

    Science.gov (United States)

    Swy, Eric R.; Schwartz-Duval, Aaron S.; Shuboni, Dorela D.; Latourette, Matthew T.; Mallet, Christiane L.; Parys, Maciej; Cormode, David P.; Shapiro, Erik M.

    2014-10-01

    Reports of molecular and cellular imaging using computed tomography (CT) are rapidly increasing. Many of these reports use gold nanoparticles. Bismuth has similar CT contrast properties to gold while being approximately 1000-fold less expensive. Herein we report the design, fabrication, characterization, and CT and fluorescence imaging properties of a novel, dual modality, fluorescent, polymer encapsulated bismuth nanoparticle construct for computed tomography and fluorescence imaging. We also report on cellular internalization and preliminary in vitro and in vivo toxicity effects of these constructs. 40 nm bismuth(0) nanocrystals were synthesized and encapsulated within 120 nm Poly(dl-lactic-co-glycolic acid) (PLGA) nanoparticles by oil-in-water emulsion methodologies. Coumarin-6 was co-encapsulated to impart fluorescence. High encapsulation efficiency was achieved ~70% bismuth w/w. Particles were shown to internalize within cells following incubation in culture. Bismuth nanocrystals and PLGA encapsulated bismuth nanoparticles exhibited >90% and >70% degradation, respectively, within 24 hours in acidic, lysosomal environment mimicking media and both remained nearly 100% stable in cytosolic/extracellular fluid mimicking media. μCT and clinical CT imaging was performed at multiple X-ray tube voltages to measure concentration dependent attenuation rates as well as to establish the ability to detect the nanoparticles in an ex vivo biological sample. Dual fluorescence and CT imaging is demonstrated as well. In vivo toxicity studies in rats revealed neither clinically apparent side effects nor major alterations in serum chemistry and hematology parameters. Calculations on minimal detection requirements for in vivo targeted imaging using these nanoparticles are presented. Indeed, our results indicate that these nanoparticles may serve as a platform for sensitive and specific targeted molecular CT and fluorescence imaging.Reports of molecular and cellular imaging using

  2. Integrated Lymphography using Fluorescence Imaging and Magnetic Resonance Imaging in Intact Mice

    Directory of Open Access Journals (Sweden)

    Yusuke Inoue

    2011-09-01

    Full Text Available We assessed lymph drainage in living mice by an integrated imaging method using fluorescence imaging (FLI and magnetic resonance imaging (MRI. Mice were subcutaneously injected with quantum dots and gadofluorine 8 into the right rear footpad. They were fixed on a transparent flat plate and underwent FLI and MRI successively. Small markers were attached to the mouse surface for spatial coregistration, and image fusion of FLIs and MRIs was performed. Two-dimensional fluorescence reflectance imaging was used for FLI. FLI and MRI provided generally consistent results and demonstrated lymphatic flow to the popliteal, sacral, and iliac lymph nodes in most mice and to the renal, inguinal, and lumbar-aortic lymph nodes in some mice. On the fusion images, the locations of the lymph nodes in the mouse trunk were in good agreement between FLI and MRI, indicating successful spatial registration even for the deep structures. The popliteal node tended to be visualized a little farther caudally in FLI than in MRI, presumably because the overlying tissues were thicker in the cranial portion. Integrated FLI/MRI lymphography with image fusion appears to be a useful tool for analysis of the murine lymphatic system.

  3. Segmentation of fluorescence microscopy cell images using unsupervised mining.

    Science.gov (United States)

    Du, Xian; Dua, Sumeet

    2010-05-28

    The accurate measurement of cell and nuclei contours are critical for the sensitive and specific detection of changes in normal cells in several medical informatics disciplines. Within microscopy, this task is facilitated using fluorescence cell stains, and segmentation is often the first step in such approaches. Due to the complex nature of cell issues and problems inherent to microscopy, unsupervised mining approaches of clustering can be incorporated in the segmentation of cells. In this study, we have developed and evaluated the performance of multiple unsupervised data mining techniques in cell image segmentation. We adapt four distinctive, yet complementary, methods for unsupervised learning, including those based on k-means clustering, EM, Otsu's threshold, and GMAC. Validation measures are defined, and the performance of the techniques is evaluated both quantitatively and qualitatively using synthetic and recently published real data. Experimental results demonstrate that k-means, Otsu's threshold, and GMAC perform similarly, and have more precise segmentation results than EM. We report that EM has higher recall values and lower precision results from under-segmentation due to its Gaussian model assumption. We also demonstrate that these methods need spatial information to segment complex real cell images with a high degree of efficacy, as expected in many medical informatics applications.

  4. In vivo imaging of Lactococcus lactis, Lactobacillus plantarum and Escherichiacoli expressing infrared fluorescent protein in mice

    OpenAIRE

    Berlec, Aleš; Štrukelj, Borut; Završnik, Janja; Turk, Boris; Butinar, Miha

    2016-01-01

    Background In vivo imaging of orally administered lactic acid bacteria (LAB) and commensal bacteria in mice is shown to provide information on the spatial and temporal distribution of bacteria in the gastrointestinal tract. The bacteria can be detected and monitored using bioluminescence or near-infrared fluorescence. Results Fluorescence imaging of bacteria was established by expressing the infrared fluorescent protein IRFP713 in Lactococcus lactis, Lactobacillus plantarum and Escherichia co...

  5. In vivo multiphoton tomography and fluorescence lifetime imaging of human brain tumor tissue.

    Science.gov (United States)

    Kantelhardt, Sven R; Kalasauskas, Darius; König, Karsten; Kim, Ella; Weinigel, Martin; Uchugonova, Aisada; Giese, Alf

    2016-05-01

    High resolution multiphoton tomography and fluorescence lifetime imaging differentiates glioma from adjacent brain in native tissue samples ex vivo. Presently, multiphoton tomography is applied in clinical dermatology and experimentally. We here present the first application of multiphoton and fluorescence lifetime imaging for in vivo imaging on humans during a neurosurgical procedure. We used a MPTflex™ Multiphoton Laser Tomograph (JenLab, Germany). We examined cultured glioma cells in an orthotopic mouse tumor model and native human tissue samples. Finally the multiphoton tomograph was applied to provide optical biopsies during resection of a clinical case of glioblastoma. All tissues imaged by multiphoton tomography were sampled and processed for conventional histopathology. The multiphoton tomograph allowed fluorescence intensity- and fluorescence lifetime imaging with submicron spatial resolution and 200 picosecond temporal resolution. Morphological fluorescence intensity imaging and fluorescence lifetime imaging of tumor-bearing mouse brains and native human tissue samples clearly differentiated tumor and adjacent brain tissue. Intraoperative imaging was found to be technically feasible. Intraoperative image quality was comparable to ex vivo examinations. To our knowledge we here present the first intraoperative application of high resolution multiphoton tomography and fluorescence lifetime imaging of human brain tumors in situ. It allowed in vivo identification and determination of cell density of tumor tissue on a cellular and subcellular level within seconds. The technology shows the potential of rapid intraoperative identification of native glioma tissue without need for tissue processing or staining.

  6. Combination of a spinning disc confocal unit with frequency-domain fluorescence lifetime imaging microscopy.

    NARCIS (Netherlands)

    van Munster, E.B.; Goedhart, J.; Kremers, G.J.; Manders, E.M.M.; Gadella, Th.W.J.

    2007-01-01

    BACKGROUND: Wide-field frequency-domain fluorescence lifetime imaging microscopy (FLIM) is an established technique to determine fluorescence lifetimes. Disadvantage of wide-field imaging is that measurements are compromised by out-of-focus blur. Conventional scanning confocal typically means long

  7. Multimodality Imaging Probe for Positron Emission Tomography and Fluorescence Imaging Studies

    Directory of Open Access Journals (Sweden)

    Suresh K. Pandey

    2014-05-01

    Full Text Available Our goal is to develop multimodality imaging agents for use in cell tracking studies by positron emission tomography (PET and optical imaging (OI. For this purpose, bovine serum albumin (BSA was complexed with biotin (histologic studies, 5(6- carboxyfluorescein, succinimidyl ester (FAM SE (OI studies, and diethylenetriamine pentaacetic acid (DTPA for chelating gallium 68 (PET studies. For synthesis of BSA-biotin-FAM-DTPA, BSA was coupled to (+-biotin N-hydroxysuccinimide ester (biotin-NHSI. BSA- biotin was treated with DTPA-anhydride and biotin-BSA-DTPA was reacted with FAM. The biotin-BSA-DTPA-FAM was reacted with gallium chloride 3 to 5 mCi eluted from the generator using 0.1 N HCl and was passed through basic resin (AG 11 A8 and 150 mCi (100 μL, pH 7–8 was incubated with 0.1 mg of FAM conjugate (100 μL at room temperature for 15 minutes to give 66Ga-BSA-biotin-DTPA-FAM. A shaved C57 black mouse was injected with FAM conjugate (50 μL at one flank and FAM-68Ga (50 μL, 30 mCi at the other. Immediately after injection, the mouse was placed in a fluorescence imaging system (Kodak In-Vivo F, Bruker Biospin Co., Woodbridge, CT and imaged (Λex: 465 nm, Λem: 535 nm, time: 8 seconds, Xenon Light Source, Kodak. The same mouse was then placed under an Inveon microPET scanner (Siemens Medical Solutions, Knoxville, TN injected (intravenously with 25 μCi of 18F and after a half-hour (to allow sufficient bone uptake was imaged for 30 minutes. Molecular weight determined using matrix-associated laser desorption ionization (MALDI for the BSA sample was 66,485 Da and for biotin-BSA was 67,116 Da, indicating two biotin moieties per BSA molecule; for biotin-BSA-DTPA was 81,584 Da, indicating an average of 30 DTPA moieties per BSA molecule; and for FAM conjugate was 82,383 Da, indicating an average of 1.7 fluorescent moieties per BSA molecule. Fluorescence imaging clearly showed localization of FAM conjugate and FAM-68Ga at respective flanks of the mouse

  8. Accurate study of FosPeg® distribution in a mouse model using fluorescence imaging technique and fluorescence white monte carlo simulations

    DEFF Research Database (Denmark)

    Xie, Haiyan; Liu, Haichun; Svenmarker, Pontus

    2010-01-01

    Fluorescence imaging is used for quantitative in vivo assessment of drug concentration. Light attenuation in tissue is compensated for through Monte-Carlo simulations. The intrinsic fluorescence intensity, directly proportional to the drug concentration, could be obtained....

  9. Hyperspectral instrumentation to image and characterize the fluorescence of materials

    Science.gov (United States)

    Bourcier, Frédéric; Walter, Philippe; Pedetti, Silvia; Faye, Delphine; Spezzigu, Piero; Infante, Fulvio; Le Nouy, Patrice; Zedda, Edoardo

    2016-09-01

    Optical instruments for space applications with improved performances (smaller pixels and spectral range extension) are becoming more and more sensitive to chemical contamination and particle sedimentation. Outgassing under vacuum conditions causes dramatic flux losses, especially in the UV bandwidth. Furthermore, it is difficult to perform physicochemical analyses of contaminated surfaces on flight models, in a clean room. Conventional analytical techniques such as FTIR (Fourier Transform Infrared interferometer) need the tool to be in contact with the studied area, which is forbidden when working on satellites. In addition, it does not give any information about the distribution of the contaminants in the field of view. The probed area is large, mono-pixel, and the sensitivity of the instrument is too low for hundred nanometer thin film deposits. A first study has shown that we could benefit from using the UV/visible fluorescence spectra to partially identify contaminants and polymer materials. The shape of the fluorescence spectra of adhesives, paints and varnishes have specific signatures that could be recorded into a designated reference database. The location of the presence of these contaminants on such sensitive optics is also relevant. To acquire both these parameters, we designed a specific compact hyperspectral instrument to remotely acquire cube images (500x500 pixels) in a 5 degree field of view, and on a wide range of continuous wavelengths from UV at 320 nm up to the near infrared at 1000 nm. This paper will present the chosen trade-off between different critical optics for a new portable version of this instrument. It is dedicated to space and cultural heritage applications and the first results on an engineering prototype will be shown.

  10. Cerebral bone subtraction CT angiography using 80 kVp and sinogram-affirmed iterative reconstruction: contrast medium and radiation dose reduction with improvement of image quality

    Energy Technology Data Exchange (ETDEWEB)

    Nagayama, Yasunori [Kumamoto City Hospital, Department of Radiology, Kumamoto (Japan); Kumamoto University, Department of Diagnostic Radiology, Chuo-ku, Kumamoto (Japan); Nakaura, Takeshi; Oda, Seitaro; Kidoh, Masafumi; Utsunomiya, Daisuke; Yamashita, Yasuyuki [Kumamoto University, Department of Diagnostic Radiology, Chuo-ku, Kumamoto (Japan); Tsuji, Akinori; Urata, Joji; Furusawa, Mitsuhiro; Yuki, Hideaki; Hirarta, Kenichiro [Kumamoto City Hospital, Department of Radiology, Kumamoto (Japan)

    2017-02-15

    The purpose of this study was to evaluate the feasibility of a contrast medium (CM), radiation dose reduction protocol for cerebral bone-subtraction CT angiography (BSCTA) using 80-kVp and sinogram-affirmed iterative reconstruction (SAFIRE). Seventy-five patients who had undergone BSCTA under the 120- (n = 37) or the 80-kVp protocol (n = 38) were included. CM was 370 mgI/kg for the 120-kVp and 296 mgI/kg for the 80-kVp protocol; the 120- and the 80-kVp images were reconstructed with filtered back-projection (FBP) and SAFIRE, respectively. We compared effective dose (ED), CT attenuation, image noise, and contrast-to-noise ratio (CNR) of two protocols. We also scored arterial contrast, sharpness, depiction of small arteries, visibility near skull base/clip, and overall image quality on a four-point scale. ED was 62% lower at 80- than 120-kVp (0.59 ± 0.06 vs 1.56 ± 0.13 mSv, p < 0.01). CT attenuation of the internal carotid artery (ICA) and middle cerebral artery (MCA) was significantly higher on 80- than 120-kVp (ICA: 557.4 ± 105.7 vs 370.0 ± 59.3 Hounsfield units (HU), p < 0.01; MCA: 551.9 ± 107.9 vs 364.6 ± 62.2 HU, p < 0.01). The CNR was also significantly higher on 80- than 120-kVp (ICA: 46.2 ± 10.2 vs 36.9 ± 7.6, p < 0.01; MCA: 45.7 ± 10.0 vs 35.7 ± 9.0, p < 0.01). Visibility near skull base and clip was not significantly different (p = 0.45). The other subjective scores were higher with the 80- than the 120-kVp protocol (p < 0.05). The 80-kVp acquisition with SAFIRE yields better image quality for BSCTA and substantial reduction in the radiation and CM dose compared to the 120-kVp with FBP protocol. (orig.)

  11. Fluorescence guided lymph node biopsy in large animals using direct image projection device

    Science.gov (United States)

    Ringhausen, Elizabeth; Wang, Tylon; Pitts, Jonathan; Akers, Walter J.

    2016-03-01

    The use of fluorescence imaging for aiding oncologic surgery is a fast growing field in biomedical imaging, revolutionizing open and minimally invasive surgery practices. We have designed, constructed, and tested a system for fluorescence image acquisition and direct display on the surgical field for fluorescence guided surgery. The system uses a near-infrared sensitive CMOS camera for image acquisition, a near-infra LED light source for excitation, and DLP digital projector for projection of fluorescence image data onto the operating field in real time. Instrument control was implemented in Matlab for image capture, processing of acquired data and alignment of image parameters with the projected pattern. Accuracy of alignment was evaluated statistically to demonstrate sensitivity to small objects and alignment throughout the imaging field. After verification of accurate alignment, feasibility for clinical application was demonstrated in large animal models of sentinel lymph node biopsy. Indocyanine green was injected subcutaneously in Yorkshire pigs at various locations to model sentinel lymph node biopsy in gynecologic cancers, head and neck cancer, and melanoma. Fluorescence was detected by the camera system during operations and projected onto the imaging field, accurately identifying tissues containing the fluorescent tracer at up to 15 frames per second. Fluorescence information was projected as binary green regions after thresholding and denoising raw intensity data. Promising results with this initial clinical scale prototype provided encouraging results for the feasibility of optical projection of acquired luminescence during open oncologic surgeries.

  12. Enhancing contrast and quantitation by spatial frequency domain fluorescence molecular imaging

    Science.gov (United States)

    Sun, Jessica; Hathi, Deep; Zhou, Haiying; Shokeen, Monica; Akers, Walter J.

    2016-03-01

    Optical imaging with fluorescent contrast agents is highly sensitive for molecular imaging but is limited in depth to a few centimeters below the skin. Planar fluorescence imaging with full-field, uniform illumination and scientific camera image capture provides a portable and robust configuration for real-time, sensitive fluorescence detection with scalable resolution, but is inherently surface weighted and therefore limited in depth to a few millimeters. At the NIR region (700-1000 nm), tissue absorption and autofluorescence are relatively reduced, increasing depth penetration and reducing background signal, respectively. Optical imaging resolution scales with depth, limiting microscopic resolution with multiphoton microscopy and optical coherence tomography to skin and peri-tumoral tissues are not uniform, varying in thickness and color, complicating subsurface fluorescence measurements. Diffuse optical imaging methods have been developed that better quantify optical signals relative to faster full-field planar reflectance imaging, but require long scan times, complex instrumentation, and reconstruction algorithms. Here we report a novel strategy for rapid measurement of subsurface fluorescence using structured light illumination to improve quantitation of deep-seated fluorescence molecular probe accumulation. This technique, in combination with highly specific, tumor-avid fluorescent molecular probes, will easily integrate noninvasive diagnostics for superficial cancers and fluorescence guided surgery.

  13. Suppression subtractive hybridization.

    Science.gov (United States)

    Ghorbel, Mohamed T; Murphy, David

    2011-01-01

    Comparing two RNA populations that differ from the effects of a single independent variable, such as a drug treatment or a specific genetic defect, can establish differences in the abundance of specific transcripts that vary in a population dependent manner. There are different methods for identifying differentially expressed genes. These methods include microarray, Serial Analysis of Gene Expression (SAGE), and quantitative Reverse-Transcriptase Polymerase Chain Reaction (qRT-PCR). Herein, the protocol describes an easy and cost-effective alternative that does not require prior knowledge of the transcriptomes under examination. It is specifically relevant when low levels of RNA starting material are available. This protocol describes the use of Switching Mechanism At RNA Termini Polymerase Chain Reaction (SMART-PCR) to amplify cDNA from small amounts of RNA. The amplified cDNA populations under comparison are then subjected to Suppression Subtractive Hybridization (SSH-PCR). SSH-PCR is a technique that couples subtractive hybridization with suppression PCR to selectively amplify fragments of differentially expressed genes. The resulting products are cDNA populations enriched for significantly overrepresented transcripts in either of the two input RNAs. These cDNA populations can then be cloned to generate subtracted cDNA library. Microarrays made with clones from the subtracted forward and reverse cDNA libraries are then screened for differentially expressed genes using targets generated from tester and driver total RNAs.

  14. NADH fluorescence imaging of isolated biventricular working rabbit hearts.

    Science.gov (United States)

    Asfour, Huda; Wengrowski, Anastasia M; Jaimes, Rafael; Swift, Luther M; Kay, Matthew W

    2012-07-24

    Since its inception by Langendorff(1), the isolated perfused heart remains a prominent tool for studying cardiac physiology(2). However, it is not well-suited for studies of cardiac metabolism, which require the heart to perform work within the context of physiologic preload and afterload pressures. Neely introduced modifications to the Langendorff technique to establish appropriate left ventricular (LV) preload and afterload pressures(3). The model is known as the isolated LV working heart model and has been used extensively to study LV performance and metabolism(4-6). This model, however, does not provide a properly loaded right ventricle (RV). Demmy et al. first reported a biventricular model as a modification of the LV working heart model(7, 8). They found that stroke volume, cardiac output, and pressure development improved in hearts converted from working LV mode to biventricular working mode(8). A properly loaded RV also diminishes abnormal pressure gradients across the septum to improve septal function. Biventricular working hearts have been shown to maintain aortic output, pulmonary flow, mean aortic pressure, heart rate, and myocardial ATP levels for up to 3 hours(8). When studying the metabolic effects of myocardial injury, such as ischemia, it is often necessary to identify the location of the affected tissue. This can be done by imaging the fluorescence of NADH (the reduced form of nicotinamide adenine dinucleotide)(9-11), a coenzyme found in large quantities in the mitochondria. NADH fluorescence (fNADH) displays a near linearly inverse relationship with local oxygen concentration(12) and provides a measure of mitochondrial redox state(13). fNADH imaging during hypoxic and ischemic conditions has been used as a dye-free method to identify hypoxic regions(14, 15) and to monitor the progression of hypoxic conditions over time(10). The objective of the method is to monitor the mitochondrial redox state of biventricular working hearts during protocols

  15. [Studies on laticifers and milk of greater celandine (Chelidonium majus L.) with fluorescence imaging and fluorescence spectroscopic methods].

    Science.gov (United States)

    Póczi, Dorottya; Böddi, Béla

    2010-01-01

    Using fluorescence imaging and fluorescence spectroscopic methods, the localisation of the laticifers and the native spectral properties of the milk were studied in various organs of greater celandine (Chelidonium majus L.). Direct measurements on tissue pieces (without the extraction and the separation of the components) provided information about the complexity of the milk and the various ratios of the alkaloid contents in the tissues. Whole plant were studied in a gel documentation system using ultraviolet light source, while the localisation of the laticifers was observed along the leaf veins in fluorescence microscope, using blue excitation light. Measuring different tissue pieces, fluorescence spectroscopic studies showed that the greater celandine alkaloids have emission bands at 469, 530-531, 553, 572-575 and 592 nm and excitation bands at 365, 370, 386 is 400 nm. These results give a possibility for conclusions about the alkaloid contents and composition or ratios of the alkaloid components in various tissue pieces directly, via comparisons with alkaloid standards.

  16. Fluorescence imaging of experimental rheumatoid arthritis in vivo using a fast flying-spot scanner

    Science.gov (United States)

    Berger, J.; Voigt, J.; Seifert, F.; Ebert, B.; Macdonald, R.; Gemeinhardt, I.; Gemeinhardt, O.; Schnorr, J.; Taupitz, M.; Vater, A.; Vollmer, S.; Licha, K.; Schirner, M.

    2007-07-01

    We have developed a flying-spot scanner for fluorescence imaging of rheumatoid arthritis in the near infrared (NIR) spectral range following intravenous administration of contrast agents. The new imaging system has been characterized with respect to linearity, dynamic range and spatial resolution with the help of fluorescent phantoms. In vivo experiments were performed on an animal model of rheumatoid arthritis. Finally, NIR-fluorescence images of early stages of joint inflammation have been compared with findings from contrast enhanced MR imaging and histology.

  17. Laser-induced fluorescence imaging of subsurface tissue structures with a volume holographic spatial-spectral imaging system.

    Science.gov (United States)

    Luo, Yuan; Gelsinger-Austin, Paul J; Watson, Jonathan M; Barbastathis, George; Barton, Jennifer K; Kostuk, Raymond K

    2008-09-15

    A three-dimensional imaging system incorporating multiplexed holographic gratings to visualize fluorescence tissue structures is presented. Holographic gratings formed in volume recording materials such as a phenanthrenquinone poly(methyl methacrylate) photopolymer have narrowband angular and spectral transmittance filtering properties that enable obtaining spatial-spectral information within an object. We demonstrate this imaging system's ability to obtain multiple depth-resolved fluorescence images simultaneously.

  18. Iodinated oil-loaded, fluorescent mesoporous silica-coated iron oxide nanoparticles for magnetic resonance imaging/computed tomography/fluorescence trimodal imaging

    Science.gov (United States)

    Xue, Sihan; Wang, Yao; Wang, Mengxing; Zhang, Lu; Du, Xiaoxia; Gu, Hongchen; Zhang, Chunfu

    2014-01-01

    In this study, a novel magnetic resonance imaging (MRI)/computed tomography (CT)/fluorescence trifunctional probe was prepared by loading iodinated oil into fluorescent mesoporous silica-coated superparamagnetic iron oxide nanoparticles (i-fmSiO4@SPIONs). Fluorescent mesoporous silica-coated superparamagnetic iron oxide nanoparticles (fmSiO4@SPIONs) were prepared by growing fluorescent dye-doped silica onto superparamagnetic iron oxide nanoparticles (SPIONs) directed by a cetyltrimethylammonium bromide template. As prepared, fmSiO4@SPIONs had a uniform size, a large surface area, and a large pore volume, which demonstrated high efficiency for iodinated oil loading. Iodinated oil loading did not change the sizes of fmSiO4@SPIONs, but they reduced the MRI T2 relaxivity (r2) markedly. I-fmSiO4@SPIONs were stable in their physical condition and did not demonstrate cytotoxic effects under the conditions investigated. In vitro studies indicated that the contrast enhancement of MRI and CT, and the fluorescence signal intensity of i-fmSiO4@SPION aqueous suspensions and macrophages, were intensified with increased i-fmSiO4@SPION concentrations in suspension and cell culture media. Moreover, for the in vivo study, the accumulation of i-fmSiO4@SPIONs in the liver could also be detected by MRI, CT, and fluorescence imaging. Our study demonstrated that i-fmSiO4@SPIONs had great potential for MRI/CT/fluorescence trimodal imaging. PMID:24904212

  19. Voxel significance mapping in epilepsy studies using subtraction ictal SPECT

    Science.gov (United States)

    Brinkmann, Benjamin H.; O'Brien, Terence J.; Webster, Desmond B.; Robins, Peter D.; Mullan, Brian P.; Robb, Richard A.

    1999-05-01

    Subtraction ictal SPECT coregistered to MRI (SISCOM) has been shown to aid epileptogenic localization and improve surgical outcomes in partial epilepsy patients. This paper reports a new method of identifying significant areas of epileptogenic activation in the SISCOM subtraction image taking into account normal variation between sequential Tc-99m Ethyl Cysteinate Diethylester SPECT scans of single individuals. The method uses the AIR 3.0 nonlinear registration software to combine a group of subtraction images into a common anatomical framework. A map of the pixel intensity standard deviation values in the subtraction images is created, and this map is nonlinearly registered to a patient's SISCOM subtraction image. Pixels in the patient subtraction image may then be evaluated based upon the statistical characteristics of corresponding pixels in the atlas. Validation experiments were performed to verify that local image variances are not constant across the image and that nonlinear registration preserves local image variances. SISCOM images created with the voxel variance method were rated higher in quality than the conventional image variance method in images from fifteen patients. No difference in localization rate was observed between the voxel variance mapping and image variance methods. The voxel significance mapping method was shown to improve the quality of clinical SISCOM images without removing localizing information.

  20. Precise diagnosis in different scenarios using photoacoustic and fluorescence imaging with dual-modality nanoparticles

    Science.gov (United States)

    Peng, Dong; Du, Yang; Shi, Yiwen; Mao, Duo; Jia, Xiaohua; Li, Hui; Zhu, Yukun; Wang, Kun; Tian, Jie

    2016-07-01

    Photoacoustic imaging and fluorescence molecular imaging are emerging as important research tools for biomedical studies. Photoacoustic imaging offers both strong optical absorption contrast and high ultrasonic resolution, and fluorescence molecular imaging provides excellent superficial resolution, high sensitivity, high throughput, and the ability for real-time imaging. Therefore, combining the imaging information of both modalities can provide comprehensive in vivo physiological and pathological information. However, currently there are limited probes available that can realize both fluorescence and photoacoustic imaging, and advanced biomedical applications for applying this dual-modality imaging approach remain underexplored. In this study, we developed a dual-modality photoacoustic-fluorescence imaging nanoprobe, ICG-loaded Au@SiO2, which was uniquely designed, consisting of gold nanorod cores and indocyanine green with silica shell spacer layers to overcome fluorophore quenching. This nanoprobe was examined by both PAI and FMI for in vivo imaging on tumor and ischemia mouse models. Our results demonstrated that the nanoparticles can specifically accumulate at the tumor and ischemic areas and be detected by both imaging modalities. Moreover, this dual-modality imaging strategy exhibited superior advantages for a precise diagnosis in different scenarios. The new nanoprobe with the dual-modality imaging approach holds great potential for diagnosis and stage classification of tumor and ischemia related diseases.Photoacoustic imaging and fluorescence molecular imaging are emerging as important research tools for biomedical studies. Photoacoustic imaging offers both strong optical absorption contrast and high ultrasonic resolution, and fluorescence molecular imaging provides excellent superficial resolution, high sensitivity, high throughput, and the ability for real-time imaging. Therefore, combining the imaging information of both modalities can provide

  1. Near-Infrared Squaraine Dye Encapsulated Micelles for in Vivo Fluorescence and Photoacoustic Bimodal Imaging.

    Science.gov (United States)

    Sreejith, Sivaramapanicker; Joseph, James; Lin, Manjing; Menon, Nishanth Venugopal; Borah, Parijat; Ng, Hao Jun; Loong, Yun Xian; Kang, Yuejun; Yu, Sidney Wing-Kwong; Zhao, Yanli

    2015-06-23

    Combined near-infrared (NIR) fluorescence and photoacoustic imaging techniques present promising capabilities for noninvasive visualization of biological structures. Development of bimodal noninvasive optical imaging approaches by combining NIR fluorescence and photoacoustic tomography demands suitable NIR-active exogenous contrast agents. If the aggregation and photobleaching are prevented, squaraine dyes are ideal candidates for fluorescence and photoacoustic imaging. Herein, we report rational selection, preparation, and micelle encapsulation of an NIR-absorbing squaraine dye (D1) for in vivo fluorescence and photoacoustic bimodal imaging. D1 was encapsulated inside micelles constructed from a biocompatible nonionic surfactant (Pluoronic F-127) to obtain D1-encapsulated micelles (D1(micelle)) in aqueous conditions. The micelle encapsulation retains both the photophysical features and chemical stability of D1. D1(micelle) exhibits high photostability and low cytotoxicity in biological conditions. Unique properties of D1(micelle) in the NIR window of 800-900 nm enable the development of a squaraine-based exogenous contrast agent for fluorescence and photoacoustic bimodal imaging above 820 nm. In vivo imaging using D1(micelle), as demonstrated by fluorescence and photoacoustic tomography experiments in live mice, shows contrast-enhanced deep tissue imaging capability. The usage of D1(micelle) proven by preclinical experiments in rodents reveals its excellent applicability for NIR fluorescence and photoacoustic bimodal imaging.

  2. A CMOS image sensor with draining only modulation pixels for fluorescence lifetime imaging

    Science.gov (United States)

    Li, Zhuo; Yasutomi, Keita; Takasawa, Taishi; Itoh, Shinya; Kawahito, Shoji

    2011-03-01

    Fluorescence lifetime imaging is becoming a powerful tool in biology. A charge-domain CMOS Fluorescence Lifetime Imaging Microscopy (FLIM) chip using a pinned photo diode (PPD) and the pinned storage diode (PSD) with different depth of potential wells has been previously developed by the authors. However, a transfer gate between PPD and PSD causes charge transfer noise due to traps at the channel surface. This paper presents a time-resolved CMOS image sensor with draining only modulation pixels for fluorescence lifetime imaging, which removes the transfer gate between PPD and PSD. The time windowing is done by draining with a draining gate only, which is attached along the carrier path from PPD to PSD. This allows us to realize a trapping less charge transfer between PPD and PSD, leading to a very low-noise time-resolved signal detection. A video-rate CMOS FLIM chip has been fabricated using 0.18μm standard CMOS pinned diode image sensor process. The pixel consists of a PPD, a PSD, a charge draining gate (TD), a readout transfer gate (TX) between the PSD and the floating diffusion (FD), a reset transistor and a source follower amplifier transistor. The pixel array has 200(Row) x 256(Column) pixels and the pixel pitch is 7.5μm. Fundamental characteristics of the implemented CMOS FLIM chip are measured. The signal intensity of the PSD as a function of the TD gate voltage is also measured. The ratio of the signal for the TD off to the signal for the TD on is 212 : 1.

  3. Preparation and Characterization of Highly Fluorescent, Glutathione-coated Near Infrared Quantum Dots for in Vivo Fluorescence Imaging

    Directory of Open Access Journals (Sweden)

    Yoshichika Yoshioka

    2008-10-01

    Full Text Available Fluorescent probes that emit in the near-infrared (NIR, 700-1,300 nm region are suitable as optical contrast agents for in vivo fluorescence imaging because of low scattering and absorption of the NIR light in tissues. Recently, NIR quantum dots (QDs have become a new class of fluorescent materials that can be used for in vivo imaging. Compared with traditional organic fluorescent dyes, QDs have several unique advantages such as size- and composition-tunable emission, high brightness, narrow emission bands, large Stokes shifts, and high resistance to photobleaching. In this paper, we report a facile method for the preparation of highly fluorescent, water-soluble glutathione (GSH-coated NIR QDs for in vivo imaging. GSH-coated NIR QDs (GSH-QDs were prepared by surface modification of hydrophobic CdSeTe/CdS (core/shell QDs. The hydrophobic surface of the CdSeTe/CdS QDs was exchanged with GSH in tetrahydrofuran-water. The resulting GSH-QDs were monodisperse particles and stable in PBS (phosphate buffered saline, pH = 7.4. The GSH-QDs (800 nm emission were highly fluorescent in aqueous solutions (quantum yield = 22% in PBS buffer, and their hydrodynamic diameter was less than 10 nm, which is comparable to the size of proteins. The cellular uptake and viability for the GSH-QDs were examined using HeLa and HEK 293 cells. When the cells were incubated with aqueous solutions of the GSH-QDs (10 nM, the QDs were taken into the cells and distributed in the perinuclear region of both cells. After 12 hrs incubation of 4 nM of GSH-QDs, the viabilities of HeLa and HEK 293 cells were ca. 80 and 50%, respectively. As a biomedical utility of the GSH-QDs, in vivo NIRfluorescence imaging of a lymph node in a mouse is presented.

  4. Near-Infrared Fluorescence Enhanced (NIR-FE) Molecular Imaging of Live Cells on Gold Substrates

    CERN Document Server

    Hong, Guosong; Welsher, Kevin; Chen, Zhuo; Robinson, Joshua T; Wang, Hailiang; Zhang, Bo; Dai, Hongjie

    2011-01-01

    Low quantum yields of near infrared (NIR) fluorophores have limited their capabilities as imaging probes in a transparent, low background imaging window. Here for the first time we reported near-infrared fluorescence enhance (NIR-FE) cell imaging using nanostructured Au substrate, which was employed as a general platform for both single-walled carbon nanotubes (SWNTs) and organic fluorescent labels in the NIR region. Fluorescence intensity, as well as cell targeting specificity, was greatly improved by this novel imaging technique. With NIR-FE imaging, we were able to image SWNT-stained cells at short exposure time of 300ms, and push the detectable limit of SWNT staining of cells down to an ultralow concentration of ~50 pM. Further, different degrees of fluorescence enhancement for endocytosed, intracellular SWNTs vs. nanotubes on the cell membrane at the cell/gold interface were observed, suggesting the possibility of using this technique to track the transmembrane behavior of NIR fluorophores.

  5. Deep-brain imaging via epi-fluorescence Computational Cannula Microscopy

    Science.gov (United States)

    Kim, Ganghun; Nagarajan, Naveen; Pastuzyn, Elissa; Jenks, Kyle; Capecchi, Mario; Shepherd, Jason; Menon, Rajesh

    2017-03-01

    Here we demonstrate widefield (field diameter = 200 μm) fluorescence microscopy and video imaging inside the rodent brain at a depth of 2 mm using a simple surgical glass needle (cannula) of diameter 0.22 mm as the primary optical element. The cannula guides excitation light into the brain and the fluorescence signal out of the brain. Concomitant image-processing algorithms are utilized to convert the spatially scrambled images into fluorescent images and video. The small size of the cannula enables minimally invasive imaging, while the long length (>2 mm) allow for deep-brain imaging with no additional complexity in the optical system. Since no scanning is involved, widefield fluorescence video at the native frame rate of the camera can be achieved.

  6. Segmenting Intracellular Distribution Images Derived by Fluorescent Dyes Using a Potts Model Hamiltonian

    CERN Document Server

    Hu, Dandan; Ronhovde, Peter; Bloch, Sharon; Achilefu, Samuel; Nussinov, Zohar

    2012-01-01

    We apply a multiresolution community detection algorithm to perform unsupervised segmentation of complex intracellular signals derived using fluorescent dyes. In our earlier work, when applying our method to benchmarks, our algorithm was shown to be one of the best and to be especially suited to the detection of camouflage images. In the current manuscript, we have explored this algorithm in a more complex scenario. The current image processing problem is framed as identifying clusters with respective average fluorescent lifetimes (FLTs) against a background or "solvent" in fluorescence lifetime imaging microscopy (FLIM) images derived using NIR fluorescent dyes. We have identified significant multiresolution structures using replica correlations in these images, where such correlations are manifested by information theoretic overlaps of the independent solutions ("replicas") attained using the proposed algorithm from different starting points. Our method is more efficient than a well-known image segmentation...

  7. Deep-brain imaging via epi-fluorescence Computational Cannula Microscopy

    Science.gov (United States)

    Kim, Ganghun; Nagarajan, Naveen; Pastuzyn, Elissa; Jenks, Kyle; Capecchi, Mario; Shepherd, Jason; Menon, Rajesh

    2017-01-01

    Here we demonstrate widefield (field diameter = 200 μm) fluorescence microscopy and video imaging inside the rodent brain at a depth of 2 mm using a simple surgical glass needle (cannula) of diameter 0.22 mm as the primary optical element. The cannula guides excitation light into the brain and the fluorescence signal out of the brain. Concomitant image-processing algorithms are utilized to convert the spatially scrambled images into fluorescent images and video. The small size of the cannula enables minimally invasive imaging, while the long length (>2 mm) allow for deep-brain imaging with no additional complexity in the optical system. Since no scanning is involved, widefield fluorescence video at the native frame rate of the camera can be achieved. PMID:28317915

  8. Spectral confocal imaging of fluorescently tagged nicotinic receptors in knock-in mice with chronic nicotine administration.

    Science.gov (United States)

    Renda, Anthony; Nashmi, Raad

    2012-02-10

    neurons via spectral confocal microscopy. The targeted fluorescent knock-in mutation is incorporated in the endogenous locus and under control of its native promoter, producing normal levels of expression and regulation of the receptor when compared to untagged receptors in wildtype mice. This knock-in approach can be extended to fluorescently tag other ion channels and offers a powerful approach of visualizing and quantifying receptors in the CNS. In this paper we describe a methodology to quantify changes in nAChR expression in specific CNS neurons after exposure to chronic nicotine. Our methods include mini-osmotic pump implantation, intracardiac perfusion fixation, imaging and analysis of fluorescently tagged nicotinic receptor subunits from α4YFP knock-in mice (Fig. 1). We have optimized the fixation technique to minimize autofluorescence from fixed brain tissue. We describe in detail our imaging methodology using a spectral confocal microscope in conjunction with a linear spectral unmixing algorithm to subtract autofluoresent signal in order to accurately obtain α4YFP fluorescence signal. Finally, we show results of chronic nicotine-induced upregulation of α4YFP receptors in the medial perforant path of the hippocampus.

  9. Cell Permeable Ratiometric Fluorescent Sensors for Imaging Phosphoinositides.

    Science.gov (United States)

    Mondal, Samsuzzoha; Rakshit, Ananya; Pal, Suranjana; Datta, Ankona

    2016-07-15

    Phosphoinositides are critical cell-signal mediators present on the plasma membrane. The dynamic change of phosphoinositide concentrations on the membrane including clustering and declustering mediates signal transduction. The importance of phosphoinositides is scored by the fact that they participate in almost all cell-signaling events, and a defect in phosphoinositide metabolism is linked to multiple diseases including cancer, bipolar disorder, and type-2 diabetes. Optical sensors for visualizing phosphoinositide distribution can provide information on phosphoinositide dynamics. This exercise will ultimately afford a handle into understanding and manipulating cell-signaling processes. The major requirement in phosphoinositide sensor development is a selective, cell permeable probe that can quantify phosphoinositides. To address this requirement, we have developed short peptide-based ratiometric fluorescent sensors for imaging phosphoinositides. The sensors afford a selective response toward two crucial signaling phosphoinositides, phosphatidylinositol-4,5-bisphosphate (PI(4,5)P2) and phosphatidylinositol-4-phosphate (PI4P), over other anionic membrane phospholipids and soluble inositol phosphates. Dissociation constant values indicate up to 4 times higher probe affinity toward PI(4,5)P2 when compared to PI4P. Significantly, the sensors are readily cell-permeable and enter cells within 15 min of incubation as indicated by multiphoton excitation confocal microscopy. Furthermore, the sensors light up signaling phosphoinositides present both on the cell membrane and on organelle membranes near the perinuclear space, opening avenues for quantifying and monitoring phosphoinositide signaling.

  10. Electron beam dispersion measurements in nitrogen using two-dimensional imaging of N2(+) fluorescence

    Science.gov (United States)

    Clapp, L. H.; Twiss, R. G.; Cattolica, R. J.

    Experimental results are presented related to the radial spread of fluorescence excited by 10 and 20 KeV electron beams passing through nonflowing rarefied nitrogen at 293 K. An imaging technique for obtaining species distributions from measured beam-excited fluorescence is described, based on a signal inversion scheme mathematically equivalent to the inversion of the Abel integral equation. From fluorescence image data, measurements of beam radius, integrated signal intensity, and spatially resolved distributions of N2(+) first-negative-band fluorescence-emitting species have been made. Data are compared with earlier measurements and with an heuristic beam spread model.

  11. Evaluation of sacroiliitis: contrast-enhanced MRI with subtraction technique

    Energy Technology Data Exchange (ETDEWEB)

    Algin, Oktay; Gokalp, Gokhan; Baran, Bulent; Ocakoglu, Gokhan; Yazici, Zeynep [Uludag University, Medical Faculty, Department of Radiology, Gorukle, Bursa (Turkey)

    2009-10-15

    The purpose of the study was to investigate the diagnostic value of contrast-enhanced MRI using the subtraction technique in the detection of active sacroiliitis. Magnetic resonance imaging was performed in 8 asymptomatic volunteers and 50 patients with clinically suspected active sacroiliitis. On precontrast MR images, T1-weighted spin-echo images with and without fat saturation (T1WFS and T1W), STIR and 3D-FLASH images with fat saturation were obtained in the semicoronal plane using a 1.5 Tesla imager. Postcontrast MRI was performed using the same T1WFS sequence as before contrast injection for all volunteers and patients. Postcontrast images were subtracted from fat-suppressed precontrast images. Enhancement within the joint space and bone marrow was considered to demonstrate active sacroiliitis. In 50 patients (100 sacroiliac joints [SIJs]), 40 (76 SIJs) were considered to have active sacroiliitis based on MR images. Bone marrow edema was present in 33 patients (62 SIJs) on STIR images. Routine MRI allowed identification of contrast enhancement in SIJs on postcontrast T1WFS images in 31 patients (49 SIJs). Contrast enhancement was observed in 40 patients (76 SIJs) who were examined by MRI using the subtraction technique. Contrast enhancement was significantly more conspicuous on subtraction images than on non-subtracted postcontrast T1WFS images (Mann-Whitney U test, p<0.001). Contrast-enhanced MRI with subtraction technique may be useful for early detection of active sacroiliitis. (orig.)

  12. A fast image registration approach of neural activities in light-sheet fluorescence microscopy images

    Science.gov (United States)

    Meng, Hui; Hui, Hui; Hu, Chaoen; Yang, Xin; Tian, Jie

    2017-03-01

    The ability of fast and single-neuron resolution imaging of neural activities enables light-sheet fluorescence microscopy (LSFM) as a powerful imaging technique in functional neural connection applications. The state-of-art LSFM imaging system can record the neuronal activities of entire brain for small animal, such as zebrafish or C. elegans at single-neuron resolution. However, the stimulated and spontaneous movements in animal brain result in inconsistent neuron positions during recording process. It is time consuming to register the acquired large-scale images with conventional method. In this work, we address the problem of fast registration of neural positions in stacks of LSFM images. This is necessary to register brain structures and activities. To achieve fast registration of neural activities, we present a rigid registration architecture by implementation of Graphics Processing Unit (GPU). In this approach, the image stacks were preprocessed on GPU by mean stretching to reduce the computation effort. The present image was registered to the previous image stack that considered as reference. A fast Fourier transform (FFT) algorithm was used for calculating the shift of the image stack. The calculations for image registration were performed in different threads while the preparation functionality was refactored and called only once by the master thread. We implemented our registration algorithm on NVIDIA Quadro K4200 GPU under Compute Unified Device Architecture (CUDA) programming environment. The experimental results showed that the registration computation can speed-up to 550ms for a full high-resolution brain image. Our approach also has potential to be used for other dynamic image registrations in biomedical applications.

  13. Photoactivation and imaging of optical highlighter fluorescent proteins.

    Science.gov (United States)

    Patterson, George H

    2011-07-01

    A major advance in the microscopic study of cells and tissues is the introduction of photoactivatable fluorescent proteins, which can specifically mark proteins of interest within a living cell. Fluorescent proteins are now available that allow a pool of molecules to be "turned on" by photoactivation. This unit discusses technical aspects for the general use of photoactivatable fluorescent proteins and introduces some specific applications in the concluding remarks.

  14. Fluorescence-enhanced imaging using a novel hand-held based optical imager: phantom studies

    Science.gov (United States)

    Ge, Jiajia; Zhu, Banghe; Regalado, Steven; Godavarty, Anuradha

    2008-02-01

    Near-infrared (NIR) optical imaging is an emerging noninvasive modality for breast cancer diagnosis. The currently available optical imaging systems towards tomography studies are limited either by instrument portability, patient comfort, or flexibility to image any given tissue volume. Hence, a novel hand-held probe based gain modulated intensified CCD camera imaging system is developed such that it can possibly overcome some of the above limitations. The unique features of this hand-held probe based optical imaging system are: (i) to perform simultaneous multiple point illumination and detection, thus decreasing the total imaging time and improving overall signal strength; (ii) to adapt to the tissue contours, thus decreasing the light leakage at contact surface; and (iii) to obtain trans-illumination measurements apart from reflectance measurements, thus improving the depth information. Phantom studies are performed to demonstrate the feasibility of performing fluorescence optical imaging under different target depths using cubical phantoms (10×6.5×10 cc). The effect of simultaneous multiple point illumination over sequential single point illumination is demonstrated from experimental phantom studies.

  15. Influence of angle's ranges for recording an X-ray fluorescence hologram on reconstructed atomic images

    Institute of Scientific and Technical Information of China (English)

    XIE Hong-Lan; CHEN Jian-Wen; GAO Hong-Yi; ZHU Hua-Feng; LI Ru-Xin; XU Zhi-Zhan

    2004-01-01

    X-ray fluorescence holography (XFH) is a novel method for three-dimensional (3D) imaging of atomic structure. Theoretically, in an XFH experiment, one has to measure the fluorescence energy on a spherical surface to get well-resolved 3D images of atoms. But in practice, the experimental system arrangement does not allow the measurement of the fluorescent intensity oscillations in the full sphere. The holographic information losses because of the limited sampling range (less than 4π) will directly result in defective reconstructed atomic images. In this work, the atomic image of a Fe single crystal (001) was reconstructed by numerically simulating X-ray fluorescence holograms of the crystal at different recording angle's ranges and step lengths. Influences of the ranges of azimuth angles and polar angles and the step length of polar angles on the reconstructed atomic images were discussed.

  16. Two-photon excited fluorescence microendoscopic imaging using a GRIN lens

    Science.gov (United States)

    Yan, Wei; Peng, Xiao; Lin, Danying; Wang, Qi; Gao, Jian; Zhou, Jie; Ye, Tong; Qu, Junle; Niu, Hanben

    2015-03-01

    With the rapid development of life sciences, there is an increasing demand for intravital fluorescence imaging of small animals. However, large dimensions and limited working distances of objective lenses in traditional fluorescence microscopes have limited the imaging applications mostly to superficial tissues. To overcome this disadvantage, researchers have developed the graded-index (GRIN) probes with small diameters for imaging internal organs of small animals in a minimally invasive fashion. Here, we present the development of a fluorescence endoscopic imaging system based on a GRIN lens using two-photon excitation. Experimental results showed that this system could perform dynamic fluorescence microendoscopic imaging and monitor the blood flow in anesthetized living mice using two-photon excitation.

  17. A detection instrument for enhanced-fluorescence and label-free imaging on photonic crystal surfaces.

    Science.gov (United States)

    Block, Ian D; Mathias, Patrick C; Ganesh, Nikhil; Jones, Sarah I; Dorvel, Brian R; Chaudhery, Vikram; Vodkin, Lila O; Bashir, Rashid; Cunningham, Brian T

    2009-07-20

    We report on the design and demonstration of an optical imaging system capable of exciting surface-bound fluorophores within the resonant evanescent electric field of a photonic crystal surface and gathering fluorescence emission that is directed toward the imaging objective by the photonic crystal. The system also has the ability to quantify shifts in the local resonance angle induced by the adsorption of biomolecules on the photonic crystal surface for label-free biomolecular imaging. With these two capabilities combined within a single detection system, we demonstrate label-free images self-registered to enhanced fluorescence images with 328x more sensitive fluorescence detection relative to a glass surface. This technique is applied to a DNA microarray where label-free quantification of immobilized capture DNA enables improved quality control and subsequent enhanced fluorescence detection of dye-tagged hybridized DNA yields 3x more genes to be detected versus commercially available microarray substrates.

  18. First X-ray fluorescence CT experimental results at the SSRF X-ray imaging beamline

    Institute of Scientific and Technical Information of China (English)

    DENG Biao; YANG Qun; XIE Hong-Lan; DU Guo-Hao; XIAO Wi-Qiao

    2011-01-01

    X-ray fluorescence CT is a non-destructive technique for detecting elemental composition and distribution inside a specimen. In this paper, the first experimental results of X-ray fluorescence CT obtained at the SSRF X-ray imaging beamline (BL13W1) are described. The test samples were investigated and the 2D elemental image was reconstructed using a filtered back-projection algorithm. In the sample the element Cd was observed. Up to now, the X-ray fluorescence CT could be carried out at the SSRF X-ray imaging beamline.

  19. Dual-Modal Nanoprobes for Imaging of Mesenchymal Stem Cell Transplant by MRI and Fluorescence Imaging

    Energy Technology Data Exchange (ETDEWEB)

    Sung, Chang Kyu; Hong, Kyung Ah; Lin, Shun Mei [Seoul Metropolitan Boramae Medical Center, Seoul (Korea, Republic of)] (and others)

    2009-12-15

    To determine the feasibility of labeling human mesenchymal stem cells (hMSCs) with bifunctional nanoparticles and assessing their potential as imaging probes in the monitoring of hMSC transplantation. The T1 and T2 relaxivities of the nanoparticles (MNP SiO{sub 2}[RITC]-PEG) were measured at 1.5T and 3T magnetic resonance scanner. Using hMSCs and the nanoparticles, labeling efficiency, toxicity, and proliferation were assessed. Confocal laser scanning microscopy and transmission electron microscopy were used to specify the intracellular localization of the endocytosed iron nanoparticles. We also observed in vitro and in vivo visualization of the labeled hMSCs with a 3T MR scanner and optical imaging. MNP SiO{sub 2}(RITC)-PEG showed both superparamagnetic and fluorescent properties. The r{sub 1} and r{sub 2} relaxivity values of the MNP SiO{sub 2}(RITC)-PEG were 0.33 and 398 mM{sup -1} s{sup -1} at 1.5T, respectively, and 0.29 and 453 mM{sup -1} s{sup -1} at 3T, respectively. The effective internalization of MNP SiO{sub 2}(RITC)-PEG into hMSCs was observed by confocal laser scanning fluorescence microscopy. The transmission electron microscopy images showed that MNP SiO{sub 2}(RITC)-PEG was internalized into the cells and mainly resided in the cytoplasm. The viability and proliferation of MNP SiO{sub 2}(RITC)-PEG-labeled hMSCs were not significantly different from the control cells. MNP SiO{sub 2}(RITC)-PEG-labeled hMSCs were observed in vitro and in vivo with optical and MR imaging. MNP SiO{sub 2}(RITC)-PEG can be a useful contrast agent for stem cell imaging, which is suitable for a bimodal detection by MRI and optical imaging.

  20. Detection of fecal residue on poultry carcasses by laser-induced fluorescence imaging.

    Science.gov (United States)

    Cho, B; Kim, M S; Chao, K; Lawrence, K; Park, B; Kim, K

    2009-04-01

    Feasibility of fluorescence imaging technique for the detection of diluted fecal matters from various parts of the digestive tract, including colon, ceca, small intestine, and duodenum, on poultry carcasses was investigated. One of the challenges for using fluorescence imaging for inspection of agricultural material is the low fluorescence yield in that fluorescence can be masked by ambient light. A laser-induced fluorescence imaging system (LIFIS) developed by our group allowed acquisition of fluorescence from feces-contaminated poultry carcasses in ambient light. Fluorescence emission images at 630 nm were captured with 415-nm laser excitation. Image processing algorithms including threshold and image erosion were used to identify fecal spots diluted up to 1: 10 by weight with double distilled water. Feces spots on the carcasses, without dilution and up to 1: 5 dilutions, could be detected with 100% accuracy regardless of feces type. Detection accuracy for fecal matters diluted up to 1: 10 was 96.6%. The results demonstrated good potential of the LIFIS for detection of diluted poultry fecal matter, which can harbor pathogens, on poultry carcasses.

  1. Evaluation of chemical fluorescent dyes as a protein conjugation partner for live cell imaging.

    Directory of Open Access Journals (Sweden)

    Yoko Hayashi-Takanaka

    Full Text Available To optimize live cell fluorescence imaging, the choice of fluorescent substrate is a critical factor. Although genetically encoded fluorescent proteins have been used widely, chemical fluorescent dyes are still useful when conjugated to proteins or ligands. However, little information is available for the suitability of different fluorescent dyes for live imaging. We here systematically analyzed the property of a number of commercial fluorescent dyes when conjugated with antigen-binding (Fab fragments directed against specific histone modifications, in particular, phosphorylated H3S28 (H3S28ph and acetylated H3K9 (H3K9ac. These Fab fragments were conjugated with a fluorescent dye and loaded into living HeLa cells. H3S28ph-specific Fab fragments were expected to be enriched in condensed chromosomes, as H3S28 is phosphorylated during mitosis. However, the degree of Fab fragment enrichment on mitotic chromosomes varied depending on the conjugated dye. In general, green fluorescent dyes showed higher enrichment, compared to red and far-red fluorescent dyes, even when dye:protein conjugation ratios were similar. These differences are partly explained by an altered affinity of Fab fragment after dye-conjugation; some dyes have less effect on the affinity, while others can affect it more. Moreover, red and far-red fluorescent dyes tended to form aggregates in the cytoplasm. Similar results were observed when H3K9ac-specific Fab fragments were used, suggesting that the properties of each dye affect different Fab fragments similarly. According to our analysis, conjugation with green fluorescent dyes, like Alexa Fluor 488 and Dylight 488, has the least effect on Fab affinity and is the best for live cell imaging, although these dyes are less photostable than red fluorescent dyes. When multicolor imaging is required, we recommend the following dye combinations for optimal results: Alexa Fluor 488 (green, Cy3 (red, and Cy5 or CF640 (far-red.

  2. Green synthesis of peptide-templated fluorescent copper nanoclusters for temperature sensing and cellular imaging.

    Science.gov (United States)

    Huang, Hong; Li, Hua; Wang, Ai-Jun; Zhong, Shu-Xian; Fang, Ke-Ming; Feng, Jiu-Ju

    2014-12-21

    A simple and green approach was developed for the preparation of fluorescent Cu nanoclusters (NCs) using the artificial peptide CLEDNN as a template. The as-synthesized Cu NCs exhibited a high fluorescence quantum yield (7.3%) and good stability, along with excitation and temperature dependent fluorescent properties, which could be employed for temperature sensing. Further investigations demonstrated low toxicity of Cu NCs for cellular imaging.

  3. Time-domain imaging with quench-based fluorescent contrast agents

    Science.gov (United States)

    Akers, Walter J.; Solomon, Metasebya; Sudlow, Gail P.; Berezin, Mikhail; Achilefu, Samuel

    2012-03-01

    Quench-based probes utilize unique characteristics of fluorescence resonance energy transfer (FRET) to enhance contrast upon de-quenching. This mechanism has been used in a variety of molecular probes for imaging of cancer related enzyme activity such as matrix metalloproteinases, cathepsins and caspases. While non-fluorescent upon administration, fluorescence can be restored by separation of donor and acceptor, resulting in higher intensity in the presence of activator. Along with decreased quantum yield, FRET also results in altered fluorescence lifetime. Time-domain imaging can further enhance contrast and information yield from quench-based probes. We present in vivo time-domain imaging for detecting activation of quench-based probes. Quench-based probes utilize unique characteristics of fluorescence resonance energy transfer (FRET) to enhance contrast upon de-quenching. This mechanism has been used in a variety of molecular probes for imaging of cancer related enzyme activity such as matrix metalloproteinases, cathepsins and caspases. While non-fluorescent upon administration, fluorescence can be restored by separation of donor and acceptor, resulting in higher intensity in the presence of activator. Along with decreased quantum yield, FRET also results in altered fluorescence lifetime. Time-domain imaging can further enhance contrast and information yield from quench-based probes. We present in vivo time-domain imaging for detecting activation of quench-based probes. Time-domain diffuse optical imaging was performed to assess the FRET and quenching in living mice with orthotopic breast cancer. Tumor contrast enhancement was accompanied by increased fluorescence lifetime after administration of quenched probes selective for matrix metalloproteinases while no significant change was observed for non-quenched probes for integrin receptors. These results demonstrate the utility of timedomain imaging for detection of cancer-related enzyme activity in vivo.

  4. Precise diagnosis in different scenarios using photoacoustic and fluorescence imaging with dual-modality nanoparticles.

    Science.gov (United States)

    Peng, Dong; Du, Yang; Shi, Yiwen; Mao, Duo; Jia, Xiaohua; Li, Hui; Zhu, Yukun; Wang, Kun; Tian, Jie

    2016-08-14

    Photoacoustic imaging and fluorescence molecular imaging are emerging as important research tools for biomedical studies. Photoacoustic imaging offers both strong optical absorption contrast and high ultrasonic resolution, and fluorescence molecular imaging provides excellent superficial resolution, high sensitivity, high throughput, and the ability for real-time imaging. Therefore, combining the imaging information of both modalities can provide comprehensive in vivo physiological and pathological information. However, currently there are limited probes available that can realize both fluorescence and photoacoustic imaging, and advanced biomedical applications for applying this dual-modality imaging approach remain underexplored. In this study, we developed a dual-modality photoacoustic-fluorescence imaging nanoprobe, ICG-loaded Au@SiO2, which was uniquely designed, consisting of gold nanorod cores and indocyanine green with silica shell spacer layers to overcome fluorophore quenching. This nanoprobe was examined by both PAI and FMI for in vivo imaging on tumor and ischemia mouse models. Our results demonstrated that the nanoparticles can specifically accumulate at the tumor and ischemic areas and be detected by both imaging modalities. Moreover, this dual-modality imaging strategy exhibited superior advantages for a precise diagnosis in different scenarios. The new nanoprobe with the dual-modality imaging approach holds great potential for diagnosis and stage classification of tumor and ischemia related diseases.

  5. Wavefront sensorless approaches to adaptive optics for in vivo fluorescence imaging of mouse retina

    Science.gov (United States)

    Wahl, Daniel J.; Bonora, Stefano; Mata, Oscar S.; Haunerland, Bengt K.; Zawadzki, Robert J.; Sarunic, Marinko V.; Jian, Yifan

    2016-03-01

    Adaptive optics (AO) is necessary to correct aberrations when imaging the mouse eye with high numerical aperture. In order to obtain cellular resolution, we have implemented wavefront sensorless adaptive optics for in vivo fluorescence imaging of mouse retina. Our approach includes a lens-based system and MEMS deformable mirror for aberration correction. The AO system was constructed with a reflectance channel for structural images and fluorescence channel for functional images. The structural imaging was used in real-time for navigation on the retina using landmarks such as blood vessels. We have also implemented a tunable liquid lens to select the retinal layer of interest at which to perform the optimization. At the desired location on the mouse retina, the optimization algorithm used the fluorescence image data to drive a modal hill-climbing algorithm using an intensity or sharpness image quality metric. The optimization requires ~30 seconds to complete a search up to the 20th Zernike mode. In this report, we have demonstrated the AO performance for high-resolution images of the capillaries in a fluorescence angiography. We have also made progress on an approach to AO with pupil segmentation as a possible sensorless technique suitable for small animal retinal imaging. Pupil segmentation AO was implemented on the same ophthalmic system and imaging performance was demonstrated on fluorescent beads with induced aberrations.

  6. Multiphoton fluorescence and second harmonic generation microscopy for imaging keratoconus

    Science.gov (United States)

    Sun, Yen; Lo, Wen; Lin, Sung-Jan; Lin, Wei-Chou; Jee, Shiou-Hwa; Tan, Hsin-Yuan; Dong, Chen-Yuan

    2006-02-01

    The purpose of this study is to assess the possible application of multiphoton fluorescence and second harmonic generation (SHG) microscopy for imaging the structural features of keratoconus cornea and to evaluate its potential as being a clinical in vivo monitoring technique. Using the near-infrared excitation source from a titanium-sapphire laser pumped by a diode-pumped, solid state (DPSS) laser system, we can induce and simultaneously acquire multiphoton autofluorescence and SHG signals from the cornea specimens with keratoconus. A home-modified commercial microscope system with specified optical components is used for optimal signal detection. Keratoconus cornea button from patient with typical clinical presentation of keratoconus was obtained at the time of penetrating keratoplasty. The specimen was also sent for the histological examination as comparison. In all samples of keratoconus, destruction of lamellar structure with altered collagen fiber orientation was observed within whole layer of the diseased stromal area. In addition, the orientation of the altered collagen fibers within the cone area shows a trend directing toward the apex of the cone, which might implicate the biomechanical response of the keratoconus stroma to the intraocular pressure. Moreover, increased autofluorescent cells were also found in the cone area, with increased density as one approaches the apical area. In conclusion, multiphoton autofluorescence and SHG microscopy non-invasively demonstrated the morphological features of keratoconus cornea, especially the structural alternations of the stromal lamellae. We believe that in the future the multiphoton microscopy can be applied in vivo as an effective, non-invasive diagnostic and monitoring technique for keratoconus.

  7. Submicron hard X-ray fluorescence imaging of synthetic elements

    Energy Technology Data Exchange (ETDEWEB)

    Jensen, Mark P., E-mail: mjensen@anl.gov [Chemical Sciences and Engineering Division, Argonne National Laboratory, Argonne, IL 60439 (United States); Aryal, Baikuntha P. [Chemical Sciences and Engineering Division, Argonne National Laboratory, Argonne, IL 60439 (United States); Department of Chemistry, University of Chicago, Chicago, IL 60637 (United States); Gorman-Lewis, Drew [Chemical Sciences and Engineering Division, Argonne National Laboratory, Argonne, IL 60439 (United States); Paunesku, Tatjana [Department of Radiation Oncology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611 (United States); Department of Radiology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611 (United States); Lai, Barry; Vogt, Stefan [X-ray Science Division, Advanced Photon Source, Argonne National Laboratory, Argonne, IL 60439 (United States); Woloschak, Gayle E. [Department of Radiation Oncology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611 (United States); Department of Radiology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611 (United States)

    2012-04-13

    Highlights: Black-Right-Pointing-Pointer Actinide elements are mapped with L-edge X-rays and better than 400 nm resolution. Black-Right-Pointing-Pointer A typical detection limit was 2.9 Multiplication-Sign 10{sup -20} moles Pu {mu}m{sup -2}. Black-Right-Pointing-Pointer XANES measurements provide chemical information in 0.1 {mu}m{sup 2} spots. Black-Right-Pointing-Pointer Selection of materials for encapsulation is important for avoiding interferences. - Abstract: Synchrotron-based X-ray fluorescence microscopy (XFM) using hard X-rays focused into sub-micron spots is a powerful technique for elemental quantification and mapping, as well as microspectroscopic measurements such as {mu}-XANES (X-ray absorption near edge structure). We have used XFM to image and simultaneously quantify the transuranic element plutonium at the L{sub 3} or L{sub 2}-edge as well as Th and lighter biologically essential elements in individual rat pheochromocytoma (PC12) cells after exposure to the long-lived plutonium isotope {sup 242}Pu. Elemental maps demonstrate that plutonium localizes principally in the cytoplasm of the cells and avoids the cell nucleus, which is marked by the highest concentrations of phosphorus and zinc, under the conditions of our experiments. The minimum detection limit under typical acquisition conditions with an incident X-ray energy of 18 keV for an average 202 {mu}m{sup 2} cell is 1.4 fg Pu or 2.9 Multiplication-Sign 10{sup -20} moles Pu {mu}m{sup -2}, which is similar to the detection limit of K-edge XFM of transition metals at 10 keV. Copper electron microscopy grids were used to avoid interference from gold X-ray emissions, but traces of strontium present in naturally occurring calcium can still interfere with plutonium detection using its L{sub {alpha}} X-ray emission.

  8. 一种免疫荧光图像减影技术在鉴定drebrin E亚型的应用%Application of an immunofluorescence image subtraction technique in identification of drebrin isoform E

    Institute of Scientific and Technical Information of China (English)

    宋明桥; 迟晓冬; 郑文旭; 刘萍

    2011-01-01

    Objective; To identify the drebrin isoform E by a novel immunofluorescence image subtraction technique without drebrin E-specific antibody. Methods; We used drebrin-non-specific antibody M2F6 and drebrin A-specific antibody DAS2 to immunostain the frozen sections of the adult rat brain by double immunofluorescence staining. Then the two immunostaining images of the same section were processed with the image subtraction technique using a computer image processing software, and the expression of drebrin E in the adult rat brain was observed. Results; We obtained the images of drebrin isoform E by the immunofluorescence image subtraction technique. The expression of drebrin E in the rostral migratory stream, the dentate gyrus of the hippocampus and the piriform cortex of the adult rat brain was identified. Conclusion; The immunofluorescence image subtraction technique can identify the drebrin isoform E, and offer a novel approach to identify the proteins which have two isoforms similar to drebrin.%目的:在缺乏drebrin E抗体的情况下利用一种新的免疫荧光计算机图像减影技术的方法鉴定drebrin E亚型.方法:应用drebrin非特异性抗体M2F6和drebrin A特异性抗体DAS2对成年大鼠脑冰冻切片进行双重免疫荧光染色,然后用计算机图像处理软件对同一切片的两种染色照片进行图像减影技术处理,观察drebrin E在成年大鼠脑中的分布情况.结果:通过免疫荧光计算机图像减影技术得到了drebrin E亚型的图像,确认了drebrin E在成年大鼠脑中吻侧辽移流、海马齿状回、梨状皮质的分布.结论:利用免疫荧光图像减影技术可以实现对drebrin E亚型的鉴定,为类似drebrin这样存在两种亚型的蛋白质鉴定提供了一种新的方法.

  9. Time-Resolved Fluorescence Spectroscopy and Fluorescence Lifetime Imaging Microscopy for Characterization of Dendritic Polymer Nanoparticles and Applications in Nanomedicine

    Directory of Open Access Journals (Sweden)

    Alexander Boreham

    2016-12-01

    Full Text Available The emerging field of nanomedicine provides new approaches for the diagnosis and treatment of diseases, for symptom relief and for monitoring of disease progression. One route of realizing this approach is through carefully constructed nanoparticles. Due to the small size inherent to the nanoparticles a proper characterization is not trivial. This review highlights the application of time-resolved fluorescence spectroscopy and fluorescence lifetime imaging microscopy (FLIM for the analysis of nanoparticles, covering aspects ranging from molecular properties to particle detection in tissue samples. The latter technique is particularly important as FLIM allows for distinguishing of target molecules from the autofluorescent background and, due to the environmental sensitivity of the fluorescence lifetime, also offers insights into the local environment of the nanoparticle or its interactions with other biomolecules. Thus, these techniques offer highly suitable tools in the fields of particle development, such as organic chemistry, and in the fields of particle application, such as in experimental dermatology or pharmaceutical research.

  10. A fluorescent probe for imaging p53-MDM2 protein-protein interaction.

    Science.gov (United States)

    Liu, Zhenzhen; Miao, Zhenyuan; Li, Jin; Fang, Kun; Zhuang, Chunlin; Du, Lupei; Sheng, Chunquan; Li, Minyong

    2015-04-01

    In this article, we describe a no-wash small-molecule fluorescent probe for detecting and imaging p53-MDM2 protein-protein interaction based on an environment-sensitive fluorescent turn-on mechanism. After extensive biological examination, this probe L1 exhibited practical activity and selectivity in vitro and in cellulo.

  11. Fluorescence imaging of single Kinesin motors on immobilized microtubules.

    Science.gov (United States)

    Korten, Till; Nitzsche, Bert; Gell, Chris; Ruhnow, Felix; Leduc, Cécile; Diez, Stefan

    2011-01-01

    Recent developments in optical microscopy and nanometer tracking have greatly improved our understanding of cytoskeletal motor proteins. Using fluorescence microscopy, dynamic interactions are now routinely observed in vitro on the level of single molecules mainly using a geometry, where fluorescently labeled motors move on surface-immobilized filaments. In this chapter, we review recent methods related to single-molecule kinesin motility assays. In particular, we aim to provide practical advice on: how to set up the assays, how to acquire high-precision data from fluorescently labeled kinesin motors and attached quantum dots, and how to analyze data by nanometer tracking.

  12. Spectral phasor analysis allows rapid and reliable unmixing of fluorescence microscopy spectral images

    NARCIS (Netherlands)

    Fereidouni, F.; Bader, A.N.; Gerritsen, H.C.

    2012-01-01

    A new global analysis algorithm to analyse (hyper-) spectral images is presented. It is based on the phasor representation that has been demonstrated to be very powerful for the analysis of lifetime imaging data. In spectral phasor analysis the fluorescence spectrum of each pixel in the image is Fou

  13. Evaluating the use of fluorescent imaging for the quantification of dental fluorosis.

    Science.gov (United States)

    McGrady, Michael G; Ellwood, Roger P; Taylor, Andrew; Maguire, Anne; Goodwin, Michaela; Boothman, Nicola; Pretty, Iain A

    2012-11-01

    The quantification of fluorosis using fluorescence imaging (QLF) hardware and stain analysis software has been demonstrated in selected populations with good correlation between fluorescent image metrics and TF Index scores from photographs. The aim of this study was to evaluate the ability of QLF to quantify fluorosis in a population of subjects (aged 11-13) participating in an epidemiological caries and fluorosis survey in fluoridated and non-fluoridated communities in Northern England. Fluorescent images of the maxillary incisors were captured together with standardized photographs were scored blind for fluorosis using the TF Index. Subjects were excluded from the analysis if there were restorations or caries on the maxillary central incisors. Data were available for 1774 subjects (n=905 Newcastle, n=869 Manchester). The data from the fluorescence method demonstrated a significant correlation with TF Index scores from photographs (Kendall's tau = 0.332 pfluorosis or at low levels of fluorosis severity had an adverse impact on tooth fluorescence and hence the outcome variable. This in conjunction with an uneven distribution of subjects across the range of fluorosis presentations may have resulted in the lower than anticipated correlations between the fluorescent imaging metrics and the photographic fluorosis scores. Nevertheless, the fluorescence imaging technique was able to discriminate between a fluoridated and non-fluoridated population (pfluorosis when used adjunctively with photographic scoring.

  14. Determination of the modulation transfer function for a time-gated fluorescence imaging system.

    Science.gov (United States)

    Gundy, Sarah; Van der Putten, Wil; Shearer, Andy; Buckton, Daniel; Ryder, Alan G

    2004-01-01

    The use of fluorescence for cancer detection is currently under investigation. Presently, steady-state fluorescence detection methods are in use, but have limitations due to poor contrast between the fluorescence of the tumor and background autofluorescence. Improved contrast can be obtained with time-resolved techniques because of the differing lifetimes between autofluorescence and exogenous photosensitizers that selectively accumulate within tumor tissue. An imaging system is constructed using a fast-gated (200-ps) charge-coupled device (CCD) camera and a pulsed 635-nm laser diode. To characterize the ability of the system to transfer object contrast to an image, the modulation transfer function (MTF) of the system is acquired by employing an extended knife-edge technique. A knife-edge target is assembled by drilling a rectangular well into a block of polymethyl methacrylate (PMMA). The imaging system records images of the photosensitizer, chloroaluminum phthalocyanine tetrasulfonate (AlPcTS), within the well. AlPcTS was chosen to test the system because of its strong absorption of 635-nm, high fluorescence yield, and relatively long fluorescence lifetime (approximately 7.5 ns). The results show that the system is capable of resolving 10(-4) M AlPcTS fluorescence as small as 1 mm. The findings of this study contribute to the development of a time-gated imaging system using fluorescence lifetimes. Copyright 2004 Society of Photo-Optical Instrumentation Engineers.

  15. B-Spline potential function for maximum a-posteriori image reconstruction in fluorescence microscopy

    Directory of Open Access Journals (Sweden)

    Shilpa Dilipkumar

    2015-03-01

    Full Text Available An iterative image reconstruction technique employing B-Spline potential function in a Bayesian framework is proposed for fluorescence microscopy images. B-splines are piecewise polynomials with smooth transition, compact support and are the shortest polynomial splines. Incorporation of the B-spline potential function in the maximum-a-posteriori reconstruction technique resulted in improved contrast, enhanced resolution and substantial background reduction. The proposed technique is validated on simulated data as well as on the images acquired from fluorescence microscopes (widefield, confocal laser scanning fluorescence and super-resolution 4Pi microscopy. A comparative study of the proposed technique with the state-of-art maximum likelihood (ML and maximum-a-posteriori (MAP with quadratic potential function shows its superiority over the others. B-Spline MAP technique can find applications in several imaging modalities of fluorescence microscopy like selective plane illumination microscopy, localization microscopy and STED.

  16. U-SPECT-BioFluo: an integrated radionuclide, bioluminescence, and fluorescence imaging platform

    NARCIS (Netherlands)

    Van Oosterom, M.N.; Kreuger, R.; Buckle, T.; Mahn, W.A.; Bunschoten, A.; Josephson, L.; Van Leeuwen, F.W.B.; Beekman, F.J.

    2014-01-01

    Background: In vivo bioluminescence, fluorescence, and single-photon emission computed tomography (SPECT) imaging provide complementary information about biological processes. However, to date these signatures are evaluated separately on individual preclinical systems. In this paper, we introduce a

  17. Endoscopic image-guided thermal therapy using targeted near infrared fluorescent gold nanorods (Conference Presentation)

    Science.gov (United States)

    Elson, Daniel S.

    2016-09-01

    We present an in vivo study of endoscopic fluorescence image-guided photothermal therapy of human oesophageal adenocarcinoma in a murine xenograft model, using intratumoural or intravenous gold nanorods functionalised with Cy5.5 and EGFR.

  18. Fluorescence cell imaging and manipulation using conventional halogen lamp microscopy.

    Directory of Open Access Journals (Sweden)

    Kazuo Yamagata

    Full Text Available Technologies for vitally labeling cells with fluorescent dyes have advanced remarkably. However, to excite fluorescent dyes currently requires powerful illumination, which can cause phototoxic damage to the cells and increases the cost of microscopy. We have developed a filter system to excite fluorescent dyes using a conventional transmission microscope equipped with a halogen lamp. This method allows us to observe previously invisible cell organelles, such as the metaphase spindle of oocytes, without causing phototoxicity. Cells remain healthy even after intensive manipulation under fluorescence observation, such as during bovine, porcine and mouse somatic cell cloning using nuclear transfer. This method does not require expensive epifluorescence equipment and so could help to reduce the science gap between developed and developing countries.

  19. Digital contrast subtraction radiography for proximal caries diagnosis

    Energy Technology Data Exchange (ETDEWEB)

    Kang, Byung Cheol; Yoon, Suk Ja [Department of Dental Radiology, Chonnam National University Hospital, Gwangju (Korea, Republic of)

    2002-06-15

    To determine whether subtraction images utilizing contrast media can improve the diagnostic performance of proximal caries diagnosis compared to conventional periapical radiographic images. Thirty-six teeth with 57 proximal surfaces were radiographied using a size no.2 RVG-ui sensor (Trophy Radiology, Marne-la-Vallee, France). The teeth immersed in water-soluble contrast media and subtraction images were taken. Each tooth was then sectioned for histologic examination. The digital radiographic images and subtraction images were examined and interpreted by three dentists for proximal caries. The results of the proximal caries diagnosis were then verified with the results of the histologic examination. The proximal caries sensitivity using digital subtraction radiography was significantly higher than simply examining a single digital radiograph. The sensitivity of the proximal dentinal carious lesion when analyzed with the subtraction radiograph and the radiograph together was higher than with the subtraction radiograph or the radiograph alone. The use of subtraction radiography with contrast media may be useful for detecting proximal dentinal carious lesions.

  20. Live imaging of Tribolium castaneum embryonic development using light-sheet-based fluorescence microscopy.

    Science.gov (United States)

    Strobl, Frederic; Schmitz, Alexander; Stelzer, Ernst H K

    2015-10-01

    Tribolium castaneum has become an important insect model organism for evolutionary developmental biology, genetics and biotechnology. However, few protocols for live fluorescence imaging of Tribolium have been reported, and little image data is available. Here we provide a protocol for recording the development of Tribolium embryos with light-sheet-based fluorescence microscopy. The protocol can be completed in 4-7 d and provides procedural details for: embryo collection, microscope configuration, embryo preparation and mounting, noninvasive live imaging for up to 120 h along multiple directions, retrieval of the live embryo once imaging is completed, and image data processing, for which exemplary data is provided. Stringent quality control criteria for developmental biology studies are also discussed. Light-sheet-based fluorescence microscopy complements existing toolkits used to study Tribolium development, can be adapted to other insect species, and requires no advanced imaging or sample preparation skills.

  1. Fabry-Perot-based Fourier-transform hyperspectral imaging allows multi-labeled fluorescence analysis.

    Science.gov (United States)

    Pisani, Marco; Zucco, Massimo

    2014-05-10

    We demonstrate the ability of our hyperspectral imaging device, based on a scanning Fabry-Perot interferometer, to obtain a single hyper-image of a sample marked with different fluorescent molecules, and to unambiguously discriminate them by observing their spectral fingerprints. An experiment carried out with cyanines, fluorescein, and quantum dots emitting in the yellow-orange region, demonstrates the feasibility of multi-labeled fluorescence microscopy without the use of multiple filter sets or dispersive means.

  2. Comparison of two detection algorithms for spot tracking in fluorescence microscopy images

    CSIR Research Space (South Africa)

    Mabaso, M

    2014-11-01

    Full Text Available for spot tracking in fluorescence microscopy images Matsilele Mabaso∗, Daniel Withey‡, Bhekisipho Twala† ∗ ‡MDS(MIAS) Council for Scientific and Industrial Research Pretoria, South Africa, Email: ∗MMabaso@csir.co.za †Department of Electrical Engineering.... The quantitative comparative results demonstrated the importance of spot detection in tracking contexts. I. INTRODUCTION In recent years, the field of fluorescence microscopy has been improved and automated, and a large volume of image data are being generated...

  3. Green Synthesis of Bifunctional Fluorescent Carbon Dots from Garlic for Cellular Imaging and Free Radical Scavenging.

    Science.gov (United States)

    Zhao, Shaojing; Lan, Minhuan; Zhu, Xiaoyue; Xue, Hongtao; Ng, Tsz-Wai; Meng, Xiangmin; Lee, Chun-Sing; Wang, Pengfei; Zhang, Wenjun

    2015-08-12

    Nitrogen and sulfur codoped carbon dots (CDs) were prepared from garlic by a hydrothermal method. The as-prepared CDs possess good water dispersibility, strong blue fluorescence emission with a fluorescent quantum yield of 17.5%, and excellent photo and pH stabilities. It is also demonstrated that the fluorescence of CDs are resistant to the interference of metal ions, biomolecules, and high ionic strength environments. Combining with low cytotoxicity properties, CDs could be used as an excellent fluorescent probe for cellular multicolor imaging. Moreover, the CDs were also demonstrated to exhibit favorable radical scavenging activity.

  4. Fluorescence imaging of tryptophan and collagen cross-links to evaluate wound closure ex vivo

    Science.gov (United States)

    Wang, Ying; Ortega-Martinez, Antonio; Farinelli, Bill; Anderson, R. R.; Franco, Walfre

    2016-02-01

    Wound size is a key parameter in monitoring healing. Current methods to measure wound size are often subjective, time-consuming and marginally invasive. Recently, we developed a non-invasive, non-contact, fast and simple but robust fluorescence imaging (u-FEI) method to monitor the healing of skin wounds. This method exploits the fluorescence of native molecules to tissue as functional and structural markers. The objective of the present study is to demonstrate the feasibility of using variations in the fluorescence intensity of tryptophan and cross-links of collagen to evaluate proliferation of keratinocyte cells and quantitate size of wound during healing, respectively. Circular dermal wounds were created in ex vivo human skin and cultured in different media. Two serial fluorescence images of tryptophan and collagen cross-links were acquired every two days. Histology and immunohistology were used to validate correlation between fluorescence and epithelialization. Images of collagen cross-links show fluorescence of the exposed dermis and, hence, are a measure of wound area. Images of tryptophan show higher fluorescence intensity of proliferating keratinocytes forming new epithelium, as compared to surrounding keratinocytes not involved in epithelialization. These images are complementary since collagen cross-links report on structure while tryptophan reports on function. HE and immunohistology show that tryptophan fluorescence correlates with newly formed epidermis. We have established a fluorescence imaging method for studying epithelialization processes during wound healing in a skin organ culture model, our approach has the potential to provide a non-invasive, non-contact, quick, objective and direct method for quantitative measurements in wound healing in vivo.

  5. Colourful FKS subtraction

    CERN Document Server

    Frixione, Stefano

    2011-01-01

    I formulate in a colour-friendly way the FKS method for the computation of QCD cross sections at the next-to-leading order accuracy. This is achieved through the definition of subtraction terms for squared matrix elements, constructed with single colour-dressed or pairs of colour-ordered amplitudes. The latter approach relies on the use of colour flows, is exact to all orders in $N$, and is thus particularly suited to being organized as a systematic expansion in 1/N.

  6. Fluorescent imaging of single nanoparticles and viruses on a smart phone.

    Science.gov (United States)

    Wei, Qingshan; Qi, Hangfei; Luo, Wei; Tseng, Derek; Ki, So Jung; Wan, Zhe; Göröcs, Zoltán; Bentolila, Laurent A; Wu, Ting-Ting; Sun, Ren; Ozcan, Aydogan

    2013-10-22

    Optical imaging of nanoscale objects, whether it is based on scattering or fluorescence, is a challenging task due to reduced detection signal-to-noise ratio and contrast at subwavelength dimensions. Here, we report a field-portable fluorescence microscopy platform installed on a smart phone for imaging of individual nanoparticles as well as viruses using a lightweight and compact opto-mechanical attachment to the existing camera module of the cell phone. This hand-held fluorescent imaging device utilizes (i) a compact 450 nm laser diode that creates oblique excitation on the sample plane with an incidence angle of ~75°, (ii) a long-pass thin-film interference filter to reject the scattered excitation light, (iii) an external lens creating 2× optical magnification, and (iv) a translation stage for focus adjustment. We tested the imaging performance of this smart-phone-enabled microscopy platform by detecting isolated 100 nm fluorescent particles as well as individual human cytomegaloviruses that are fluorescently labeled. The size of each detected nano-object on the cell phone platform was validated using scanning electron microscopy images of the same samples. This field-portable fluorescence microscopy attachment to the cell phone, weighing only ~186 g, could be used for specific and sensitive imaging of subwavelength objects including various bacteria and viruses and, therefore, could provide a valuable platform for the practice of nanotechnology in field settings and for conducting viral load measurements and other biomedical tests even in remote and resource-limited environments.

  7. In vivo tomographic imaging with fluorescence and MRI using tumor-targeted dual-labeled nanoparticles

    Directory of Open Access Journals (Sweden)

    Zhang Y

    2013-12-01

    Full Text Available Yue Zhang,1 Bin Zhang,1 Fei Liu,1,2 Jianwen Luo,1,3 Jing Bai1 1Department of Biomedical Engineering, School of Medicine, 2Tsinghua-Peking Center for Life Sciences, 3Center for Biomedical Imaging Research, Tsinghua University, Beijing, People's Republic of China Abstract: Dual-modality imaging combines the complementary advantages of different modalities, and offers the prospect of improved preclinical research. The combination of fluorescence imaging and magnetic resonance imaging (MRI provides cross-validated information and direct comparison between these modalities. Here, we report on the application of a novel tumor-targeted, dual-labeled nanoparticle (NP, utilizing iron oxide as the MRI contrast agent and near infrared (NIR dye Cy5.5 as the fluorescent agent. Results of in vitro experiments verified the specificity of the NP to tumor cells. In vivo tumor targeting and uptake of the NPs in a mouse model were visualized by fluorescence and MR imaging collected at different time points. Quantitative analysis was carried out to evaluate the efficacy of MRI contrast enhancement. Furthermore, tomographic images were also acquired using both imaging modalities and cross-validated information of tumor location and size between these two modalities was revealed. The results demonstrate that the use of dual-labeled NPs can facilitate the dual-modal detection of tumors, information cross-validation, and direct comparison by combing fluorescence molecular tomography (FMT and MRI. Keywords: dual-modality, fluorescence molecular tomography (FMT, magnetic resonance imaging (MRI, nanoparticle

  8. In-vivo optical detection of cancer using chlorin e6 – polyvinylpyrrolidone induced fluorescence imaging and spectroscopy

    Directory of Open Access Journals (Sweden)

    Soo Khee

    2009-01-01

    Full Text Available Abstract Background Photosensitizer based fluorescence imaging and spectroscopy is fast becoming a promising approach for cancer detection. The purpose of this study was to examine the use of the photosensitizer chlorin e6 (Ce6 formulated in polyvinylpyrrolidone (PVP as a potential exogenous fluorophore for fluorescence imaging and spectroscopic detection of human cancer tissue xenografted in preclinical models as well as in a patient. Methods Fluorescence imaging was performed on MGH human bladder tumor xenografted on both the chick chorioallantoic membrane (CAM and the murine model using a fluorescence endoscopy imaging system. In addition, fiber optic based fluorescence spectroscopy was performed on tumors and various normal organs in the same mice to validate the macroscopic images. In one patient, fluorescence imaging was performed on angiosarcoma lesions and normal skin in conjunction with fluorescence spectroscopy to validate Ce6-PVP induced fluorescence visual assessment of the lesions. Results Margins of tumor xenografts in the CAM model were clearly outlined under fluorescence imaging. Ce6-PVP-induced fluorescence imaging yielded a specificity of 83% on the CAM model. In mice, fluorescence intensity of Ce6-PVP was higher in bladder tumor compared to adjacent muscle and normal bladder. Clinical results confirmed that fluorescence imaging clearly captured the fluorescence of Ce6-PVP in angiosarcoma lesions and good correlation was found between fluorescence imaging and spectral measurement in the patient. Conclusion Combination of Ce6-PVP induced fluorescence imaging and spectroscopy could allow for optical detection and discrimination between cancer and the surrounding normal tissues. Ce6-PVP seems to be a promising fluorophore for fluorescence diagnosis of cancer.

  9. Non-fused phospholes as fluorescent probes for imaging of lipid droplets in living cells

    Science.gov (United States)

    Öberg, Elisabet; Appelqvist, Hanna; Nilsson, K. Peter R.

    2017-04-01

    Molecular tools for fluorescent imaging of specific compartments in cells are essential for understanding the function and activity of cells. Here, we report the synthesis of a series of pyridyl- and thienyl-substituted phospholes and the evaluation of these dyes for fluorescent imaging of cells. The thienyl-substituted phospholes proved to be successful for staining of cultured normal and malignant cells due to their fluorescent properties and low toxicity. Co-staining experiments demonstrated that these probes target lipid droplets, which are, lipid-storage organelles found in the cytosol of nearly all cell types. Our findings confirm that thienyl-substituted phospholes can be utilized as fluorescent tools for vital staining of cells, and we foresee that these fluorescent dyes might be used in studies to unravel the roles that lipid droplets play in cellular physiology and their role in diseases.

  10. In Vivo Imaging of Far-red Fluorescent Proteins after DNA Electrotransfer to Muscle Tissue

    Directory of Open Access Journals (Sweden)

    Hojman Pernille

    2009-04-01

    Full Text Available Abstract DNA electrotransfer to muscle tissue yields long-term, high levels of gene expression; showing great promise for future gene therapy. We want to characterize the novel far-red fluorescent protein Katushka as a marker for gene expression using time domain fluorescence in vivo imaging. Highly efficient transgenic expression was observed after DNA electrotransfer with 100-fold increase in fluorescent intensity. The fluorescent signal peaked 1 week after transfection and returned to background level within 4 weeks. Katushka expression was not as stable as GFP expression, which was detectable for 8 weeks. Depth and 3D analysis proved that the expression was located in the target muscle. In vivo bio-imaging using the novel Katushka fluorescent protein enables excellent evaluation of the transfection efficacy, and spatial distribution, but lacks long-term stability.

  11. Silica nanocapsules of fluorescent conjugated polymers and superparamagnetic nanocrystals for dual-mode cellular imaging.

    Science.gov (United States)

    Tan, Happy; Wang, Miao; Yang, Chang-Tong; Pant, Shilpa; Bhakoo, Kishore Kumar; Wong, Siew Yee; Chen, Zhi-Kuan; Li, Xu; Wang, John

    2011-06-01

    We describe here a facile and benign synthetic strategy to integrate the fluorescent behavior of conjugated polymers and superparamagnetic properties of iron oxide nanocrystals into silica nanocapsules, forming a new type of bifunctional magnetic fluorescent silica nanocapsule (BMFSN). The resultant BMFSNs are uniform, colloidally stable in aqueous medium, and exhibit the desired dual functionality of fluorescence and superparamagnetism in a single entity. Four conjugated polymers with different emissions were used to demonstrate the versatility of employing this class of fluorescent materials for the preparation of BMFSNs. The applicability of BMFSNs in cellular imaging was studied by incubating them with human liver cancer cells, the result of which demonstrated that the cells could be visualized by dual-mode fluorescence and magnetic resonance imaging. Furthermore, the superparamagnetic behavior of the BMFSNs was exploited for in vitro magnetic-guided delivery of the nanocapsules into the cancer cells, thereby highlighting their potential for targeting biomedical applications.

  12. In Vivo Imaging of Far-red Fluorescent Proteins after DNA Electrotransfer to Muscle Tissue

    DEFF Research Database (Denmark)

    Hojman, Pernille; Eriksen, Jens; Gehl, Julie

    2009-01-01

    DNA electrotransfer to muscle tissue yields long-term, high levels of gene expression; showing great promise for future gene therapy. We want to characterize the novel far-red fluorescent protein Katushka as a marker for gene expression using time domain fluorescence in vivo imaging. Highly...... weeks. Depth and 3D analysis proved that the expression was located in the target muscle. In vivo bio-imaging using the novel Katushka fluorescent protein enables excellent evaluation of the transfection efficacy, and spatial distribution, but lacks long-term stability....

  13. A Novel Molecular Fluorescent Technique for Imaging the Somatostatin Receptor 2, Using a DOTATOC Lanthanide Conjugate

    DEFF Research Database (Denmark)

    Andersen, Rune Wiik; Prakash, Vineet; Stensballe, Allan

    to easily obtain commercial receptor antibodies. We propose an alternative with the novel use of lanthanide fluorescent DOTATOC imaging.Purpose is to prove that it is feasible to combine the fluorescent nuclear imaging of neuroendocrine tumors with histopathological correlates using the same bio......-functional DOTATOC component.                       METHOD AND MATERIALS            The chelation of Europium and Samarium to DOTATOC was proven using MALDI-TOF Mass Spectrometry. The rise in quantum yield between unchelated lanthanides and those bound by DOTATOC was examined using fluorescence spectroscopy...

  14. Bright field microscopy as an alternative to whole cell fluorescence in automated analysis of macrophage images.

    Directory of Open Access Journals (Sweden)

    Jyrki Selinummi

    Full Text Available BACKGROUND: Fluorescence microscopy is the standard tool for detection and analysis of cellular phenomena. This technique, however, has a number of drawbacks such as the limited number of available fluorescent channels in microscopes, overlapping excitation and emission spectra of the stains, and phototoxicity. METHODOLOGY: We here present and validate a method to automatically detect cell population outlines directly from bright field images. By imaging samples with several focus levels forming a bright field -stack, and by measuring the intensity variations of this stack over the -dimension, we construct a new two dimensional projection image of increased contrast. With additional information for locations of each cell, such as stained nuclei, this bright field projection image can be used instead of whole cell fluorescence to locate borders of individual cells, separating touching cells, and enabling single cell analysis. Using the popular CellProfiler freeware cell image analysis software mainly targeted for fluorescence microscopy, we validate our method by automatically segmenting low contrast and rather complex shaped murine macrophage cells. SIGNIFICANCE: The proposed approach frees up a fluorescence channel, which can be used for subcellular studies. It also facilitates cell shape measurement in experiments where whole cell fluorescent staining is either not available, or is dependent on a particular experimental condition. We show that whole cell area detection results using our projected bright field images match closely to the standard approach where cell areas are localized using fluorescence, and conclude that the high contrast bright field projection image can directly replace one fluorescent channel in whole cell quantification. Matlab code for calculating the projections can be downloaded from the supplementary site: http://sites.google.com/site/brightfieldorstaining.

  15. Applying fluorescence lifetime imaging microscopy to evaluate the efficacy of anticancer drugs

    Science.gov (United States)

    Kawanabe, Satoshi; Araki, Yoshie; Uchimura, Tomohiro; Imasaka, Totaro

    2015-06-01

    Fluorescence lifetime imaging microscopy was applied to evaluate the efficacy of anticancer drugs. A decrease in the fluorescence lifetime of the nucleus in apoptotic cancer cells stained by SYTO 13 dye was detected after treatment with antitumor antibiotics such as doxorubicin or epirubicin. It was confirmed that the change in fluorescence lifetime occurred earlier than morphological changes in the cells. We found that the fluorescence lifetime of the nucleus in the cells treated with epirubicin decreased more rapidly than that of the cells treated with doxorubicin. This implies that epirubicin was more efficacious than doxorubicin in the treatment of cancer cells. The change in fluorescence lifetime was, however, not indicated when the cells were treated with cyclophosphamide. The decrease in fluorescence lifetime was associated with the processes involving caspase activation and chromatin condensation. Therefore, this technique would provide useful information about apoptotic cells, particularly in the early stages.

  16. An endoscopic fluorescence imaging system for simultaneous visual examination and photodetection of cancers

    Science.gov (United States)

    Wagnières, Georges A.; Studzinski, André P.; van den Bergh, Hubert E.

    1997-01-01

    We describe the design and performance tested during six years of clinical trials of a fluorescence endoscope for the detection and delineation of cancers in several hollow organs. The apparatus is based on the imaging of the laser-induced fluorescence that differs between a tumor and its surrounding normal tissue. The tests are carried out in the upper aerodigestive tract, the tracheobronchial tree, the esophagus, and the colon. In the three former cases an exogenous dye is used (Photofrin II), whereas in the latter case fluorescein molecules conjugated with monoclonal antibodies directed against carcinoembryonic antigen are injected. The decrease of native tissue autofluorescence observed in early cancers is also used for detecting lesions in the tracheobronchial tree. The fluorescence contrast between the tumor and surrounding normal tissue is enhanced by real time image processing. This is done by simultaneously recording the fluorescence image in two spectral domains, after which these two images are digitized and manipulated with a mathematical operator (look-up table) at video frequency. Moreover, the device that is described below allows for an immediate observation of the endoscopic area under white light illumination during fluorescence detection in order to localize the origin of the "positive" fluorescence signals. Typical results obtained in the tracheobronchial tree and in the colon are presented and the sources of false positives and false negatives are evaluated in terms of the fluorescent dye, tissue optical properties, and illumination optics.

  17. Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors.

    Science.gov (United States)

    Lee, Sungmoo; Piao, Hong Hua; Sepheri-Rad, Masoud; Jung, Arong; Sung, Uhna; Song, Yoon-Kyu; Baker, Bradley J

    2016-02-04

    Genetically encoded voltage indicators (GEVIs) have improved to the point where they are beginning to be useful for in vivo recordings. While the ultimate goal is to image neuronal activity in vivo, one must be able to image activity of a single cell to ensure successful in vivo preparations. This procedure will describe how to image membrane potential in a single cell to provide a foundation to eventually image in vivo. Here we describe methods for imaging GEVIs consisting of a voltage-sensing domain fused to either a single fluorescent protein (FP) or two fluorescent proteins capable of Förster resonance energy transfer (FRET) in vitro. Using an image splitter enables the projection of images created by two different wavelengths onto the same charge-coupled device (CCD) camera simultaneously. The image splitter positions a second filter cube in the light path. This second filter cube consists of a dichroic and two emission filters to separate the donor and acceptor fluorescent wavelengths depending on the FPs of the GEVI. This setup enables the simultaneous recording of both the acceptor and donor fluorescent partners while the membrane potential is manipulated via whole cell patch clamp configuration. When using a GEVI consisting of a single FP, the second filter cube can be removed allowing the mirrors in the image splitter to project a single image onto the CCD camera.

  18. Fluorescence detection and imaging of amino-functionalized organic monolayer

    Energy Technology Data Exchange (ETDEWEB)

    Shirahata, Naoto [National Institute for Materials Science (NIMS), 1-2-1 Sengen, Tsukuba, Ibaraki 305-0047 (Japan)], E-mail: SHIRAHATA.naoto@nims.go.jp; Furumi, Seiichi [National Institute for Materials Science (NIMS), 1-2-1 Sengen, Tsukuba, Ibaraki 305-0047 (Japan); Masuda, Yoshitake; Hozumi, Atsushi [National Institute of Advanced Industrial Science and Technology (AIST), 2266-98 Anagahora, Shimo-shidami, Moriyama, Nagoya 463-8560 (Japan); Sakka, Yoshio [National Institute for Materials Science (NIMS), 1-2-1 Sengen, Tsukuba, Ibaraki 305-0047 (Japan)

    2008-03-03

    Amino-terminated organic monolayer formed on silicon covered with native oxide (SiO{sub 2}/Si) was directly visualized under observation with fluorescent microscopy. This successful fluorescence visualization was achieved by a combination of fluorescamine method and photopatterning of the amino-terminated surface. As a typical example, an amino-terminated self-assembled monolayer (SAM) was formed on SiO{sub 2}/Si substrate in a vapor of 12.5 vol.% solution of N-(6-aminohexyl)-3-aminopropyltrimethoxysilane [H{sub 2}N(CH{sub 2}){sub 6}NH(CH{sub 2}){sub 3}Si(OCH{sub 3}){sub 3}, AHAPS] diluted with absolute toluene. A micropattern of AHAPS-SAM was photolithographycally prepared using 172 nm vacuum ultraviolet (VUV) light under a reduced pressure of 10 Pa for 30 min through a photomask. The resultant micropattern composed of AHAPS- and SiOH-covered regions was provided to fluorescamine method. Due to a nonluminescence of fluorescamine itself under UV/visible irradiation, a fluorescent emission could not be observed on SiOH regions of the micropattern. In contrast, fluorescamine reacted with the outermost amino group of the AHAPS-SAM to give a fluorescent emission. A comprehensible fluorescence method for verifying formation of an amino-terminated organic monolayer has been developed.

  19. Monitoring photosensitizer uptake using two photon fluorescence lifetime imaging microscopy.

    Science.gov (United States)

    Yeh, Shu-Chi Allison; Diamond, Kevin R; Patterson, Michael S; Nie, Zhaojun; Hayward, Joseph E; Fang, Qiyin

    2012-01-01

    Photodynamic Therapy (PDT) provides an opportunity for treatment of various invasive tumors by the use of a cancer targeting photosensitizing agent and light of specific wavelengths. However, real-time monitoring of drug localization is desirable because the induction of the phototoxic effect relies on interplay between the dosage of localized drug and light. Fluorescence emission in PDT may be used to monitor the uptake process but fluorescence intensity is subject to variability due to scattering and absorption; the addition of fluorescence lifetime may be beneficial to probe site-specific drug-molecular interactions and cell damage. We investigated the fluorescence lifetime changes of Photofrin(®) at various intracellular components in the Mat-LyLu (MLL) cell line. The fluorescence decays were analyzed using a bi-exponential model, followed by segmentation analysis of lifetime parameters. When Photofrin(®) was localized at the cell membrane, the slow lifetime component was found to be significantly shorter (4.3 ± 0.5 ns) compared to those at other locations (cytoplasm: 7.3 ± 0.3 ns; mitochondria: 7.0 ± 0.2 ns, p < 0.05).

  20. Monitoring Photosensitizer Uptake Using Two Photon Fluorescence Lifetime Imaging Microscopy

    Directory of Open Access Journals (Sweden)

    Shu-Chi Allison Yeh, Kevin R. Diamond, Michael S. Patterson, Zhaojun Nie, Joseph E. Hayward, Qiyin Fang

    2012-01-01

    Full Text Available Photodynamic Therapy (PDT provides an opportunity for treatment of various invasive tumors by the use of a cancer targeting photosensitizing agent and light of specific wavelengths. However, real-time monitoring of drug localization is desirable because the induction of the phototoxic effect relies on interplay between the dosage of localized drug and light. Fluorescence emission in PDT may be used to monitor the uptake process but fluorescence intensity is subject to variability due to scattering and absorption; the addition of fluorescence lifetime may be beneficial to probe site-specific drug-molecular interactions and cell damage. We investigated the fluorescence lifetime changes of Photofrin® at various intracellular components in the Mat-LyLu (MLL cell line. The fluorescence decays were analyzed using a bi-exponential model, followed by segmentation analysis of lifetime parameters. When Photofrin® was localized at the cell membrane, the slow lifetime component was found to be significantly shorter (4.3 ± 0.5 ns compared to those at other locations (cytoplasm: 7.3 ± 0.3 ns; mitochondria: 7.0 ± 0.2 ns, p < 0.05.

  1. 基于人工蜂群算法的彩色铁谱图像背景减除%Color Ferrography Image Background Subtraction Based on Artificial Bee Colony Algorithm

    Institute of Scientific and Technical Information of China (English)

    刘万龙; 王静秋

    2016-01-01

    Aiming at the ferrography image background and wear particle segmentation, this paper proposes the color ferrography image background ( ABCTO) . based on the artificial bee colony algorithm which is used to respectively segment L, a, b three chan-nel images in CIE L∗a∗b color spaces. Then it is operated according to each channel segmentation result, to get the binary image of ferrography image and realize the separation of the wear particle and the background. In order to get the color ferrography image background subtraction, the background pixels of the original image is let to be white. The experimental results prove that this meth-od can be used to precisely and effectively subtract the ferrography image background.%针对铁谱图像背景与磨粒的分割,提出了基于人工蜂群算法的彩色铁谱图像背景减除方法( ABCTO)。该方法是在CIE L∗a∗b∗颜色空间,利用人工蜂群算法分别对L、a、b三通道图进行分割;再通过对各通道分割结果进行操作,得到铁谱图像二值图,实现磨粒与背景的分离;最后将原始图像的背景像素点设置为白色,从而得到减除背景的彩色铁谱图像。实验证明该方法可以精确减除铁谱图像背景,是一种有效的背景减除方法。

  2. Functional brain fluorescence plurimetry in rat by implantable concatenated CMOS imaging system.

    Science.gov (United States)

    Kobayashi, Takuma; Masuda, Hiroyuki; Kitsumoto, Chikara; Haruta, Makito; Motoyama, Mayumi; Ohta, Yasumi; Noda, Toshihiko; Sasagawa, Kiyotaka; Tokuda, Takashi; Shiosaka, Sadao; Ohta, Jun

    2014-03-15

    Measurement of brain activity in multiple areas simultaneously by minimally invasive methods contributes to the study of neuroscience and development of brain machine interfaces. However, this requires compact wearable instruments that do not inhibit natural movements. Application of optical potentiometry with voltage-sensitive fluorescent dye using an implantable image sensor is also useful. However, the increasing number of leads required for the multiple wired sensors to measure larger domains inhibits natural behavior. For imaging broad areas by numerous sensors without excessive wiring, a web-like sensor that can wrap the brain was developed. Kaleidoscopic potentiometry is possible using the imaging system with concatenated sensors by changing the alignment of the sensors. This paper describes organization of the system, evaluation of the system by a fluorescence imaging, and finally, functional brain fluorescence plurimetry by the sensor. The recorded data in rat somatosensory cortex using the developed multiple-area imaging system compared well with electrophysiology results.

  3. Sub-diffraction-limit imaging using mode multiplexing

    Science.gov (United States)

    Wang, Nan; Miyazaki, Jun; He, Jinping; Seto, Keisuke; Kobayashi, Takayoshi

    2015-05-01

    Pixel-by-pixel processed fluorescence difference microscopy is experimentally demonstrated by multiplexing excitation laser beams with Gaussian and donut spot shapes and then demultiplexing the fluorescent signals using lock-in amplifiers. With this scheme, a fixed sample of fluorescent spheres and a slice of mouse brain tissue are imaged with resolutions that exceed the diffraction limit. Compared to previously reported subtraction imaging techniques, this pixel-by-pixel scan can be applied to improve the resolution of a moving sample without introducing subtraction errors. The synchronized signal detection feature makes this method extendible to various applications.

  4. Removal of Out-of-Plane Fluorescence for Single Cell Visualization and Quantification in Cryo-Imaging

    OpenAIRE

    Steyer, Grant J.; Roy, Debashish; Salvado, Olivier; Stone, Meredith E.; Wilson, David L.

    2009-01-01

    We developed a cryo-imaging system, which alternates between sectioning (10–40 μm) and imaging bright field and fluorescence block-face image volumes with micronscale-resolution. For applications requiring single-cell detection of fluorescently labeled cells anywhere in a mouse, we are developing software for reduction of out-of-plane fluorescence. In mouse experiments, we imaged GFP-labeled cancer and stem cells, and cell-sized fluorescent microspheres. To remove out-of-plane fluorescence, w...

  5. Compact multispectral fluorescence imaging system with spectral multiplexed volume holographic grating

    Science.gov (United States)

    Lv, Yanlu; Cai, Chuangjian; Bai, Jing; Luo, Jianwen

    2016-12-01

    Traditional spectral imaging systems mainly rely on spatial scanning or spectral scanning methods to acquire spatial and spectral features. The acquisition is time-consuming and cannot fully satisfy the need of monitoring dynamic phenomenon and observing different structures of the specimen simultaneously. To overcome these barriers, we develop a video-rate simultaneous multispectral imaging system built with a spectral multiplexed volume holographic grating (VHG) and few optical components. Four spectral multiplexed volume holograms optimized for four discrete spectral bands (centered at 488 nm, 530 nm, 590 nm and 620 nm) are recorded into an 8×12 mm photo-thermal refractive glass. The diffraction efficiencies of all the holograms within the multiplexed VHG are greater than 80%. With the high throughout multiplexed VHG, the system can work with both reflection and fluorescence modes and allow simultaneous acquisition of spectral and spatial information with a single exposure. Imaging experiments demonstrate that the multispectral images of the target illuminated with white light source can be obtained. Fluorescence images of multiple fluorescence objects (two glass beads filled with 20 uL 1.0 mg/mL quantum dots solutions that emit 530 +/- 15 nm and 620 +/- 15 nm fluorescence, respectively) buried 3 mm below the surface of a tissue mimicking phantom are acquired. The results demonstrate that the system can provide complementary information in fluorescence imaging. The design diagram of the proposed system is given to explain the advantage of compactness and flexibility in integrating with other imaging platforms.

  6. Fluorescence microscopy imaging of cells with a plasmonic dish integrally molded

    Science.gov (United States)

    Tawa, Keiko; Sasakawa, Chisato; Fujita, Tsuyoshi; Kiyosue, Kazuyuki; Hosokawa, Chie; Nishii, Junji; Oike, Makoto; Kakinuma, Norihiro

    2016-03-01

    A plastic dish with a wavelength-scale periodic structure at a bottom panel was integrally molded and coated with thin metal films. The integrally molded dish called plasmonic dish was applied to bioimaging under a fluorescence microscope. On the plasmonic substrate, the enhanced electric field based on a grating-coupled surface plasmon resonance (GC-SPR) can provide an enhanced fluorescence. In this study, two kinds of cells, human embryonic kidney (HEK) cells and neuronal cells, were observed in our plasmonic dish. Fluorescence images of HEK cells were above 10 times brighter than those obtained on a conventional glass-bottomed dish. Neuronal cells were successfully cultured for 10 d on the plasmonic dish integrally molded, and in fluorescence images with transmitted light, a higher contrast was obtained than in epifluorescence images. The plasmonic dish integrally molded, as well as that fabricated by the UV nanoimprint method, was also found to be useful for sensitive bioimaging.

  7. Fluorescence Modified Chitosan-Coated Magnetic Nanoparticles for High-Efficient Cellular Imaging

    Directory of Open Access Journals (Sweden)

    Nie Fang

    2009-01-01

    Full Text Available Abstract Labeling of cells with nanoparticles for living detection is of interest to various biomedical applications. In this study, novel fluorescent/magnetic nanoparticles were prepared and used in high-efficient cellular imaging. The nanoparticles coated with the modified chitosan possessed a magnetic oxide core and a covalently attached fluorescent dye. We evaluated the feasibility and efficiency in labeling cancer cells (SMMC-7721 with the nanoparticles. The nanoparticles exhibited a high affinity to cells, which was demonstrated by flow cytometry and magnetic resonance imaging. The results showed that cell-labeling efficiency of the nanoparticles was dependent on the incubation time and nanoparticles’ concentration. The minimum detected number of labeled cells was around 104by using a clinical 1.5-T MRI imager. Fluorescence and transmission electron microscopy instruments were used to monitor the localization patterns of the magnetic nanoparticles in cells. These new magneto-fluorescent nanoagents have demonstrated the potential for future medical use.

  8. CEA-targeted nanoparticles allow specific in vivo fluorescent imaging of colorectal cancer models.

    Science.gov (United States)

    Tiernan, James P; Ingram, Nicola; Marston, Gemma; Perry, Sarah L; Rushworth, Jo V; Coletta, P Louise; Millner, Paul A; Jayne, David G; Hughes, Thomas A

    2015-01-01

    Fluorescent imaging of colorectal tumor cells would improve tumor localization and allow intra-operative staging, facilitating stratification of surgical resections thereby improving patient outcomes. We aimed to develop and test fluorescent nanoparticles capable of allowing this in vivo. Dye-doped silica nanoparticles were synthesized. Anti-CEA (carcinoembryonic antigen) or control IgGs were conjugated to nanoparticles using various chemical strategies. Binding of CEA-targeted or control nanoparticles to colorectal cancer cells was quantified in vitro, and in vivo after systemic-delivery to murine xenografts. CEA-targeted, polyamidoamine dendrimer-conjugated, nanoparticles, but not control nanoparticles, allowed strong tumor-specific imaging. We are the first to demonstrate live, specific, in vivo imaging of colorectal cancer cells using antibody-targeted fluorescent nanoparticles. These nanoparticles have potential to allow intra-operative fluorescent visualization of tumor cells.

  9. Imaging Primary Mouse Sarcomas After Radiation Therapy Using Cathepsin-Activatable Fluorescent Imaging Agents

    Energy Technology Data Exchange (ETDEWEB)

    Cuneo, Kyle C. [Department of Radiation Oncology, Duke University School of Medicine, Durham, North Carolina (United States); Mito, Jeffrey K.; Javid, Melodi P. [Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, North Carolina (United States); Ferrer, Jorge M. [Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States); Kim, Yongbaek [Department of Clinical Pathology, College of Veterinary Medicine, Seoul National University, Seoul (Korea, Republic of); Lee, W. David [The David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States); Bawendi, Moungi G. [Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States); Brigman, Brian E. [Department of Orthopedic Surgery, Duke University School of Medicine, Durham, North Carolina (United States); Kirsch, David G., E-mail: david.kirsch@duke.edu [Department of Radiation Oncology, Duke University School of Medicine, Durham, North Carolina (United States); Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, North Carolina (United States)

    2013-05-01

    Purpose: Cathepsin-activated fluorescent probes can detect tumors in mice and in canine patients. We previously showed that these probes can detect microscopic residual sarcoma in the tumor bed of mice during gross total resection. Many patients with soft tissue sarcoma (STS) and other tumors undergo radiation therapy (RT) before surgery. This study assesses the effect of RT on the ability of cathepsin-activated probes to differentiate between normal and cancerous tissue. Methods and Materials: A genetically engineered mouse model of STS was used to generate primary hind limb sarcomas that were treated with hypofractionated RT. Mice were injected intravenously with cathepsin-activated fluorescent probes, and various tissues, including the tumor, were imaged using a hand-held imaging device. Resected tumor and normal muscle samples were harvested to assess cathepsin expression by Western blot. Uptake of activated probe was analyzed by flow cytometry and confocal microscopy. Parallel in vitro studies using mouse sarcoma cells were performed. Results: RT of primary STS in mice and mouse sarcoma cell lines caused no change in probe activation or cathepsin protease expression. Increasing radiation dose resulted in an upward trend in probe activation. Flow cytometry and immunofluorescence showed that a substantial proportion of probe-labeled cells were CD11b-positive tumor-associated immune cells. Conclusions: In this primary murine model of STS, RT did not affect the ability of cathepsin-activated probes to differentiate between tumor and normal muscle. Cathepsin-activated probes labeled tumor cells and tumor-associated macrophages. Our results suggest that it would be feasible to include patients who have received preoperative RT in clinical studies evaluating cathepsin-activated imaging probes.

  10. Hyperspectral imaging of endogenous fluorescent metabolic molecules to identify pain states in central nervous system tissue

    Science.gov (United States)

    Staikopoulos, Vasiliki; Gosnell, Martin E.; Anwer, Ayad G.; Mustafa, Sanam; Hutchinson, Mark R.; Goldys, Ewa M.

    2016-12-01

    Fluorescence-based bio-imaging methods have been extensively used to identify molecular changes occurring in biological samples in various pathological adaptations. Auto-fluorescence generated by endogenous fluorescent molecules within these samples can interfere with signal to background noise making positive antibody based fluorescent staining difficult to resolve. Hyperspectral imaging uses spectral and spatial imaging information for target detection and classification, and can be used to resolve changes in endogenous fluorescent molecules such as flavins, bound and free NADH and retinoids that are involved in cell metabolism. Hyperspectral auto-fluorescence imaging of spinal cord slices was used in this study to detect metabolic differences within pain processing regions of non-pain versus sciatic chronic constriction injury (CCI) animals, an established animal model of peripheral neuropathy. By using an endogenous source of contrast, subtle metabolic variations were detected between tissue samples, making it possible to distinguish between animals from non-injured and injured groups. Tissue maps of native fluorophores, flavins, bound and free NADH and retinoids unveiled subtle metabolic signatures and helped uncover significant tissue regions with compromised mitochondrial function. Taken together, our results demonstrate that hyperspectral imaging provides a new non-invasive method to investigate central changes of peripheral neuropathic injury and other neurodegenerative disease models, and paves the way for novel cellular characterisation in health, disease and during treatment, with proper account of intrinsic cellular heterogeneity.

  11. Breast Cancer Imaging Using the Near-Infrared Fluorescent Agent, CLR1502

    Directory of Open Access Journals (Sweden)

    Melissa L. Korb

    2015-01-01

    Full Text Available Positive margins after breast conservation surgery represent a significant problem in the treatment of breast cancer. The near-infrared fluorescence agent CLR1502 (Cellectar Biosciences, Madison, WI was studied in a preclinical breast cancer model to determine imaging properties and ability to detect small islands of malignancy. Nude mice bearing human breast cancer flank xenografts were given a systemic injection of CLR1502, and imaging was performed using LUNA (Novadaq Technologies Inc., Richmond, BC and Pearl Impulse (LI-COR Biosciences, Lincoln, NE devices. Normal tissues were examined for fluorescence signal, and conventional and fluorescence histology was performed using the Odyssey scanner. Peak tumor to background ratio occurred 2 days after injection with CLR1502. The smallest amount of tumor that was imaged and detected using these devices was 1.9 mg, equivalent to 1.9 × 106 cells. The highest fluorescence signal was seen in tumor and normal lymph node tissue, and the lowest fluorescence signal was seen in muscle and plasma. Human breast cancer tumors can be imaged in vivo with multiple optical imaging platforms using CLR1502. This pilot study supports further investigations of this fluorescent agent for improving surgical resection of malignancies, with the goal of eventual clinical translation.

  12. Evaluation of Mobile Phone Performance for Near-Infrared Fluorescence Imaging.

    Science.gov (United States)

    Ghassemi, Pejhman; Wang, Bohan; Wang, Jianting; Wang, Quanzeng; Chen, Yu; Pfefer, T Joshua

    2016-08-19

    We have investigated the potential for contrast-enhanced near-infrared fluorescence imaging of tissue on a mobile phone platform. CCD- and phone-based cameras were used to image molded and 3Dprinted tissue phantoms, and an ex vivo animal model. Quantitative and qualitative evaluations of image quality demonstrate the viability of this approach and elucidate variations in performance due to wavelength, pixel color and image processing.

  13. Clinical multi-colour fluorescence imaging of malignant tumours - initial experience

    Energy Technology Data Exchange (ETDEWEB)

    Svanberg, K.; Wang, I. [Lund Univ. Hospital (Sweden). Lund Medical Laser Centre]|[Lund Univ. Hospital (Sweden). Dept. of Oncology; Colleen, S. [Lund Univ. Hospital (Sweden). Lund Medical Laser Centre]|[Lund Univ. Hospital (Sweden). Dept. of Urology; Idvall, I. [Lund Univ. Hospital (Sweden). Lund Medical Laser Centre]|[Lund Univ. Hospital (Sweden). Dept. of Pathology; Ingvar, C. [Lund Univ. Hospital (Sweden). Lund Medical Laser Centre]|[Lund Univ. Hospital (Sweden). Dept. of Surgery; Rydell, R. [Lund Univ. Hospital (Sweden). Lund Medical Laser Centre]|[Lund Univ. Hospital (Sweden). Dept. of Oto-Rhino-Laryngology; Jocham, D. [Luebeck Univ. (Germany). Abt. fuer Urologie; Diddens, H. [Luebeck Univ. (Germany). Medizinisches Laser Zentrum; Bown, S.; Gregory, G. [National Medical Laser Centre, Dept. of Surgery, Rayne Inst., London (United Kingdom); Montan, S. [Spectraphos AB, Ideon, Lund (Sweden); Andersson-Engels, S.; Svanberg, S. [Lund Univ. Hospital (Sweden). Lund Medical Laser Centre]|[Lund Inst. of Technology (Sweden). Dept. of Physics

    1998-01-01

    The purpose of this study was to present a new technique for non-invasive tumour detection based on tissue fluorescence imaging. A clinically adapted multi-colour fluorescence system was employed in the real-time imaging of malignant tumours of the skin, breast, head and neck region, and urinary bladder. Tumour detection was based on the contrast displayed in fluorescence between normal and malignant tissue, related to the selective uptake of tumour-marking agents and natural chromophore differences between various tissues. In order to demarcate basal cell carcinomas of the skin, ALA was applied topically 4-6 h before the fluorescence investigation. For urinary bladder tumour visualisation, ALA was instilled into the bladder 1-2 h prior to the study. Malignant and premalignant lesions in the head and neck region were imaged after i.v. injection of HPD (Photofrin). The tumour imaging system was coupled to an endoscope. Fluorescence light emission from the tissue surface was induced with 100-ns-long optical pulses at 390 nm, generated from a frequency-doubled alexandrite laser. With the use of special image-splitting optics, the tumour fluorescence, intensified in a micro-channel plate, was imaged in 3 selected wavelength bands. These 3 images were processed together to form a new optimised-contrast image of the tumour. This image, updated at a rate of about 3 frames/s was mixed with a normal colour video image of the tissue. A clear demarcation from normal surrounding tissue was found during in vivo measurements of superficial bladder carcinoma, basal cell carcinoma of the skin, and leukoplakia with dysplasia of the lip, and in vitro investigations of resected breast cancer. (orig./MG).

  14. Quantitative method to assess caries via fluorescence imaging from the perspective of autofluorescence spectral analysis

    Science.gov (United States)

    Chen, Q. G.; Zhu, H. H.; Xu, Y.; Lin, B.; Chen, H.

    2015-08-01

    A quantitative method to discriminate caries lesions for a fluorescence imaging system is proposed in this paper. The autofluorescence spectral investigation of 39 teeth samples classified by the International Caries Detection and Assessment System levels was performed at 405 nm excitation. The major differences in the different caries lesions focused on the relative spectral intensity range of 565-750 nm. The spectral parameter, defined as the ratio of wavebands at 565-750 nm to the whole spectral range, was calculated. The image component ratio R/(G + B) of color components was statistically computed by considering the spectral parameters (e.g. autofluorescence, optical filter, and spectral sensitivity) in our fluorescence color imaging system. Results showed that the spectral parameter and image component ratio presented a linear relation. Therefore, the image component ratio was graded as 1.62 to quantitatively classify sound, early decay, established decay, and severe decay tissues, respectively. Finally, the fluorescence images of caries were experimentally obtained, and the corresponding image component ratio distribution was compared with the classification result. A method to determine the numerical grades of caries using a fluorescence imaging system was proposed. This method can be applied to similar imaging systems.

  15. Multiparameter fluorescence imaging for quantification of TH-1 and TH-2 cytokines at the single-cell level

    Science.gov (United States)

    Fekkar, Hakim; Benbernou, N.; Esnault, S.; Shin, H. C.; Guenounou, Moncef

    1998-04-01

    procedure of the original image using a structuring element. The opened image was therefore subtracted from the original one, and the gray intensities were subsequently measured. Fluorescence intensities are mapped in MD representation using Matlab software. Consequently, quantitative comparative expression of intracellular cytokines and cell membrane markers was achieved. Using this technique, we showed that CD4+ and CD8+T lymphocytes expressed a large panel of cytokines, and that protein kinase A (PKA) activation pathway induced a polarization of activated human T cells to the TH-2 type profile. Data also showed different sensitivities of CD45 RO/CD45RA lymphocytes to the activation of PKA, thus suggesting the implication of memory CD4+- and CD8+-T cells in the T cell specific immune and inflammatory processes and their control by PKA activation pathway. Finally, this method represents a powerful tool for the detection and quantification of intracellular cytokine expression and the analysis of the functional properties of T lymphocytes during immune responses.

  16. Functional imaging in bulk tissue specimens using optical emission tomography: fluorescence preservation during optical clearing

    Science.gov (United States)

    Sakhalkar, H. S.; Dewhirst, M.; Oliver, T.; Cao, Y.; Oldham, M.

    2007-04-01

    Optical emission computed tomography (optical-ECT) is a technique for imaging the three-dimensional (3D) distribution of fluorescent probes in biological tissue specimens with high contrast and spatial resolution. In optical-ECT, functional information can be imaged by (i) systemic application of functional labels (e.g. fluorophore labelled proteins) and/or (ii) endogenous expression of fluorescent reporter proteins (e.g. red fluorescent protein (RFP), green fluorescent protein (GFP)) in vivo. An essential prerequisite for optical-ECT is optical clearing, a procedure where tissue specimens are made transparent to light by sequential perfusion with fixing, dehydrating and clearing agents. In this study, we investigate clearing protocols involving a selection of common fixing (4% buffered paraformaldehyde (PFA), methanol and ethanol), dehydrating (methanol and ethanol) and clearing agents (methyl salicylate and benzyl-alcohol-benzyl-benzoate (BABB)) in order to determine a 'fluorescence friendly' clearing procedure. Cell culture experiments were employed to optimize the sequence of chemical treatments that best preserve fluorescence. Texas red (TxRed), fluorescein isothiocyanate (FITC), RFP and GFP were tested as fluorophores and fluorescent reporter proteins of interest. Fluorescent and control cells were imaged on a microscope using a DSred2 and FITC filter set. The most promising clearing protocols of cell culture experiments were applied to whole xenograft tumour specimens, to test their effectiveness in large unsectioned samples. Fluorescence of TxRed/FITC fluorophores was not found to be significantly affected by any of the test clearing protocols. RFP and GFP fluorescence, however, was found to be significantly greater when cell fixation was in ethanol. Fixation in either PFA or methanol resulted in diminished fluorescence. After ethanol fixation, the RFP and GFP fluorescence proved remarkably robust to subsequent exposure to either methyl salicylate or BABB

  17. Benzothiadiazole Derivatives as Fluorescence Imaging Probes: Beyond Classical Scaffolds.

    Science.gov (United States)

    Neto, Brenno A D; Carvalho, Pedro H P R; Correa, Jose R

    2015-06-16

    This Account describes the origins, features, importance, and trends of the use of fluorescent small-molecule 2,1,3-benzothiadiazole (BTD) derivatives as a new class of bioprobes applied to bioimaging analyses of several (live and fixed) cell types. BTDs have been successfully used as probes for a plethora of biological analyses for only a few years, and the impressive responses obtained by using this important class of heterocycle are fostering the development of new fluorescent BTDs and expanding the biological applications of such derivatives. The first use of a fluorescent small-molecule BTD derivative as a selective cellular probe dates back to 2010, and since then impressive advances have been described by us and others. The well-known limitations of classical scaffolds urged the development of new classes of bioprobes. Although great developments have been achieved by using classical scaffolds such as coumarins, BODIPYs, fluoresceins, rhodamines, cyanines, and phenoxazines, there is still much to be done, and BTDs aim to succeed where these dyes have shown their limitations. Important organelles and cell components such as nuclear DNA, mitochondria, lipid droplets, and others have already been successfully labeled by fluorescent small-molecule BTD derivatives. New technological systems that use BTDs as the fluorophores for bioimaging experiments have been described in recent scientific literature. The successful application of BTDs as selective bioprobes has led some groups to explore their potential for use in studying membrane pores or tumor cells under hypoxic conditions. Finally, BTDs have also been used as fluorescent tags to investigate the action mechanism of some antitumor compounds. The attractive photophysical data typically observed for π-extended BTD derivatives is fostering interest in the use of this new class of bioprobes. Large Stokes shifts, large molar extinction coefficients, high quantum yields, high stability when stored in solution or

  18. Intraoperative fluorescence imaging for personalized brain tumor resection: Current state and future directions

    Directory of Open Access Journals (Sweden)

    Evgenii Belykh

    2016-10-01

    Full Text Available Introduction: Fluorescence-guided surgery is one of the rapidly emerging methods of surgical theranostics. In this review, we summarize current fluorescence techniques used in neurosurgical practice for brain tumor patients, as well as future applications of recent laboratory and translational studies.Methods: Review of the literature.Results: A wide spectrum of fluorophores that have been tested for brain surgery is reviewed. Beginning with a fluorescein sodium application in 1948 by Moore, fluorescence guided brain tumor surgery is either routinely applied in some centers or is under active study in clinical trials. Besides the trinity of commonly used drugs (fluorescein sodium, 5-ALA and ICG, less studied fluorescent stains, such as tetracyclines, cancer-selective alkylphosphocholine analogs, cresyl violet, acridine orange, and acriflavine can be used for rapid tumor detection and pathological tissue examination. Other emerging agents such as activity-based probes and targeted molecular probes that can provide biomolecular specificity for surgical visualization and treatment are reviewed. Furthermore, we review available engineering and optical solutions for fluorescent surgical visualization. Instruments for fluorescent-guided surgery are divided into wide-field imaging systems and hand-held probes. Recent advancements in quantitative fluorescence-guided surgery are discussed.Conclusion: We are standing on the doorstep of the era of marker-assisted tumor management. Innovations in the fields of surgical optics, computer image analysis, and molecular bioengineering are advancing fluorescence-guided tumor resection paradigms, leading to cell-level approaches to visualization and resection of brain tumors.

  19. Fluorescence endoscopic imaging study of anastomotic recurrence of Crohn's disease after right ileocolonic resection

    Science.gov (United States)

    Mordon, Serge R.; Maunoury, Vincent; Klein, Olivier; Colombel, Jean-Frederic

    1995-12-01

    Crohn's disease is an inflammatory bowel disease of unknown etiology. Vasculitis is hypothesized but it was never demonstrated in vivo. This study aimed to evaluate the vascular mucosa perfusion using fluorescence imaging in 13 patients who had previously undergone eileocolonic resection and who agreed to participate in a prospective endoscopic study of anastomotic recurrence. This anastomotic recurrence rate is known to be high (73% after 1 year follow-up) and is characterized by ulcerations. The fluorescence study was started with an I.V. bolus injection of sodium fluorescein. The pre-anastomotic mucosa was endoscopically examined with blue light that stimulates fluorescein fluorescence. Fluorescence emission was recorded with an ultra-high-sensitivity camera connected to the endoscope via an interference filter (520 - 560 nm). A uniform fluorescence was observed a few seconds after the injection and lasted for 15 min in healthy subjects. In case of recurrence, the centers of the ulcerations displayed a very low fluorescence indicating localized ischemia. In contrast, the rims of the ulcers revealed brighter fluorescent images than those of normal mucosa. The anastomotic ulcerations of Crohn's disease recurrence exhibit a high fluorescence intensity at their margins indicating an increased mucosal blood flow and/or enhanced transcapillary diffusion. These findings support the hypothesis of a primary vasculitis in Crohn's disease.

  20. Cyanine-loaded lipid nanoparticles for improved in vivo fluorescence imaging

    Science.gov (United States)

    Texier, Isabelle; Goutayer, Mathieu; da Silva, Anabela; Guyon, Laurent; Djaker, Nadia; Josserand, Véronique; Neumann, Emmanuelle; Bibette, Jérôme; Vinet, Françoise

    2009-09-01

    Fluorescence is a very promising radioactive-free technique for functional imaging in small animals and, in the future, in humans. However, most commercial near-infrared dyes display poor optical properties, such as low fluorescence quantum yields and short fluorescence lifetimes. In this paper, we explore whether the encapsulation of infrared cyanine dyes within the core of lipid nanoparticles (LNPs) could improve their optical properties. Lipophilic dialkylcarbocyanines DiD and DiR are loaded very efficiently in 30-35-nm-diam lipid droplets stabilized in water by surfactants. No significant fluorescence autoquenching is observed up to 53 dyes per particle. Encapsulated in LNP, which are stable for more than one year at room temperature in HBS buffer (HEPES 0.02 M, EDTA 0.01 M, pH 5.5), DiD and DiR display far improved fluorescence quantum yields Φ (respectively, 0.38 and 0.25) and longer fluorescence lifetimes τ (respectively, 1.8 and 1.1 ns) in comparison to their hydrophilic counterparts Cy5 (φ=0.28, τ=1.0 ns) and Cy7 (φ=0.13, τ=0.57 ns). Moreover, dye-loaded LNPs are able to accumulate passively in various subcutaneous tumors in mice, thanks to the enhanced permeability and retention effect. These new fluorescent nanoparticles therefore appear as very promising labels for in vivo fluorescence imaging.

  1. Stepwise multi-photon activation fluorescence reveals a new method of melanoma imaging for dermatologists

    Science.gov (United States)

    Lai, Zhenhua; Lian, Christine; Ma, Jie; Yu, Jingyi; Gu, Zetong; Rajadhyaksha, Milind; DiMarzio, Charles A.

    2014-02-01

    Previous research has shown that the stepwise multi-photon activated fluorescence (SMPAF) of melanin, activated by a continuous-wave (CW) mode near infrared (NIR) laser, is a low cost and reliable method of detecting melanin. SMPAF images of melanin in a mouse hair and a formalin fixed mouse melanoma were compared with conventional multiphoton fluorescence microscopy (MPFM) images and confocal reflectance microscopy (CRM) images, all of which were acquired at an excitation wavelength of 920 nm, to further prove the effectiveness of SMPAF in detecting melanin. SMPAF images add specificity for melanin detection to MPFM images and CRM images. Melanin SMPAF can be a promising technology to enable melanoma imaging for dermatologists.

  2. Development of in vivo confocal microscope for reflection and fluorescence imaging simultaneously

    Science.gov (United States)

    Ahn, MyoungKi; Chun, ByungSeon; Song, Cheol; Gweon, DaeGab

    2010-02-01

    In-vivo confocal microscope technology can be applied to the medical imaging diagnosis and new drug development. We present an in-vivo confocal microscope that can acquire a reflection image and a fluorescence image simultaneously and independently. To obtain reflection confocal images, we used a linearly polarized diode laser with the wavelength of 830 nm. To acquire fluorescence confocal images, we used two diode lasers with the wavelength of 488 nm and 660 nm, respectively. Because of a broad wavelength bandwidth from visible (488 nm) to near-IR (830 nm), we designed and optimized the optical system to reduce various optical aberrations. With the developed in-vivo confocal microscope, we performed ex-vivo cell imaging and in-vivo imaging of the human skin.

  3. Hybrid fluorescence and electron cryo-microscopy for simultaneous electron and photon imaging.

    Science.gov (United States)

    Iijima, Hirofumi; Fukuda, Yoshiyuki; Arai, Yoshihiro; Terakawa, Susumu; Yamamoto, Naoki; Nagayama, Kuniaki

    2014-01-01

    Integration of fluorescence light and transmission electron microscopy into the same device would represent an important advance in correlative microscopy, which traditionally involves two separate microscopes for imaging. To achieve such integration, the primary technical challenge that must be solved regards how to arrange two objective lenses used for light and electron microscopy in such a manner that they can properly focus on a single specimen. To address this issue, both lateral displacement of the specimen between two lenses and specimen rotation have been proposed. Such movement of the specimen allows sequential collection of two kinds of microscopic images of a single target, but prevents simultaneous imaging. This shortcoming has been made up by using a simple optical device, a reflection mirror. Here, we present an approach toward the versatile integration of fluorescence and electron microscopy for simultaneous imaging. The potential of simultaneous hybrid microscopy was demonstrated by fluorescence and electron sequential imaging of a fluorescent protein expressed in cells and cathodoluminescence imaging of fluorescent beads. Copyright © 2013 Elsevier Inc. All rights reserved.

  4. Folic acid-targeted magnetic Tb-doped CeF3 fluorescent nanoparticles as bimodal probes for cellular fluorescence and magnetic resonance imaging.

    Science.gov (United States)

    Ma, Zhi-Ya; Liu, Yu-Ping; Bai, Ling-Yu; An, Jie; Zhang, Lin; Xuan, Yang; Zhang, Xiao-Shuai; Zhao, Yuan-Di

    2015-10-07

    Magnetic fluorescent nanoparticles (NPs) have great potential applications for diagnostics, imaging and therapy. We developed a facile polyol method to synthesize multifunctional Fe3O4@CeF3:Tb@CeF3 NPs with small size (magnetic resonance imaging (MRI) of HeLa cells with overexpressed folate receptors (FR). The results indicated that these NPs had strong luminescence and enhanced T2-weighted MR contrast and would be promising candidates as multimodal probes for both fluorescence and MRI imaging.

  5. Breast MRI at very short TE (minTE). Image analysis of minTE sequences on non-fat-saturated, subtracted T1-weighted images

    Energy Technology Data Exchange (ETDEWEB)

    Wenkel, Evelyn; Janka, Rolf; Kaemmerer, Nadine; Uder, Michael; Hammon, Matthias; Brand, Michael [Univ. Hospital Erlangen (Germany). Dept. of Radiology; Geppert, Christian [Siemens Healthcare GmbH, Erlangen (Germany); Hartmann, Arndt [Univ. Hospital Erlangen (Germany). Dept. of Pathology

    2017-02-15

    The aim was to evaluate a minimum echo time (minTE) protocol for breast magnetic resonance imaging (MRI) in patients with breast lesions compared to a standard TE (nTE) time protocol. Breasts of 144 women were examined with a 1.5 Tesla MRI scanner. Additionally to the standard gradient-echo sequence with nTE (4.8 ms), a variant with minimum TE (1.2 ms) was used in an interleaved fashion which leads to a better temporal resolution and should reduce the scan time by approximately 50%. Lesion sizes were measured and the signal-to-noise ratio (SNR) as well as the contrast-to-noise ratio (CNR) were calculated. Subjective confidence was evaluated using a 3-point scale before looking at the nTE sequences (1 = very sure that I can identify a lesion and classify it, 2 = quite sure that I can identify a lesion and classify it, 3 = definitely want to see nTE for final assessment) and the subjective image quality of all examinations was evaluated using a four-grade scale (1 = sharp, 2 = slight blur, 3 = moderate blur and 4 = severe blur/not evaluable) for lesion and skin sharpness. Lesion morphology and contrast enhancement were also evaluated. With minTE sequences, no lesion was rated with ''definitely want to see nTE sequences for final assessment''. The difference of the longitudinal and transverse diameter did not differ significantly (p>0.05). With minTE, lesions and skin were rated to be significantly more blurry (p<0.01 for lesions and p<0.05 for skin). There was no difference between both sequences with respect to SNR, CNR, lesion morphology, contrast enhancement and detection of multifocal disease. Dynamic breast MRI with a minTE protocol is feasible without a major loss of information (SNR, CNR, lesion morphology, contrast enhancement and lesion sizes) and the temporal resolution can be increased by a factor of 2 using minTE sequences.

  6. Current Concepts and Future Perspectives on Intraoperative Fluorescence Imaging in Cancer : Clinical Need

    NARCIS (Netherlands)

    van Dam, Gooitzen M.; Ntziachristos, Vasilis

    Progress with technology and regulatory approvals has recently allowed the successful clinical translation of fluorescence molecular imaging to intra-operative applications. Initial studies have demonstrated a promising outlook for imaging cancer micro-foci, margins and lymph-nodes. However, not all

  7. Image overlay solution based on threshold detection for a compact near infrared fluorescence goggle system.

    Science.gov (United States)

    Gao, Shengkui; Mondal, Suman B; Zhu, Nan; Liang, RongGuang; Achilefu, Samuel; Gruev, Viktor

    2015-01-01

    Near infrared (NIR) fluorescence imaging has shown great potential for various clinical procedures, including intraoperative image guidance. However, existing NIR fluorescence imaging systems either have a large footprint or are handheld, which limits their usage in intraoperative applications. We present a compact NIR fluorescence imaging system (NFIS) with an image overlay solution based on threshold detection, which can be easily integrated with a goggle display system for intraoperative guidance. The proposed NFIS achieves compactness, light weight, hands-free operation, high-precision superimposition, and a real-time frame rate. In addition, the miniature and ultra-lightweight light-emitting diode tracking pod is easy to incorporate with NIR fluorescence imaging. Based on experimental evaluation, the proposed NFIS solution has a lower detection limit of 25 nM of indocyanine green at 27 fps and realizes a highly precise image overlay of NIR and visible images of mice in vivo. The overlay error is limited within a 2-mm scale at a 65-cm working distance, which is highly reliable for clinical study and surgical use.

  8. Shading correction and calibration in bacterial fluorescence measurement by image processing system

    NARCIS (Netherlands)

    Wilkinson, M.H.F.

    1994-01-01

    An image processing system with applications in bacterial (immuno-)fluorescence measurement has been developed. To reach quantitative results, correction for non-uniformities in system sensitivity, both as a function of time (calibration for drifts) and as a function of image coordinates (shading co

  9. Shading correction and calibration in bacterial fluorescence measurement by image processing system

    NARCIS (Netherlands)

    Wilkinson, M.H.F.

    1994-01-01

    An image processing system with applications in bacterial (immuno-)fluorescence measurement has been developed. To reach quantitative results, correction for non-uniformities in system sensitivity, both as a function of time (calibration for drifts) and as a function of image coordinates (shading co

  10. A Validation Study of Near-Infrared Fluorescence Imaging of Lymphatic Vessels in Humans

    DEFF Research Database (Denmark)

    Groenlund, Jacob Hinnerup; Telinius, Niklas; Skov, Soeren Nielsen

    2017-01-01

    BACKGROUND: Near-infrared fluorescence (NIRF) imaging is a new imaging technique that is used to visualize lymphatic vessels in humans. It has a high spatial and temporal resolution, allowing real-time visualization of lymphatic flow. METHODS AND RESULTS: The current study investigated the intra...

  11. A portable UV-fluorescence multispectral imaging system for the analysis of painted surfaces.

    Science.gov (United States)

    Comelli, Daniela; Valentini, Gianluca; Nevin, Austin; Farina, Andrea; Toniolo, Lucia; Cubeddu, Rinaldo

    2008-08-01

    A portable fluorescence multispectral imaging system was developed and has been used for the analysis of artistic surfaces. The imaging apparatus exploits two UV lamps for fluorescence excitation and a liquid crystal tunable filter coupled to a low-noise charge coupled device as the image detector. The main features of the system are critically presented, outlining the assets, drawbacks, and practical considerations of portability. A multivariate statistical treatment of spectral data is further considered. Finally, the in situ analysis with the new apparatus of recently restored Renaissance wall paintings is presented.

  12. Near-infrared fluorescence imaging of mammalian cells and xenograft tumors with SNAP-tag.

    Directory of Open Access Journals (Sweden)

    Haibiao Gong

    Full Text Available Fluorescence in the near-infrared (NIR spectral region is suitable for in vivo imaging due to its reduced background and high penetration capability compared to visible fluorescence. SNAP(f is a fast-labeling variant of SNAP-tag that reacts with a fluorescent dye-conjugated benzylguanine (BG substrate, leading to covalent attachment of the fluorescent dye to the SNAP(f. This property makes SNAP(f a valuable tool for fluorescence imaging. The NIR fluorescent substrate BG-800, a conjugate between BG and IRDye 800CW, was synthesized and characterized in this study. HEK293, MDA-MB-231 and SK-OV-3 cells stably expressing SNAP(f-Beta-2 adrenergic receptor (SNAP(f-ADRβ2 fusion protein were created. The ADRβ2 portion of the protein directs the localization of the protein to the cell membrane. The expression of SNAP(f-ADRβ2 in the stable cell lines was confirmed by the reaction between BG-800 substrate and cell lysates. Microscopic examination confirmed that SNAP(f-ADRβ2 was localized on the cell membrane. The signal intensity of the labeled cells was dependent on the BG-800 concentration. In vivo imaging study showed that BG-800 could be used to visualize xenograph tumors expressing SNAP(f-ADRβ2. However, the background signal was relatively high, which may be a reflection of non-specific accumulation of BG-800 in the skin. To address the background issue, quenched substrates that only fluoresce upon reaction with SNAP-tag were synthesized and characterized. Although the fluorescence was successfully quenched, in vivo imaging with the quenched substrate CBG-800-PEG-QC1 failed to visualize the SNAP(f-ADRβ2 expressing tumor, possibly due to the reduced reaction rate. Further improvement is needed to apply this system for in vivo imaging.

  13. The application of anti-ESAT-6 monoclonal antibody fluorescent probe in ex vivo near-infrared fluorescence imaging in mice with pulmonary tuberculosis.

    Science.gov (United States)

    Feng, Feng; Zhang, Haoling; Zhu, Zhaoqin; Li, Cong; Shi, Yuxin; Zhang, Zhiyong

    2014-09-01

    Here, we aimed to assess the feasibility of anti-ESAT-6 monoclonal antibody (mAb) coupling with IR783 and rhodamine fluorescent probe in the detection of ESAT-6 expression in tuberculosis tissue of mice using near-infrared fluorescence imaging. IR783 and rhodamine were conjugated to the anti-ESAT-6 mAb or IgG. Mice in the experimental group were injected with fluorescence-labeled mAb probe, and mice in the control group were injected with fluorescence-labeled non-specific IgG antibody. Twenty-four hours later, the lung tissue of mice was examined using ex vivo near-infrared fluorescence imaging. In addition, the contrast-to-noise ratio (CNR) was calculated by measuring the signal intensities of the pulmonary lesions, normal lung tissue and background noise. The frozen lung tissue section was examined under fluorescence microscopy and compared with hemoxylin and eosin (HE) staining. The ex vivo near-infrared fluorescence imaging showed that the fluorescence signal in the lung tuberculosis lesions in the experimental group was significantly enhanced, whereas there was only a weak fluorescence signal or even no fluorescence signal in the control group. CNR values were 64.40 ± 7.02 (n = 6) and 8.75 ± 3.87 (n = 6), respectively (t = 17.01, p fluorescence accumulation distribution detected under fluorescence microscopy was consistent with HE staining of the tuberculosis region. In conclusion, anti-ESAT-6 mAb fluorescent probe could target and be applied in specific ex vivo imaging of mice tuberculosis, and may be of further use in tuberculosis in living mice. Copyright © 2013 John Wiley & Sons, Ltd.

  14. Cell tracking with gadophrin-2: a bifunctional contrast agent for MR imaging, optical imaging, and fluorescence microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Daldrup-Link, Heike E. [Department of Radiology, UCSF Medical Center, University of California in San Francisco, 513 Parnassus Ave, CA 94143, San Francisco (United States); Rudelius, Martina; Piontek, Guido; Schlegel, Juergen [Institute of Pathology, Technical University, Munich (Germany); Metz, Stephan; Settles, Marcus; Rummeny, Ernst J. [Department of Radiology, Technical University, Munich (Germany); Pichler, Bernd [Department of Biomedical Engineering, University of California Davis, Davis (United States); Heinzmann, Ulrich [National Research Center for Environment and Health, Technical University, Munich (Germany); Oostendorp, Robert A.J. [3. Clinic of Internal Medicine, Laboratory of Stem Cell Physiology, Technical University, Munich (Germany)

    2004-09-01

    The purpose of this study was to assess the feasibility of use of gadophrin-2 to trace intravenously injected human hematopoietic cells in athymic mice, employing magnetic resonance (MR) imaging, optical imaging (OI), and fluorescence microscopy. Mononuclear peripheral blood cells from GCSF-primed patients were labeled with gadophrin-2 (Schering AG, Berlin, Germany), a paramagnetic and fluorescent metalloporphyrin, using established transfection techniques with cationic liposomes. The labeled cells were evaluated in vitro with electron microscopy and inductively coupled plasma atomic emission spectrometry. Then, 1 x 10{sup 6}-3 x 10{sup 8} labeled cells were injected into 14 nude Balb/c mice and the in vivo cell distribution was evaluated with MR imaging and OI before and 4, 24, and 48 h after intravenous injection (p.i.). Five additional mice served as controls: three mice were untreated controls and two mice were investigated after injection of unlabeled cells. The contrast agent effect was determined quantitatively for MR imaging by calculating signal-to-noise-ratio (SNR) data. After completion of in vivo imaging studies, fluorescence microscopy of excised organs was performed. Intracellular cytoplasmatic uptake of gadophrin-2 was confirmed by electron microscopy. Spectrometry determined an uptake of 31.56 nmol Gd per 10{sup 6} cells. After intravenous injection, the distribution of gadophrin-2 labeled cells in nude mice could be visualized by MR, OI, and fluorescence microscopy. At 4 h p.i., the transplanted cells mainly distributed to lung, liver, and spleen, and 24 h p.i. they also distributed to the bone marrow. Fluorescence microscopy confirmed the distribution of gadophrin-2 labeled cells to these target organs. Gadophrin-2 is suited as a bifunctional contrast agent for MR imaging, OI, and fluorescence microscopy and may be used to combine the advantages of each individual imaging modality for in vivo tracking of intravenously injected hematopoietic cells

  15. The possibilities of improvement in the sensitivity of cancer fluorescence diagnostics by computer image processing

    Science.gov (United States)

    Ledwon, Aleksandra; Bieda, Robert; Kawczyk-Krupka, Aleksandra; Polanski, Andrzej; Wojciechowski, Konrad; Latos, Wojciech; Sieron-Stoltny, Karolina; Sieron, Aleksander

    2008-02-01

    Background: Fluorescence diagnostics uses the ability of tissues to fluoresce after exposition to a specific wavelength of light. The change in fluorescence between normal and progression to cancer allows to see early cancer and precancerous lesions often missed by white light. Aim: To improve by computer image processing the sensitivity of fluorescence images obtained during examination of skin, oral cavity, vulva and cervix lesions, during endoscopy, cystoscopy and bronchoscopy using Xillix ONCOLIFE. Methods: Function of image f(x,y):R2 --> R 3 was transformed from original color space RGB to space in which vector of 46 values refers to every point labeled by defined xy-coordinates- f(x,y):R2 --> R 46. By means of Fisher discriminator vector of attributes of concrete point analalyzed in the image was reduced according to two defined classes defined as pathologic areas (foreground) and healthy areas (background). As a result the highest four fisher's coefficients allowing the greatest separation between points of pathologic (foreground) and healthy (background) areas were chosen. In this way new function f(x,y):R2 --> R 4 was created in which point x,y corresponds with vector Y, H, a*, c II. In the second step using Gaussian Mixtures and Expectation-Maximisation appropriate classificator was constructed. This classificator enables determination of probability that the selected pixel of analyzed image is a pathologically changed point (foreground) or healthy one (background). Obtained map of probability distribution was presented by means of pseudocolors. Results: Image processing techniques improve the sensitivity, quality and sharpness of original fluorescence images. Conclusion: Computer image processing enables better visualization of suspected areas examined by means of fluorescence diagnostics.

  16. Fluorescent Probes for Analysis and Imaging of Monoamine Oxidase Activity

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Dokyoung; Jun, Yong Woong; Ahn, Kyo Han [POSTECH, Pohang (Korea, Republic of)

    2014-05-15

    Monoamine oxidases catalyze the oxidative deamination of dietary amines and amine neurotransmitters, and assist in maintaining the homeostasis of the amine neurotransmitters in the brain. Dysfunctions of these enzymes can cause neurological and behavioral disorders including Parkinson's and Alzheimer's diseases. To understand their physiological roles, efficient assay methods for monoamine oxidases are essential. Reviewed in this Perspective are the recent progress in the development of fluorescent probes for monoamine oxidases and their applications to enzyme assays in cells and tissues. It is evident that still there is strong need for a fluorescent probe with desirable substrate selectivity and photophysical properties to challenge the much unsolved issues associated with the enzymes and the diseases.

  17. Intraoperative Near-infrared Imaging for Parathyroid Gland Identification by Auto-fluorescence: A Feasibility Study.

    Science.gov (United States)

    De Leeuw, Frederic; Breuskin, Ingrid; Abbaci, Muriel; Casiraghi, Odile; Mirghani, Haïtham; Ben Lakhdar, Aïcha; Laplace-Builhé, Corinne; Hartl, Dana

    2016-09-01

    Parathyroid glands (PGs) can be particularly hard to distinguish from surrounding tissue and thus can be damaged or removed during thyroidectomy. Postoperative hypoparathyroidism is the most common complication after thyroidectomy. Very recently, it has been found that the parathyroid tissue shows near-infrared (NIR) auto-fluorescence which could be used for intraoperative detection, without any use of contrast agents. The work described here presents a histological validation ex vivo of the NIR imaging procedure and evaluates intraoperative PG detection by NIR auto-fluorescence using for the first time to our knowledge a commercially available clinical NIR imaging device. Ex vivo study on resected operative specimens combined with a prospective in vivo study of consecutive patients who underwent total or partial thyroid, or parathyroid surgery at a comprehensive cancer center. During surgery, any tissue suspected to be a potential PG by the surgeon was imaged with the Fluobeam 800 (®) system. NIR imaging was compared to conventional histology (ex vivo) and/or visual identification by the surgeon (in vivo). We have validated NIR auto-fluorescence with an ex vivo study including 28 specimens. Sensitivity and specificity were 94.1 and 80 %, respectively. Intraoperative NIR imaging was performed in 35 patients and 81 parathyroids were identified. In 80/81 cases, the fluorescence signal was subjectively obvious on real-time visualization. We determined that PG fluorescence is 2.93 ± 1.59 times greater than thyroid fluorescence in vivo. Real-time NIR imaging based on parathyroid auto-fluorescence is fast, safe, and non-invasive and shows very encouraging results, for intraoperative parathyroid identification.

  18. Parameter estimation method for blurred cell images from fluorescence microscope

    Science.gov (United States)

    He, Fuyun; Zhang, Zhisheng; Luo, Xiaoshu; Zhao, Shulin

    2016-10-01

    Microscopic cell image analysis is indispensable to cell biology. Images of cells can easily degrade due to optical diffraction or focus shift, as this results in low signal-to-noise ratio (SNR) and poor image quality, hence affecting the accuracy of cell analysis and identification. For a quantitative analysis of cell images, restoring blurred images to improve the SNR is the first step. A parameter estimation method for defocused microscopic cell images based on the power law properties of the power spectrum of cell images is proposed. The circular radon transform (CRT) is used to identify the zero-mode of the power spectrum. The parameter of the CRT curve is initially estimated by an improved differential evolution algorithm. Following this, the parameters are optimized through the gradient descent method. Using synthetic experiments, it was confirmed that the proposed method effectively increased the peak SNR (PSNR) of the recovered images with high accuracy. Furthermore, experimental results involving actual microscopic cell images verified that the superiority of the proposed parameter estimation method for blurred microscopic cell images other method in terms of qualitative visual sense as well as quantitative gradient and PSNR.

  19. Ratiometric fluorescence imaging of free Zn2+ in brain

    Science.gov (United States)

    Thompson, Richard B.; Suh, Sang W.; Frederickson, Christopher J.

    2001-05-01

    Recently, the function of zinc in the axonal boutons of hippocampal neurons has come under increased scrutiny as evidence has emerged of a putative role for this metal ion in neural damage following insults such as ischemia, blunt force trauma, and seizure. Indeed, the nonpathological role of free zinc in the brain remains cryptic after more than 40 years. We have used a biosensing approach to determine free zinc ion concentrations by fluorescence lifetime, intensity, intensity ratio, or anisotropy changes caused by binding of zinc to variants of a protein, apocarbonic anhydrase II (apo-CA). This approach permits real time measurement of zinc down to picomolar levels, with no perceptible interference from other divalent metal ions abundant in serum and tissue, such as calcium and magnesium. Recently, we used apo-CA together with a fluorescent ligand whose binding is metal-dependent to obtain the first fluorescence micrographs of zinc release from a rat hippocampus model in response to electrical stimulus. In our view, elucidation of the zinc fluxes in neural tissue ultimately requires quantitation, as in the case of calcium. Recent results will be shown.

  20. Fluorescence endoscopic imaging for evaluation of gastric mucosal blood flow: a preliminary study

    Science.gov (United States)

    Bocquillon, Nicolas; Mordon, Serge R.; Mathieu, D.; Maunoury, Vincent; Marechal, Xavier-Marie; Neviere, Remi; Wattel, Francis; Chopin, Claude

    1999-02-01

    Microcirculatory disorders of the gastrointestinal tract appear to be a major compound of the multiple organ dysfunction syndrome secondary to sepsis or septic shock. A better analysis of mucosal hypoperfusion in critically ill patients with sepsis may be helpful for the comprehension of this high mortality-associated syndrome. Fluorescence endoscopy has been recognized as a non-invasive method for both spatial and temporal evaluation of gastrointestinal mucosal perfusion. We performed this imaging technique during routine gastric endoscopy in patients with sepsis criteria. The study included gastric observation and appearance time of gastric fluorescence after an intravenous 10% sodium - fluorescein bolus. Qualitative analysis of high fluorescence areas was compared with mucosal blood flow measurements by laser - Doppler flowmetry. We concluded that the fluorescence endoscopic imaging in critically ill patients with sepsis may reveal spacial and temporal differences in the mucosal microcirculation distribution.

  1. Noninvasive measurement of pharmacokinetics by near-infrared fluorescence imaging in the eye of mice

    Science.gov (United States)

    Dobosz, Michael; Strobel, Steffen; Stubenrauch, Kay-Gunnar; Osl, Franz; Scheuer, Werner

    2014-01-01

    Purpose: For generating preclinical pharmacokinetics (PKs) of compounds, blood is drawn at different time points and levels are quantified by different analytical methods. In order to receive statistically meaningful data, 3 to 5 animals are used for each time point to get serum peak-level and half-life of the compound. Both characteristics are determined by data interpolation, which may influence the accuracy of these values. We provide a method that allows continuous monitoring of blood levels noninvasively by measuring the fluorescence intensity of labeled compounds in the eye and other body regions of anesthetized mice. Procedures: The method evaluation was performed with four different fluorescent compounds: (i) indocyanine green, a nontargeting dye; (ii) OsteoSense750, a bone targeting agent; (iii) tumor targeting Trastuzumab-Alexa750; and (iv) its F(-alxea750 fragment. The latter was used for a direct comparison between fluorescence imaging and classical blood analysis using enzyme-linked immunosorbent assay (ELISA). Results: We found an excellent correlation between blood levels measured by noninvasive eye imaging with the results generated by classical methods. A strong correlation between eye imaging and ELISA was demonstrated for the F( fragment. Whole body imaging revealed a compound accumulation in the expected regions (e.g., liver, bone). Conclusions: The combination of eye and whole body fluorescence imaging enables the simultaneous measurement of blood PKs and biodistribution of fluorescent-labeled compounds.

  2. Slanted channel microfluidic chip for 3D fluorescence imaging of cells in flow.

    Science.gov (United States)

    Jagannadh, Veerendra Kalyan; Mackenzie, Mark D; Pal, Parama; Kar, Ajoy K; Gorthi, Sai Siva

    2016-09-19

    Three-dimensional cellular imaging techniques have become indispensable tools in biological research and medical diagnostics. Conventional 3D imaging approaches employ focal stack collection to image different planes of the cell. In this work, we present the design and fabrication of a slanted channel microfluidic chip for 3D fluorescence imaging of cells in flow. The approach employs slanted microfluidic channels fabricated in glass using ultrafast laser inscription. The slanted nature of the microfluidic channels ensures that samples come into and go out of focus, as they pass through the microscope imaging field of view. This novel approach enables the collection of focal stacks in a straight-forward and automated manner, even with off-the-shelf microscopes that are not equipped with any motorized translation/rotation sample stages. The presented approach not only simplifies conventional focal stack collection, but also enhances the capabilities of a regular widefield fluorescence microscope to match the features of a sophisticated confocal microscope. We demonstrate the retrieval of sectioned slices of microspheres and cells, with the use of computational algorithms to enhance the signal-to-noise ratio (SNR) in the collected raw images. The retrieved sectioned images have been used to visualize fluorescent microspheres and bovine sperm cell nucleus in 3D while using a regular widefield fluorescence microscope. We have been able to achieve sectioning of approximately 200 slices per cell, which corresponds to a spatial translation of ∼ 15 nm per slice along the optical axis of the microscope.

  3. Improved detection of soma location and morphology in fluorescence microscopy images of neurons.

    Science.gov (United States)

    Kayasandik, Cihan Bilge; Labate, Demetrio

    2016-12-01

    Automated detection and segmentation of somas in fluorescent images of neurons is a major goal in quantitative studies of neuronal networks, including applications of high-content-screenings where it is required to quantify multiple morphological properties of neurons. Despite recent advances in image processing targeted to neurobiological applications, existing algorithms of soma detection are often unreliable, especially when processing fluorescence image stacks of neuronal cultures. In this paper, we introduce an innovative algorithm for the detection and extraction of somas in fluorescent images of networks of cultured neurons where somas and other structures exist in the same fluorescent channel. Our method relies on a new geometrical descriptor called Directional Ratio and a collection of multiscale orientable filters to quantify the level of local isotropy in an image. To optimize the application of this approach, we introduce a new construction of multiscale anisotropic filters that is implemented by separable convolution. Extensive numerical experiments using 2D and 3D confocal images show that our automated algorithm reliably detects somas, accurately segments them, and separates contiguous ones. We include a detailed comparison with state-of-the-art existing methods to demonstrate that our algorithm is extremely competitive in terms of accuracy, reliability and computational efficiency. Our algorithm will facilitate the development of automated platforms for high content neuron image processing. A Matlab code is released open-source and freely available to the scientific community. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. In vivo time-gated fluorescence imaging with biodegradable luminescent porous silicon nanoparticles.

    Science.gov (United States)

    Gu, Luo; Hall, David J; Qin, Zhengtao; Anglin, Emily; Joo, Jinmyoung; Mooney, David J; Howell, Stephen B; Sailor, Michael J

    2013-01-01

    Fluorescence imaging is one of the most versatile and widely used visualization methods in biomedical research. However, tissue autofluorescence is a major obstacle confounding interpretation of in vivo fluorescence images. The unusually long emission lifetime (5-13 μs) of photoluminescent porous silicon nanoparticles can allow the time-gated imaging of tissues in vivo, completely eliminating shorter-lived (50-fold in vitro and by >20-fold in vivo when imaging porous silicon nanoparticles. Time-gated imaging of porous silicon nanoparticles accumulated in a human ovarian cancer xenograft following intravenous injection is demonstrated in a live mouse. The potential for multiplexing of images in the time domain by using separate porous silicon nanoparticles engineered with different excited state lifetimes is discussed.

  5. Systemic Fluorescence Imaging of Zebrafish Glycans with Bioorthogonal Chemistry.

    Science.gov (United States)

    Agarwal, Paresh; Beahm, Brendan J; Shieh, Peyton; Bertozzi, Carolyn R

    2015-09-21

    Vertebrate glycans constitute a large, important, and dynamic set of post-translational modifications that are notoriously difficult to manipulate and image. Although the chemical reporter strategy has been used in conjunction with bioorthogonal chemistry to image the external glycosylation state of live zebrafish and detect tumor-associated glycans in mice, the ability to image glycans systemically within a live organism has remained elusive. Here, we report a method that combines the metabolic incorporation of a cyclooctyne-functionalized sialic acid derivative with a ligation reaction of a fluorogenic tetrazine, allowing for the imaging of sialylated glycoconjugates within live zebrafish embryos.

  6. CMOS time-resolved, contact, and multispectral fluorescence imaging for DNA molecular diagnostics.

    Science.gov (United States)

    Guo, Nan; Cheung, Kawai; Wong, Hiu Tong; Ho, Derek

    2014-10-31

    Instrumental limitations such as bulkiness and high cost prevent the fluorescence technique from becoming ubiquitous for point-of-care deoxyribonucleic acid (DNA) detection and other in-field molecular diagnostics applications. The complimentary metal-oxide-semiconductor (CMOS) technology, as benefited from process scaling, provides several advanced capabilities such as high integration density, high-resolution signal processing, and low power consumption, enabling sensitive, integrated, and low-cost fluorescence analytical platforms. In this paper, CMOS time-resolved, contact, and multispectral imaging are reviewed. Recently reported CMOS fluorescence analysis microsystem prototypes are surveyed to highlight the present state of the art.

  7. Spectral Properties of a Water-Soluble Squaraine Dye and Its Application in Cell Fluorescent Imaging

    Science.gov (United States)

    Hu, L.; Yuan, H.; Li, Q. Q.; Jin, J. C.; Chang, W. G.; Yan, Z. Q.

    2015-09-01

    A water-soluble bis-1,3,5-trihydroxybenzene squaraine dye (t-OH-SQ) with a D-π-A-π-D conjugated structure was identified and prepared. After its structure was characterized by FTIR, 1H NMR and elemental analysis, the UV-Vis absorption and fluorescent spectra of the target dye were studied in detail. The results showed that t-OH-SQ combining multi-hydroxyl groups possessed excellent optical properties changing with pH and solvents. In aqueous solution under physiological pH ~ 7-8, it had especially high near-infrared fluorescence, which might be a latent application for cell fluorescent imaging.

  8. CMOS Time-Resolved, Contact, and Multispectral Fluorescence Imaging for DNA Molecular Diagnostics

    Directory of Open Access Journals (Sweden)

    Nan Guo

    2014-10-01

    Full Text Available Instrumental limitations such as bulkiness and high cost prevent the fluorescence technique from becoming ubiquitous for point-of-care deoxyribonucleic acid (DNA detection and other in-field molecular diagnostics applications. The complimentary metal-oxide-semiconductor (CMOS technology, as benefited from process scaling, provides several advanced capabilities such as high integration density, high-resolution signal processing, and low power consumption, enabling sensitive, integrated, and low-cost fluorescence analytical platforms. In this paper, CMOS time-resolved, contact, and multispectral imaging are reviewed. Recently reported CMOS fluorescence analysis microsystem prototypes are surveyed to highlight the present state of the art.

  9. Fluorescent metal nanoshell and CK19 detection on single cell image

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Jian, E-mail: jian@cfs.biomet.umaryland.edu [Center for Fluorescence Spectroscopy, University of Maryland School of Medicine, Department of Biochemistry and Molecular Biology, 725 West Lombard Street, Baltimore, MD 21201 (United States); Fu, Yi [Center for Fluorescence Spectroscopy, University of Maryland School of Medicine, Department of Biochemistry and Molecular Biology, 725 West Lombard Street, Baltimore, MD 21201 (United States); Li, Ge [Division of Molecular Pathology, Department of Pathology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201 (United States); Lakowicz, Joseph R. [Center for Fluorescence Spectroscopy, University of Maryland School of Medicine, Department of Biochemistry and Molecular Biology, 725 West Lombard Street, Baltimore, MD 21201 (United States); Zhao, Richard Y., E-mail: rzhao@som.umaryland.edu [Division of Molecular Pathology, Department of Pathology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201 (United States); Department of Microbiology-Immunology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201 (United States); Institute of Human Virology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201 (United States)

    2011-09-16

    Highlights: {yields} Novel metal nanoshell as fluorescence imaging agent. {yields} Fluorescent mAb-metal complex with enhanced intensity and shortened lifetime. {yields} Immuno-interactions of mAb-metal complexes with CK19 molecules on CNCAP and HeLa cell surfaces. {yields} Isolation of conjugated mAb-metal complexes from cellular autofluorescence on cell image. -- Abstract: In this article, we report the synthesis strategy and optical properties of a novel type of fluorescence metal nanoshell when it was used as imaging agent for fluorescence cell imaging. The metal nanoshells were made with 40 nm silica cores and 10 nm silver shells. Unlike typical fluorescence metal nanoshells which contain the organic dyes in the cores, novel metal nanoshells were composed of Cy5-labelled monoclonal anti-CK19 antibodies (mAbs) on the external surfaces of shells. Optical measurements to the single nanoparticles showed that in comparison with the metal free labelled mAbs, the mAb-Ag complexes displayed significantly enhanced emission intensity and dramatically shortened lifetime due to near-field interactions of fluorophores with metal. These metal nanoshells were found to be able to immunoreact with target cytokeratin 19 (CK19) molecules on the surfaces of LNCAP and HeLa cells. Fluorescence cell images were recorded on a time-resolved confocal microscope. The emissions from the metal nanoprobes could be clearly isolated from the cellular autofluorescence backgrounds on the cell images as either individuals or small clusters due to their stronger emission intensities and shorter lifetimes. These emission signals could also be precisely counted on single cell images. The count number may provide an approach for quantifying the target molecules in the cells.

  10. Use of multiphoton tomography and fluorescence lifetime imaging to investigate skin pigmentation in vivo

    Science.gov (United States)

    Dancik, Yuri; Favre, Amandine; Loy, Chong Jin; Zvyagin, Andrei V.; Roberts, Michael S.

    2013-02-01

    There is a growing body of literature showing the usefulness of multiphoton tomography (MPT) and fluorescence lifetime imaging for in situ characterization of skin constituents and the ensuing development of noninvasive diagnostic tools against skin diseases. Melanin and pigmentation-associated skin cancers constitute some of the major applications. We show that MPT and fluorescence lifetime imaging can be used to measure changes in cutaneous melanin concentration and that these can be related to the visible skin color. Melanin in the skin of African, Indian, Caucasian, and Asian volunteers is detected on the basis of its emission wavelength and fluorescence lifetimes in solution and in a melanocyte-keratinocyte cell culture. Fluorescence intensity is used to characterize the melanin content and distribution as a function of skin type and depth into the skin (stratum granulosum and stratum basale). The measured fluorescence intensities in given skin types agree with melanin amounts reported by others using biopsies. Our results suggest that spatial distribution of melanin in skin can be studied using MPT and fluorescence lifetime imaging, but further studies are needed to ascertain that the method can resolve melanin amount in smaller depth intervals.

  11. Highly confined, enhanced surface fluorescence imaging with two-dimensional silver nanoparticle sheets

    Energy Technology Data Exchange (ETDEWEB)

    Usukura, Eiji; Shinohara, Shuhei; Okamoto, Koichi; Tamada, Kaoru, E-mail: tamada@ms.ifoc.kyushu-u.ac.jp [Institute for Materials Chemistry and Engineering, Kyushu University, Fukuoka 812-8581 (Japan); Lim, Jaehoon; Char, Kookheon [The National Creative Research Center for Intelligent Hybrid, School of Chemical and Biological Engineering, Seoul National University, Seoul 151-744 (Korea, Republic of)

    2014-03-24

    A method of obtaining highly confined, enhanced surface fluorescence imaging is proposed using two-dimensional (2D) silver nanoparticle (AgMy) sheets. This technique is based on the localized surface plasmon resonance excited homogeneously on a 2D silver nanoparticle sheet. The AgMy sheets are fabricated at the air–water interface by self-assembly and transferred onto hydrophobic glass substrates. These sheets can enhance the fluorescence only when the excitation wavelength overlaps with the plasmon resonance wavelength. To confirm the validity of this technique, two separate test experiments are performed. One is the epifluorescence microscope imaging of a quantum dot 2D sheet on the AgMy 2D sheet with a SiO{sub 2} spacer layer, where the fluorescence is maximized with the 20 nm SiO{sub 2} layer, determined by the Förster resonance energy transfer distances. The second experiment is the imaging of a single fluorescence bead with a total internal reflection fluorescent microscope. We confirmed that the AgMy sheet provides a 4-fold increase in fluorescence with a 160-nm spatial resolution at 30 ms/frame snapshot. The AgMy sheet will be a powerful tool for high sensitivity and high-resolution real time bioimaging at nanointerfaces.

  12. Improving Surgical Resection of Metastatic Liver Tumors With Near-Infrared Optical-Guided Fluorescence Imaging.

    Science.gov (United States)

    Barabino, Gabriele; Porcheron, Jack; Cottier, Michèle; Cuilleron, Muriel; Coutard, Jean-Guillaume; Berger, Michel; Molliex, Serge; Beauchesne, Brigitte; Phelip, Jean Marc; Grichine, Alexei; Coll, Jean-Luc

    2016-08-01

    Objective The aim of this study was to investigate the feasibility and future clinical applications of near-infrared (NIR) fluorescence imaging to guide liver resection surgery for metastatic cancer to improve resection margins. Summary Background Data A subset of patients with metastatic hepatic tumors can be cured by surgery. The degree of long-term and disease-free survival is related to the quality of surgery, with the best resection defined as "R0" (complete removal of all tumor cells, as evidenced by microscopic examination of the margins). Although intraoperative ultrasonography can evaluate the surgical margins, surgeons need a new tool to perfect the surgical outcome. Methods A preliminary study was performed on 3 patients. We used NIR imaging postoperatively "ex vivo" on the resected liver tissue. The liver tumors were preoperatively labelled by intravenously injecting the patient with indocyanine green (ICG), a NIR fluorescent agent (24 hours before surgery, 0.25 mg/kg). Fluorescent images were obtained using a miniaturized fluorescence imaging system (FluoStic, Fluoptics, Grenoble, France). Results After liver resection, the surgical specimens from each patient were sliced into 10-mm sections in the operating room and analyzed with the FluoStic. All metastatic tumors presented rim-type fluorescence. Two specimens had incomplete rim fluorescence. The pathologist confirmed the presence of R1 margins (microscopic residual resection), even though the ultrasonographic analysis indicated that the result was R0. Conclusions Surgical liver resection guided by NIR fluorescence can help detect potentially uncertain anatomical areas that may be missed by preoperative imaging and by ultrasonography during surgery. These preliminary results will need to be confirmed in a larger prospective patient series.

  13. Application of subtraction CT perfusion imaging in early avascular necrosis of femoral head%数字减影CT灌注成像在股骨头早期缺血性坏死中的应用

    Institute of Scientific and Technical Information of China (English)

    刘海明; 梁辉清; 刘万新; 刘日新; 黄锦钊; 袁国奇

    2013-01-01

    Objective To investigate the application of subtraction CT perfusion imaging (SCTP) in early avascular necrosis of femoral head (ANFH).Methods Nineteen patients of ANFH were performed plain CT scan before the perfusion,and then performed perfusion scan at the same levels.The perfusion images were processed by digital subtraction perfusion software to get a group of new perfusion images.Then the images were post-processed by workstation perfusion software,and blood flow (BF),blood volume (BV),mean transit time (MTT) and the corresponding color mapping were calculated for the diagnosis of ANFH.Results The BF value of femoral head at the affected side of early ANFH (n=27) were significantly reduced,compared with that at the healthy side (n=11),P<0.05.There were no statistically significant differences in BV or MTT between the two sides (P>0.05).Conclusion BF,BV,MTT can demonstrate the microcirculation perfusion information of ANFH,which provides an important basis for the clinical diagnosis of ANFH.%目的 探讨数字减影CT灌注成像(Subtraction CT perfusion imaging,SCTP)在股骨头早期缺血坏死(Avascular necrosis of femoral head,ANFH)中的应用.方法 股骨头早期缺血性坏死19例,股骨头灌注前先行平扫,再进行同层面灌注扫描,将灌注图像经数字减影后得到一组新灌注图像.再经工作站灌注软件后处理,计算出血流量(BF)、血容量(BV)、对比剂平均通过时间(MTT)及相应色阶图,用于ANFH诊断.结果 早期ANFH患侧(27侧)股骨头BF数值较健侧(11侧)BF显著减少,差异有统计学意义(P<0.05).患侧股骨头BV、MTT数值对比健侧组间差异无统计学意义(P>0.05).结论 SCTP计算出的BF、BV、MTT能反映早期股骨头缺血的微循环灌注信息,可以为临床诊断ANFH提供重要的诊断依据.

  14. In-vivo optical detection of cancer using chlorin e6 – polyvinylpyrrolidone induced fluorescence imaging and spectroscopy

    OpenAIRE

    Soo Khee; Bhuvaneswari Ramaswamy; Thong Patricia SP; Chin William WL; Heng Paul WS; Olivo Malini

    2009-01-01

    Abstract Background Photosensitizer based fluorescence imaging and spectroscopy is fast becoming a promising approach for cancer detection. The purpose of this study was to examine the use of the photosensitizer chlorin e6 (Ce6) formulated in polyvinylpyrrolidone (PVP) as a potential exogenous fluorophore for fluorescence imaging and spectroscopic detection of human cancer tissue xenografted in preclinical models as well as in a patient. Methods Fluorescence imaging was performed on MGH human...

  15. Improving Automated Annotation of Benthic Survey Images Using Wide-band Fluorescence

    Science.gov (United States)

    Beijbom, Oscar; Treibitz, Tali; Kline, David I.; Eyal, Gal; Khen, Adi; Neal, Benjamin; Loya, Yossi; Mitchell, B. Greg; Kriegman, David

    2016-03-01

    Large-scale imaging techniques are used increasingly for ecological surveys. However, manual analysis can be prohibitively expensive, creating a bottleneck between collected images and desired data-products. This bottleneck is particularly severe for benthic surveys, where millions of images are obtained each year. Recent automated annotation methods may provide a solution, but reflectance images do not always contain sufficient information for adequate classification accuracy. In this work, the FluorIS, a low-cost modified consumer camera, was used to capture wide-band wide-field-of-view fluorescence images during a field deployment in Eilat, Israel. The fluorescence images were registered with standard reflectance images, and an automated annotation method based on convolutional neural networks was developed. Our results demonstrate a 22% reduction of classification error-rate when using both images types compared to only using reflectance images. The improvements were large, in particular, for coral reef genera Platygyra, Acropora and Millepora, where classification recall improved by 38%, 33%, and 41%, respectively. We conclude that convolutional neural networks can be used to combine reflectance and fluorescence imagery in order to significantly improve automated annotation accuracy and reduce the manual annotation bottleneck.

  16. Fluorescence and confocal imaging of mammalian cells using conjugated oligoelectrolytes with phenylenevinylene core

    Energy Technology Data Exchange (ETDEWEB)

    Milczarek, Justyna; Pawlowska, Roza; Zurawinski, Remigiusz; Lukasik, Beata; Garner, Logan E.; Chworos, Arkadiusz

    2017-05-01

    Over the last few years, considerable efforts are taken, in order to find a molecular fluorescent probe fulfilling their applicability requirements. Due to a good optical properties and affinity to biological structures conjugated oligoelectrolytes (COEs) can be considered as a promising dyes for application in fluorescence-based bioimaging. In this work, we synthetized COEs with phenylenevinylene core (PV-COEs) and applied as fluorescent membranous-specific probes. Cytotoxicity effects of each COE were probed on cancerous and non-cancerous cell types and little to no toxicity effects were observed at the high range of concentrations. The intensity of cell fluorescence following the COE staining was determined by the photoluminescence analysis and fluorescence activated cell sorting method (FACS). Intercalation of tested COEs into mammalian cell membranes was revealed by fluorescent and confocal microscopy colocalization with commercial dyes specific for cellular structures including mitochondria, Golgi apparatus and endoplasmic reticulum. The phenylenevinylene conjugated oligoelectrolytes have been found to be suitable for fluorescent bioimaging of mammalian cells and membrane-rich organelles. Due to their water solubility coupled with spontaneous intercalation into cells, favorable photophysical features, ease of cell staining, low cytotoxicity and selectivity for membranous structures, PV-COEs can be applied as markers for fluorescence imaging of a variety of cell types.

  17. Fluorescence imaging for a noninvasive in vivo toxicity-test using a transgenic silkworm expressing green fluorescent protein.

    Science.gov (United States)

    Inagaki, Yoshinori; Matsumoto, Yasuhiko; Ishii, Masaki; Uchino, Keiro; Sezutsu, Hideki; Sekimizu, Kazuhisa

    2015-06-10

    In drug development, the toxicity of candidate chemicals must be carefully examined in an animal model. Here we developed a live imaging technique using silkworms for a noninvasive toxicity test applicable for drug screening. Injection of carbon tetrachloride, a tissue-injuring chemical, into transgenic silkworms expressing green fluorescent protein (GFP) induced leakage of GFP from the tissues into the hemolymph. The leakage of GFP was suppressed by pre-administration of either cimetidine, a cytochrome P450 inhibitor, or N-acetyl cysteine, a free-radical scavenger. The transgenic silkworm was made transparent by feeding a diet containing chemicals that inhibit uric acid deposition in the epithelial cells. In the transparent silkworms, GFP fluorescence in the fat body could be observed from outside the body. Injection of salicylic acid or iron sulfate, tissue-injuring chemicals, into the transparent silkworms decreased the fluorescence intensity of the GFP in the fat body. These findings suggest that the transparent GFP-expressing silkworm model is useful for evaluating the toxicity of chemicals that induce tissue injury.

  18. Augmented microscopy: real-time overlay of bright-field and near-infrared fluorescence images.

    Science.gov (United States)

    Watson, Jeffrey R; Gainer, Christian F; Martirosyan, Nikolay; Skoch, Jesse; Lemole, G Michael; Anton, Rein; Romanowski, Marek

    2015-10-01

    Intraoperative applications of near-infrared (NIR) fluorescent contrast agents can be aided by instrumentation capable of merging the view of surgical field with that of NIR fluorescence. We demonstrate augmented microscopy, an intraoperative imaging technique in which bright-field (real) and electronically processed NIR fluorescence (synthetic) images are merged within the optical path of a stereomicroscope. Under luminance of 100,000 lx, representing typical illumination of the surgical field, the augmented microscope detects 189 nM concentration of indocyanine green and produces a composite of the real and synthetic images within the eyepiece of the microscope at 20 fps. Augmentation described here can be implemented as an add-on module to visualize NIR contrast agents, laser beams, or various types of electronic data within the surgical microscopes commonly used in neurosurgical, cerebrovascular, otolaryngological, and ophthalmic procedures.

  19. High-speed confocal fluorescence lifetime imaging microscopy by analog mean-delay method

    Science.gov (United States)

    Won, Youngjae; Kim, Donguk; Yang, Wenzhong; Kim, Dug Y.

    2010-02-01

    We have demonstrated the high-speed confocal fluorescence lifetime imaging microscopy (FLIM) by analog mean-delay (AMD) method. The AMD method is a new signal processing technique for calculation of fluorescence lifetime and it is very suitable for the high-speed confocal FLIM with good accuracy and photon economy. We achieved the acquisition speed of 7.7 frames per second for confocal FLIM imaging. Here, the highest photon detection rate for one pixel was larger than 125 MHz and averaged photon detection rate was more than 62.5 MHz. Based on our system, we successfully obtained a sequence of confocal fluorescence lifetime images of RBL-2H3 cell labeled with Fluo-3/AM and excited by 4αPDD (TRPV channel agonist) within one second.

  20. Full-angle optical imaging of near-infrared fluorescent probes implanted in small animals

    Institute of Scientific and Technical Information of China (English)

    Gang Hu; Junjie Yao; Jing Bai

    2008-01-01

    To provide a valuable experimental platform for in vivo biomedical research of small animal model with fluorescence mediated approach, we developed a whole-body near-infrared fluorescence molecular imaging system as described in this paper. This system is based on a sensitive CCD camera and has the ability to achieve 360° full-angle source illuminations and projections capture of the targets to obtain the dense sampling by performing rotational scan. The measurement accuracy is validated from cylinder phantom experiments by the comparison between the experimental data and theoretical predictions. Finally, we also present typical in vivo images of fluorescent tube implanted into the mouse body. The results are promising and have proved the system imaging performance for macroscopic optical biomedical research.

  1. Neutron, fluorescence, and optical imaging: An in situ combination of complementary techniques

    Energy Technology Data Exchange (ETDEWEB)

    Wagner, D.; Egelhaaf, S. U.; Hermes, H. E. [Condensed Matter Physics Laboratory, Heinrich Heine University, 40225 Düsseldorf (Germany); Börgardts, M.; Müller, T. J. J. [Institute for Organic and Macromolecular Chemistry, Heinrich Heine University, 40225 Düsseldorf (Germany); Grünzweig, C.; Lehmann, E. [Neutron Imaging and Activation Group, Paul Scherrer Institute, 5232 Villigen (Switzerland)

    2015-09-15

    An apparatus which enables the simultaneous combination of three complementary imaging techniques, optical imaging, fluorescence imaging, and neutron radiography, is presented. While each individual technique can provide information on certain aspects of the sample and their time evolution, a combination of the three techniques in one setup provides a more complete and consistent data set. The setup can be used in transmission and reflection modes and thus with optically transparent as well as opaque samples. Its capabilities are illustrated with two examples. A polymer hydrogel represents a transparent sample and the diffusion of fluorescent particles into and through this polymer matrix is followed. In reflection mode, the absorption of solvent by a nile red-functionalized mesoporous silica powder and the corresponding change in fluorescent signal are studied.

  2. Assessment of the quality of durum wheat products by spectrofluorometry and fluorescence video image analysis

    Science.gov (United States)

    Novales, Bruno; Abecassis, Joel; Bertrand, Dominique; Devaux, Marie-Francoise; Robert, Paul

    1995-01-01

    Because assessment of Durum wheat semolina purity by standard ash-test has been widely criticized, we attempted to characterize products of a semolina mill by spectrofluorometry and fluorescence imaging. A collection of milled wheat products ranging from very pure semolina to brans were chosen for this study. Multidimensional statistical analyses (Principal component analyses) were applied to the spectral and image data. Maps showing a classification of the products according to purity were obtained without biochemical calibration. Principal component regression was applied to the data in order to test the relationship of aleurone fluorescence to ash content. Both spectrofluorometry and fluorescence imaging gave similar results with good determination coefficients (r2 equals 0.97 and 0.92) for the study of a single wheat variety. Products obtained from different wheat varieties were more difficult to compare.

  3. Evaluating the use of fluorescent imaging for the quantification of dental fluorosis

    Directory of Open Access Journals (Sweden)

    McGrady Michael G

    2012-11-01

    Full Text Available Abstract Background The quantification of fluorosis using fluorescence imaging (QLF hardware and stain analysis software has been demonstrated in selected populations with good correlation between fluorescent image metrics and TF Index scores from photographs. The aim of this study was to evaluate the ability of QLF to quantify fluorosis in a population of subjects (aged 11–13 participating in an epidemiological caries and fluorosis survey in fluoridated and non-fluoridated communities in Northern England. Methods Fluorescent images of the maxillary incisors were captured together with standardized photographs were scored blind for fluorosis using the TF Index. Subjects were excluded from the analysis if there were restorations or caries on the maxillary central incisors. Results Data were available for 1774 subjects (n=905 Newcastle, n=869 Manchester. The data from the fluorescence method demonstrated a significant correlation with TF Index scores from photographs (Kendall’s tau = 0.332 p Conclusions Despite confounding factors the fluorescence imaging system may provide a useful objective, blinded system for the assessment of enamel fluorosis when used adjunctively with photographic scoring.

  4. High-Speed Fluorescence Microscopy: Lifetime Imaging in the Biomedical Sciences

    Science.gov (United States)

    Periasamy, Ammasi; Wang, Xue F.; Wodnick, Pawel; Gordon, Gerald W.; Kwon, Seongwook; Diliberto, Pamela A.; Herman, Brian

    1995-02-01

    The ability to observe the behavior of living cells and tissues provides unparalleled access to information regarding the organization and dynamics of complex cellular structures. While great strides have been made over the past 30 to 40 years in the design and application of a variety of novel optical microscopic techniques, until recently, it has not been possible to image biological phenomena that occur over very short time periods (nanosecond to millisecond) or over short distances (10 to 1000 [Angstrom capital A, ring]). However, the recent combination of (1) very rapidly gated and sensitive image intensifiers and (2) the ability to deliver fluorescence excitation energy to intact living biological specimens in a pulsed or sinusoidally modulated fashion has allowed such measurements to become a reality through the imaging of the lifetimes of fluorescent molecules. This capability has resulted in the ability to observe the dynamic organization and interaction of cellular components on a spatial and temporal scale previously not possible using other microscopic techniques. This paper discusses the implementation of a fluorescence lifetime imaging microscope (FLIM) and provides a review of some of the applications of such an instrument. These include measurements of receptor topography and subunit interactions using fluorescence resonance energy transfer (FRET), fluorescence anisotropy of phospholipids in cell membranes, cytosolic free calcium (Ca2+)i and the detection of human papillomavirus (HPV) infection in clinical cervicovaginal smears.

  5. An all-fiber-optic endoscopy platform for simultaneous OCT and fluorescence imaging.

    Science.gov (United States)

    Mavadia, Jessica; Xi, Jiefeng; Chen, Yongping; Li, Xingde

    2012-11-01

    We present an all-fiber-optically based endoscope platform for simultaneous optical coherence tomography (OCT) and fluorescence imaging. This design entails the use of double-clad fiber (DCF) in the endoscope for delivery of OCT source and fluorescence excitation light while collecting the backscattered OCT signal through the single-mode core and fluorescence emission through the large inner cladding of the DCF. Circumferential beam scanning was performed by rotating a 45° reflector using a miniature DC motor at the distal end of the endoscope. Additionally, a custom DCF coupler and a wavelength division multiplexer (WDM) were utilized to seamlessly integrate both imaging modalities to achieve an entirely fiber-optically based dual-modality imaging system. We demonstrated simultaneous intraluminal 3D OCT and 2D (surface) fluorescence imaging in ex vivo rabbit esophagus using the dual-modal endomicroscopy system. Structural morphologies (provided by OCT) and fluorophore distribution (provided by the fluorescence module) could be clearly visualized, suggesting the potential of the dual-modality system for future in vivo and clinical applications.

  6. Differentiation of ocular fundus fluorophores by fluorescence lifetime imaging using multiple excitation and emission wavelengths

    Science.gov (United States)

    Hammer, M.; Schweitzer, D.; Schenke, S.; Becker, W.; Bergmann, A.

    2006-10-01

    Ocular fundus autofluorescence imaging has been introduced into clinical diagnostics recently. It is in use for the observation of the age pigment lipofuscin, a precursor of age - related macular degeneration (AMD). But other fluorophores may be of interest too: The redox pair FAD - FADH II provides information on the retinal energy metabolism, advanced glycation end products (AGE) indicate protein glycation associated with pathologic processes in diabetes as well as AMD, and alterations in the fluorescence of collagen and elastin in connective tissue give us the opportunity to observe fibrosis by fluorescence imaging. This, however, needs techniques able to differentiate particular fluorophores despite limited permissible ocular exposure as well as excitation wavelength (limited by the transmission of the human ocular lens to >400 nm). We present an ophthalmic laser scanning system (SLO), equipped with picosecond laser diodes (FWHM 100 ps, 446 nm or 468 nm respectively) and time correlated single photon counting (TCSPC) in two emission bands (500 - 560 nm and 560 - 700 nm). The decays were fitted by a bi-exponential model. Fluorescence spectra were measured by a fluorescence spectrometer fluorolog. Upon excitation at 446 nm, the fluorescence of AGE, FAD, and lipofuscin were found to peak at 503 nm, 525 nm, and 600 nm respectively. Accordingly, the statistical distribution of the fluorescence decay times was found to depend on the different excitation wavelengths and emission bands used. The use of multiple excitation and emission wavelengths in conjunction with fluorescence lifetime imaging allows us to discriminate between intrinsic fluorophores of the ocular fundus. Taken together with our knowledge on the anatomical structure of the fundus, these findings suggest an association of the short, middle and long fluorescence decay time to the retinal pigment epithelium, the retina, and connective tissue respectively.

  7. A Single-Photon Avalanche Diode Array for Fluorescence Lifetime Imaging Microscopy.

    Science.gov (United States)

    Schwartz, David Eric; Charbon, Edoardo; Shepard, Kenneth L

    2008-11-21

    We describe the design, characterization, and demonstration of a fully integrated single-photon avalanche diode (SPAD) imager for use in time-resolved fluorescence imaging. The imager consists of a 64-by-64 array of active SPAD pixels and an on-chip time-to-digital converter (TDC) based on a delay-locked loop (DLL) and calibrated interpolators. The imager can perform both standard time-correlated single-photon counting (TCSPC) and an alternative gated-window detection useful for avoiding pulse pile-up when measuring bright signal levels. To illustrate the use of the imager, we present measurements of the decay lifetimes of fluorescent dyes of several types with a timing resolution of 350 ps.

  8. Fusion and subtraction post-processing in body MRI

    Energy Technology Data Exchange (ETDEWEB)

    Watson, Tom A.; Olsen, Oeystein E. [Great Ormond Street Hospital for Children NHS Foundation Trust, Department of Radiology, London (United Kingdom)

    2014-09-02

    Interpreting complex paediatric body MRI studies requires the integration of information from multiple sequences. Image processing software, some freely available, allows the radiologist to use simple and rapid post-processing techniques that may aid diagnosis. We demonstrate the use of fusion and subtraction post-processing techniques with examples from four areas of application: enterography, oncological imaging, musculoskeletal imaging and MR fistulography. (orig.)

  9. Characterizing fluorescent imaging properties of antibodies conjugated to IRDye800CW for use in imaging of head and neck cancer

    Science.gov (United States)

    Foster, Robert C.; Krell, Asher M.; Chung, Thomas K.; Warram, Jason M.; Zinn, Kurt R.; Rosenthal, Eben L.

    2014-03-01

    Introduction: Proteins conjugated to the near infrared (NIR) moieties for detection of head and neck cancers are being translated to the clinic. However, little is known about the fluorescent properties of IRDye800CW after conjugation to antibodies. We investigated factors that may alter the real-time observed fluorescence of antibody conjugated dye and the rate of fluorescent signal loss. Methods: Signal loss was examined using three FDA approved monoclonal antibodies conjugated to IRDye800CW (LICOR) over a period of 15 days. Temperature effects on fluorescence were examined for conjugated dye in both solution and a mouse tumor model. Samples were cooled to -20°C then warmed to predetermined temperatures up to 60°C with imaging performed using the PEARL Impulse (LI-COR) and LUNA (Novadaq) systems. Results: Short term fluorescent signal loss (decreasing temperature with statistically significant increases seen at -20°C and 4°C (p=0.0015, p=0.03). Conclusions: TBR is increased with decreasing sample temperature, suggesting that the clinical exam of fluorescently labeled tissues may be improved at cooler temperatures. Our results indicate that both the rate of signal loss and the change in fluorescence with temperature observed for IRDye800CW are independent of the conjugating antibody.

  10. Development of a noncontact 3-D fluorescence tomography system for small animal in vivo imaging

    Science.gov (United States)

    Zhang, Xiaofeng; Badea, Cristian; Jacob, Mathews; Johnson, G. Allan

    2009-02-01

    Fluorescence imaging is an important tool for tracking molecular-targeting probes in preclinical studies. It offers high sensitivity, but nonetheless low spatial resolution compared to other leading imaging methods such CT and MRI. We demonstrate our methodological development in small animal in vivo whole-body imaging using fluorescence tomography. We have implemented a noncontact fluid-free fluorescence diffuse optical tomography system that uses a raster-scanned continuous-wave diode laser as the light source and an intensified CCD camera as the photodetector. The specimen is positioned on a motorized rotation stage. Laser scanning, data acquisition, and stage rotation are controlled via LabVIEW applications. The forward problem in the heterogeneous medium is based on a normalized Born method, and the sensitivity function is determined using a Monte Carlo method. The inverse problem (image reconstruction) is performed using a regularized iterative algorithm, in which the cost function is defined as a weighted sum of the L-2 norms of the solution image, the residual error, and the image gradient. The relative weights are adjusted by two independent regularization parameters. Our initial tests of this imaging system were performed with an imaging phantom that consists of a translucent plastic cylinder filled with tissue-simulating liquid and two thin-wall glass tubes containing indocyanine green. The reconstruction is compared to the output of a finite element method-based software package NIRFAST and has produced promising results.

  11. Ultrasmall near-infrared gold nanoclusters for tumor fluorescence imaging in vivo

    Science.gov (United States)

    Wu, Xu; He, Xiaoxiao; Wang, Kemin; Xie, Can; Zhou, Bing; Qing, Zhihe

    2010-10-01

    In this paper, we explore the possibility of using ultrasmall near-infrared (NIR) gold nanoclusters (AuNCs) as novel contrast imaging agents for tumor fluorescence imaging in vivo. The fluorescence imaging signal of the tail vein administrated AuNCs in living organisms can spectrally be well distinguished from the background with maximum emission wavelength at about 710 nm, and the high photostability of AuNCs promises continuous imaging in vivo. The uptake of AuNCs by the reticuloendothelial system is relatively low in comparison with other nanoparticle-based contrast imaging agents due to their ultrasmall hydrodynamic size (~2.7 nm). Through the body weight change analysis, the results show that the body weight of the mice administrated with AuNCs has not been changed obviously in comparison with that of the control mice injected with PBS. Furthermore, using MDA-MB-45 and Hela tumor xenograft models, in vivo and ex vivo imaging studies show that the ultrasmall NIR AuNCs are able to be highly accumulated in the tumor areas, thanks to the enhanced permeability and retention (EPR) effects. And the tumor-to-background ratio is about 15 for 6 h postinjection. The results indicate that the ultrasmall NIR AuNCs appear as very promising contrast imaging agents for in vivo fluorescence tumor imaging.

  12. Imaging of Bacterial and Fungal Cells Using Fluorescent Carbon Dots Prepared from Carica papaya Juice.

    Science.gov (United States)

    Kasibabu, Betha Saineelima B; D'souza, Stephanie L; Jha, Sanjay; Kailasa, Suresh Kumar

    2015-07-01

    In this paper, we have described a simple hydrothermal method for preparation of fluorescent carbon dots (C-dots) using Carica papaya juice as a precursor. The synthesized C-dots show emission peak at 461 nm with a quantum yield of 7.0 %. The biocompatible nature of C-dots was confirmed by a cytotoxicity assay on E. coli. The C-dots were used as fluorescent probes for imaging of bacterial (Bacillus subtilis) and fungal (Aspergillus aculeatus) cells and emitted green and red colors under different excitation wavelengths, which indicates that the C-dots can be used as a promising material for cell imaging.

  13. 减影CT灌注成像技术及对犬股骨头坏死观察应用%Subtraction computed tomographic perfusion imaging and observation of ischemic necrosis of femoral head on dog

    Institute of Scientific and Technical Information of China (English)

    杨秀军; 任其乐; 李巍; 胥文娟

    2009-01-01

    目的 探讨减影计算机体层摄影灌注成像(sCTP)技术及观察犬缺血性股骨头坏死的方法与可行性.方法 对16只犬于旋股动脉内结扎术前后行股骨头CTP扫描,以观察股骨头坏死.在AW 4.2工作站利用减影软件对CTP源影像进行减影处理,再以Perfusion 3软件对减影图像数据和源影像数据分别作血流量(BF)、血容量(BV)、平均通过时间(MTT)色阶图分析并测量兴趣区其参数值. 结果 ①对CTP源影像减影所得新图像数据行灌注成像软件后处理成功率为100%,均获得了BF、BV、MTT色阶图及其数值,sCTP后处理时间约需1~5 h;②sCTP提供的BF、BV、MTT数值及色阶图能对比显示股骨头坏死,常规骨CTP难以揭示这些改变. 结论 sCTP技术可行,适用于骨的CT灌注成像、定量诊断骨坏死.%Objective To investigate the technical protocols and feasibility of subtraction computed tomography perfusion imaging (sCTP) on observation of ischemic necrosis of femoral head (ANFH) on dog. Methods Sixteen laboratory canines underwent CT perfusion imaging (CTP) of femoral head before and after selective femoral circumflex artery embolization, and ANFH were observed. Then new sequence imaging data, created by sources imaging of CT perfusion scan using subtraction software, were analyzed at workstation (AW 4.2) with CT perfusion 3 analysis program, and data of sCTP were obtained. The parametric maps and indexes of capillary-level hemodynamics including blood volume (BV), blood flow (BF) and mean transit time (MTT) of CTP and sCTP were compared. Results ①The technical success rate of sCTP post-processing created from CTP sources imaging data was 100%. The values and mappings of BF, BV and MTT of region of interest (ROI) were all obtained from subtraction sequence images data. The post-processing time of sCTP was about 1-5 h. ② sCTP depicted ANFH well, though the values and mappings of BF, BV and MTT were different from those obtained with

  14. Rational design of a monomeric and photostable far-red fluorescent protein for fluorescence imaging in vivo.

    Science.gov (United States)

    Yu, Dan; Dong, Zhiqiang; Gustafson, William Clay; Ruiz-González, Rubén; Signor, Luca; Marzocca, Fanny; Borel, Franck; Klassen, Matthew P; Makhijani, Kalpana; Royant, Antoine; Jan, Yuh-Nung; Weiss, William A; Guo, Su; Shu, Xiaokun

    2016-02-01

    Fluorescent proteins (FPs) are powerful tools for cell and molecular biology. Here based on structural analysis, a blue-shifted mutant of a recently engineered monomeric infrared fluorescent protein (mIFP) has been rationally designed. This variant, named iBlueberry, bears a single mutation that shifts both excitation and emission spectra by approximately 40 nm. Furthermore, iBlueberry is four times more photostable than mIFP, rendering it more advantageous for imaging protein dynamics. By tagging iBlueberry to centrin, it has been demonstrated that the fusion protein labels the centrosome in the developing zebrafish embryo. Together with GFP-labeled nucleus and tdTomato-labeled plasma membrane, time-lapse imaging to visualize the dynamics of centrosomes in radial glia neural progenitors in the intact zebrafish brain has been demonstrated. It is further shown that iBlueberry can be used together with mIFP in two-color protein labeling in living cells and in two-color tumor labeling in mice.

  15. Comparison between the indocyanine green fluorescence and blue dye methods for sentinel lymph node biopsy using novel fluorescence image-guided resection equipment in different types of hospitals.

    Science.gov (United States)

    He, Kunshan; Chi, Chongwei; Kou, Deqiang; Huang, Wenhe; Wu, Jundong; Wang, Yabing; He, Lifang; Ye, Jinzuo; Mao, Yamin; Zhang, Guo-Jun; Wang, Jiandong; Tian, Jie

    2016-12-01

    Sentinel lymph node biopsy (SLNB) has become a standard of care to detect axillary lymph metastasis in early-stage breast cancer patients with clinically negative axillary lymph nodes. Current SLNB detection modalities comprising a blue dye, a radioactive tracer, or a combination of both have advantages as well as disadvantages. Thus, near-infrared fluorescence imaging using indocyanine green (ICG) has recently been regarded as a novel method that has generated interest for SLNB around the world. However, the lack of appropriate fluorescence imaging systems has hindered further research and wide application of this method. Therefore, we developed novel fluorescence image-guided resection equipment (FIRE) to detect sentinel lymph nodes (SLNs). Moreover, to compare the ICG fluorescence imaging method with the blue dye method and to explore the universal feasibility of the former, a different type of hospital study was conducted. Ninety-nine eligible patients participated in the study at 3 different types of hospitals. After subcutaneous ICG allergy testing, all the patients were subcutaneously injected with methylene blue and ICG into the subareolar area. Consequently, 276 SLNs (range 1-7) were identified in 98 subjects (detection rate: 99%) by using the ICG fluorescence imaging method. In contrast, the blue dye method only identified 202 SLNs (range 1-7) in 91 subjects (detection rate: 91.92%). Besides, the results of the fluorescence imaging method were similar in the 3 hospitals. Our findings indicate the universal feasibility of the ICG fluorescence imaging method for SLNB using the fluorescence image-guided resection equipment in early breast cancer detection. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Background subtraction theory and practice

    CERN Document Server

    Elgammal, Ahmed

    2014-01-01

    Background subtraction is a widely used concept for detection of moving objects in videos. In the last two decades there has been a lot of development in designing algorithms for background subtraction, as well as wide use of these algorithms in various important applications, such as visual surveillance, sports video analysis, motion capture, etc. Various statistical approaches have been proposed to model scene backgrounds. The concept of background subtraction also has been extended to detect objects from videos captured from moving cameras. This book reviews the concept and practice of back

  17. CMOS image sensor with lateral electric field modulation pixels for fluorescence lifetime imaging with sub-nanosecond time response

    Science.gov (United States)

    Li, Zhuo; Seo, Min-Woong; Kagawa, Keiichiro; Yasutomi, Keita; Kawahito, Shoji

    2016-04-01

    This paper presents the design and implementation of a time-resolved CMOS image sensor with a high-speed lateral electric field modulation (LEFM) gating structure for time domain fluorescence lifetime measurement. Time-windowed signal charge can be transferred from a pinned photodiode (PPD) to a pinned storage diode (PSD) by turning on a pair of transfer gates, which are situated beside the channel. Unwanted signal charge can be drained from the PPD to the drain by turning on another pair of gates. The pixel array contains 512 (V) × 310 (H) pixels with 5.6 × 5.6 µm2 pixel size. The imager chip was fabricated using 0.11 µm CMOS image sensor process technology. The prototype sensor has a time response of 150 ps at 374 nm. The fill factor of the pixels is 5.6%. The usefulness of the prototype sensor is demonstrated for fluorescence lifetime imaging through simulation and measurement results.

  18. Motion Tracking with Fast Adaptive Background Subtraction

    Institute of Scientific and Technical Information of China (English)

    Xiao; De-Gui; Yu; Sheng-sheng; 等

    2003-01-01

    To extract and track moving objects is usually one of the most important tasks of intelligent video surveillance systems. This paper presents a fast and adaptive background subtraction algorithm and the motion tracking process using this algorithm. The algorithm uses only luminance components of sampled image sequence pixels and models every pixel in a statistical model.The algorithm is characterized by its ability of real time detecting sudden lighting changes, and extracting and tracking motion objects faster. It is shown that our algorithm can be realized with lower time and space complexity and adjustable object detection error rate with comparison to other background subtraction algorithms. Making use of the algorithm, an indoor monitoring system is also worked out and the motion tracking process is presented in this paper.Experimental results testify the algorithms' good performances when used in an indoor monitoring system.

  19. Interferometry with Photon-Subtracted Thermal Light

    CERN Document Server

    Rafsanjani, Seyed Mohammad Hashemi; Magana-Loaiza, Omar S; Gard, Bryan T; Birrittella, Richard; Koltenbah, B E; Parazzoli, C G; Capron, Barbara A; Gerry, Christopher C; Dowling, Jonathan P; Boyd, Robert W

    2016-01-01

    We propose and implement a quantum procedure for enhancing the sensitivity with which one can determine the phase shift experienced by a weak light beam possessing thermal statistics in passing through an interferometer. Our procedure entails subtracting exactly one (which can be generalized to m) photons from the light field exiting an interferometer containing a phase-shifting element in one of its arms. As a consequence of the process of photon subtraction, and somewhat surprisingly, the mean photon number and signal-to-noise ratio of the resulting light field are thereby increased, leading to enhanced interferometry. This method can be used to increase measurement sensitivity in a variety of practical applications, including that of forming the image of an object illuminated only by weak thermal light.

  20. Fluorescent metal nanoshell and CK19 detection on single cell image.

    Science.gov (United States)

    Zhang, Jian; Fu, Yi; Li, Ge; Lakowicz, Joseph R; Zhao, Richard Y

    2011-09-16

    In this article, we report the synthesis strategy and optical properties of a novel type of fluorescence metal nanoshell when it was used as imaging agent for fluorescence cell imaging. The metal nanoshells were made with 40 nm silica cores and 10nm silver shells. Unlike typical fluorescence metal nanoshells which contain the organic dyes in the cores, novel metal nanoshells were composed of Cy5-labelled monoclonal anti-CK19 antibodies (mAbs) on the external surfaces of shells. Optical measurements to the single nanoparticles showed that in comparison with the metal free labelled mAbs, the mAb-Ag complexes displayed significantly enhanced emission intensity and dramatically shortened lifetime due to near-field interactions of fluorophores with metal. These metal nanoshells were found to be able to immunoreact with target cytokeratin 19 (CK19) molecules on the surfaces of LNCAP and HeLa cells. Fluorescence cell images were recorded on a time-resolved confocal microscope. The emissions from the metal nanoprobes could be clearly isolated from the cellular autofluorescence backgrounds on the cell images as either individuals or small clusters due to their stronger emission intensities and shorter lifetimes. These emission signals could also be precisely counted on single cell images. The count number may provide an approach for quantifying the target molecules in the cells.

  1. Near-field measurements of vegetation by laser-induced fluorescence imaging

    Science.gov (United States)

    Sowinska, Malgorzata; Cunin, Bernard; Deruyver, Aline; Heisel, Francine; Miehe, Joseph-Albert; Langsdorf, Gabriele; Lichtenthaler, Hartmut K.

    1999-12-01

    In this paper, a validation of a new UV-A laser-induced fluorescence imaging system implemented in an all-road car for near-field remote sensing of vegetation will be presented. It has been developed as a part of a European Community Program INTERREG II and is consisting of three main parts: excitation, detection and control units. The excitation source is a frequency tripled Nd:YAG laser and the laser spot size is adjusted via a variable beam expander. Fluorescence images are recorded at four characteristic fluorescence bands: 440, 520, 690 and 740 nm with a gated intensified digital CCD camera. The laser head and camera are situated on a directed in site and azimuth platform which can be high up to 6 meters. The platform positioning, localization and distance detection, spot size determination and adjustment, focus, sharpness, selection of the filter, laser and camera synchronization, gain of the intensifier, real time visualization of images, acquisition time are controlled by a newly developed software which allows also image storage, analysis and treatment. Examples of remote sensing fluorescence images from several plant species recorded at a distance of 10 - 30 m will be given and discussed further in this paper.

  2. Endoscopic fluorescence imaging for early assessment of anastomotic recurrence of Crohn's disease

    Science.gov (United States)

    Mordon, Serge R.; Maunoury, Vincent; Geboes, K.; Klein, Olivier; Desreumaux, P.; Debaert, A.; Colombel, Jean-Frederic

    1999-02-01

    Crohn's disease is an inflammatory bowel disease of unknown etiology. The mechanism of the initial mucosal alterations is still unclear: ulcerations overlying lymphoid follicles and/or vasculitis have been proposed as the early lesions. We have developed a new and original method combining endoscopy of fluorescence angiography for identifying the early pathological lesions, occurring in the neo-terminal ileum after right ileocolonic resection. The patient population consisted of 10 subjects enrolled in a prospective protocol of endoscopic follow-up at 3 and 12 months after surgery. Fluorescence imaging showed small spots giving a bright fluorescence distributed singly in mucosa which appeared normal in routine endoscopy. Histopathological examination demonstrated that the fluorescence of small spots originated from small, usually superficial, erosive lesions. In several cases, these erosive lesions occurred over lymphoid follicles. Endoscopic fluorescence imaging provides a suitable means of investigating the initial aspect of the Crohn's disease process in displaying some correlative findings between fluorescent aspects and early pathological mucosal alterations.

  3. Evaluation of PpIX formation in Cervical Intraepithelial Neoplasia I (CIN) using widefield fluorescence images

    Science.gov (United States)

    Carbinatto, Fernanda M.; Inada, Natalia M.; Fortunato, Thereza C.; Lombardi, Welington; da Silva, Eduardo V.; Vollet Filho, José D.; Kurachi, Cristina; Pratavieira, Sebastião.; Bagnato, Vanderlei S.

    2016-03-01

    Optical techniques has been described as auxiliary technology for screening of neoplasia because shows the potential for tissues differentiation in real-time and it is a noninvasive detection and safe. However, only endogenous fluorophores presents the lesion may be insufficient and needed of the administration of the fluorophores synthesized, such as, precursor molecule of protoporphyrin IX (PpIX) induced by 5- aminolevulinic acid and your derivatives. Topical application of methylaminolevulinate (MAL), induces formation of the endogenous photosensitizer, PpIX in tissues where carcinogenesis has begun. The PpIX tend to accumulate in premalignant and malignant tissues and the illumination with light with appropriate wavelength beginning to excitation of PpIX fluorescence, which helps to localize PpIX-rich areas and identify potentially malignant tissues. The aim of the study is to evaluate the production of PpIX in the cervix with CIN I through of the fluorescence