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Sample records for hexylaminolaevulinate fluorescence cystoscopy

  1. Role of hexaminolevulinate-guided fluorescence cystoscopy in bladder cancer

    DEFF Research Database (Denmark)

    Malmström, Per-Uno; Grabe, Magnus; Haug, Erik Skaaheim;

    2012-01-01

    Hexaminolevulinate (HAL) is an optical imaging agent used with fluorescence cystoscopy (FC) for the detection of non-muscle-invasive bladder cancer (NMIBC). Guidelines from the European Association of Urology (EAU) and a recent, more detailed European expert consensus statement agree that HAL......-FC has a role in improving detection of NMIBC and provide recommendations on situations for its use. Since the publication of the EAU guidelines and the European consensus statement, new evidence on the efficacy of HAL-FC in reducing recurrence of NMIBC, compared with white light cystoscopy (WLC), have...

  2. Comparison of hexaminolevulinate based flexible and rigid fluorescence cystoscopy with rigid white light cystoscopy in bladder cancer: results of a prospective Phase II study.

    NARCIS (Netherlands)

    Witjes, J.A.; Moonen, P.M.J.; Heijden, A.G. van der

    2005-01-01

    INTRODUCTION AND OBJECTIVE: Several studies have shown that rigid fluorescence cystoscopy (RFC) with hexaminolevulinate (HAL) is superior to standard rigid white light (RWLC) cystoscopy in diagnosing bladder tumours, with a clinically relevant impact on the patient's management. These studies, howev

  3. Fluorescence and white light cystoscopy for detection of carcinoma in situ of the urinary bladder.

    NARCIS (Netherlands)

    Lerner, S.P.; Liu, H.; Wu, M.F.; Thomas, Y.K.; Witjes, J.A.

    2012-01-01

    OBJECTIVES: To understand the additional benefits of HAL compared with conventional cystoscopy at the patient level and to explore relationships of urine cytology and CIS. METHODS: We reanalyzed pooled data from 3 phase III studies comparing hexaminolevulinate (HAL, Hexvix) fluorescence cystoscopy w

  4. An optical method for reducing green fluorescence from urine during fluorescence-guided cystoscopy

    DEFF Research Database (Denmark)

    Lindvold, Lars René; Hermann, Gregers G

    2016-01-01

    Photodynamic diagnosis (PDD) of bladder tumour tissue significantly improves endoscopic diagnosis and treatment of bladder cancer in rigid cystoscopes in the operating theatre and thus reduces tumour recurrence. PDD comprises the use of blue light, which unfortunately excites green fluorescence...... from urine. As this green fluorescence confounds the desired red fluorescence of the PDD, methods for avoiding this situation particularly in cystoscopy using flexible cystoscopes are desirable. In this paper we demonstrate how a tailor made high power LED light source at 525 nm can be used...... for fluorescence assisted tumour detection using both a flexible and rigid cystoscope used in the outpatient department (OPD) and operating room (OR) respectively. It is demonstrated both in vitro and in vivo how this light source can significantly reduce the green fluorescence problem with urine. At the same time...

  5. Fluorescence cystoscopy in patients with non-muscle invasive bladder cancer

    Directory of Open Access Journals (Sweden)

    I. G. Rusakov

    2015-01-01

    Full Text Available The main challenge of treating non-muscle invasive bladder cancer is multifocal tumors. Current methods of diagnosis are failed to detect all superficial flat tumor lesions in bladder mucosa. The use of fluorescence imaging with 5-aminolevulinic acid (5-ALA allows to improve the sensibility of routine cystoscopy, but low specificity decreases its diagnostic accuracy. The method of fluorescence imaging combined with local fluorescence spectroscopy developed in P.A. Herzen MCRI has been shown to increase the specificity from 71% to 84%. Thus, local fluorescence spectroscopy in visible fluorescence of 5-ALA-induced protoporphyrin allows to perform guided biopsy and decrease the rate of diagnostic mistakes. 

  6. The role of urine markers, white light cystoscopy and fluorescence cystoscopy in recurrence, progression and follow-up of non-muscle invasive bladder cancer

    NARCIS (Netherlands)

    Karaoglu, I.; Heijden, A.G. van der; Witjes, J.A.

    2014-01-01

    Non-muscle invasive bladder cancer (NMIBC) accounts for approximately 70 % of all bladder cancer cases and represents a heterogeneous pathological entity, characterized by a variable natural history and oncological outcome. The combination of cystoscopy and urine cytology is considered the gold stan

  7. Hexaminolevulinate-guided fluorescence cystoscopy in the diagnosis and follow-up of patients with non-muscle-invasive bladder cancer: review of the evidence and recommendations.

    NARCIS (Netherlands)

    Witjes, J.A.; Redorta, J.P.; Jacqmin, D.; Sofras, F.; Malmstrom, P.U.; Riedl, C.; Jocham, D.; Conti, G.; Montorsi, F.; Arentsen, H.C.; Zaak, D.; Mostafid, A.H.; Babjuk, M.

    2010-01-01

    CONTEXT: Compared with standard white-light cystoscopy, photodynamic diagnosis with blue light and the photosensitiser hexaminolevulinate has been shown to improve the visualisation of bladder tumours, reduce residual tumour rates by at least 20%, and improve recurrence-free survival. There is curre

  8. Virtual computed tomography cystoscopy in bladder pathologies

    Directory of Open Access Journals (Sweden)

    Halil Arslan

    2006-04-01

    Full Text Available OBJECTIVE: Assessed the usefulness of virtual cystoscopy performed with multidetector computed tomography (CT in patients with different urinary bladder pathologies compared to the conventional cystoscopy. MATERIALS AND METHODS: Eighteen patients with different bladder pathologies, which consisted of 11 tumors, 3 diverticula, 2 trabecular changes and 2 stones, were assessed with conventional cystoscopy and virtual CT cystoscopy. The results of virtual CT cystoscopy were compared with the findings of conventional cystoscopy. We determined the detection rate and positive predictive value of CT imaging based virtual cystoscopy in the diagnosis of urinary bladder lesions. RESULTS: CT scanning was well tolerated by all patients, and no complications occurred. Images in 16 (88% of the 18 virtual cystoscopic examinations were either of excellent or good quality. All tumors except one, 2 trabecular changes and 2 stones were characterized with similar findings in the both of methods. The masses ranged from 0.4 to 7.0 cm in diameter. While conventional cystoscopy could not evaluate interior part of the diverticulum, virtual CT cystoscopy could demonstrate clearly within it. There were no false-positive findings in our series. CONCLUSION: Virtual CT cystoscopy is a promising technique to be used in the detection of bladder lesions. It should be considered especially at the evaluation of bladder diverticula. In the future, it may be possible or even advantageous to incorporate into the imaging algorithm for evaluation of bladder lesion.

  9. Virtual computed tomography cystoscopy in bladder pathologies

    Energy Technology Data Exchange (ETDEWEB)

    Arslan, Halil; Ceylan, Kadir; Harman, Mustafa; Yilmaz, Yuksel; Temizoz, Osman; Can, Saban [Yuzuncu Yil University School of Medicine, Van (Turkey). Dept. of Radiology and Urology]. E-mail: drhalilarslan@hotmail.com

    2006-03-15

    Objective: assessed the usefulness of virtual cystoscopy performed with multidetector computed tomography (CT) in patients with different urinary bladder pathologies compared to the conventional cystoscopy.Materials and methods: eighteen patients with different bladder pathologies, which consisted of 11 tumors, 3 diverticula, 2 trabecular changes and 2 stones, were assessed with conventional cystoscopy and virtual CT cystoscopy. The results of virtual CT cystoscopy were compared with the findings of conventional cystoscopy. We determined the detection rate and positive predictive value of CT imaging based virtual cystoscopy in the diagnosis of urinary bladder lesions. Results: CT scanning was well tolerated by all patients, and no complications occurred. Images in 16 (88%) of the 18 virtual cystoscopic examinations were either of excellent or good quality. All tumors except one, 2 trabecular changes and 2 stones were characterized with similar findings in the both of methods. The masses ranged from 0.4 to 7.0 cm in diameter. While conventional cystoscopy could not evaluate interior part of the diverticulum, virtual CT cystoscopy could demonstrate clearly within it. There were no false-positive findings in our series. Conclusion: virtual CT cystoscopy is a promising technique to be used in the detection of bladder lesions. It should be considered especially at the evaluation of bladder diverticula. In the future, it may be possible or even advantageous to incorporate into the imaging algorithm for evaluation of bladder lesion. (author)

  10. Cystoscopy

    Science.gov (United States)

    ... Performed The test is done to: Check for cancer of the bladder or urethra Diagnose the cause of blood in the urine ... Elsevier; 2017:chap 73. Read More Bladder ... bladder and urethra Urethral stricture Urge incontinence Urination - painful Review Date ...

  11. Bladder carcinoma: MDCT cystography and virtual cystoscopy.

    Science.gov (United States)

    Panebianco, Valeria; Sciarra, Alessandro; Di Martino, Michele; Bernardo, Silvia; Vergari, Valeria; Gentilucci, Alessandro; Catalano, Carlo; Passariello, Roberto

    2010-06-01

    Bladder carcinoma is the most common tumor among the low urinary tract, accounting for 90% of cancer cases. Conventional cystoscopy represents the gold standard for diagnosis and local management of bladder carcinoma. As the prevalence of transitional cell carcinoma is four-fold greater in men than in women, the endoscopic procedure presents objective difficulties related to the length and bending of male urethra. The most important problems are represented by intense discomfort for the patient and bleeding; furthermore, the high cost, invasivity, and local complications such as infections and mechanical lesions are well-known drawbacks. Additionally, conventional cystoscopy does not provide information about extravescical extensions of the tumor. CT cystography, combined with virtual cystoscopy, is mandatory for TNM staging of the tumor and also is useful when conventional cystoscopy is inconclusive or cannot be performed. We presents the CT cystography findings with virtual endoscopy correlation and bladder carcinoma appearance.

  12. Appearance of a colovesical fistula at cystoscopy

    OpenAIRE

    2015-01-01

    Key Clinical Message Colovesical fistulae typically present with pneumaturia and/or fecaluria. Diverticulitis, inflammatory bowel disease, and malignancies of the colon are the commonest causes. The fistulous tract and adjacent organs are best demonstrated by contrast-enhanced CT scan with rectal contrast or MRI. Biopsy at cystoscopy/colonoscopy is necessary for complete evaluation and treatment planning.

  13. Appearance of a colovesical fistula at cystoscopy.

    Science.gov (United States)

    Aiken, William D; Reid, Gareth; Powell, Leo-Paul

    2015-11-01

    Colovesical fistulae typically present with pneumaturia and/or fecaluria. Diverticulitis, inflammatory bowel disease, and malignancies of the colon are the commonest causes. The fistulous tract and adjacent organs are best demonstrated by contrast-enhanced CT scan with rectal contrast or MRI. Biopsy at cystoscopy/colonoscopy is necessary for complete evaluation and treatment planning.

  14. Factors Predictive of Pain During Cystoscopy: A Prospective Study

    Directory of Open Access Journals (Sweden)

    Ahmet Ozdemir

    2013-10-01

    Full Text Available Aim: To determine the patient characteristics and surgical conditions which is associated with patient pain and discomfort during cystoscopic procedures.Material and Method: Consecutive patients who had a cystoscopy for cancer-related and benign indications were included. Patients were randomized to undergo either rigid or flexible cystoscopy. Procedures were performed in a teaching setting that included residents and attending physicians to assess the impact of experience on pain/discomfort. Patients were further subdivided into the groups that were received detailed verbal explanation during the cystoscopy and allowed to watch the video screen together with the urologist during the procedure or not. Patients were asked to assess their pain on a 10-mm VAS after the procedure the location of the pain was noted. Results: A total of 146 outpatient cystoscopies (99 rigid, 47 flexible were evaluated. There was no correlation between patients age and VAS scores. Mean VAS during flexible cystoscopy was lesser than rigid one. Univariate analysis showed that lower BMI, real time detailed verbal explanation by the physician, lesser physician experience and initial cystoscopies were significantly associated with painful cystoscopies. Lineer regression analysis confirmed that type of instrument, detailed verbal information, physicians experience-more than 3 years and history of previous cystoscopy were independent factors related to VAS scores. The most painful localizations were lidocaine injections during flexible cystoscopy whereas visualization of the side wall of the bladder for rigid technique. Discussion: Instrument type (flexible or rigid, explaining the procedure, experience of the physician, number of previos cystoscopies may have an effect on the amount of pain and discomfort during cystoscopy.

  15. The effect of office based flexible and rigid cystoscopy on pain experience in female patients.

    Science.gov (United States)

    Casteleijn, Niek F; Vriesema, Jessica L; Stomps, Saskia P; van Balen, Olav L W B; Cornel, Erik B

    2017-01-01

    Rigid and flexible cystoscopies are both routinely used in female patients. Literature is conflicting whether flexible cystoscopy is less painful compared to rigid cystoscopy. The aim of this study was therefore to investigate whether using flexible cystoscopy leads to less discomfort and pain compared to rigid cystoscopy in female patients who underwent first time cystoscopy. One hundred eighty-nine female patients, who never had undergone cystoscopy, were randomized into 2 groups: 92 patients underwent rigid cystoscopy and 97 patients flexible cystoscopy. Directly after the cystoscopy procedure all patients were asked to fill out their pain experience on a 100-mm visual analogue pain scale (VAS). Median VAS score was significantly lower for women undergoing flexible cystoscopy (0 [0-20]) compared to rigid cystoscopy (15 [0-38], pflexible cystoscopy was associated with a decrease in VAS score and remained significant after adjustment for age, sex of urologist, performing urologist and indication (standardized β=-0.17, p=0.048). The use of flexible cystoscopy resulted in a significantly lower pain experience compared to rigid cystoscopy. Based on patient's pain experience during cystoscopy, this study implicates to use flexible cystoscopy in female patients who undergo first time cystoscopy.

  16. Low incidence of bacteriuria with outpatient flexible cystoscopy.

    Science.gov (United States)

    Lo, Tsia-Shu; Khanuengkitkong, Siwatchaya; Tan, Yiap Loong; Dass, Anil Krishna; Wu, Pei-Ying; Al-Kharabsheh, Ahlam Mahmoud

    2014-08-01

    To evaluate the incidence of bacteriuria after outpatient diagnostic flexible cystoscopy utilising sterile catheter urine collection technique in women. This is a prospective observational study on 78 women who had outpatient flexible cystoscopy for various indications between March and December 2011. Urine for culture was collected by transurethral catheterisation three to five days before cystoscopy, and a second urine sample was collected five to seven days after the procedure. Antibiotic prophylaxis was not administered. Significant bacteriuria was defined as 10(2)  cfu/mL or more of a single organism cultured. Five (6.4%) had significant bacteriuria and were asymptomatic. The most common organism cultured was Escherichia coli in 4 (80%) of the infected women. No other cystoscopic complication was observed. The mean age was 54.7 years (range 27-80), mean parity was 2.8 (range 0-7). In the study group, 54 (69.2%) women were postmenopausal, 9 (11.5%) had diabetes mellitus, 42 (53.8%) had pelvic organ prolapse and 22 (28.2%) had a prior hysterectomy. Most common indications for cystoscopy were hematuria and recurrent UTI, 48 (61.5%) and 22 (28.2%) women respectively. The cystoscopic findings were normal in 59 (75.6%) women. The procedure was completed within five minutes in all participants. The incidence of bacteriuria following outpatient flexible cystoscopy is low. © 2014 The Royal Australian and New Zealand College of Obstetricians and Gynaecologists.

  17. Prospective evaluation of the efficacy of antibiotic prophylaxis before cystoscopy

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    Kamil Cam

    2009-01-01

    Full Text Available Background: The aim of this study was to prospectively compare single-dose intravenous antibiotic prophylaxis vs. no prophylaxis before minor cystoscopic procedures, including punch biopsy and transurethral resection (TUR of small bladder tumors. Materials and Methods: A total of 200 patients with a mean age of 47.3 years old (range: 19-84 years old with initial negative urine cultures were recruited. All patients underwent a diagnostic cystoscopy. Patients were then randomized into 2 groups: o0 ne group that did not receive antibiotics (100 patients and the other group that received antibiotic treatment (100 patients with a single intravenous dose of cefoperazone. All patients had urine analysis and urine cultures on the second day after the operation. Additionally, clinical parameters including fever and dysuria were recorded. In 15% of the patients, incidental additional interventions such as punch biopsy or TUR of a small bladder tumor that were similarly distributed in both groups were performed. Results: In 1 patient from the antibiotic group and 2 patients from the no prophylaxis group, the urine cultures after cystoscopy were positive. No statistically significant difference was observed between these groups based on the microbiological and clinical parameters. Conclusion: The current study provides evidence that no antibiotic prophylaxis is required before diagnostic cystoscopy in patients without bacteriuria. But, the absolute risk of infection was small, suggesting that a much larger study is required.

  18. Randomized trial comparing office flexible to rigid cystoscopy in women.

    Science.gov (United States)

    Quiroz, Lieschen H; Shobeiri, S Abbas; Nihira, Mikio A; Brady, Jordan; Wild, Robert A

    2012-11-01

    The objective of the study was to compare office rigid cystoscopy (RC) versus flexible cystoscopy (FC) in women. This was a prospective randomized trial comparing FC to RC. Aims were to assess 1-week post-procedural complications, compare procedure pain scores, and to assess physician perception of patient discomfort. Pain scores were assessed by visual analogue scale (VAS) and 5-point verbal descriptor scale (VDS). Chi-square was used for categorical comparison and t tests or Wilcoxon test for continuous variables. One hundred women were enrolled. The mean age of participants was 59.7 years (± SD 14.6), and 91 % were Caucasian. This was the first cystoscopy for 86 % of participants. On the 1-week post-procedure questionnaire (85 % response rate), participants in the FC group reported urinary frequency more often than in the RC group (p = 0.041). The FC group reported urgency with urination lasting 1-2 days (p = 0.030) and burning with urination lasting >3 days (p = 0.026), more than the RC group. These symptoms did not persist at 7 days. The duration of the procedure was slightly faster for the FC group (4.6 ± 1.8 min vs 5.7 ± 3.4 min, p = 0.046). Median VAS scores were 0.9 (0.1-2.72) for the FC group and 0.5 (0-2.4) for the RC group (p = 0.505). There were no significant differences between patient or physician perception of pain in either group. Urinary frequency and duration of urinary burning post procedure occurred more frequently in the FC group, although these symptoms were transient. Both office FC and RC are generally well tolerated in women with overall low morbidity.

  19. The effect on pain experienced by male patients of watching their office-based flexible cystoscopy.

    NARCIS (Netherlands)

    Cornel, E.B.; Oosterwijk, E.; Kiemeney, L.A.L.M.

    2008-01-01

    OBJECTIVE: To confirm the recently published positive effect on visual analogue pain (VAS) scale levels for men watching their flexible cystoscopy. PATIENTS AND METHODS: From June 2007 to September 2007, 154 men had a flexible cystoscopy for various indications, all carried out by one urologist.

  20. Virtual cystoscopy--a surgical planning and guidance tool.

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    Braticevici, Bogdan; Onu, Mihaela; Bengus, Florentin

    2006-03-01

    Image guided-surgery systems facilitates surgical planning phases of endoscopic procedures. In this paper, we used a software package for 3D surface model generation and vizualization of the urinary bladder, based on magnetic resonance (MR) cross sectional images of the pelvis. The patients group consisted in 6 patients diagnosed with urinary bladder tumour. They were submitted to MRI exam. Twelve consecutive cross sectional images of the pelvis were aquired (TR (repetition time) = 600 msec, TE (echo time) = 19 msec, slice thickness = 6 divided by 7 mm, FOV (field of view) = 36 cm. All these images were transferred to a personal computer running the 3DSlicer software. We obtained, for each patient, a 3D model of the pelvis including the urinary bladder. In This way, the surgical enviroment was simulated and we are able to investigate the bladder by virtual cystoscopy. The virtual endoscopy may be used as a tool in the preoperative training and in surgical planning.

  1. Prospective evaluation of anxiety, pain, and embarrassment associated with cystoscopy and urodynamic testing in clinical practice.

    Science.gov (United States)

    Biardeau, Xavier; Lam, Ornella; Ba, Van; Campeau, Lysanne; Corcos, Jacques

    2017-01-01

    We sought to prospectively assess anxiety, pain, and embarrassment associated with diagnostic cystoscopy and multi-channel urodynamic study (UDS). All consecutive patients undergoing diagnostic cystoscopy or UDS in our department over a period of nine months were asked to participate. Two anonymous auto-administered questionnaires were specifically designed to collect basic epidemiological data, document medical history, and assess the quality of information provided, along with prevalence and level (0-10 numerical visual analog rating scale) of anxiety, pain, and embarrassment experienced before and/or during the procedures. Statistical analysis was carried out to identify underlying factors that could have influenced patients' experience and ascertain potential correlations between anxiety, pain, and embarrassment. 101 and 185 patients were respectively evaluated immediately after cystoscopy and UDS. Multivariate analysis repeatedly showed statistical correlations between anxiety, pain, and embarrassment, with regard to prevalence and level of intensity in both cystoscopy and UDS populations. Males and young patients were more likely to present anxiety, pain, or embarrassment during cystoscopy and UDS. Interestingly, patients who reported having received complete information before cystoscopy were significantly more likely to experience anxiety (62.6% vs. 20.0%; p=0.009). The present study demonstrated the major impact of gender and age on patients' experience. Interestingly, information provided before cystoscopy was reported to have a negative impact on patients' perception of anxiety; this could be partly prevented by optimizing the way information is provided to patients.

  2. Effect of diclofenac suppository on pain control during flexible cystoscopy-A randomized controlled trial

    National Research Council Canada - National Science Library

    Nadeem, Mehwash; Ather, M Hammad

    2016-01-01

    TRIAL DESIGN: To compare the difference in pain score during flexible cystoscopy between patients undergoing the procedure with plain lubricating gel only and plain gel with diclofenac suppository in a randomized control trial. METHODS...

  3. Prospective trial comparing intraoperative flexible, rigid, and no cystoscopy after ultrasound-guided transperineal permanent seed prostate brachytherapy

    Directory of Open Access Journals (Sweden)

    John Sylvester

    2016-06-01

    Conclusion: There was no significant difference in dysuria in the first four urinations post-PB between patients in the rigid, flexible, and no cystoscopy groups. Larger blood clots that may have been difficult to void, seeds in the bladder and/or urethra, and other abnormalities were found in 7% of patients who had cystoscopy. This may suggest that cystoscopy may be worthwhile post-PB. The incidence of AUR was not significantly different between the three cohorts.

  4. Evaluation of urine dipstick and cystoscopy in bovine enzootic haematuria

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    Mariana Pavelski

    2014-06-01

    Full Text Available Enzootic bovine haematuria is mainly caused by the chronic ingestion of bracken fern and is still considered an important cause of economic losses in beef herds, particularly in large metropolitan areas of Brazil that are not suitable for agriculture (weak soil, steep hills.The aim of this paper was evaluate if there is a correlation between the degree of haematuria and the presence of lesions in the urinary bladder of cows with bovine enzootic haematuria (BEH.We evaluated the bladder of twenty Nelore cows with bracken fern intoxication using a flexible endoscope to describe the lesions present and we performed a rapid urine test with a chemical multiple-reagent commercial strips (Combur test Roche® – Swiss to check the degree of haematuria; then we correlated both results with the Spearman statistical test. All cows presented lesions in their bladder walls and the urine was visually reddish and positive for haematuria in the strips. Urine dipstick has shown to correlate with bladder lesion number. In the bladders of the euthanised animals, multiple sub-mucosal reddish nodules and multiple ecchymotic haemorrhages were seen macroscopically, and one bladder had a larger tumour with a 3cm diameter, which was determined to be a haemangioma by analysing the histopathology. We concluded that urine dipstick is a valuable device for bovine enzootic haematuria prognostic purposes and to evaluate the severity of lesions from the urinary bladder. Moreover, cystoscopy is an important diagnostic tool to assess bladder lesions in cattle. Further studies are necessary to evaluate the involvement of papillomaviruses and bladder tumour occurrence in these cows.

  5. Do lubricants with 2% lidocaine gel have an effect on patient comfort in diagnostic cystoscopy?

    Science.gov (United States)

    Goktug, Hasan Nedim Goksel; Ozturk, Ufuk; Sener, Nevzat Can; Tuygun, Can; Bakırtas, Hasan; Imamoglu, M Abdurrahim

    2014-01-01

    Patients undergoing both rigid and flexible cystoscopic evaluation suffer from a great deal of pain and discomfort. In this study, we aimed to investigate the effect of lidocaine gel anestesia on patient comfort on diagnostic rigid cystoscopy. 11 mL of lubricant gel applied to each patient via the external meatus in 10 s. Patients were randomized into three groups. In group 1, liquid glycerine was applied and cystoscopy was immediately performed, in group 2 lidocaine gel (Aqua Touch™: İstem Tıbbi Cihaz Ve Sanayi Ltd.Şti, Ostim, Ankara, Türkiye) was applied and the procedure undergone immediately and in group 3, lidocaine gel was applied and penis was clemped for 10 minutes before the procedure. VAS forms were filled to determine the discomfort and pain during cystoscopy and the first micturation after. After the evaluation between groups, VAS scores were significantly lower in Group II and III than Group I and in Group III than in Group II (p ocal anesthetic lidocaine gel in rigid cystoscopy, is a practical, safe and efficient method to improve patient comfort when applied in appropriate dose and waiting duration.

  6. Effectiveness of the UroMentor virtual reality simulator in the skill acquisition of flexible cystoscopy

    Institute of Scientific and Technical Information of China (English)

    ZHANG Yi; LIU Jin-shun; WANG Gang; YU Cheng-fan; ZHU He; NA Yan-qun

    2013-01-01

    Background Virtual reality (VR) has been recognized as a useful modality in the training of surgical skills.With respect to basic endoscopic skill training of urology,we sought to investigate the effectiveness of the UroMentorTM virtual reality simulator (VRS) in the skill acquisition of flexible cystoscopy.Methods Urologists familiar with rigid cystoscopy procedures were selected to take part in a virtual training course of flexible cystoscopy.Changes in total operating time,frequency of injury,number of digital markers inside the bladder,and the global rating scale (GRS) scores were assessed following eight repeated training sessions on the UroMentorTM.Results Eighteen urologists voluntarily took part in the study.Total operating time was significantly lower after eight sessions of training by comparison ((111+10) seconds and (511+67) seconds,respectively; P<0.001).Additionally,the frequency of injury decreased with training from (12+2) times to (5+1) times (P<0.001),while the number of digital markers observed increased from 9+0 to 10+1 (P=0.005).Finally,training with the UroMentorTM resulted in a GRS increase from (1.3+0.2) points to (3.9+0.2) points (P<0.001).Conclusion the VRS UroMentorTM can improve urologists' ability to perform flexible cystoscopy and could be used as an effective training tool for trainees.

  7. Effect of diclofenac suppository on pain control during flexible cystoscopy-A randomized controlled trial

    Science.gov (United States)

    Nadeem, Mehwash; Ather, M Hammad

    2016-01-01

    TRIAL DESIGN: To compare the difference in pain score during flexible cystoscopy between patients undergoing the procedure with plain lubricating gel  only and plain gel with diclofenac suppository in a randomized control trial. METHODS:  A total of 60 male patients with an indication of flexible cystoscopy were enrolled in a prospective, randomized controlled study. Patients were randomized in two groups. In group “A”, patients received diclofenac suppository one hour prior to the procedure while group “B” did not receive diclofenac suppository. Both groups received 10 ml of intra-urethral  plain gel for lubrication during flexible cystoscopy. Pain score was recorded immediately after the procedure using the visual analogue scale (VAS). Pre- and post-procedure pulse rate and systolic blood pressure was also recorded. Statistical analyses were performed using chi-square test and student t-test. Regression analysis was performed to address the confounding variables. RESULTS: Both groups were comparable for variables including age, duration of procedure, level of operating surgeon and indication of procedure. Most common indication for flexible cystoscopy was removal of double J stent. There was a statistically significant difference in the mean pain score between two groups ( p = 0.012).  The difference in post-procedure mean pulse rate in the two groups was statistically significant ( p= 0.01) however there was no difference observed in mean post procedure systolic blood pressure. Regression analysis showed that none of the confounding variables were significantly affecting pain perception. CONCLUSIONS: Intra rectal diclofenac suppository is simple and effective pre-emptive analgesia. We recommend its routine use during flexible cystoscopy for better pain control. PMID:28299180

  8. Cystoscopy-assisted laparoscopy for bladder endometriosis: modified light-to-light technique for bladder preservation

    Science.gov (United States)

    Stopiglia, Rafael Mamprin; Ferreira, Ubirajara; Faundes, Daniel Gustavo; Petta, Carlos Alberto

    2017-01-01

    ABSTRACT Introduction Endometriosis is a disease with causes still unclear, affecting approximately 15% of women of reproductive age, and in 1%-2% of whom it may involve the urinary tract. The bladder is the organ most frequently affected by endometriosis, observed around 85% of the cases. In such cases, the most effective treatment is partial cystectomy, especially via videolaparoscopy. Study Objective, Design, Size and Duration In order to identify and delimit the extent of the intravesical endometriosis lesion, to determine the resection limits, as well as to perform an optimal reconstruction of the organ aiming for its maximum preservation, we performed a cystoscopy simultaneously with the surgery, employing a modified light-to-light technique in 25 consecutive patients, from September 2006 to May 2012. Setting Study performed at Campinas Medical Center – Campinas – Sao Paulo – Brazil.Participants/materials, setting and methods: Patients aged 27 to 47 (average age: 33.4 years) with deep endometriosis with total bladder involvement were selected for the study. The technique used was conventional laparoscopy with a transvaginal uterine manipulator and simultaneous cystoscopy (the light-to-light technique). A partial videolaparoscopic cystectomy was performed with cystoscopy-assisted vesical reconstruction throughout the entire surgical time. The lesions had an average size of 2.75cm (ranging from 1.5 to 5.5cm). The average surgical time was 137.7 minutes, ranging from 110 to 180 minutes. Main Results Postoperative follow-up time was 32.4 months (12-78 months), with clinical evaluation and a control cystoscopy performed every six months. No relapse was observed during the follow-up period. Conclusions A cystoscopy-assisted partial laparoscopic cystectomy with a modified light-to-light technique is a method that provides adequate identification of the lesion limits, intra or extravesically. It also allows a safe reconstruction of the organ aiming for its

  9. Cystoscopy-assisted laparoscopy for bladder endometriosis: modified light-to-light technique for bladder preservation

    Directory of Open Access Journals (Sweden)

    Rafael Mamprin Stopiglia

    Full Text Available ABSTRACT Introduction Endometriosis is a disease with causes still unclear, affecting approximately 15% of women of reproductive age, and in 1%-2% of whom it may involve the urinary tract. The bladder is the organ most frequently affected by endometriosis, observed around 85% of the cases. In such cases, the most effective treatment is partial cystectomy, especially via videolaparoscopy. Study Objective, Design, Size and Duration In order to identify and delimit the extent of the intravesical endometriosis lesion, to determine the resection limits, as well as to perform an optimal reconstruction of the organ aiming for its maximum preservation, we performed a cystoscopy simultaneously with the surgery, employing a modified light-to-light technique in 25 consecutive patients, from September 2006 to May 2012. Setting Study performed at Campinas Medical Center – Campinas – Sao Paulo – Brazil.Participants/materials, setting and methods: Patients aged 27 to 47 (average age: 33.4 years with deep endometriosis with total bladder involvement were selected for the study. The technique used was conventional laparoscopy with a transvaginal uterine manipulator and simultaneous cystoscopy (the light-to-light technique. A partial videolaparoscopic cystectomy was performed with cystoscopy-assisted vesical reconstruction throughout the entire surgical time. The lesions had an average size of 2.75cm (ranging from 1.5 to 5.5cm. The average surgical time was 137.7 minutes, ranging from 110 to 180 minutes. Main Results Postoperative follow-up time was 32.4 months (12-78 months, with clinical evaluation and a control cystoscopy performed every six months. No relapse was observed during the follow-up period. Conclusions A cystoscopy-assisted partial laparoscopic cystectomy with a modified light-to-light technique is a method that provides adequate identification of the lesion limits, intra or extravesically. It also allows a safe reconstruction of the organ

  10. Listening to music during cystoscopy decreases anxiety, pain, and dissatisfaction in patients: a pilot randomized controlled trial.

    Science.gov (United States)

    Yeo, Jeong Kyun; Cho, Dae Yeon; Oh, Mi Mi; Park, Seok San; Park, Min Gu

    2013-04-01

    To determine whether listening to music during cystoscopy decreases anxiety, pain, and dissatisfaction among patients and results in a more comfortable and better-tolerated procedure. Seventy male patients who underwent rigid cystoscopy between May 2011 and December 2011 were randomized into the following: no music (Group I, n=35) or classical music during procedure (Group II, n=35). Before cystoscopy, lidocaine gel was instilled in the urethra, and both groups viewed their procedures on a video monitor. Anxiety levels were quantified according to the State-Trait Anxiety Inventory. A visual analog scale (0-10) was used for a self-assessment of satisfaction, discomfort, and willingness among patients to repeat the cystoscopy. Demographic characteristics, mean age, procedure duration, and procedure indications were statistically similar between the two groups. The mean anxiety level and mean pain score of Group II were significantly lower than those of Group I (pmusic during rigid cystoscopy significantly reduces feelings of pain, discomfort, and dissatisfaction. Music can serve as a simple, inexpensive, and effective adjunct to sedation during cystoscopy. We recommend the application of music during rigid cystoscopy for clinical use.

  11. NaHCO3-Buffered Lidocaine Gel for Outpatient Rigid Cystoscopy in Men.

    Science.gov (United States)

    Li, Hong; Cheng, Yanxin; Li, Jun; Chen, Yongxue; Yuan, Jinge; Yang, Shuhong; Shi, Huiqiao; Li, Wenpin; Yang, Shijie; Wang, Wei; Xu, Guanjie; Zhao, Senming

    2016-04-01

    The purpose of this study was to explore the effect of NaHCO3-buffered lidocaine gel as a topical anesthetic agent for pain relief for rigid cystoscopy. Prospective randomized controlled trial. ASA I-II male patients undergoing rigid cystoscopy randomly received 10 mL 2% Carbocaine lidocaine gel with 1 mL 0.9% saline (group 1) or 1 mL 5% NaHCO3 solution (group 2). After 3 minutes exposure, the cystoscope was inserted into the urethra. On receiving the gel, cystoscope insertion, and intravesical observation, pain score was recorded using the visual analog scale. The gel pH with or without NaHCO3 was 7.20 and 6.41, respectively. The concentration of soluble lidocaine in the gel was stable for 24 hours or more. The visual analog scale score in group 2 was significantly lower (1.3 ± 0.9) than in group 1 (5.28 ± 1.99). No adverse effects were recorded. Alkalized lidocaine gel resulted in successful analgesia for rigid cystoscopy in men without adverse effects. Copyright © 2016 American Society of PeriAnesthesia Nurses. Published by Elsevier Inc. All rights reserved.

  12. Music reduces panic: an initial study of listening to preferred music improves male patient discomfort and anxiety during flexible cystoscopy.

    Science.gov (United States)

    Zhang, Zhen-Sheng; Wang, Xiao-Lin; Xu, Chuan-Liang; Zhang, Chao; Cao, Zhi; Xu, Wei-Dong; Wei, Rong-Chao; Sun, Ying-Hao

    2014-06-01

    To assess the impact of listening to preferred music on relieving male patients' pain and anxiety during flexible cystoscopy. A total of 124 male patients were admitted to our hospital for flexile cystoscopy by a single urologist between January 2013 and September 2013 and randomized to two equal groups. Group 2 included 62 patients who could select and listen to their preferred music during flexible cystoscopy. Group 1 included 62 patients who were unable to listen to the music. All patients were administered the same amount of lidocaine (10 mL) for 3 minutes for local anesthesia before flexible cystoscopy. A visual analog scale (VAS) ranging from 0 to 10 was used to assess patients' pain feeling after the cystoscopy procedure. Anxiety levels were calculated according to the State Instrument of State-Trait Anxiety Inventory (STAI-S), and the pulse rate were recorded 5 minutes before and immediately after the procedure. The duration of the procedure of each group were also analyzed. Statistically significant differences were detected between group 1 and group 2 in the mean pain score on VAS (2.53 ± 1.34 vs 1.63 ± 1.09, P=0.002, Mann-Whitney U test), mean postprocedural State Anxiety Inventory pain score (39.4 ± 6.5 vs 34.5 ± 5.8), and postprocedural pulse rate (79.8 ± 5.5 vs 76.0 ± 7.3) (Ppreferred music experienced less discomfort and lower anxiety at cystoscopy. Patient age, duration of the procedure, preprocedural STAI-S, and preprocedural pulse rate of each group were comparable. Listening to preferred music during flexible cystoscopy is an easy way to improves male patients' comfort and reduce their anxiety. It could be recommended for male patients.

  13. Virtual multislice computed tomography cystoscopy for evaluation of urinary bladder lesions; Virtuelle MDCT-Zystoskopie bei der Detektion von Blasenlaesionen

    Energy Technology Data Exchange (ETDEWEB)

    Heinz-Peer, G.; Happel, B.; Memarsadeghi, M.; Mang, T. [Medizinische Universitaet Wien (Austria). Klinik fuer Radiodiagnostik

    2005-10-01

    The introduction of multislice computed tomography (MDCT) with the possibility of acquiring isotropic datasets has been an ideal prerequisite for development of virtual MDCT cystoscopy. Remarkable technical progress regarding post-processing of high-resolution 3D datasets as well as a considerable reduction of the time required for post-processing made it possible to introduce virtual MDCT cystoscopy into the clinical routine. 3D post-processing that often required 7-8 h when virtual endoscopy techniques were first developed can now be performed in less than 5 min after transfer of data to the 3D workstation. With the limitations and contraindications of conventional cystoscopy in mind, virtual MDCT cystoscopy may be seen as a valuable alternative to conventional cystoscopy for evaluation of hematuria. (orig.) [German] Mit der Einfuehrung der Multidetektorcomputertomographie (MDCT) und der damit verbundenen Moeglichkeit, isotrope Datensaetze zu akquirieren, ergaben sich ideale Voraussetzungen zur Entwicklung der virtuellen MDCT-Zystoskopie. Beachtliche technische Fortschritte bei der Nachbearbeitung der hochaufloesenden 3D-Datensaetze sowie eine deutliche Reduktion der Nachbearbeitungszeit haben den Einsatz der virtuellen MDCT-Zystoskopie in der klinischen Routine ermoeglicht. Waren zu Beginn der Entwicklung virtueller Endoskopietechniken 3D-Nachbearbeitungszeiten von 7-8 h keine Seltenheit, kann diese heute nach dem Datentransfer auf die Workstation in weniger als 5 min erfolgen. Unter Betrachtung der Limitationen und Kontraindikationen der konventionellen Zystoskopie kann die virtuelle MDCT-Zystoskopie als wertvolle Alternative bei der Haematurieabklaerung gesehen werden. (orig.)

  14. Comparison of virtual cystoscopy and ultrasonography for bladder cancer detection: A meta-analysis

    Energy Technology Data Exchange (ETDEWEB)

    Qu Xinhua; Huang Xiaolu [Department of Nuclear Medicine, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200127 (China); Department of Ultrasonic, Ninth People' s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011 (China); Wu Lianming [Department of Radiology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200127 (China); Huang Gang [Department of Nuclear Medicine, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200127 (China); Ping Xiong, E-mail: pxiong6@126.com [Department of Ultrasonic, Ninth People' s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011 (China); Yan Weili, E-mail: wl_yan67@126.com [Department of Nuclear Medicine, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200127 (China)

    2011-11-15

    Background and purpose: Bladder cancer is the most commonly diagnosed malignancy in patients presenting with haematuria. Early detection is crucial for improving patient prognosis. We therefore performed a meta-analysis to evaluate and compare the detection validity (sensitivity and specificity) of virtual cystoscopy (VC) and ultrasonography (US). Methods: We searched MEDLINE, EMBASE, PubMed and the Cochrane Library for studies evaluating diagnosis validity of VC and US between January 1966 and December 2009. Meta-analysis methods were used to pool sensitivity and specificity and to construct a summary receiver-operating characteristic (SROC) curve. Results: A total of 26 studies that included 3084 patients who fulfilled all of the inclusion criteria were considered for inclusion in the analysis. The pooled sensitivity for bladder cancer detection using CT virtual cystoscopy (CTVC), MR virtual cystoscopy (MRVC) and US was 0.939 (95% CI, 0.919-0.956), 0.908 (95% CI, 0.827-0.959) and 0.779 (95% CI, 0.744-0.812), respectively. The pooled specificity for bladder cancer detection using CTVC, MRVC and US was 0.981 (95% CI, 0.973-0.988), 0.948 (95% CI, 0.884-0.983) and 0.962 (95% CI, 0.953-0.969), respectively. The pooled diagnostic odd ratio (DOR) estimate for CTVC (604.22) were significantly higher than for MRVC (144.35, P < 0.001) and US (72.472, P < 0.001). Conclusion: Our results showed that both CTVC and MRVC are better imaging methods for diagnosing bladder cancer than US. CTVC has higher diagnostic value (sensitivity, specificity and DOR) for the detection of bladder cancer than either MRCT or US.

  15. Virtual 3D bladder reconstruction for augmented medical records from white light cystoscopy (Conference Presentation)

    Science.gov (United States)

    Lurie, Kristen L.; Zlatev, Dimitar V.; Angst, Roland; Liao, Joseph C.; Ellerbee, Audrey K.

    2016-02-01

    Bladder cancer has a high recurrence rate that necessitates lifelong surveillance to detect mucosal lesions. Examination with white light cystoscopy (WLC), the standard of care, is inherently subjective and data storage limited to clinical notes, diagrams, and still images. A visual history of the bladder wall can enhance clinical and surgical management. To address this clinical need, we developed a tool to transform in vivo WLC videos into virtual 3-dimensional (3D) bladder models using advanced computer vision techniques. WLC videos from rigid cystoscopies (1280 x 720 pixels) were recorded at 30 Hz followed by immediate camera calibration to control for image distortions. Video data were fed into an automated structure-from-motion algorithm that generated a 3D point cloud followed by a 3D mesh to approximate the bladder surface. The highest quality cystoscopic images were projected onto the approximated bladder surface to generate a virtual 3D bladder reconstruction. In intraoperative WLC videos from 36 patients undergoing transurethral resection of suspected bladder tumors, optimal reconstruction was achieved from frames depicting well-focused vasculature, when the bladder was maintained at constant volume with minimal debris, and when regions of the bladder wall were imaged multiple times. A significant innovation of this work is the ability to perform the reconstruction using video from a clinical procedure collected with standard equipment, thereby facilitating rapid clinical translation, application to other forms of endoscopy and new opportunities for longitudinal studies of cancer recurrence.

  16. Hexaminolevulinate blue-light cystoscopy in non-muscle-invasive bladder cancer: review of the clinical evidence and consensus statement on appropriate use in the USA

    NARCIS (Netherlands)

    Daneshmand, S.; Schuckman, A.K.; Bochner, B.H.; Cookson, M.S.; Downs, T.M.; Gomella, L.G.; Grossman, H.B.; Kamat, A.M.; Konety, B.R.; Lee, C.T.; Pohar, K.S.; Pruthi, R.S.; Resnick, M.J.; Smith, N.D.; Witjes, J.A.; Schoenberg, M.P.; Steinberg, G.D.

    2014-01-01

    Hexaminolevulinate (HAL) is a tumour photosensitizer that is used in combination with blue-light cystoscopy (BLC) as an adjunct to white-light cystoscopy (WLC) in the diagnosis and management of non-muscle-invasive bladder cancer (NMIBC). Since being licensed in Europe in 2005, HAL has been used in

  17. A Comparative Study between Cystoscopy and Cystography in Detecting Bladder Diverticulae in Patients with Benign Prostatic Hyperplasia

    Directory of Open Access Journals (Sweden)

    Manish Swarnkar

    2016-04-01

    Full Text Available Background: Bladder diverticulae are mucosal outpunching and majority are acquired and secondary to either benign prostatic hypertrophy or the upper motor type of neurogenic bladder. Aim & Objectives: This study was undertaken to increase knowledge and understanding the presumed role of Benign Prostatic Hyperplasia (BPH in the development of bladder diverticulae and to compare cystoscopy and cystography in detecting bladder diverticulae. Material and Methods: During a one year period, 74 patients with BPH, who were admitted in the hospital for surgery, were examined by cystoscopy and cystography to detect bladder diverticulae. Results: In this study, prevalence of bladder diverticulae was found to be 61%. There was a slight increase of bladder diverticulae with increasing age. We could not find any statistical difference between cystoscopy and cystography in detecting bladder diverticulae 2 (c =1.86, p value=.60, NS, p<0.05.Malignancy within diverticulae seen in 6.8% of cases. Conclusion: This study shows that the prevalence of bladder diverticulae is considerably high. Considering this high prevalence and potential complications, we suggest cystoscopy for the early diagnosis and possible treatment of bladder diverticulae in older patients with lower urinary tract symptoms

  18. The effects of visual fluorescence marking induced by 5-aminolevulinic acid for endoscopic diagnosis of urinary bladder cancer

    Science.gov (United States)

    Daniltchenko, Dmitri I.; Koenig, Frank; Schnorr, Dietmar; Valdman, Alexander; Al-Shukri, Salman; Loening, Stefan A.

    2003-10-01

    During cystoscopy procedure, fluorescence diagnostics induced by 5-ALA improves visual detection of the bladder cancer. Macroscopic ALA-fluorescence allows visualizing of small flat tumors, carcinoma in situ, true neoplasm margins and dysplasias of the bladder. Following ALA instillation, cystoscopy has been performed under both standard and blue light illumination. Totally, 153 biopsies have been carried out at 53 patients with suspicion of bladder cancer. The results were compared to ALA-fluorescence data. In 13% of the patients, bladder cancer and dysplasia were found out in addition, due to red fluorescence. The sensitivity and specificity of ALA-fluorescence technique aggregated 96% and 52% respectively. The sensitivity and specificity of 5-ALA-fluorescent detection exceeded standard endoscopy under white light on 20%. The new method does not exclude a false positive and a false negative fluorescent luminescence. The ALA-based fluorescence detection system enhances the diagnosis of malignant/dysplastic bladder lesions significantly.

  19. Effect of diclofenac suppository on pain control during flexible cystoscopy-A randomized controlled trial [version 1; referees: 2 approved

    Directory of Open Access Journals (Sweden)

    Mehwash Nadeem

    2016-12-01

    Full Text Available TRIAL DESIGN: To compare the difference in pain score during flexible cystoscopy between patients undergoing the procedure with plain lubricating gel  only and plain gel with diclofenac suppository in a randomized control trial. METHODS:  A total of 60 male patients with an indication of flexible cystoscopy were enrolled in a prospective, randomized controlled study. Patients were randomized in two groups. In group “A”, patients received diclofenac suppository one hour prior to the procedure while group “B” did not receive diclofenac suppository. Both groups received 10 ml of intra-urethral  plain gel for lubrication during flexible cystoscopy. Pain score was recorded immediately after the procedure using the visual analogue scale (VAS. Pre- and post-procedure pulse rate and systolic blood pressure was also recorded. Statistical analyses were performed using chi-square test and student t-test. Regression analysis was performed to address the confounding variables. RESULTS: Both groups were comparable for variables including age, duration of procedure, level of operating surgeon and indication of procedure. Most common indication for flexible cystoscopy was removal of double J stent. There was a statistically significant difference in the mean pain score between two groups (p = 0.012.  The difference in post-procedure mean pulse rate in the two groups was statistically significant (p= 0.01 however there was no difference observed in mean post procedure systolic blood pressure. Regression analysis showed that none of the confounding variables were significantly affecting pain perception. CONCLUSIONS: Intra rectal diclofenac suppository is simple and effective pre-emptive analgesia. We recommend its routine use during flexible cystoscopy for better pain control.

  20. PHOTODYNAMIC DIAGNOSIS AND FLUORESCENCE SPECTROSCOPY IN SUPERFICIAL BLADDER CANCER

    Directory of Open Access Journals (Sweden)

    I. G. Rusakov

    2009-01-01

    Full Text Available A comprehensive fluorescence technique has been developed to study the urinary bladder mucosa in patients with superficial bladder cancer (BC, by using alasense, white light cystoscopy, fluorescence cytoscopy, and local fluorescence spectroscopy in vivo. Quantification of urothelium fluorescence in the red emission foci of 5-ALA-induced protophorphyrin, with the local autofluorescence intensity being borne in mind, has been shown to increase the specificity of photodynamic diagnosis of superficial BC from 70 to 85% (p ≤ 0.05 and the total accuracy of the technique from 80 to 86%.  

  1. Use of Lidocaine 2% Gel Does Not Reduce Pain during Flexible Cystoscopy and Is Not Cost-Effective.

    Science.gov (United States)

    Cano-Garcia, Maria Del Carmen; Casares-Perez, Rosario; Arrabal-Martin, Miguel; Merino-Salas, Sergio; Arrabal-Polo, Miguel Angel

    2015-11-14

    To compare the use of lubricant gel with lidocaine versus lubricant gel without anesthetic in flexible cystoscopy in terms of pain and tolerability. In this observational non-randomized study, 72 patients were divided into two groups. Group 1 included 38 patients in whom lidocaine gel 2% was used and group 2 included 34 patients in whom lubricant gel without anesthetic was administered. The main variables analyzed were score in visual analogue scale (VAS) and score in Spanish Pain Questionnaire (SPQ). Student's t-test and Chi-square test were used to compare differences between 2 groups. The P values lidocaine is 1.25 euro and gel without anesthetic 0.22 euro. The use of lidocaine gel does not produce benefit in terms of pain relief in flexible cystoscopy and also is costly.

  2. Diagnosis of bladder tumours in patients with macroscopic haematuria: a prospective comparison of split-bolus computed tomography urography, magnetic resonance urography and flexible cystoscopy.

    Science.gov (United States)

    Gandrup, Karen L; Løgager, Vibeke B; Bretlau, Thomas; Nordling, Jørgen; Thomsen, Henrik S

    2015-06-01

    The aim of this study was to compare split-bolus computed tomography urography (CTU), magnetic resonance urography (MRU) and flexible cystoscopy in patients with macroscopic haematuria regarding the diagnosis of bladder tumours. In this prospective study, 150 patients underwent CTU, MRU and flexible cystoscopy. Two uroradiologists individually reviewed the images without any clinical information, using a questionnaire. Patient records and pathology reports were also reviewed. At flexible cystoscopy, MRU and CTU, 32, 19 and 15 bladder lesions were identified, respectively. Histopathology showed that 13 of the 29 biopsied lesions were transitional cell carcinomas. Compared with the histopathology, the sensitivity and specificity for detection of tumours by CTU and MRU were 61.5% and 94.9%, and 79.9% and 93.4%, respectively. False-positive detection of bladder tumours, compared with histopathology, was reported in seven CTUs and nine MRUs, whereas the number of false-negative findings was five for CTUs and three for MRUs. Split-bolus CTU or MRU cannot replace cystoscopy in cases of macroscopic haematuria. MRU has a higher sensitivity than split-bolus CTU, and is a potential alternative to flexible cystoscopy. Patients with a low risk of bladder cancer may forgo flexible cystoscopy if a bladder tumour is identified by either CTU or MRU, and proceed straight to transuretheral resection of the bladder.

  3. Comparison of Nuclear Matrix Protein (NMP22 test with cystoscopy and urine cytology in follow-up of patients with superficial bladder cancer

    Directory of Open Access Journals (Sweden)

    Ali Ozudogru

    2012-06-01

    Conclusion: NMP22 was found more sensitive than urinary cytology. NMP22 were not influenced by intracavitary immunotherapy. As long as tumor grade increased, NMP22 values increased. But no significant relation with tumor stage was detected. The NMP22 test remains incapable for the supplanting of control cystoscopy. Until a better tumor marker was acquired, NMP22 test can be used. In patients with negative NMP22 test, control cystoscopy intervals may prolonged. [J Contemp Med 2012; 2(2.000: 64-68

  4. Use of Flexible Cystoscopy to Insert a Foley Catheter over a Guide Wire in Spinal Cord Injury Patients: Special Precautions to be Observed

    Directory of Open Access Journals (Sweden)

    Subramanian Vaidyanathan

    2011-01-01

    Full Text Available When urethral catheterisation is difficult or impossible in spinal cord injury patients, flexible cystoscopy and urethral catheterisation over a guide wire can be performed on the bedside, thus obviating the need for emergency suprapubic cystostomy. Spinal cord injury patients, who undergo flexible cystoscopy and urethral catheterisation over a guide wire, may develop potentially serious complications. (1 Persons with lesion above T-6 are susceptible to develop autonomic dysreflexia during cystoscopy and urethral catheterisation over a guide wire; nifedipine 5–10 milligrams may be administered sublingually just prior to the procedure to prevent autonomic dysreflexia. (2 Spinal cord injury patients are at increased risk for getting urine infections as compared to able-bodied individuals. Therefore, antibiotics should be given to patients who get haematuria or urethral bleeding following urethral catheterisation over a guide wire. (3 Some spinal cord injury patients may have a small capacity bladder; in these patients, the guide wire, which is introduced into the urinary bladder, may fold upon itself with the tip of guide wire entering the urethra. If this complication is not recognised and a catheter is inserted over the guide wire, the Foley catheter will then be misplaced in urethra despite using cystoscopy and guide wire.

  5. Patients with a negative cystoscopy and negative Nmp22® Bladderchek® test are at low risk of missed transitional cell carcinoma of the bladder: a prospective evaluation

    Directory of Open Access Journals (Sweden)

    John D. Terrell

    2011-12-01

    Full Text Available OBJECTIVES: Urine based tumor markers have uncertain utility in diagnosis or surveillance of patients with bladder cancer while cytology is commonly used. We evaluated whether cytology provides additional diagnostic information in patients with a negative NMP22® BladderChek® test (BladderChek and negative cystoscopy. MATERIALS AND METHODS: We performed subset analyses of 2 large prospective multi-center databases evaluating BladderChek for UCB detection and surveillance. These cohorts were analyzed for presence of cancer and result of urine cytology in setting of a negative cystoscopy and negative BladderChek. Subsequently, we prospectively performed cystoscopy, cytology and BladderChek on 434 patients at our institution being evaluated for UCB. RESULTS: In the detection database (n = 1331, 1065 patients had a negative cystoscopy and BladderChek. There were 3 cancers (stages Ta, Tis and T1 and cytology was atypical in one and reactive in two. In the surveillance cohort (n = 668 patients, 437 patients had negative cystoscopy and BladderChek. Cancer was found in 2 patients (stages Tis and Ta. The patient with Tis has dysplastic cytology and Ta tumor had reactive cytology. In our cohort of 434 patients, 288 pts had negative cystoscopy and BladderChek. One cancer was missed, a Ta ureteral urothelial carcinoma with a reactive cytology. CONCLUSIONS: In patients with negative cystoscopy and BladderChek, very few cancers are missed and cytology was not effective in detection. Use of a point-of-care test in conjunction with cystoscopy in lieu of cytology could decrease cost, provide immediate results, improve negative predictive value and reduce the uncertainty that results from inconclusive cytologic results.

  6. Bladder tumors:dynamic contrast-enhanced axial imaging, multiplanar reformation, three-dimensional reconstruction and virtual cystoscopy using helical CT

    Institute of Scientific and Technical Information of China (English)

    王东; 张挽时; 熊明辉; 喻敏; 徐家兴

    2004-01-01

    Background There have been few studies to evaluate the effects of helical CT on bladder tumor. This study was to evaluate the clinical applications of helical CT dynamic contrast-enhanced axial imaging, multiplanar reformation (MPR), three-dimensional (3D) reconstruction and virtual cystoscopy (CTVC) in bladder tumors. Methods The precontrast and four-phase postcontrast helical CT scans were performed in 42 patients with bladder tumors confirmed by conventional cystoscopy and pathology. MPR, 3D and CTVC images were generated from the volumetric data of the excretory phase. The results were then compared with the findings of conventional cystoscopy and surgery in a double-blinded mode. Results The sensitivity of the axial, 3D and CTVC images in detecting the bladder tumors were 90.8%, 76.9% and 95.4% respectively. The dynamic contrast-enhanced axial images could provide excellent intramural and extravesical information, and the accuracy in preoperative tumor staging was 87.7%. MPR could directly demonstrate the origin and extravesical invasions of the tumors and their relation to the ureter. 3D and CTVC images were useful for displaying the surface morphology of the tumor and the relationship between the tumor and the ureteric orifices, whereas CTVC could depict the tumors smaller than 5 mm that were not seen on the axial images.Conclusions The combination of axial, MPR, 3D and CTVC images with helical CT can provide comprehensive information on bladder tumor.

  7. An Investigation of the Basic Physics of Irrigation in Urology and the Role of Automated Pump Irrigation in Cystoscopy

    Directory of Open Access Journals (Sweden)

    Dwayne Chang

    2012-01-01

    Full Text Available Objective. To investigate the effects of height, external pressure, and bladder fullness on the flow rate in continuous, non-continuous cystoscopy and the automated irrigation fluid pumping system (AIFPS. Materials. Each experiment had two 2-litre 0.9% saline bags connected to a continuous, non-continuous cystoscope or AIFPS via irrigation tubing. Other equipment included height-adjustable drip poles, uroflowmetry devices, and model bladders. Methods. In Experiment 1, saline bags were elevated to measure the increment in flow rate. In Experiment 2, saline bags were placed under external pressures to evaluate the effect on flow rate. In Experiment 3, flow rate changes in response to variable bladder fullness were measured. Results. Elevating saline bags caused an increase in flow rates, however the increment slowed down beyond a height of 80 cm. Increase in external pressure on saline bags elevated flow rates, but inconsistently. A fuller bladder led to a decrease in flow rates. In all experiments, the AIFPS posted consistent flow rates. Conclusions. Traditional irrigation systems were susceptible to changes in height of irrigation solution, external pressure application, and bladder fullness thus creating inconsistent flow rates. The AIFPS produced consistent flow rates and was not affected by any of the factors investigated in the study.

  8. An Investigation of the Basic Physics of Irrigation in Urology and the Role of Automated Pump Irrigation in Cystoscopy

    Science.gov (United States)

    Chang, Dwayne; Manecksha, Rustom P.; Syrrakos, Konstantinos; Lawrentschuk, Nathan

    2012-01-01

    Objective. To investigate the effects of height, external pressure, and bladder fullness on the flow rate in continuous, non-continuous cystoscopy and the automated irrigation fluid pumping system (AIFPS). Materials. Each experiment had two 2-litre 0.9% saline bags connected to a continuous, non-continuous cystoscope or AIFPS via irrigation tubing. Other equipment included height-adjustable drip poles, uroflowmetry devices, and model bladders. Methods. In Experiment 1, saline bags were elevated to measure the increment in flow rate. In Experiment 2, saline bags were placed under external pressures to evaluate the effect on flow rate. In Experiment 3, flow rate changes in response to variable bladder fullness were measured. Results. Elevating saline bags caused an increase in flow rates, however the increment slowed down beyond a height of 80 cm. Increase in external pressure on saline bags elevated flow rates, but inconsistently. A fuller bladder led to a decrease in flow rates. In all experiments, the AIFPS posted consistent flow rates. Conclusions. Traditional irrigation systems were susceptible to changes in height of irrigation solution, external pressure application, and bladder fullness thus creating inconsistent flow rates. The AIFPS produced consistent flow rates and was not affected by any of the factors investigated in the study. PMID:22666130

  9. Cystoscopy and Ureteroscopy

    Science.gov (United States)

    ... the bladder. A sterile liquid—water or salt water, called saline—flows through the cystoscope or ureteroscope to slowly fill the bladder and stretch it so the urologist has a better view of the bladder wall. As the bladder fills with liquid, the patient ...

  10. Fluorescence spectroscopy

    DEFF Research Database (Denmark)

    Bagatolli, Luis

    2016-01-01

    Fluorescence spectroscopy is a powerful experimental tool used by scientists from many disciplines. During the last decades there have been important developments on distinct fluorescence methods, particularly those related to the study of biological phenomena. This chapter discusses...

  11. A Comparison of the Progression and Recurrence Risk Index in Non-Muscle-Invasive Bladder Tumors Detected by Narrow-Band Imaging Versus White Light Cystoscopy, Based on the EORTC Scoring System

    Directory of Open Access Journals (Sweden)

    Shadpour

    2016-01-01

    Full Text Available Background Transitional cell carcinoma of the bladder, the second most common urologic malignancy, is amenable to early diagnosis. This study presents the potential prognostic benefit for a less invasive modification to the standard endoscopic approach. Objectives To evaluate the risk index for the progression and recurrence of additional tumors detected with narrow-band imaging (NBI cystoscopy compared to standard white light imaging (WLI cystoscopy in non-muscle-invasive bladder cancer (NMIBC, based on the European organization for research and treatment of cancer (EORTC scoring system. Patients and Methods Patients with NMIBC, who were scheduled for resection between May 2012 and May 2013, were studied and mapped under NBI and WLI cystoscopy by independent surgeons prior to resection. Detection rates and tumor characteristics, including EORTC progression and the recurrence risk index, were compared. Results Fifty patients, aged 63.86 ± 10.05 years, were enrolled. The overall detection rate was 98.9% for NBI vs. 89.4% for WLI (P = 0.001, and the false-positive rates were 9.6% and 5.8%, respectively (P = 0.051. Ten tumors were detected by NBI alone, including four grade I tumors, four grade III tumors, and two carcinomas in situ. The tumor progression index was not significantly reduced with NBI compared to WLI (P > 0.05; however, the recurrence index was significantly lower in the NBI group (P < 0.05. Conclusions NBI cystoscopy improved the detection rate. Although false positives were more common with NBI, this was not statistically significant. NBI found additional aggressive tumors, which underscores the impact of detection in EORTC recurrence risk scoring.

  12. Fluorescent microspheres

    Science.gov (United States)

    Rembaum, A.

    1978-01-01

    Latex particles with attached antibodies have potential biochemical and environmental applications. Human red blood cells and lymphocytes have been labeled with fluorescent microspheres by either direct or indirect immunological technique. Immunolatex spheres can also be used for detecting and localizing specific cell surface receptors. Hormones and toxins may also be bondable.

  13. 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.

  14. Automatic segmentation of lesions for the computer-assisted detection in fluorescence urology

    Science.gov (United States)

    Kage, Andreas; Legal, Wolfgang; Kelm, Peter; Simon, Jörg; Bergen, Tobias; Münzenmayer, Christian; Benz, Michaela

    2012-03-01

    Bladder cancer is one of the most common cancers in the western world. The diagnosis in Germany is based on the visual inspection of the bladder. This inspection performed with a cystoscope is a challenging task as some kinds of abnormal tissues do not differ much in their appearance from their surrounding healthy tissue. Fluorescence Cystoscopy has the potential to increase the detection rate. A liquid marker introduced into the bladder in advance of the inspection is concentrated in areas with high metabolism. Thus these areas appear as bright "glowing". Unfortunately, the fluorescence image contains besides the glowing of the suspicious lesions no more further visual information like for example the appearance of the blood vessels. A visual judgment of the lesion as well as a precise treatment has to be done using white light illumination. Thereby, the spatial information of the lesion provided by the fluorescence image has to be guessed by the clinical expert. This leads to a time consuming procedure due to many switches between the modalities and increases the risk of mistreatment. We introduce an automatic approach, which detects and segments any suspicious lesion in the fluorescence image automatically once the image was classified as a fluorescence image. The area of the contour of the detected lesion is transferred to the corresponding white light image and provide the clinical expert the spatial information of the lesion. The advantage of this approach is, that the clinical expert gets the spatial and the visual information of the lesion together in one image. This can save time and decrease the risk of an incomplete removal of a malign lesion.

  15. Fundamentals of fluorescence and fluorescence microscopy.

    Science.gov (United States)

    Wolf, David E

    2013-01-01

    This chapter discusses the fundamental physics of fluorescence. The application of fluorescence to microscopy represents an important transition in the development of microscopy, particularly as it applies to biology. It enables quantitating the amounts of specific molecules within a cell, determining whether molecules are complexing on a molecular level, measuring changes in ionic concentrations within cells and organelles, and measuring molecular dynamics. This chapter also discusses the issues important to quantitative measurement of fluorescence and focuses on four of quantitative measurements of fluorescence--boxcar-gated detection, streak cameras, photon correlation, and phase modulation. Although quantitative measurement presents many pitfalls to the beginner, it also presents significant opportunities to one skilled in the art. This chapter also examines how fluorescence is measured in the steady state and time domain and how fluorescence is applied in the modern epifluorescence microscope.

  16. Reviews in fluorescence 2010

    CERN Document Server

    Geddes, Chris D

    2011-01-01

    ""Reviews in Fluorescence 2010"", the seventh volume of the book serial from Springer, serves as a comprehensive collection of current trends and emerging hot topics in the field of fluorescence and closely related disciplines. It summarizes the year's progress in fluorescence and its applications, with authoritative analytical reviews specialized enough to be attractive to professional researchers, yet also appealing to the wider audience of scientists in related disciplines of fluorescence. ""Reviews in Fluorescence"" offers an essential reference material for any lab working in the fluoresc

  17. Principles of fluorescence techniques

    CERN Document Server

    2016-01-01

    Fluorescence techniques are being used and applied increasingly in academics and industry. The Principles of Fluorescence Techniques course will outline the basic concepts of fluorescence techniques and the successful utilization of the currently available commercial instrumentation. The course is designed for students who utilize fluorescence techniques and instrumentation and for researchers and industrial scientists who wish to deepen their knowledge of fluorescence applications. Key scientists in the field will deliver theoretical lectures. The lectures will be complemented by the direct utilization of steady-state and lifetime fluorescence instrumentation and confocal microscopy for FLIM and FRET applications provided by leading companies.

  18. Fluorescence correlation spectroscopy

    NARCIS (Netherlands)

    Hink, M.A.; Verveer, P.J.

    2015-01-01

    Fluorescence fluctuation spectroscopy techniques allow the quantification of fluorescent molecules present at the nanomolar concentration level. After a brief introduction to the technique, this chapter presents a protocol including background information in order to measure and quantify the molecul

  19. Safe biodegradable fluorescent particles

    Science.gov (United States)

    Martin, Sue I.; Fergenson, David P.; Srivastava, Abneesh; Bogan, Michael J.; Riot, Vincent J.; Frank, Matthias

    2010-08-24

    A human-safe fluorescence particle that can be used for fluorescence detection instruments or act as a safe simulant for mimicking the fluorescence properties of microorganisms. The particle comprises a non-biological carrier and natural fluorophores encapsulated in the non-biological carrier. By doping biodegradable-polymer drug delivery microspheres with natural or synthetic fluorophores, the desired fluorescence can be attained or biological organisms can be simulated without the associated risks and logistical difficulties of live microorganisms.

  20. Fluorescence antibunching microscopy

    CERN Document Server

    Schwartz, Osip

    2011-01-01

    Breaking the diffraction limit in microscopy by utilizing quantum properties of light has been the goal of intense research in the recent years. We propose a quantum superresolution technique based on non-classical emission statistics of fluorescent markers, routinely used as contrast labels for bio-imaging. The technique can be readily implemented using standard fluorescence microscopy equipment.

  1. Fluorescence of atopic allergens

    NARCIS (Netherlands)

    Berrens, L.

    1967-01-01

    Purified atopic allergens have been found to emit flue fluorescence upon irradiation with ultraviolet light of 365 mμ wavelength. The maximum of fluorescence is in the region 445–490 mμ and the intensity is of the same order of magnitude for different atopic allergens. Synthetic model compounds, inc

  2. Fluorescent Lamp Replacement Study

    Science.gov (United States)

    2017-07-01

    Light-emitting diode, LED, lighting, fluorescent, waste reduction, energy conservation, net zero , mercury 16. SECURITY CLASSIFICATION OF: 17. LIMITATION...Center (ATC) to assess the benefits of converting fluorescent tube lighting to light-emitting diode (LED) technology. The report documents the waste ...1-15 SECTION 2. SUBTESTS 2.1 HAZARDOUS WASTE REDUCTION

  3. LEDs for fluorescence microscopy

    NARCIS (Netherlands)

    Young, I.T.; Garini, Y.; Dietrich, H.R.C.; Van Oel, W.; Liqui Lung, G.

    2004-01-01

    Traditional light sources for fluorescence microscopy have been mercury lamps, xenon lamps, and lasers. These sources have been essential in the development of fluorescence microscopy but each can have serious disadvantages: lack of near monochromaticity, heat generation, cost, lifetime of the light

  4. 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.

  5. Highly thermostable fluorescent proteins

    Science.gov (United States)

    Bradbury, Andrew M [Santa Fe, NM; Waldo, Geoffrey S [Santa Fe, NM; Kiss, Csaba [Los Alamos, NM

    2012-05-01

    Thermostable fluorescent proteins (TSFPs), methods for generating these and other stability-enhanced proteins, polynucleotides encoding such proteins, and assays and method for using the TSFPs and TSFP-encoding nucleic acid molecules are provided. The TSFPs of the invention show extremely enhanced levels of stability and thermotolerance. In one case, for example, a TSFP of the invention is so stable it can be heated to 99.degree. C. for short periods of time without denaturing, and retains 85% of its fluorescence when heated to 80.degree. C. for several minutes. The invention also provides a method for generating stability-enhanced variants of a protein, including but not limited to fluorescent proteins.

  6. Highly thermostable fluorescent proteins

    Science.gov (United States)

    Bradbury, Andrew M.; Waldo, Geoffrey S.; Kiss, Csaba

    2011-03-22

    Thermostable fluorescent proteins (TSFPs), methods for generating these and other stability-enhanced proteins, polynucleotides encoding such proteins, and assays and method for using the TSFPs and TSFP-encoding nucleic acid molecules are provided. The TSFPs of the invention show extremely enhanced levels of stability and thermotolerance. In one case, for example, a TSFP of the invention is so stable it can be heated to 99.degree. C. for short periods of time without denaturing, and retains 85% of its fluorescence when heated to 80.degree. C. for several minutes. The invention also provides a method for generating stability-enhanced variants of a protein, including but not limited to fluorescent proteins.

  7. Introduction to fluorescence

    CERN Document Server

    Jameson, David M

    2014-01-01

    "An essential contribution to educating scientists in the principles of fluorescence. It will also be an important addition to the libraries of practitioners applying the principles of molecular fluorescence."-Ken Jacobson, Kenan Distinguished Professor of Cell Biology and Physiology, University of North Carolina at Chapel Hill"An exquisite compendium of fluorescence and its applications in biochemistry enriched by a very exciting historical perspective. This book will become a standard text for graduate students and other scientists."-Drs. Zygmunt (Karol) Gryczynski and Ignacy Gryczynski, University of North Texas Health Science Center"… truly a masterwork, combining clarity, precision, and good humor. The reader, novice or expert, will be pleased with the text and will not stop reading. It is a formidable account of the fluorescence field, which has impacted the life sciences so considerably in the last 60 years."-Jerson L. Silva, M.D., Ph.D., Professor and Director, National Institute of Science and Tech...

  8. Functional Fluorescent Organic Nanoparticles

    OpenAIRE

    Campioli, Elisa

    2013-01-01

    This thesis presents an extensive study on fluorescent organic nanoparticles and fluorescent organic binary and ternary nanoassemblies. In particular the attention is focused on the preparation and characterization of organic nanoparticles and new nanocomposites obtained from different types of small organic molecules, their stabilization and the use of these materials for biological and optoelectronics applications. The work deals at the beginning with the description of some methods used...

  9. Nine New Fluorescent Probes

    Science.gov (United States)

    Lin, Tsung-I.; Jovanovic, Misa V.; Dowben, Robert M.

    1989-06-01

    Absorption and fluorescence spectroscopic studies are reported here for nine new fluorescent probes recently synthesized in our laboratories: four pyrene derivatives with substituents of (i) 1,3-diacetoxy-6,8-dichlorosulfonyl, (ii) 1,3-dihydroxy-6,8-disodiumsulfonate, (iii) 1,3-disodiumsulfonate, and (iv) l-ethoxy-3,6,8-trisodiumsulfonate groups, and five [7-julolidino] coumarin derivatives with substituents of (v) 3-carboxylate-4-methyl, (vi) 3- methylcarboxylate, (vii) 3-acetate-4-methyl, (viii) 3-propionate-4-methyl, and (ix) 3-sulfonate-4-methyl groups. Pyrene compounds i and ii and coumarin compounds v and vi exhibit interesting absorbance and fluorescence properties: their absorption maxima are red shifted compared to the parent compound to the blue-green region, and the band width broadens considerably. All four blue-absorbing dyes fluoresce intensely in the green region, and the two pyrene compounds emit at such long wavelengths without formation of excimers. The fluorescence properties of these compounds are quite environment-sensitive: considerable spectral shifts and fluorescence intensity changes have been observed in the pH range from 3 to 10 and in a wide variety of polar and hydrophobic solvents with vastly different dielectric constants. The high extinction and fluorescence quantum yield of these probes make them ideal fluorescent labeling reagents for proteins, antibodies, nucleic acids, and cellular organelles. The pH and hydrophobicity-dependent fluorescence changes can be utilized as optical pH and/or hydrophobicity indicators for mapping environmental difference in various cellular components in a single cell. Since all nine probes absorb in the UV, but emit at different wavelengths in the visible, these two groups of compounds offer an advantage of utilizing a single monochromatic light source (e.g., a nitrogen laser) to achieve multi-wavelength detection for flow cytometry application. As a first step to explore potential application in

  10. 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.

  11. Nanosecond fluorescence spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Leskovar, B.

    1985-03-01

    This article is a summary of a short course lecture given in conjunction with the 1984 Nuclear Science Symposium. Measuring systems for nanosecond fluorescence spectroscopy using single-photon counting techniques are presented. These involve systems based on relaxation-type spark gap light pulser and synchronously pumped mode-locked dye lasers. Furthermore, typical characteristics and optimization of operating conditions of the critical components responsible for the system time resolution are discussed. A short comparison of the most important deconvolution methods for numerical analysis of experimental data is given particularly with respect to the signal-to-noise ratio of the fluorescence signal. 22 refs., 8 figs.

  12. Fluorescence Experiments with Quinine

    Science.gov (United States)

    O'Reilly, James E.

    1975-01-01

    Describes a series of experiments which illustrate the analytical capabilities of fluorescence, and outlines two straightforward analyses involving real analyses. These experiments are suitable for an undergraduate instrumental analysis course and require approximately six to seven hours of laboratory time. (MLH)

  13. FLEX: fluorescence explorer

    NARCIS (Netherlands)

    Stoll, M.Ph.; Court, A.J.; Smorenburg, C.; Visser, H.; Crocco, L.; Heilimo, J.; Honig, A.

    1999-01-01

    FLEX is a scientifically driven space mission to provide demonstration/validation of the instrumentation and technique for measuring the natural fluorescence of vegetation in the Fraunhofer lines. The payload consists of high spectral resolution (0.1-0.3 nm) CCD imaging grating spectrometer with two

  14. Random bladder biopsy and hydrodistention with cystoscopy under anesthesia play an important diagnostic and therapeutic role in bladder pain syndrome/interstitial cystitis%膀胱镜随机活检及麻醉下水扩张对膀胱疼痛综合征/间质性膀胱炎诊治的价值

    Institute of Scientific and Technical Information of China (English)

    朱绪辉; 杜鹏; 善辉; 李涛; 张鹏; 杨勇

    2012-01-01

    Objective To evaluate the diagnostic and therapeutic role in bladder pain syndrome/interstitial cystitis (BPS/IC) of random bladder biopsy and hydrodistention with cystoscopy under anesthesia.Methods A retrospective review of cases in our BPS/IC center was performed from 2005 to 2010. One hundred and nineteen patients were included who are diagnosed as the bladder pain syndrome/interstitial cystitis (BPS/IC).There were 32 male patients,aged 47 to 64 years,and 56 years on average; 87 female cases,aged 23 to 67 years,49 years on average.Patients with bladder pain symptoms underwent a thorough evaluation which include voiding diary,pelvic pain、urgency and frequency questionnaire,urine culture,cytology,acid fast bacilli and upper tract imaging.Cystoscopy and random bladder biopsy had been undertaken with general anesthesia.Before and after hydrodistention with cystoscopy,the patients daily frequency of urination,the maximum urine volume,pain scores,symptom scores,QOL score were observed to understand whether there are significant improvement. Results One hundred and nineteen BPS/IC cases underwent random bladder biopsy and hydrodistention with cystoscopy under anesthesia,and finally 102 cases were confirmed of IC; 17 cases were not IC. Eight patients who were previously diagnosed as BPS/IC were found bladder transitional cell carcinoma as the cause of bladder pain symptoms( including 4 cases carcinoma in situ,1 case Low grade non-invasive bladder cancer,3 cases High grade invasive bladder cancers),and 4 of whom had no hematuria. Mean time from the occurrence of BPS to diagnosis of transitional cell carcinoma was 10.8 months. Three patients previously diagnosed as BPS/IC were found tuberculous cystitis as the cause of symptoms,and one eosinophilic cystitis,three chemical cystitis,two radiation cystitis were also detected.Before hydrodistention with cystoscopy under anesthesia,the daily frequency of urination in 102 cases diagnosed for IC patients was 42.1 ±5.6; the

  15. Synthesis and Fluorescence Spectra of Triazolylcoumarin Fluorescent Dyes

    Institute of Scientific and Technical Information of China (English)

    PENG Xian-fu; LI Hong-qi

    2009-01-01

    Much attention is devoted to fluorescent dyes especially those with potential in versatile applications. Reactions under "click" conditions between nonfluorescent 3 - azidocoumarins and terminal alkynes produced 3 -(1, 2, 3- triazol- 1 - yl)cournarins, a novel type of fluorescent dyes with intense fluorescence. The structures of the new coumarins were characterized by 1H NMR, MS, and IR spectra. Fluorescence spectra measurement demonstrated excellent fluorescence performance of the triazolylcoumarins and this click reaction is a promising candidate for bioconjugation and bioimaging applications since both azide and alkynes are quite inert to biological systems.

  16. Fluorescence nanoscopy. Methods and applications

    OpenAIRE

    Requejo-Isidro, Jose

    2013-01-01

    Fluorescence nanoscopy refers to the experimental techniques and analytical methods used for fluorescence imaging at a resolution higher than conventional, diffraction-limited, microscopy. This review explains the concepts behind fluorescence nanoscopy and focuses on the latest and promising developments in acquisition techniques, labelling strategies to obtain highly detailed super-resolved images and in the quantitative methods to extract meaningful information from them.

  17. Delayed fluorescence in photosynthesis.

    Science.gov (United States)

    Goltsev, Vasilij; Zaharieva, Ivelina; Chernev, Petko; Strasser, Reto J

    2009-01-01

    Photosynthesis is a very efficient photochemical process. Nevertheless, plants emit some of the absorbed energy as light quanta. This luminescence is emitted, predominantly, by excited chlorophyll a molecules in the light-harvesting antenna, associated with Photosystem II (PS II) reaction centers. The emission that occurs before the utilization of the excitation energy in the primary photochemical reaction is called prompt fluorescence. Light emission can also be observed from repopulated excited chlorophylls as a result of recombination of the charge pairs. In this case, some time-dependent redox reactions occur before the excitation of the chlorophyll. This delays the light emission and provides the name for this phenomenon-delayed fluorescence (DF), or delayed light emission (DLE). The DF intensity is a decreasing polyphasic function of the time after illumination, which reflects the kinetics of electron transport reactions both on the (electron) donor and the (electron) acceptor sides of PS II. Two main experimental approaches are used for DF measurements: (a) recording of the DF decay in the dark after a single turnover flash or after continuous light excitation and (b) recording of the DF intensity during light adaptation of the photosynthesizing samples (induction curves), following a period of darkness. In this paper we review historical data on DF research and recent advances in the understanding of the relation between the delayed fluorescence and specific reactions in PS II. An experimental method for simultaneous recording of the induction transients of prompt and delayed chlorophyll fluorescence and decay curves of DF in the millisecond time domain is discussed.

  18. Value of carbon dioxide for cystoscopy and its clinical application in the diagnosis of adult patients with hematuria%二氧化碳气膀胱镜在成人血尿患者诊断中的临床应用价值

    Institute of Scientific and Technical Information of China (English)

    陈友干; 孙健; 李光; 张邓新; 向从明; 邓圆圆; 丁锡奇

    2013-01-01

    目的 探讨二氧化碳(CO2)气膀胱镜在成人血尿患者诊断中的临床应用价值.方法 以CO2为介质,对入选的215例成人血尿患者进行CO2气膀胱镜检查,其中男141例,女74例,肉眼血尿180例,镜下血尿35例.观察膀胱内壁、前列腺及尿道内等情况,必要时活检病理检查.结果 所有患者均顺利完成检查或活检,手术时间13~24 min,术中无1例因出血而导致操作中断.患者术中术后均无明显CO2相关并发症.本组最终诊断为膀胱炎73例,膀胱癌46例,前列腺增生症27例,膀胱结石9例,上尿路出血17例,尿道炎10例,其中33例未见异常改变.结论 CO2气膀胱镜安全、可靠,操作视野清晰,对成人血尿患者尤其是严重血尿者的临床诊治有一定优势,值得临床推广应用.%Objective To investigate the value of carbon dioxide for cystoscopy and its clinical application in the diagnosis of adult patients with hematuria. Methods The study group consisted of 215 adult patients with hematuria requiring CO2 cystoscopy. Among them, there were 141 male patients and 74 female patients. The number of patients with gross hematuria was 180, and patients with microscopic hematuria was 35. Results All examinations were successful with an average examination duration of 13-24 min. No examination was stopped as a result of bleeding. There were no CO2 - related complications during preoperative and postoperative periods. Among 215 adult patients with hematuria, cystitis occurred in 73, bladder cancer in 46, benign prostate hy-perplasia (BPH) in 27, bladder stones in 9, upper urinary tract hemorrhage in 17, urethritis in 10, and no abnormalities in 33. Conclusion Carbon dioxide for cystoscopy is a safe and effective technique that provides a good endoscopic view, and this technique would seem to have considerable advantages in the clinical diagnosis of adult patients with hematuria.

  19. Retroperitoneal laparoscopic pyelolithotomy combined with holmium laser lithotripsy under flexible cystoscopy in treatment of complicated nephrolithiasis%后腹腔镜肾盂切开取石联合膀胱软镜钬激光碎石取石术治疗复杂性肾结石

    Institute of Scientific and Technical Information of China (English)

    肖亚; 付卫华; 张艮甫; 王平贤; 范明齐; 冯嘉瑜; 钟晓; 黄赤兵

    2014-01-01

    Objective To evaluate the efficacy and safety of retroperitoneal laparoscopic pyelolithotomy ( RLP) combined with holmium laser lithotripsy under flexible cystoscopy in the treatment of complicated nephrolithiasis. Methods The retrospective analysis was made on the clinical data of 37 patients who underwent RLP and holmium laser lithotripsy under flexible cystoscopy for complicated nephrolithiasis from January 2013 to January 2014. The clinic parameters involved basic data of patients,operational time,blood loss,post-operative hospital stay,the status of stone-free,perioperative complications,and the follow-up data of patients were observed. Results No patient was converted to open surgery. The mean stone size was (2. 8 ± 0. 9) cm in diameter,operational time was (89 ± 24) min,blood loss was (21. 3 ± 7. 7) mL,post-operative hospital stay was (6. 8 ± 1. 7) d,the stone removal rate in one session was 94. 6%. One case occurred urinary leakage,1 case occurred fever after operation,who were all recovered through conservative treatment. All cases were followed up at the sixth months after operation. Conclusion RLP combined with holmium laser lithotripsy under flexible cystoscopy is effective and safe for the treatment of com-plicated nephrolithiasis.%目的:分析后腹腔镜肾盂切开取石联合膀胱软镜钬激光碎石取石术治疗复杂性肾结石的有效性和安全性。方法2012年1月至2014年1月对37例复杂性肾结石患者行后腹腔镜下肾盂切开取石联合膀胱软镜钬激光碎石取石术;统计并分析患者基本资料、手术时间、术中出血量、术后住院时间、一期清石率、围手术期并发症、术后复查等情况。结果37例患者顺利完成手术。平均结石直径(2.8±0.9) cm,手术时间(89±24) min,术中出血量(21.3±7.7) mL,术后住院时间(6.8±1.7) d,一期清石率为94.6%,围手术期1例发热,1例尿漏,经保守治疗均痊愈。术后6个月对所有患者进行随访,肾功能

  20. Fluorescence-Based Sensors

    Science.gov (United States)

    Orellana, Guillermo

    The natural luminescent phenomena (from the Latin words "lumen" and "essentia", i.e., "made of light") such as northern lights (aurora borealis), marine brightness, glow-worms, shining putrid fish scales, "bluish"- appearing water when contained in certain wooden cups (quinine fluorescence), some stones heated at high temperatures with reducing agents (BaS phosphorescence), or light emitted while crushing sugar (triboluminescence) already fascinated our ancestors. Nowadays we understand that ultraviolet and visible emission of light originates from a competitive deactivation pathway of the lowest electronic excited state of atoms and molecules that produces the so called luminescence (the sub-terms fluorescence and phosphorescence just designate whether the return of the excited to the ground state is an "allowed" or "forbidden" process, namely it is fast or slow, the loosely-defined border between them being a 1-μs-1 rate constant). Actually, luminescence is the only method to generate light in the known Universe regardless it is powered by the nuclear reactions in the stars, the ohmical heating in bulbs, an electric discharge, the absorption of light or a (bio)chemical reaction (chemiluminescence).

  1. The TALE Fluorescence Detectors

    Science.gov (United States)

    Jui, Charles

    2009-05-01

    The TALE fluorescence detectors are designed to extend the threshold for fluorescence observation by TA down to 3x10^16 eV. It will comprise two main components. The first is a set of 24 telescopes working in stereo, with an existing TA FD station at ˜6 km separation. These will cover between 3-31 degrees in elevation and have azimuthal coverage maximizing the stereo aperture in the 10^18-10^19 eV energy range. The second component consists of 15 telescopes equipped with 4m diameter mirrors and covering the sky between 31 and 73 degrees in elevation. The larger mirror size pushes the physics threshold down to 3x10^16 eV, and provides view of the shower maximum for the lower energy events. The Tower detector will cover one quadrant in azimuth and operate in hybrid mode with the TALE infill array to provide redundant composition measurements from both shower maximum information and muon-to-electron ratio.

  2. A fluorescence scanning electron microscope

    Directory of Open Access Journals (Sweden)

    Takaaki Kanemaru

    2010-01-01

    Full Text Available Fluorescence techniques are widely used in biological research to examine molecular localization, while electron microscopy can provide unique ultrastructural information. To date, correlative images from both fluorescence and electron microscopy have been obtained separately using two different instruments, i.e. a fluorescence microscope (FM and an electron microscope (EM. In the current study, a scanning electron microscope (SEM (JEOL JXA8600 M was combined with a fluorescence digital camera microscope unit and this hybrid instrument was named a fluorescence SEM (FL-SEM. In the labeling of FL-SEM samples, both Fluolid, which is an organic EL dye, and Alexa Fluor, were employed. We successfully demonstrated that the FL-SEM is a simple and practical tool for correlative fluorescence and electron microscopy.

  3. Fluorescence of ceramic color standards.

    Science.gov (United States)

    Koo, Annette; Clare, John F; Nield, Kathryn M; Deadman, Andrew; Usadi, Eric

    2010-04-20

    Fluorescence has been found in color standards available for use in calibration and verification of color measuring instruments. The fluorescence is excited at wavelengths below about 600?nm and emitted above 700?nm, within the response range of silicon photodiodes, but at the edge of the response of most photomultipliers and outside the range commonly scanned in commercial colorimeters. The degree of fluorescence on two of a set of 12 glossy ceramic tiles is enough to introduce significant error when those tiles have been calibrated in one mode of measurement and are used in another. We report the nature of the fluorescence and the implications for color measurement.

  4. Fluorescence dynamics of green fluorescent protein in AOT reversed micelles

    NARCIS (Netherlands)

    Uskova, M.A.; Borst, J.W.; Hink, M.A.; Hoek, van A.; Schots, A.; Klyachko, N.L.; Visser, A.J.W.G.

    2000-01-01

    We have used the enhanced green fluorescent protein (EGFP) to investigate the properties of surfactant-entrapped water pools in organic solvents (reversed micelles) with steady-state and time-resolved fluorescence methods. The surfactant used was sodium bis(2-ethylhexyl)sulfosuccinate (AOT) and the

  5. 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…

  6. X-ray Fluorescence Sectioning

    CERN Document Server

    Cong, Wenxiang

    2012-01-01

    In this paper, we propose an x-ray fluorescence imaging system for elemental analysis. The key idea is what we call "x-ray fluorescence sectioning". Specifically, a slit collimator in front of an x-ray tube is used to shape x-rays into a fan-beam to illuminate a planar section of an object. Then, relevant elements such as gold nanoparticles on the fan-beam plane are excited to generate x-ray fluorescence signals. One or more 2D spectral detectors are placed to face the fan-beam plane and directly measure x-ray fluorescence data. Detector elements are so collimated that each element only sees a unique area element on the fan-beam plane and records the x-ray fluorescence signal accordingly. The measured 2D x-ray fluorescence data can be refined in reference to the attenuation characteristics of the object and the divergence of the beam for accurate elemental mapping. This x-ray fluorescence sectioning system promises fast fluorescence tomographic imaging without a complex inverse procedure. The design can be ad...

  7. Optical Properties of Fluorescent Dyes

    Institute of Scientific and Technical Information of China (English)

    李戎; 陈东辉

    2001-01-01

    Fluorescent dyes have been widely used these years.Because of the special optical performance, conventional CCM systems seem to be unable to predict the recipes of fabrics dyed with fluorescent dyes. In order to enhance the functions of CCM systems, the optical properties of fluorescent dyes in their absorption region were investigated. It has been found that there was a fixed maximum absorption wavelength for each fluorescent dyes whatever its concentration is. Both absorption region and maximum absorption wavelength of the dyes in solution are the same to those in fabric, and that the absorption is directly proportional to the concentration of the dye. So the optical properties obtained in solutions cna be applied for describing the optics performance of fluorescent dyes in fabrics.

  8. Fluorescence calibration method for single-particle aerosol fluorescence instruments

    Science.gov (United States)

    Shipley Robinson, Ellis; Gao, Ru-Shan; Schwarz, Joshua P.; Fahey, David W.; Perring, Anne E.

    2017-05-01

    Real-time, single-particle fluorescence instruments used to detect atmospheric bioaerosol particles are increasingly common, yet no standard fluorescence calibration method exists for this technique. This gap limits the utility of these instruments as quantitative tools and complicates comparisons between different measurement campaigns. To address this need, we have developed a method to produce size-selected particles with a known mass of fluorophore, which we use to calibrate the fluorescence detection of a Wideband Integrated Bioaerosol Sensor (WIBS-4A). We use mixed tryptophan-ammonium sulfate particles to calibrate one detector (FL1; excitation = 280 nm, emission = 310-400 nm) and pure quinine particles to calibrate the other (FL2; excitation = 280 nm, emission = 420-650 nm). The relationship between fluorescence and mass for the mixed tryptophan-ammonium sulfate particles is linear, while that for the pure quinine particles is nonlinear, likely indicating that not all of the quinine mass contributes to the observed fluorescence. Nonetheless, both materials produce a repeatable response between observed fluorescence and particle mass. This procedure allows users to set the detector gains to achieve a known absolute response, calculate the limits of detection for a given instrument, improve the repeatability of the instrumental setup, and facilitate intercomparisons between different instruments. We recommend calibration of single-particle fluorescence instruments using these methods.

  9. Single Molecule Spectroscopy of Fluorescent Proteins

    NARCIS (Netherlands)

    Blum, Christian; Subramaniam, Vinod

    2009-01-01

    The discovery and use of fluorescent proteins has revolutionized cellular biology. Despite the widespread use of visible fluorescent proteins as reporters and sensors in cellular environments the versatile photophysics of fluorescent proteins is still subject to intense research. Understanding the

  10. FLEX: fluorescence explorer

    Science.gov (United States)

    Stoll, Marc-Ph.; Court, Andrew; Smorenburg, Kees; Visser, Huib; Crocco, Luiggi; Heilimo, Jyro; Honig, Andre

    1999-12-01

    FLEX is a scientifically driven space mission to provide demonstration/validation of the instrumentation and technique for measuring the natural fluorescence of vegetation in the Fraunhofer lines. The payload consists of high spectral resolution (0.1 - 0.3 nm) CCD imaging grating spectrometer with two channels: one in the red (648 - 664 nm) and one in the blue (391 - 438 nm) for working with several Fraunhofer lines. The across track FOV is 8.4 degrees; ground spatial resolution is better than 0.5 X 0.5 km2. To increase the S/N ratio a steering mirror will be used, if necessary, to 'freeze' the image and also to provide plus or minus 4 degrees across track depointing. Calibration is made by viewing the sun via a diffuser plate switched into the telescope field of view. A separate CCD camera will allow cloud detection and scene identification. A TIR radiometer will provide simultaneous surface temperature measurements. The spacecraft, overall mass estimated at 200 kg, is derived from the ASI-MITA bus which provides all the necessary subsystems and stabilized platform. By use of on-board storage, ground requirements for satellite control and data link are minimized; the possibility of local stations for real time reception/distribution is also envisaged. Provisional orbit characteristics are: LEO sun synchronous, 500 - 900 km altitude. Priority will be given to highest revisit frequency on a sufficient number of selected test sites.

  11. Fluorescent blood cell angiography

    Science.gov (United States)

    Ben-nun, Joshua; Constable, Ian J.

    1994-06-01

    Fluorescein angiography is currently the main method for evaluation of the retinal vascular patency. Ashton noted that capillary patency to the small fluorescein molecule may differ from that of the larger red blood cells. He concluded that fluorescein angiography is not able to demonstrate a developing stenosis, that might be the precipitating cause of a later capillary closure in various microvasculopathies. Sarelius et al have shown, in hamster cheek pouch and cremaster muscle, that fluorescently labeled erythrocytes in known concentrations can be used for the direct measurement of capillary flow parameters. The only assumption that this method relies on, is that the labeled cells are rheologically normal and therefore reflect the behavior of the total cell population. We have developed a new method for an in-vivo, real-time demonstration of the blood cell flow in the retinal capillary net. Based on the assumption presented by Sarelius et al, measurement and analysis of the retinal capillary blood cell flow is also possible from the results achieved by the new method.

  12. Fluorescence endoscopy and photodynamic therapy.

    Science.gov (United States)

    Messmann, H; Endlicher, E; Gelbmann, C M; Schölmerich, J

    2002-10-01

    Fluorescence endoscopy is a new technique which allows a better detection of non-visible malignant or premalignant lesions or, those which are difficult to detect. Exogenously applied sensitisers accumulate selectively in malignant lesions and induce fluorescence after illumination with light of adequate wavelength. However, also endogenous fluorophores, different located in malignant or benign lesions, induce a different autofluorescence in these lesions. Tissue fluorescence can be detected by optical sampling of the mucosa using fluorescence spectroscopy or by generating real time fluorescence images with specialised camera systems. Compared to point fluorescence spectroscopy the latter technique enables the screening of large surface areas of mucosa. Meanwhile, fluorescence endoscopy is a widely used technique in urology employing 5-aminolaevulinic acid sensitisation. In gastroenterology, this technique seems promising for the detection of early cancers or dysplasia in patients with Barrett's oesophagus or ulcerative colitis. Using different sensitisers, photodynamic therapy seems to be a promising option for patients with advanced oesophageal cancer and in the palliative treatment of non-resectable bile duct cancer, furthermore for patients with early gastric cancer and dysplasia in Barrett's oesophagus. Probably, by laser light fractionation or a combination of different sensitisers, an enhanced effect can be expected.

  13. Modular generation of fluorescent phycobiliproteins.

    Science.gov (United States)

    Wu, Xian-Jun; Chang, Kun; Luo, Juan; Zhou, Ming; Scheer, Hugo; Zhao, Kai-Hong

    2013-06-01

    Phycobiliproteins are brightly-fluorescent light-harvesting pigments for photosynthesis in cyanobacteria and red algae. They are also of interest as fluorescent biomarkers, but their heterologous generation in vivo has previously required multiple transformations. We report here a modular approach that requires only two DNA segments. The first codes for the apo-protein. The second codes for fusions capable of chromophore biosynthesis and its covalent attachment to the apo-protein; it contains the genes of heme oxygenase, a bilin reductase, and a chromophore lyase. Phycobiliproteins containing phycoerythrobilin (λ(fluor) ~ 560 nm), phycourobilin (λ(fluor) ~ 500 nm), phycocyanobilin (λ(fluor) ~ 630 nm) or phycoviolobilin (λ(fluor) ~ 580 nm) were obtained in high yield in E. coli. This approach facilitates chromophorylation studies of phycobiliproteins, as well as their use for fluorescence labeling based on their high fluorescence.

  14. Fluorescence diagnosis in tissue injury

    Science.gov (United States)

    Maciel, Vitória H.; Ferreira, Juliana; Bagnato, Vanderlei S.

    2009-06-01

    Background and Objectives: The paper aim was to evaluate the efficacy of the fluorescence spectroscopy in the detection of UV-induced skin change of Wistar rats. Study Design/ Materials and Methods: In a group male Wistar rats, the skin damage was produced by an UV-C lamp, periodically monitored using the laser-induced fluorescence, until complete healing process. After determining a characteristic emission band present in the fluorescence spectra of the induced injuries, the amplitude band monitoring allowed the follow up on the injury and the recovery. Results: We observed the appearance of two new emission bands more evident at the injury spectra when compared to the spectrums from normal non-exposed tissue. Following such spectral bands was possible to observe the establishment and recovery. Conclusions: The fluorescence spectroscopy is a promising technique in distinguishing between normal and UV induced skin change helping the evaluation of changes which are irreversible cancer tissue characteristics.

  15. Fluorescent Sensors for Biological Applications

    Directory of Open Access Journals (Sweden)

    Hui-wang Ai

    2014-09-01

    Full Text Available Fluorescence is one of the most important analytical methods used in biological studies. In the past decade or two, instrumentation in this field has greatly advanced, and now it is possible to detect single photons or fluorescent molecules [1,2], or break the Abbe diffraction limit to distinguish two points spaced less than 50 nm apart [3]. Concurrently, the development of improved fluorescent probes, which can be coupled with state-of-the-art instruments, has been equally important. This special issue on “fluorescent biosensors” in Sensors reports recent results from eight research groups in the field of sensor development. It includes three review articles, and six research articles reporting original results. [...

  16. X-ray fluorescence holography

    CERN Document Server

    Hayashi, K; Takahashi, Y

    2003-01-01

    X-ray fluorescence holography (XFH) is a new structural analysis method of determining a 3D atomic arrangement around fluorescing atoms. We developed an XFH apparatus using advanced X-ray techniques and succeeded in obtaining high-quality hologram data. Furthermore, we introduced applications to the structural analysis of a thin film and the environment around dopants and, discussed the quantitative analysis of local lattice distortion. (author)

  17. Fluorescence lifetimes: fundamentals and interpretations.

    Science.gov (United States)

    Noomnarm, Ulai; Clegg, Robert M

    2009-01-01

    Fluorescence measurements have been an established mainstay of photosynthesis experiments for many decades. Because in the photosynthesis literature the basics of excited states and their fates are not usually described, we have presented here an easily understandable text for biology students in the style of a chapter in a text book. In this review we give an educational overview of fundamental physical principles of fluorescence, with emphasis on the temporal response of emission. Escape from the excited state of a molecule is a dynamic event, and the fluorescence emission is in direct kinetic competition with several other pathways of de-excitation. It is essentially through a kinetic competition between all the pathways of de-excitation that we gain information about the fluorescent sample on the molecular scale. A simple probability allegory is presented that illustrates the basic ideas that are important for understanding and interpreting most fluorescence experiments. We also briefly point out challenges that confront the experimenter when interpreting time-resolved fluorescence responses.

  18. Fluorescence detection of esophageal neoplasia

    Science.gov (United States)

    Borisova, E.; Vladimirov, B.; Avramov, L.

    2008-06-01

    White-light endoscopy is well-established and wide used modality. However, despite the many technological advances that have been occurred, conventional endoscopy is suboptimal and usually detects advanced stage lesions. The limitations of standard endoscopy initiate development of spectroscopic techniques, additional to standard endoscopic equipment. One of the most sensitive approaches is fluorescence spectroscopy of gastrointestinal mucosa for neoplasia detection. In the recent study delta-aminolevulinic acid/Protoporphyrin IX (5-ALA/PpIX) is used as fluorescent marker for dysplasia and tumor detection in esophagus. The 5-ALA is administered per os six hours before measurements at dose 20 mg/kg weight. Excitation source has max of emission at 405 nm and light is delivered by the standard light guide of the endoscopic equipment. Through endoscopic instrumental channel a fiber is applied to return information about fluorescence to microspectrometer. Spectral features observed during endoscopic investigations could be distinct as the next regions: 450-630 nm region, where tissue autofluorescence is observed; 630-710 nm region, where fluorescence of PpIX is clearly pronounced; 530-580 nm region, where minima in the autofluorescence signal are observed, related to reabsorption of blood. The lack of fluorescence peaks in the red spectral area for normal mucosa is an indication for selective accumulation of 5-ALA/PpIX only in abnormal sites Very good correlation between fluorescence signals and histology examination of the lesions investigated is achieved.

  19. Combining fluorescence and bioluminescence microscopy.

    Science.gov (United States)

    Goda, Kazuhito; Hatta-Ohashi, Yoko; Akiyoshi, Ryutaro; Sugiyama, Takashi; Sakai, Ikuko; Takahashi, Takeo; Suzuki, Hirobumi

    2015-08-01

    Bioluminescence microscopy has revealed that gene expression in individual cells can respond differently to the same stimulus. To understand this phenomenon, it is important to sequentially observe the series of events from cellular signal transduction to gene expression regulated by specific transcription factors derived from signaling cascades in individual cells. However, these processes have been separately analyzed with fluorescence and bioluminescence microscopy. Furthermore, in culture medium, the background fluorescence of luciferin-a substrate of luciferase in promoter assays of gene expression in cultured cells-confounds the simultaneous observation of fluorescence and bioluminescence. Therefore, we optimized conditions for optical filter sets based on spectral properties and the luciferin concentration based on cell permeability for fluorescence observation combined with bioluminescence microscopy. An excitation and emission filter set (492-506 nm and 524-578 nm) was suitable for green fluorescent protein and yellow fluorescent protein imaging of cells, and >100 μM luciferin was acceptable in culture medium based on kinetic constants and the estimated intracellular concentration. Using these parameters, we present an example of sequential fluorescence and bioluminescence microscopic observation of signal transduction (translocation of protein kinase C alpha from the cytoplasm to the plasma membrane) coupled with activation of gene expression by nuclear factor of kappa light polypeptide B in individual cells and show that the gene expression response is not completely concordant with upstream signaling following stimulation with phorbol-12-myristate-13-acetate. Our technique is a powerful imaging tool for analysis of heterogeneous gene expression together with upstream signaling in live single cells.

  20. Gallstone identification by fluorescence spectroscopy

    Science.gov (United States)

    Pradhan, Asima; Laxmi, B. V.; Jena, Sidhartha S.; Khulbe, P. K.; Bist, Hari D.; Agarwal, Asha

    1998-04-01

    Gallstones have been classified as being cholesterol type and pigment type. The classification is important for diet control of the patient to avoid recurrence of the stone. Spectroscopy is a sensitive technique to determine the composition of the gallstone both in-vitro and in-vivo. this work deals with the fluorescence spectroscopy of gallstone. For fluorescence spectroscopic studies of gallstone, samples were excited with 5 mw of 488 nm line of argon-ion laser and spectra were recorded with a SPEX 1877E triplemate attached with a cooled PMT and DM3000R data acquisition system. Fluorescence spectra from pure cholesterol and bilirubin were also recorded for comparison. Different types of gallstones: mixed, cholesterol, pigment type were studied. All spectra exhibited a very broad band, 500 to 800 nm and sometimes two bands, depending on type of stone. Pure cholesterol shows three prominent fluorescence peaks at 513, 550, 583 nm along with two peaks at approximately 568 and 586 nm. Pure bilirubin shows prominent peak at 628 nm, without any Raman line. From fluorescence spectra different types of stones are identified. Different gallstones studied show a mixture of cholesterol and bilirubin types and the ratio of the two varies from one sample type to another.

  1. Plasmonics Enhanced Smartphone Fluorescence Microscopy

    KAUST Repository

    Wei, Qingshan

    2017-05-12

    Smartphone fluorescence microscopy has various applications in point-of-care (POC) testing and diagnostics, ranging from e.g., quantification of immunoassays, detection of microorganisms, to sensing of viruses. An important need in smartphone-based microscopy and sensing techniques is to improve the detection sensitivity to enable quantification of extremely low concentrations of target molecules. Here, we demonstrate a general strategy to enhance the detection sensitivity of a smartphone-based fluorescence microscope by using surface-enhanced fluorescence (SEF) created by a thin metal-film. In this plasmonic design, the samples are placed on a silver-coated glass slide with a thin spacer, and excited by a laser-diode from the backside through a glass hemisphere, generating surface plasmon polaritons. We optimized this mobile SEF system by tuning the metal-film thickness, spacer distance, excitation angle and polarization, and achieved ~10-fold enhancement in fluorescence intensity compared to a bare glass substrate, which enabled us to image single fluorescent particles as small as 50 nm in diameter and single quantum-dots. Furthermore, we quantified the detection limit of this platform by using DNA origami-based brightness standards, demonstrating that ~80 fluorophores per diffraction-limited spot can be readily detected by our mobile microscope, which opens up new opportunities for POC diagnostics and sensing applications in resource-limited-settings.

  2. Fluorescence applications in molecular neurobiology.

    Science.gov (United States)

    Taraska, Justin W; Zagotta, William N

    2010-04-29

    Macromolecules drive the complex behavior of neurons. For example, channels and transporters control the movements of ions across membranes, SNAREs direct the fusion of vesicles at the synapse, and motors move cargo throughout the cell. Understanding the structure, assembly, and conformational movements of these and other neuronal proteins is essential to understanding the brain. Developments in fluorescence have allowed the architecture and dynamics of proteins to be studied in real time and in a cellular context with great accuracy. In this review, we cover classic and recent methods for studying protein structure, assembly, and dynamics with fluorescence. These methods include fluorescence and luminescence resonance energy transfer, single-molecule bleaching analysis, intensity measurements, colocalization microscopy, electron transfer, and bimolecular complementation analysis. We present the principles of these methods, highlight recent work that uses the methods, and discuss a framework for interpreting results as they apply to molecular neurobiology.

  3. Lasing from fluorescent protein crystals.

    Science.gov (United States)

    Oh, Heon Jeong; Gather, Malte C; Song, Ji-Joon; Yun, Seok Hyun

    2014-12-15

    We investigated fluorescent protein crystals for potential photonic applications, for the first time to our knowledge. Rod-shaped crystals of enhanced green fluorescent protein (EGFP) were synthesized, with diameters of 0.5-2 μm and lengths of 100-200 μm. The crystals exhibit minimal light scattering due to their ordered structure and generate substantially higher fluorescence intensity than EGFP or dye molecules in solutions. The magnitude of concentration quenching in EGFP crystals was measured to be about 7-10 dB. Upon optical pumping at 485 nm, individual EGFP crystals located between dichroic mirrors generated laser emission with a single-mode spectral line at 513 nm. Our results demonstrate the potential of protein crystals as novel optical elements for self-assembled, micro- or nano-lasers and amplifiers in aqueous environment.

  4. Going Viral with Fluorescent Proteins.

    Science.gov (United States)

    Costantini, Lindsey M; Snapp, Erik L

    2015-10-01

    Many longstanding questions about dynamics of virus-cell interactions can be answered by combining fluorescence imaging techniques with fluorescent protein (FP) tagging strategies. Successfully creating a FP fusion with a cellular or viral protein of interest first requires selecting the appropriate FP. However, while viral architecture and cellular localization often dictate the suitability of a FP, a FP's chemical and physical properties must also be considered. Here, we discuss the challenges of and offer suggestions for identifying the optimal FPs for studying the cell biology of viruses.

  5. Fluorescence for high school students

    CERN Document Server

    Schultheiss, Niek G

    2012-01-01

    In a not obligatory series of lessons for high school students in the Netherlands we discuss the fluorescence aspects of anthracene. These lessons were developed because HiSPARC (High school Project on Astrophysics Research with Cosmics) detection of cosmic rays are available for different secondary schools. With the help of special designed scintillator detection stations, containing anthracene, cosmic rays can be detected. Fluorescence of anthracene is one of the topics discussed in these series of extra curricular lessons aimed at excellent pupils working on cosmic radiation within the HiSPARC - project.

  6. Multichromophoric sugar for fluorescence photoswitching

    Directory of Open Access Journals (Sweden)

    Stéphane Maisonneuve

    2014-06-01

    Full Text Available A multichromophoric glucopyranoside 2 bearing three dicyanomethylenepyran (DCM fluorophores and one diarylethene (DAE photochrome has been prepared by Cu(I-catalyzed alkyne–azide cycloaddition reaction. The fluorescence of 2 was switched off upon UV irradiation, in proportion with the open to closed form (OF to CF conversion extent of the DAE moiety. A nearly 100% Förster-type resonance energy transfer (FRET from all three DCM moieties to a single DAE (in its CF moiety was achieved. Upon visible irradiation, the initial fluorescence intensity was recovered. The observed photoswiching is reversible, with excellent photo resistance.

  7. Biomolecule-to-fluorescent-color encoder: modulation of fluorescence emission via DNA structural changes.

    Science.gov (United States)

    Nishimura, Takahiro; Ogura, Yusuke; Yamada, Kenji; Ohno, Yuko; Tanida, Jun

    2014-07-01

    A biomolecule-to-fluorescent-color (B/F) encoder for optical readout of biomolecular information is proposed. In the B/F encoder, a set of fluorescence wavelengths and their intensity levels are used for coding of a biomolecular signal. A hybridization chain reaction of hairpin DNAs labeled with fluorescent reporters was performed to generate the fluorescence color codes. The fluorescence is modulated via fluorescence resonance energy transfer, which is controlled by DNA structural changes. The results demonstrate that fluorescent color codes can be configured based on two wavelengths and five intensities using the B/F encoder, and the assigned codes can be retrieved via fluorescence measurements.

  8. Fluorescence for high school students

    NARCIS (Netherlands)

    Schultheiss, N.G.; Kool, T.W.

    2012-01-01

    In a not obligatory series of lessons for high school students in the Netherlands we discuss the fluorescence aspects of anthracene. These lessons were developed because HiSPARC (High school Project on Astrophysics Research with Cosmics) detection of cosmic rays are available for different secondary

  9. Development of Sealed Fluorescence Spectrometer

    Institute of Scientific and Technical Information of China (English)

    QIAN; Hong-juan; ZHANG; Li-hua; LIU; Huan-liang; FAN; De-jun

    2012-01-01

    <正>In nuclear fuel reprocessing, the fluorescent analytical instrument can be used to analyze various trace elements, such as boron and thorium in uranium product. Due to the high radioactivity, strong acidity, fatal toxic and complex components of spent nuclear fuel reprocessing sample, analytical works become more difficult and instruments used can be damaged easier.

  10. A fluorescent probe for ecstasy.

    Science.gov (United States)

    Masseroni, D; Biavardi, E; Genovese, D; Rampazzo, E; Prodi, L; Dalcanale, E

    2015-08-18

    A nanostructure formed by the insertion in silica nanoparticles of a pyrene-derivatized cavitand, which is able to specifically recognize ecstasy in water, is presented. The absence of effects from interferents and an efficient electron transfer process occurring after complexation of ecstasy, makes this system an efficient fluorescent probe for this popular drug.

  11. Studying Photosynthesis by Measuring Fluorescence

    Science.gov (United States)

    Sanchez, Jose Francisco; Quiles, Maria Jose

    2006-01-01

    This paper describes an easy experiment to study the absorption and action spectrum of photosynthesis, as well as the inhibition by heat, high light intensity and the presence of the herbicide 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) on the photosynthetic process. The method involves measuring the chlorophyll fluorescence emitted by intact…

  12. In vivo fluorescence lifetime tomography

    Science.gov (United States)

    Nothdurft, Ralph E.; Patwardhan, Sachin V.; Akers, Walter; Ye, Yunpeng; Achilefu, Samuel; Culver, Joseph P.

    2009-03-01

    Local molecular and physiological processes can be imaged in vivo through perturbations in the fluorescence lifetime (FLT) of optical imaging agents. In addition to providing functional information, FLT methods can quantify specific molecular events and multiplex diagnostic and prognostic information. We have developed a fluorescence lifetime diffuse optical tomography (DOT) system for in vivo preclinical imaging. Data is captured using a time-resolved intensified charge coupled device (ICCD) system to measure fluorescence excitation and emission in the time domain. Data is then converted to the frequency domain, and we simultaneously reconstruct images of yield and lifetime using an extension to the normalized Born approach. By using differential phase measurements, we demonstrate DOT imaging of short lifetimes (from 350 ps) with high precision (+/-5 ps). Furthermore, this system retains the efficiency, speed, and flexibility of transmission geometry DOT. We demonstrate feasibility of FLT-DOT through a progressive series of experiments. Lifetime range and repeatability are first measured in phantoms. Imaging of subcutaneous implants then verifies the FLT-DOT approach in vivo in the presence of inhomogeneous optical properties. Use in a common research scenario is ultimately demonstrated by imaging accumulation of a targeted near-infrared (NIR) fluorescent-labeled peptide probe (cypate-RGD) in a mouse with a subcutaneous tumor.

  13. Fluorescence diagnostics in oncological gynecology

    Science.gov (United States)

    Belyaeva, Ludmila A.; Adamyan, Leila V.; Kozachenko, Vladimir P.; Stratonnikov, Alexander A.; Stranadko, Eugene F.; Loschenov, Victor B.

    2003-10-01

    The method of fluorescent diagnostics (FD) of tumors is a promising tool that may allow to increase sensitivity of tumor detection especially at initial stages. One of the most promising photosensitizers today is 5-aminolevulinic acid (5-ALA) that, actually, is not photosensitizer itself but precursor of protoporphyrin IX (PpIX). This paper deals with cancer diagnostics in gynecology by means of ALA-induced Pp IX laser-fluorescence spectroscopy. The tissue fluorescence spectra in vivo were studied in patients with various pathologies of ovaries, uterine and vulva after 5-aminolevulinic acid administration. It was shown that different pathologies varies in accumulation of Pp IX. Coefficient of fluorescence kf for normal tissue is not high, but exceptions are endometrium and mucous membrane of uterine tubes. Benign tumors of uterus and ovary have low values of kf, but polyps of endometrium exhibit high kf. Optical express-biopsy is important for diagnosis of ovarian cancer and micrometastatic spread. Coefficients of diagnostic contrast were determined for cancer of endometrium, cervical cancer, vulvar cancer.

  14. Fluorescent Labeling of Nanometer Hydroxyapatite

    Institute of Scientific and Technical Information of China (English)

    Yuan ZHANG; Yuan YUAN; Changsheng LIU

    2008-01-01

    A novel surface treatment method using 3-aminopropyltriethoxysilane (AMPTES), was developed to immobilize the fluorescein molecule on nano-HAP (nanometer hydroxyapatite) powders. By pretreating the nano-HAP powders surface with AMPTES, fluorescein, chosen on the basis of the chemical structure of the nano- HAP powders, could be bound to the nano-HAP powders surface. The chemical compositions of nano-HAP before and after being labeled were characterized by Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS). The morphology, phase composition, and the fluorescence characteristics of the nano-HAP powders with and without staining were also investigated. The FTIR and XPS results revealed that fiuorescein had been successfully immobilized on the surface of AMPTES-bound nano-HAP powders via the acylamide bond formation between the -COOH of fluorescein and the -NH2 of AMPTES. The labeled nano-HAP powders possessed strong fluorescent intensity with a little deviation from the maximum emission wavelength of fluorescein. But the morphology and phase composition had no obvious alteration. Under fluorescence microscopy, the labeled nano-HAP powders., even after 24 h cell incubation, exhibited strong fluorescence.

  15. Fluorescent sensors based on bacterial fusion proteins

    Science.gov (United States)

    Prats Mateu, Batirtze; Kainz, Birgit; Pum, Dietmar; Sleytr, Uwe B.; Toca-Herrera, José L.

    2014-06-01

    Fluorescence proteins are widely used as markers for biomedical and technological purposes. Therefore, the aim of this project was to create a fluorescent sensor, based in the green and cyan fluorescent protein, using bacterial S-layers proteins as scaffold for the fluorescent tag. We report the cloning, expression and purification of three S-layer fluorescent proteins: SgsE-EGFP, SgsE-ECFP and SgsE-13aa-ECFP, this last containing a 13-amino acid rigid linker. The pH dependence of the fluorescence intensity of the S-layer fusion proteins, monitored by fluorescence spectroscopy, showed that the ECFP tag was more stable than EGFP. Furthermore, the fluorescent fusion proteins were reassembled on silica particles modified with cationic and anionic polyelectrolytes. Zeta potential measurements confirmed the particle coatings and indicated their colloidal stability. Flow cytometry and fluorescence microscopy showed that the fluorescence of the fusion proteins was pH dependent and sensitive to the underlying polyelectrolyte coating. This might suggest that the fluorescent tag is not completely exposed to the bulk media as an independent moiety. Finally, it was found out that viscosity enhanced the fluorescence intensity of the three fluorescent S-layer proteins.

  16. Application of multicolor fluorescence in situ hybridization in postoperative follow-up monitoring of bladder cancer patients with negative urine cytology%多色荧光原位杂交技术在尿脱落细胞学阴性膀胱癌患者术后随访监测中的应用

    Institute of Scientific and Technical Information of China (English)

    王南雄; 王尉; 吕军; 王健; 叶纯

    2015-01-01

    目的 探讨多色荧光原位杂交技术( M-FISH)在尿脱落细胞学阴性膀胱癌患者术后随访监测中的应用价值. 方法 收集接受经尿道膀胱肿瘤切除术后的膀胱尿路上皮癌患者76例,术前、术后随访尿脱落细胞学检查均阴性,分别采用膀胱镜、M-FISH进行术后随访监测.结果 76例患者中14例(18.4%)复发,M-FISH仅检出3例(3.9%)复发,漏诊11例(14.5%). 膀胱镜检查结果阴性54例,复发1例(低级别),M-FISH检查结果阳性. 膀胱镜检查结果可疑12例,复发4例(低级别2例、高级别2例) ,M-FISH检查结果阳性1例(低级别). 膀胱镜检查结果阳性10例,复发9例(低级别5例、高级别4例) ,M-FISH检查结果阳性1例(高级别). 膀胱镜检查结果阴性、M-FISH检查结果为阳性预测复发的敏感性为100%、特异性94 .3%,阳性预测值25%,阴性预测值100%. 膀胱镜检查结果可疑、M-FISH检查结果为阳性预测复发的敏感性为25%、特异性87.5%,阳性预测值50%,阴性预测值70%. 膀胱镜、M-FISH检查结果均为阳性预测复发的敏感性为11.1%、特异性100%,阳性预测值100%,阴性预测值11 .1%. 结论 M-FISH检测在尿脱落细胞学阴性膀胱癌患者术后随访复发中的检出率不高,经济效益低,不考虑作为膀胱癌术后随访的常规监测项目.%Objective To evaluate the application value of multicolor fluorescence in situ hybridization ( M-FISH) in postoperative follow-up monitoring of bladder cancer patients with negative urine cytology .Methods A total of 78 patients with urothelial cancer who were treated by transurethral resection of the bladder were included in this study .All of them had negative urine cytology .The cystoscopy and M-FISH were used to monitor the recurrence of the disease , and the results were compared.Results In 76 patients, 14 cases recurred (18.4%).M-FISH only detected 3 cases of recurrence (4%), and 11 cases of missed diagnosis (14.5%).Cystoscopy showed 54 cases with

  17. NOVEL FLUORESCENT PROBES FOR THE DOPAMINE TRANSPORTER

    DEFF Research Database (Denmark)

    Cha, J; Vægter, Christian Bjerggaard; Adkins, Erica

    To enable visualization of the dopamine transporter (DAT) through fluorescence technologies we have synthesized a novel series of fluorescently tagged analogs of cocaine. Previous structure-activity relationship (SAR) studies have demonstrated that the dopamine transporter (DAT) can tolerate...

  18. NOVEL FLUORESCENT PROBES FOR THE DOPAMINE TRANSPORTER

    DEFF Research Database (Denmark)

    Cha, J; Vægter, Christian Bjerggaard; Adkins, Erica

    To enable visualization of the dopamine transporter (DAT) through fluorescence technologies we have synthesized a novel series of fluorescently tagged analogs of cocaine. Previous structure-activity relationship (SAR) studies have demonstrated that the dopamine transporter (DAT) can tolerate...

  19. Fluorescent Protein Approaches in Alpha Herpesvirus Research

    Directory of Open Access Journals (Sweden)

    Ian B. Hogue

    2015-11-01

    Full Text Available In the nearly two decades since the popularization of green fluorescent protein (GFP, fluorescent protein-based methodologies have revolutionized molecular and cell biology, allowing us to literally see biological processes as never before. Naturally, this revolution has extended to virology in general, and to the study of alpha herpesviruses in particular. In this review, we provide a compendium of reported fluorescent protein fusions to herpes simplex virus 1 (HSV-1 and pseudorabies virus (PRV structural proteins, discuss the underappreciated challenges of fluorescent protein-based approaches in the context of a replicating virus, and describe general strategies and best practices for creating new fluorescent fusions. We compare fluorescent protein methods to alternative approaches, and review two instructive examples of the caveats associated with fluorescent protein fusions, including describing several improved fluorescent capsid fusions in PRV. Finally, we present our future perspectives on the types of powerful experiments these tools now offer.

  20. Preparation and Application of Fluorescent Carbon Dots

    Directory of Open Access Journals (Sweden)

    Jun Zuo

    2015-01-01

    Full Text Available Fluorescent carbon dots (CDs are a novel type of fluorescent nanomaterials, which not only possess the specific quantum confinement effects of nanomaterials due to the small size of nanomaterials, but also have good biocompatibility and high fluorescence. Meanwhile, fluorescence CDs overcome the shortcomings of high toxicity of traditional nanomaterials. Moreover, the preparation procedure of fluorescent CDs is simple and easy. Therefore, fluorescent CDs have great potential applied in photocatalysis, biochemical sensing, bioimaging, drug delivery, and other related areas. In this paper, recent hot researches on fluorescent CDs are reviewed and some problems in the progress of fluorescent CDs are also summarized. At last, a future outlook in this direction is presented.

  1. Photochromicity and fluorescence lifetimes of green fluorescent protein

    OpenAIRE

    1999-01-01

    The green fluorescent protein (GFP) of the bioluminescent jellyfish Aequorea and its mutants have gained widespread usage as an indicator of structure and function within cells. Proton transfer has been implicated in the complex photophysics of the wild-type molecule, exhibiting a protonated A species excited at 400 nm, and two deprotonated excited-state species I* and B* with red-shifted excitation similar to 475 nm. Photochromicity between the protonated and deprotonated species has been re...

  2. Synthesis and characterization of new fluorescent nanoparticles

    Institute of Scientific and Technical Information of China (English)

    Liang Tao; Xu Hun; Zhu Jun Zhang

    2008-01-01

    A novel kind of fluorescent nanoparticles (FNPs) has been prepared using a precipitation polymerization method.Methacrylic acid,trimethylolpropane trimethacrylate and azobisisobutyronitrile were used as functional-monomer,cross-linker and initiator,respectively.Compared with other fluorescent nanoparticles,the FNPs have the characteristics including low dye leakage and good photostability.The fluorescence microscopy imaging indicates that the FNPs can be used as fluorescent labels in bioanalysis.

  3. Laser-Stimulated Fluorescence in Paleontology

    OpenAIRE

    Kaye, Thomas G.; Falk, Amanda R.; Michael Pittman; Sereno, Paul C.; Martin, Larry D.; Burnham, David A.; Enpu Gong; Xing Xu; Yinan Wang

    2015-01-01

    Fluorescence using ultraviolet (UV) light has seen increased use as a tool in paleontology over the last decade. Laser-stimulated fluorescence (LSF) is a next generation technique that is emerging as a way to fluoresce paleontological specimens that remain dark under typical UV. A laser's ability to concentrate very high flux rates both at the macroscopic and microscopic levels results in specimens fluorescing in ways a standard UV bulb cannot induce. Presented here are five paleontological c...

  4. Highlights of the optical highlighter fluorescent proteins.

    Science.gov (United States)

    Patterson, G H

    2011-07-01

    The development of super-resolution microscopy techniques using molecular localization, such as photoactivated localization microscopy, fluorescence photoactivated localization microscopy, stochastic optical reconstruction microscopy, photoactivated localization microscopy with independent running acquisition and many others, has heightened interest in molecules that will be grouped here into a category referred to as 'optical highlighter' fluorescent proteins. This review will survey many of the advances in development of fluorescent proteins for optically highlighting sub-populations of fluorescently labelled molecules.

  5. Quantification of fluorescent reporters in plant cells.

    Science.gov (United States)

    Pound, Michael; French, Andrew P; Wells, Darren M

    2015-01-01

    Fluorescent reporters are powerful tools for plant research. Many studies require accurate determination of fluorescence intensity and localization. Here, we describe protocols for the quantification of fluorescence intensity in plant cells from confocal laser scanning microscope images using semiautomated software and image analysis techniques.

  6. Demonstrating Fluorescence with Neon Paper and Plastic

    Science.gov (United States)

    Birriel, Jennifer J.; Roe, Clarissa

    2015-01-01

    Several papers in this journal have dealt with the fluorescence in orange neon plastic, olive oil, and soda. In each case, the fluorescent emission was excited by either green or violet-blue laser light. In this paper, we examine the fluorescent emission spectra of so-called neon colored papers and plastic clipboards available in department and…

  7. Characterization of natural fluorescence in mice

    Science.gov (United States)

    Djeziri, Salim; Ma, Guobin; Mincu, Niculae; Benyamin Seeyar, Anader; Khayat, Mario

    2008-02-01

    One important challenge for in-vivo imaging fluorescence in cancer research and related pharmaceutical studies is to discriminate the exogenous fluorescence signal of the specific tagged agents from the natural fluorescence. For mice, natural fluorescence is composed of endogenous fluorescence from organs like the skin, the bladder, etc. and from ingested food. The discrimination between the two kinds of fluorescence makes easy monitoring the targeted tissues. Generally, the amplitude of the fluorescence signal depends on the location and on the amount of injected fluorophore, which is limited in in-vivo experiments. This paper exposes some results of natural fluorescence analysis from in-vivo mice experiments using a time domain small animal fluorescence imaging system: eXplore Optix TM. Fluorescence signals are expressed by a Time Point Spread Function (TPSF) at each scan point. The study uses measures of similarity applied purposely to the TPSF to evaluate the discrepancy and/or the homogeneity of scanned regions of a mouse. These measures allow a classification scheme to be performed on the TPSF's based on their temporal shapes. The work ends by showing how the exogenous fluorescence can be distinguished from natural fluorescence by using the TPSF temporal shape.

  8. Demonstrating Fluorescence with Neon Paper and Plastic

    Science.gov (United States)

    Birriel, Jennifer J.; Roe, Clarissa

    2015-01-01

    Several papers in this journal have dealt with the fluorescence in orange neon plastic, olive oil, and soda. In each case, the fluorescent emission was excited by either green or violet-blue laser light. In this paper, we examine the fluorescent emission spectra of so-called neon colored papers and plastic clipboards available in department and…

  9. The light bulb, cystoscopy, and Thomas Alva Edison.

    Science.gov (United States)

    Moran, Michael E

    2010-09-01

    Thomas Alva Edison was an icon of American achievement who literally invented the 20th century. Although best known as the inventor of the electric light bulb, the phonograph, and motion pictures, he also left a lasting legacy via peripheral developmental applications, such as endoscopes. A review of published urologic writings about incandescent cystoscopes was cross-referenced to writings about or from Edison. Important events that allowed transference of technology from the Edison laboratory to clinical practice were emphasized. Edison was born in 1847 while Lincoln was serving in Congress; he died in 1931 when Hoover struggled with the Great Depression. Edison's life spanned the formative period of America that Henry Adams called the "coming of age." Edison received a Sprengel vacuum device in late 1879, and as usual, he was able to tweak the machine to better performance. For 5 days in October, 16 to 21, he improved the vacuum from 1/100,000 to 1/1,000,000 atm, and his first incandescent bulb burned softly. On December 21, 1879, he leaked the story to N.Y. Herald journalist Marshall Fox, and the world was notified of the light bulb. Special Christmas light visits started in Menlo Park just 4 days later. Edison patented the screw cap for easy changes, and the first bulbs sold for 40 cents (cost $1.40). 100,000 bulbs sold in 1882, 4 million by 1892, and 45 million in 1903. Immediately, competitors and specialty manufacturers entered the market. Dr. Henry Koch and Charles Preston in Rochester, N.Y., developed a smaller, low amperage bulb that could be fitted to medical devices. No discussion of electricity and modern applications would be complete without some discussion of Thomas Alva Edison and his sentinel contributions. The first church, post office, and ship were illuminated in 1892. The first hotel, theater, and electric sign were in 1893. The rapidity of dispersal and secondary applications of Edison's inventions is typified by the rise of cystoscopes. Nitze used a modified Edison bulb in his second and third generation scopes by 1887-1888 within 8 years of discovery.

  10. Molecular-sized fluorescent nanodiamonds

    Science.gov (United States)

    Vlasov, Igor I.; Shiryaev, Andrey A.; Rendler, Torsten; Steinert, Steffen; Lee, Sang-Yun; Antonov, Denis; Vörös, Márton; Jelezko, Fedor; Fisenko, Anatolii V.; Semjonova, Lubov F.; Biskupek, Johannes; Kaiser, Ute; Lebedev, Oleg I.; Sildos, Ilmo; Hemmer, Philip. R.; Konov, Vitaly I.; Gali, Adam; Wrachtrup, Jörg

    2014-01-01

    Doping of carbon nanoparticles with impurity atoms is central to their application. However, doping has proven elusive for very small carbon nanoparticles because of their limited availability and a lack of fundamental understanding of impurity stability in such nanostructures. Here, we show that isolated diamond nanoparticles as small as 1.6 nm, comprising only ~400 carbon atoms, are capable of housing stable photoluminescent colour centres, namely the silicon vacancy (SiV). Surprisingly, fluorescence from SiVs is stable over time, and few or only single colour centres are found per nanocrystal. We also observe size-dependent SiV emission supported by quantum-chemical simulation of SiV energy levels in small nanodiamonds. Our work opens the way to investigating the physics and chemistry of molecular-sized cubic carbon clusters and promises the application of ultrasmall non-perturbative fluorescent nanoparticles as markers in microscopy and sensing.

  11. Ruby fluorescence pressure scale: Revisited

    Science.gov (United States)

    Liu, Lei; Bi, Yan; Xu, Ji-An

    2013-05-01

    Effect of non-hydrostatic stress on X-ray diffraction in a diamond anvil cell (DAC) is studied. The pressure gradient in the sample chamber leads to the broadening of the diffraction peaks, which increase with the hkl index of the crystal. It is found that the difference between the determined d-spacing compressive ratio d/d0 and the real d-spacing compressive ratio dr/d0 is determined by the yield stress of the pressure transmitting media (if used) and the shear modulus of the sample. On the basis of the corrected experiment data of Mao et al. (MXB86), which was used to calibrate the most widely used ruby fluorescence scale, a new relationship of ruby fluorescence pressure scale is corrected, i.e., P = (1904/9.827)[(1 + Δλ/λ0)9.827-1].

  12. New Fluorescence Probes for Biomolecules

    Directory of Open Access Journals (Sweden)

    Katarzyna Jurek

    2015-07-01

    Full Text Available Steady state fluorescence measurements have been used for the investigation of interaction between the bovine serum albumin (BSA and fluorescence probes: 3-hydroxy-2,4- bis[(3-methyl-1,3-benzoxazol-2(3H-ylidenemethyl]cyclobut-2-en-1-one (SQ6, 3-hydroxy- 2,4-bis[(3-methyl-1,3-benzothiazol-2(3H-ylidenemethyl]cyclobut-2-en-1-one (SQ7 and 3-hydroxy-2,4-bis[(1,3,3-trimethyl-1,3-dihydro-2H-indol-2-ylidenemethyl]cyclobut-2-en-1-one (SQ8. The binding constant between bovine serum albumin and squarine dyes has been determined by using both the Benesi-Hildebrand and Stern-Volmer equations. The negative value of free energy change indicates the existence of a spontaneous complexation process of BSA with squarine dyes.

  13. Fluorescence spectroscopy for neoplasms control

    Science.gov (United States)

    Bratchenko, I. A.; Kristoforova, Yu. A.; Myakinin, O. O.; Artemyev, D. N.; Kozlov, S. V.; Moryatov, A. A.; Zakharov, V. P.

    2016-04-01

    Investigation of malignant skin tumors diagnosis was performed involving two setups for native tissues fluorescence control in visible and near infrared regions. Combined fluorescence analysis for skin malignant melanomas and basal cell carcinomas was performed. Autofluorescence spectra of normal skin and oncological pathologies stimulated by 457 nm and 785 nm lasers were registered for 74 skin tissue samples. Spectra of 10 melanomas and 27 basal cell carcinomas were registered ex vivo. Skin tumors analysis was made on the basis of autofluorescence spectra intensity and curvature for analysis of porphyrins, lipo-pigments, flavins and melanin. Separation of melanomas and basal cell carcinomas was performed on the basis of discriminant analysis. Overall accuracy of basal cell carcinomas and malignant melanomas separation in current study reached 86.5% with 70% sensitivity and 92.6% specificity.

  14. Sorting fluorescent nanocrystals with DNA

    Energy Technology Data Exchange (ETDEWEB)

    Gerion, Daniele; Parak, Wolfgang J.; Williams, Shara C.; Zanchet, Daniela; Micheel, Christine M.; Alivisatos, A. Paul

    2001-12-10

    Semiconductor nanocrystals with narrow and tunable fluorescence are covalently linked to oligonucleotides. These biocompounds retain the properties of both nanocrystals and DNA. Therefore, different sequences of DNA can be coded with nanocrystals and still preserve their ability to hybridize to their complements. We report the case where four different sequences of DNA are linked to four nanocrystal samples having different colors of emission in the range of 530-640 nm. When the DNA-nanocrystal conjugates are mixed together, it is possible to sort each type of nanoparticle using hybridization on a defined micrometer -size surface containing the complementary oligonucleotide. Detection of sorting requires only a single excitation source and an epifluorescence microscope. The possibility of directing fluorescent nanocrystals towards specific biological targets and detecting them, combined with their superior photo-stability compared to organic dyes, opens the way to improved biolabeling experiments, such as gene mapping on a nanometer scale or multicolor microarray analysis.

  15. 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.

  16. FAMOUS. The fluorescence telescope prototype

    Energy Technology Data Exchange (ETDEWEB)

    Schumacher, Johannes; Bretz, Thomas; Hebbeker, Thomas; Lauscher, Markus; Middendorf, Lukas; Niggemann, Tim; Peters, Christine; Sommer, Dominik; Stephan, Maurice [III. Physikalisches Institut A, RWTH Aachen University (Germany); Auffenberg, Jan; Schaufel, Merlin [III. Physikalisches Institut B, RWTH Aachen University (Germany)

    2015-07-01

    One of the most successful techniques for the detection of air showers produced by ultra-high-energy cosmic rays are fluorescence telescopes. The light produced by de-exciting nitrogen in the atmosphere is typically detected by photomultiplier tubes (PMTs). This technique has been successfully used by the Pierre Auger Observatory in Argentina for many years. Silicon photomultipliers (SiPMs) promise higher photon detection efficiencies than PMTs. This and other advantages motivate the construction of the fluorescence telescope prototype FAMOUS (First Auger Multi-pixel photon counter camera for the Observation of Ultra-high-energy air Showers) which makes use of SiPMs. In this talk we discuss the FAMOUS telescope with a new 64-pixel camera including power supply and DAQ.

  17. Correlative fluorescence and electron microscopy.

    Science.gov (United States)

    Schirra, Randall T; Zhang, Peijun

    2014-10-01

    Correlative fluorescence and electron microscopy (CFEM) is a multimodal technique that combines dynamic and localization information from fluorescence methods with ultrastructural data from electron microscopy, to give new information about how cellular components change relative to the spatiotemporal dynamics within their environment. In this review, we will discuss some of the basic techniques and tools of the trade for utilizing this attractive research method, which is becoming a very powerful tool for biology labs. The information obtained from correlative methods has proven to be invaluable in creating consensus between the two types of microscopy, extending the capability of each, and cutting the time and expense associated with using each method separately for comparative analysis. The realization of the advantages of these methods in cell biology has led to rapid improvement in the protocols and has ushered in a new generation of instruments to reach the next level of correlation--integration.

  18. FLUORESCENCE LIFETIME DISTRIBUTIONS IN PROTEINS

    OpenAIRE

    ALCALA, JR; Gratton, E; PRENDERGAST, FG

    1987-01-01

    The fluorescence lifetime value of tryptophan residues varies by more than a factor of 100 in different proteins and is determined by several factors, which include solvent exposure and interactions with other elements of the protein matrix. Because of the variety of different elements that can alter the lifetime value and the sensitivity to the particular environment of the tryptophan residue, it is likely that non-unique lifetime values result in protein systems. The emission decay of most ...

  19. Fluorescent compounds present in food

    OpenAIRE

    Soto Serrano, Axel

    2014-01-01

    Póster The food industry demands fast, reliable, cheap and reproducible methods for quality and process control. This bibliographic review work investigates florescence spectroscopy, a method that couldn’t be used in food until the recent technological advances, concretely front-face fluorescence and chemometric tools. This technology presents advantages as compared to classical methods like HPLC or capillary electrophoresis, which require qualified staff, sample preparation and are time-c...

  20. 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.

  1. Interaction of fluorescent phospholipids with cyclodextrins.

    Science.gov (United States)

    Denz, Manuela; Haralampiev, Ivan; Schiller, Sabine; Szente, Lajos; Herrmann, Andreas; Huster, Daniel; Müller, Peter

    2016-01-01

    Fluorescent analogs of phospholipids are often employed to investigate the structure and dynamics of lipids in membranes. Some of those studies have used cyclodextrins e.g., to modulate the lipid phase. However, the role of the fluorescence moiety of analogs for the interaction between cyclodextrins and fluorescent lipids has not been investigated so far in detail. Therefore, in the present study the interaction of various fluorescent phospholipid analogs with methylated α-, β- and γ- cyclodextrins was investigated. The analogs differed in their structure, in the length of the fatty acyl chain, in the position of the fluorescence group, and in the attached fluorescence moiety (7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD) or dipyrrometheneboron difluoride (BODIPY)). In aqueous buffer, cyclodextrins bind fluorescent lipids disturbing the organization of the analogs. When incorporated into lipid vesicles, analogs are selectively extracted from the membrane upon addition of cyclodextrins. The results show that the interaction of cyclodextrins with fluorescent phospholipids depends on the cyclodextrin species, the fluorescence moiety and the phospholipid structure. The presented data should be of interest for studies using fluorescent phospholipids and cyclodextrins, since the interaction between the fluorescence group and the cyclodextrin may interfere with the process(es) under study.

  2. Fluorescence Studies of Selected 2-Alkylaminopyrimidines

    Directory of Open Access Journals (Sweden)

    B. K. Low

    2004-06-01

    Full Text Available The reactions of 2-chloropyrimidine with methylamine, ethylamine and piperidine gave the corresponding 2-N-methylamino-, 2-N-ethylamino- and 2N- piperidinopyrimidines, respectively. The fluorescence properties of these alkylamino derivatives in chloroform, ethyl acetate, carbon tetrachloride, acetone, ether, ethanol and methanol were studied. All the alkylamino derivatives showed the highest fluorescence intensity in polar protic solvents; thus 2-N-methylaminopyrimidine (highest fluorescence intensity at 377 nm when excited at 282 nm and 2-N-ethylaminopyrimidine (highest fluorescence intensity at 375 nm, when excited at 286 nm showed the highest fluorescence in methanol. In ethanol, 2-N-piperidinopyrimidine showed a fluorescence peak at 403 nm when excited at 360 nm and in chloroform it fluoresced at 392 nm when excited at 356 nm.

  3. 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).

  4. Bioaerosol Analysis by Online Fluorescence Detection and Fluorescence Microscopy

    Science.gov (United States)

    Huffman, Alex; Pöhlker, Christopher; Treutlein, Bärbel; Pöschl, Ulrich

    2010-05-01

    Primary biological aerosol particles (PBAPs), including bacteria, spores and pollen, are essential for the spread of organisms and disease in the biosphere, and numerous studies have suggested that they may be important for atmospheric processes, including the formation of clouds and precipitation. The atmospheric abundance and size distribution of PBAPs, however, are largely unknown. At a semi-urban site in Mainz, Germany, we used an ultraviolet aerodynamic particle sizer (UV-APS) to measure fluorescent biological aerosol particles (FBAPs), which can be regarded as viable bioaerosol particles representing a lower limit for the actual abundance of PBAPs. Fluorescence of non-biological aerosol components are likely to influence the measurement results obtained for fine particles (concentration of coarse FBAPs was 3x10-2 cm-3, corresponding to 4% of total coarse particle number [1]. The mean mass concentration of FBAPs was 1 ?g m-3, corresponding to 20% of total coarse particle mass. The FBAP number size distributions exhibited alternating patterns with peaks at various diameters, though a pronounced peak at 3 μm was essentially always observed. This peak is likely due to fungal spores or agglomerated bacteria, and it exhibited a pronounced diel cycle with maximum intensity during early/mid-morning. FBAP peaks around 1.5 μm, 5 μm, and 13 μm were also observed, but less pronounced and less frequent. These may be explained by single bacterial cells, larger fungal spores, and pollen grains, respectively. The observed number concentrations and characteristic sizes of FBAPs are consistent with microscopic, biological and chemical analyses of PBAPs in aerosol filter samples. To our knowledge, however, this is the first study reporting continuous online measurements of bioaerosol particles over several months, a range of characteristic size distribution patterns, and a persistent bioaerosol peak at 3 μm. The measurement results confirm that PBAPs account for a

  5. Fluorescent nanoparticles for intracellular sensing: A review

    Energy Technology Data Exchange (ETDEWEB)

    Ruedas-Rama, Maria J., E-mail: mjruedas@ugr.esmailto [Department of Physical Chemistry, Faculty of Pharmacy, University of Granada, Campus Cartuja, 18071, Granada (Spain); Walters, Jamie D. [Department of Chemical Engineering and Biotechnology, University of Cambridge, Tennis Court Road, Cambridge, UK CB2 1QT (United Kingdom); Orte, Angel [Department of Physical Chemistry, Faculty of Pharmacy, University of Granada, Campus Cartuja, 18071, Granada (Spain); Hall, Elizabeth A.H., E-mail: lisa.hall@biotech.cam.ac.uk [Department of Chemical Engineering and Biotechnology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QT (United Kingdom)

    2012-11-02

    Highlights: Black-Right-Pointing-Pointer Analytical applications of fluorescent nanoparticles (NPs) in intracellular sensing. Black-Right-Pointing-Pointer Critical review on performance of QDots, metal NPs, silica NPs, and polymer NPs. Black-Right-Pointing-Pointer Highlighted potential of fluorescence lifetime imaging microscopy (FLIM). - Abstract: Fluorescent nanoparticles (NPs), including semiconductor NPs (Quantum Dots), metal NPs, silica NPs, polymer NPs, etc., have been a major focus of research and development during the past decade. The fluorescent nanoparticles show unique chemical and optical properties, such as brighter fluorescence, higher photostability and higher biocompatibility, compared to classical fluorescent organic dyes. Moreover, the nanoparticles can also act as multivalent scaffolds for the realization of supramolecular assemblies, since their high surface to volume ratio allow distinct spatial domains to be functionalized, which can provide a versatile synthetic platform for the implementation of different sensing schemes. Their excellent properties make them one of the most useful tools that chemistry has supplied to biomedical research, enabling the intracellular monitoring of many different species for medical and biological purposes. In this review, we focus on the developments and analytical applications of fluorescent nanoparticles in chemical and biological sensing within the intracellular environment. The review also points out the great potential of fluorescent NPs for fluorescence lifetime imaging microscopy (FLIM). Finally, we also give an overview of the current methods for delivering of fluorescent NPs into cells, where critically examine the benefits and liabilities of each strategy.

  6. Red and Green Fluorescence from Oral Biofilms

    Science.gov (United States)

    Hoogenkamp, Michel A.; Krom, Bastiaan P.; Janus, Marleen M.; ten Cate, Jacob M.; de Soet, Johannes J.; Crielaard, Wim; van der Veen, Monique H.

    2016-01-01

    Red and green autofluorescence have been observed from dental plaque after excitation by blue light. It has been suggested that this red fluorescence is related to caries and the cariogenic potential of dental plaque. Recently, it was suggested that red fluorescence may be related to gingivitis. Little is known about green fluorescence from biofilms. Therefore, we assessed the dynamics of red and green fluorescence in real-time during biofilm formation. In addition, the fluorescence patterns of biofilm formed from saliva of eight different donors are described under simulated gingivitis and caries conditions. Biofilm formation was analysed for 12 hours under flow conditions in a microfluidic BioFlux flow system with high performance microscopy using a camera to allow live cell imaging. For fluorescence images dedicated excitation and emission filters were used. Both green and red fluorescence were linearly related with the total biomass of the biofilms. All biofilms displayed to some extent green and red fluorescence, with higher red and green fluorescence intensities from biofilms grown in the presence of serum (gingivitis simulation) as compared to the sucrose grown biofilms (cariogenic simulation). Remarkably, cocci with long chain lengths, presumably streptococci, were observed in the biofilms. Green and red fluorescence were not found homogeneously distributed within the biofilms: highly fluorescent spots (both green and red) were visible throughout the biomass. An increase in red fluorescence from the in vitro biofilms appeared to be related to the clinical inflammatory response of the respective saliva donors, which was previously assessed during an in vivo period of performing no-oral hygiene. The BioFlux model proved to be a reliable model to assess biofilm fluorescence. With this model, a prediction can be made whether a patient will be prone to the development of gingivitis or caries. PMID:27997567

  7. Red and Green Fluorescence from Oral Biofilms.

    Science.gov (United States)

    Volgenant, Catherine M C; Hoogenkamp, Michel A; Krom, Bastiaan P; Janus, Marleen M; Ten Cate, Jacob M; de Soet, Johannes J; Crielaard, Wim; van der Veen, Monique H

    2016-01-01

    Red and green autofluorescence have been observed from dental plaque after excitation by blue light. It has been suggested that this red fluorescence is related to caries and the cariogenic potential of dental plaque. Recently, it was suggested that red fluorescence may be related to gingivitis. Little is known about green fluorescence from biofilms. Therefore, we assessed the dynamics of red and green fluorescence in real-time during biofilm formation. In addition, the fluorescence patterns of biofilm formed from saliva of eight different donors are described under simulated gingivitis and caries conditions. Biofilm formation was analysed for 12 hours under flow conditions in a microfluidic BioFlux flow system with high performance microscopy using a camera to allow live cell imaging. For fluorescence images dedicated excitation and emission filters were used. Both green and red fluorescence were linearly related with the total biomass of the biofilms. All biofilms displayed to some extent green and red fluorescence, with higher red and green fluorescence intensities from biofilms grown in the presence of serum (gingivitis simulation) as compared to the sucrose grown biofilms (cariogenic simulation). Remarkably, cocci with long chain lengths, presumably streptococci, were observed in the biofilms. Green and red fluorescence were not found homogeneously distributed within the biofilms: highly fluorescent spots (both green and red) were visible throughout the biomass. An increase in red fluorescence from the in vitro biofilms appeared to be related to the clinical inflammatory response of the respective saliva donors, which was previously assessed during an in vivo period of performing no-oral hygiene. The BioFlux model proved to be a reliable model to assess biofilm fluorescence. With this model, a prediction can be made whether a patient will be prone to the development of gingivitis or caries.

  8. DNA nanotechnology and fluorescence applications.

    Science.gov (United States)

    Schlichthaerle, Thomas; Strauss, Maximilian T; Schueder, Florian; Woehrstein, Johannes B; Jungmann, Ralf

    2016-06-01

    Structural DNA nanotechnology allow researchers to use the unique molecular recognition properties of DNA strands to construct nanoscale objects with almost arbitrary complexity in two and three dimensions. Abstracted as molecular breadboards, DNA nanostructures enable nanometer-precise placement of guest molecules such as proteins, fluorophores, or nanoparticles. These assemblies can be used to study biological phenomena with unprecedented control over number, spacing, and molecular identity. Here, we give a general introduction to structural DNA nanotechnology and more specifically discuss applications of DNA nanostructures in the field of fluorescence and plasmonics.

  9. FAA Fluorescent Penetrant Laboratory Inspections

    Energy Technology Data Exchange (ETDEWEB)

    WINDES,CONNOR L.; MOORE,DAVID G.

    2000-08-02

    The Federal Aviation Administration Airworthiness Assurance NDI Validation Center currently assesses the capability of various non-destructive inspection (NDI) methods used for analyzing aircraft components. The focus of one such exercise is to evaluate the sensitivity of fluorescent liquid penetrant inspection. A baseline procedure using the water-washable fluorescent penetrant method defines a foundation for comparing the brightness of low cycle fatigue cracks in titanium test panels. The analysis of deviations in the baseline procedure will determine an acceptable range of operation for the steps in the inspection process. The data also gives insight into the depth of each crack and which step(s) of the inspection process most affect penetrant sensitivities. A set of six low cycle fatigue cracks produced in 6.35-mm thick Ti-6Al-4V specimens was used to conduct the experiments to produce sensitivity data. The results will document the consistency of the crack readings and compare previous experiments to find the best parameters for water-washable penetrant.

  10. Single-primer fluorescent sequencing

    Energy Technology Data Exchange (ETDEWEB)

    Ruth, J.L.; Morgan, C.A.; Middendorf, L.R.; Grone, D.L.; Brumbaugh, J.A.

    1987-05-01

    Modified linker arm oligonucleotides complementary to standard M13 priming sites were synthesized, labelled with either one, two, or three fluoresceins, and purified by reverse-phase HPLC. When used as primers in standard dideoxy M13 sequencing with /sup 32/P-dNTPs, normal autoradiographic patterns were obtained. To eliminate the radioactivity, direct on-line fluorescence detection was achieved by the use of a scanning 10 mW Argon laser emitting 488 nm light. Fluorescent bands were detected directly in standard 0.2 or 0.35 mm thick polyacrylamide gels at a distance of 24 cm from the loading wells by a photomultiplier tube filtered at 520 nm. Horizontal and temporal location of each band was displayed by computer as a band in real time, providing visual appearance similar to normal 4-lane autoradiograms. Using a single primer labelled with two fluoresceins, sequences of between 500 and 600 bases have been read in a single loading with better than 98% accuracy; up to 400 bases can be read reproducibly with no errors. More than 50 sequences have been determined by this method. This approach requires only 1-2 ug of cloned template, and produces continuous sequence data at about one band per minute.

  11. Lifetime Resolved Fluorescence Fluctuation Spectroscopy

    Science.gov (United States)

    Guo, Peng; Berland, Keith

    2009-11-01

    Fluorescence correlation spectroscopy (FCS) has been widely used investigate molecular dynamics and interactions in biological systems. FCS typically resolves the component species of a sample either through differences in diffusion coefficient or molecular brightness. Diffusion based assays currently have a major limitation which requires that the diffusion coefficients of component species in a sample must be substantially different in order to be resolved. This criterion is not met in many important cases, such as when molecules of similar molecular weight bind to each other. This limitation can be overcome, and resolution of FCS measurements enhanced, by combining FCS measurements with measurements of fluorescence lifetimes. By using of global analysis on simultaneously acquired FCS and lifetime data we show that we can dramatically enhance resolution in FCS measurements, and accurately resolve the concentration and diffusion coefficients of multiple sample components even when their diffusion coefficients are identical provided there is a difference in the lifetime of the component species. We show examples of this technique using both simulations and experiments. It is expected that this method will be of significance for binding assays studying molecular interactions.

  12. Mitochondrially targeted fluorescent redox sensors.

    Science.gov (United States)

    Yang, Kylie; Kolanowski, Jacek L; New, Elizabeth J

    2017-04-06

    The balance of oxidants and antioxidants within the cell is crucial for maintaining health, and regulating physiological processes such as signalling. Consequently, imbalances between oxidants and antioxidants are now understood to lead to oxidative stress, a physiological feature that underlies many diseases. These processes have spurred the field of chemical biology to develop a plethora of sensors, both small-molecule and fluorescent protein-based, for the detection of specific oxidizing species and general redox balances within cells. The mitochondrion, in particular, is the site of many vital redox reactions. There is therefore a need to target redox sensors to this particular organelle. It has been well established that targeting mitochondria can be achieved by the use of a lipophilic cation-targeting group, or by utilizing natural peptidic mitochondrial localization sequences. Here, we review how these two approaches have been used by a number of researchers to develop mitochondrially localized fluorescent redox sensors that are already proving useful in providing insights into the roles of reactive oxygen species in the mitochondria.

  13. Detection of Counterfeit Tequila by Fluorescence Spectroscopy

    Directory of Open Access Journals (Sweden)

    José Manuel de la Rosa Vázquez

    2015-01-01

    Full Text Available An ultraviolet (UV light induced fluorescence study to discriminate fake tequila from genuine ones is presented. A portable homemade system based on four light emitting diodes (LEDs from 255 to 405 nm and a miniature spectrometer was used. It has been shown that unlike fake and silver tequila, which produce weak fluorescence signal, genuine mixed, rested, and aged tequilas show high fluorescence emission in the range from 400 to 750 nm. The fluorescence intensity grows with aging in 100% agave tequila. Such fluorescence differences can even be observed with naked eyes. The presented results demonstrate that the fluorescence measurement could be a good method to detect counterfeit tequila.

  14. 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.

  15. Thermochromism and fluorescence in dyed PEO films

    Energy Technology Data Exchange (ETDEWEB)

    Kamath, Archana; S, Raghu; V, Mini; C, Sharanappa; H, Devendrappa, E-mail: dehu2010@gmail.com [Dept. of Physics, Mangalore University, Managalagangothri, Mangalore--574199 (India)

    2015-06-24

    The optical absorbance spectra of solution casted pure & methyl blue (MB) dyed polyethylene oxide (PEO) films were recorded in a wavelength range from 190-1100nm at different temperatures. The absorbance was found to increases with increasing temperature. Fluorescence micrographs confirmed the interaction between polymer and dye and also revealed decreased crystallinity of the sample. Fluorescence quantum yield has been calculated with the help of fluorescence spectra.

  16. Dye Fluorescence Analysis from Bacterial Metabolism.

    Science.gov (United States)

    1984-04-01

    M were reported for the cell-free extracts of the cultured mouse lymphoma cells mentioned above and an in vitAo solution of porcine pancreas lipase ...fluorescence Fluorescent product Diacetyl fluorescein Lipase Bacterial metabolism 20. ABTRACT fCauhw a o de dif rNooeel md ~Id1)fp by block number) A...nonfluorescing dye is metabolized intracel- lularly by an organism through an enzyme-specific reaction . This produces a fluorescent product which when

  17. Fluorescence fluctuation spectroscopy (FFS), part A

    CERN Document Server

    Tetin, Sergey

    2013-01-01

    This new volume of Methods in Enzymology continues the legacy of this premier serial by containing quality chapters authored by leaders in the field. This volume covers Fluorescence Fluctuation SpectroscopyContains chapters on such topics as Time-integrated fluorescence cumulant analysis, Pulsed Interleaved Excitation, and raster image correlation spectroscopy and number and brightness analysis.Continues the legacy of this premier serial with quality chapters authored by leaders in the fieldCovers fluorescence fluctuation spectroscopyContains chapte

  18. Detection of Counterfeit Tequila by Fluorescence Spectroscopy

    OpenAIRE

    José Manuel de la Rosa Vázquez; Diego Adrián Fabila-Bustos; Luis Felipe de Jesús Quintanar-Hernández; Alma Valor; Suren Stolik

    2015-01-01

    An ultraviolet (UV) light induced fluorescence study to discriminate fake tequila from genuine ones is presented. A portable homemade system based on four light emitting diodes (LEDs) from 255 to 405 nm and a miniature spectrometer was used. It has been shown that unlike fake and silver tequila, which produce weak fluorescence signal, genuine mixed, rested, and aged tequilas show high fluorescence emission in the range from 400 to 750 nm. The fluorescence intensity grows with aging in 100% ag...

  19. Fluorescent Quantum Dots for Biological Labeling

    Science.gov (United States)

    McDonald, Gene; Nadeau, Jay; Nealson, Kenneth; Storrie-Lomardi, Michael; Bhartia, Rohit

    2003-01-01

    Fluorescent semiconductor quantum dots that can serve as "on/off" labels for bacteria and other living cells are undergoing development. The "on/off" characterization of these quantum dots refers to the fact that, when properly designed and manufactured, they do not fluoresce until and unless they come into contact with viable cells of biological species that one seeks to detect. In comparison with prior fluorescence-based means of detecting biological species, fluorescent quantum dots show promise for greater speed, less complexity, greater sensitivity, and greater selectivity for species of interest. There are numerous potential applications in medicine, environmental monitoring, and detection of bioterrorism.

  20. [Fluorescence spectroscopy study of synthetic food colors].

    Science.gov (United States)

    Chen, Guo-qing; Wu, Ya-min; Wang, Jun; Zhu, Tuo; Gao, Shu-mei

    2009-09-01

    According to the characteristic of synthetic food colors molecule and the relationship between fluorescence and molecular structure, and through analyzing, it has been concluded that synthetic food colors is fluorescent material. By using SP-2558 multifunctional spectral measuring system, the three-dimensional fluorescence spectra of ponceau 4R, amaranth, tartrazine, sunset yellow and brilliant blue were measured. The results show that ponceau 4R excited by light at the wavelength of 330-430 nm can generate a strong fluorescence at the 621 nm peak wavelength with its best excitation wavelength being 376 nm, amaranth excited by light at the wavelength of 300-440 nm can generate a strong fluorescence at the 643 nm peak wavelength with its best excitation wavelength being 370 nm, tartrazine excited by light at the wavelength of 280-380 nm can generate a strong fluorescence at the 565 nm peak wavelength with its best excitation wavelength being 315 nm, sunset yellow excited by light with wavelength of 310-410 nm can generate a strong fluorescence at the 592 nm peak wavelength with its best excitation wavelength being 348 nm, and brilliant blue excited by light at the wavelength of 320-390 nm can generate a strong fluorescence at the 456 nm peak wavelength with its best excitation wavelength being 350 nm. Moreover, the fluorescence spectra of the five kinds of synthetic food colors were discussed. These results can provide helps for testing of food colors and food safety.

  1. Fluorescence properties of meso-tetrafurylporphyrins

    Indian Academy of Sciences (India)

    Iti Gupta; M Ravikanth

    2005-03-01

    Fluorescence properties of meso-tetrafurylporphyrins with N4, N3S and N2S2 porphyrin cores are studied by both steady-state and time-resolved fluorescence techniques and compared with the corresponding meso-tetraarylporphyrins. The study shows that the replacement of six-membered aryl groups with five-membered furyl groups at meso-positions alter the fluorescence properties considerably, as reflected in the large red shifts and broadening of fluorescence bands with reduction in quantum yields and singlet excited-state lifetimes. However, zinc(II) derivatives of meso-tetrafurylporphyrin and mesotetraarylporphyrin did not show significant differences in their emission properties.

  2. One-year monitoring of an oligonucleotide fluorescence in situ hybridization probe panel laboratory-developed test for bladder cancer detection

    Directory of Open Access Journals (Sweden)

    Tinawi-Aljundi R

    2015-04-01

    Full Text Available Rima Tinawi-Aljundi,1 Lauren King,2 Shannon T Knuth,2 Michael Gildea,2 Carrie Ng,2 Josh Kahl,2 Jacqueline Dion,2 Chris Young,2 Edward W Schervish,1 J Rene Frontera,1 Jason Hafron,1 Kenneth M Kernen,1 Robert Di Loreto,1 Joan Aurich-Costa21Michigan Institute of Urology, St Claire Shores, MI, USA; 2Cellay, Inc., Cambridge, MA, USA Background: Previously, we had developed and manufactured an oligonucleotide fluorescence in situ hybridization (OligoFISH probe panel based on the most clinically sensitive chromosomes found in a reference set of bladder carcinoma cases. The panel was clinically validated for use as a diagnostic and monitoring assay for bladder cancer, reaching 100% correlation with the results of the UroVysion test. After 1 year of using this probe panel, we present here the comparison of cytology, cystoscopy, and pathology findings to the OligoFISH probe panel results to calculate its clinical performance. Materials and methods: In order to calculate clinical performance, we compared the OligoFISH results to the cytology and cystoscopy/pathology findings for 147 initial diagnoses and 399 recurrence monitorings. Finally, we compared clinical performance to published values for the UroVysion test, including both low- and high-grade tumors. Results: Chromosomes 3, 6, 7, and 20 were highly involved in bladder carcinoma aneuploidy. At the initial diagnosis, we obtained 90.5% (95% confidence interval [CI]: 84.5%–94.7% accuracy, 96.8% sensitivity (95% CI: 91.0%–99.3%, 79.2% specificity (95% CI: 65.9%–87.8%, 89.2% positive predictive value (PPV; 95% CI: 81.5%–94.5%, and 93.3% negative predictive value (NPV; 95% CI: 81.7%–97.3%. When monitoring for recurrence, we obtained 85.2% accuracy (95% CI: 81.3%–88.5%, 82.0% sensitivity (95% CI: 76.0%–87.1%, 88.4% specificity (95% CI: 83.2%–92.5%, 87.7% PPV (95% CI: 82.1%–92.0%, and 83.0% NPV (95% CI: 77.3%–87.8%. When looking at low- and high-grade tumors, the test showed 100

  3. Evidence for covalent binding of epicocconone with proteins from synchronous fluorescence spectra and fluorescence lifetimes

    Indian Academy of Sciences (India)

    Debashis Panda; Anindya Datta

    2007-03-01

    Synchronous fluorescence and time-resolved fluorescence spectroscopic studies that reveal the interaction of epicocconone with human serum albumin is significantly different from its interaction with surfactant assemblies. This observation, along with steady-state fluorescence data, indicates groundstate interaction between the fluorophore epicocconone and the protein. Similarity in fluorescence properties with the adduct of the fluorophore with -butylamine indicates that bonding occurs at the Nterminus of the protein.

  4. Molecules for Fluorescence Detection of Specific Chemicals

    Science.gov (United States)

    Fedor, Steve

    2008-01-01

    A family of fluorescent dye molecules has been developed for use in on-off fluorescence detection of specific chemicals. By themselves, these molecules do not fluoresce. However, when exposed to certain chemical analytes in liquid or vapor forms, they do fluoresce (see figure). These compounds are amenable to fixation on or in a variety of substrates for use in fluorescence-based detection devices: they can be chemically modified to anchor them to porous or non-porous solid supports or can be incorporated into polymer films. Potential applications for these compounds include detection of chemical warfare agents, sensing of acidity or alkalinity, and fluorescent tagging of proteins in pharmaceutical research and development. These molecules could also be exploited for use as two-photon materials for photodynamic therapy in the treatment of certain cancers and other diseases. A molecule in this family consists of a fluorescent core (such as an anthracene or pyrene) attached to two end groups that, when the dye is excited by absorption of light, transfer an electron to the core, thereby quenching the fluorescence. The end groups can be engineered so that they react chemically with certain analytes. Upon reaction, electrons on the end groups are no longer available for transfer to the core and, consequently, the fluorescence from the core is no longer quenched. The chemoselectivity of these molecules can be changed by changing the end groups. For example, aniline end groups afford a capability for sensing acids or acid halides (including those contained in chemical warfare agents). Pyridine or bipyridyl end groups would enable sensing of metal ions. Other chemicals that can be selectively detected through suitable choice of end groups include glucose and proteins. Moreover, the fluorescent cores can be changed to alter light-absorption and -emission characteristics: anthracene cores fluoresce at wavelengths around 500 nm, whereas perylene cores absorb and emit at

  5. ULTRAFINE FLUORESCENT DIAMONDS IN NANOTECHNOLOGY

    Directory of Open Access Journals (Sweden)

    Kanyuk M. I.

    2014-07-01

    Full Text Available The purpose of the work is to summarize the literature data concerning ultrafine diamonds, namely their industrial production, as well as considerable photostability and biocompatibility that promote their use in modern visualization techniques. It is shown that due to the unique physical properties, they are promising materials for using in nanotechnology in the near future. Possibility of diverse surface modification, small size and large absorption surface are the basis for their use in different approaches for drug and gene delivery into a cell. The changes in the properties of nanodiamond surface modification methods of their creation, stabilization and applications are described. It can be said that fluorescent surface-modified nanodiamonds are a promising target in various research methods that would be widely used for labeling of living cells, as well as in the processes of genes and drugs delivery into a cell.

  6. Fluorescence nanoscopy in cell biology.

    Science.gov (United States)

    Sahl, Steffen J; Hell, Stefan W; Jakobs, Stefan

    2017-09-06

    Fluorescence nanoscopy uniquely combines minimally invasive optical access to the internal nanoscale structure and dynamics of cells and tissues with molecular detection specificity. While the basic physical principles of 'super-resolution' imaging were discovered in the 1990s, with initial experimental demonstrations following in 2000, the broad application of super-resolution imaging to address cell-biological questions has only more recently emerged. Nanoscopy approaches have begun to facilitate discoveries in cell biology and to add new knowledge. One current direction for method improvement is the ambition to quantitatively account for each molecule under investigation and assess true molecular colocalization patterns via multi-colour analyses. In pursuing this goal, the labelling of individual molecules to enable their visualization has emerged as a central challenge. Extending nanoscale imaging into (sliced) tissue and whole-animal contexts is a further goal. In this Review we describe the successes to date and discuss current obstacles and possibilities for further development.

  7. Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein

    NARCIS (Netherlands)

    Shaner, Nathan C; Campbell, Robert E; Steinbach, Paul A; Giepmans, Ben N G; Palmer, Amy E; Tsien, Roger Y

    2004-01-01

    Fluorescent proteins are genetically encoded, easily imaged reporters crucial in biology and biotechnology. When a protein is tagged by fusion to a fluorescent protein, interactions between fluorescent proteins can undesirably disturb targeting or function. Unfortunately, all wild-type yellow-to-red

  8. Multi-Photon Fluorescence Spectroscopy of Fluorescent Bio-Probes and Bio-Molecules

    Science.gov (United States)

    2000-07-01

    the set-up of a multi-photon fluorescence microscope. The information can also be useful in the detection of multi-photon fluorescence in bio -chip...technology. In addition, we have investigated a few highly fluorescent bio -molecules commonly found in plant cells.

  9. Fluorescence and Thermostability of Nanometer Porphyrin Trimer

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    A nanometer porphyrin trimer was firstly synthesized with 1,3-dibromopropane as a bridge-linked agent and the fluorescence property and thermostability were studied. The results show that the fluorescence property and thermostability of the trimer are different from those of monoporphyrin. The effects of the molecule structure on the optical property and the thermostability were also studied in detail.

  10. Classroom Activity Connections: Lessons from Fluorescence

    Science.gov (United States)

    MacCormac, Aoife; O'Brien, Emma; O'Kennedy, Richard

    2010-01-01

    This Classroom Activity Connections paper describes an extension to the "JCE" Classroom Activity #68 "Turning on the Light". A number of additional common items that display fluorescence under UV light are described, including fruits, vegetables, and seashells. Two classroom extensions on fluorescence are also described. From these activities,…

  11. Classroom Activity Connections: Lessons from Fluorescence

    Science.gov (United States)

    MacCormac, Aoife; O'Brien, Emma; O'Kennedy, Richard

    2010-01-01

    This Classroom Activity Connections paper describes an extension to the "JCE" Classroom Activity #68 "Turning on the Light". A number of additional common items that display fluorescence under UV light are described, including fruits, vegetables, and seashells. Two classroom extensions on fluorescence are also described. From these activities,…

  12. 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

  13. FLUORESCENCE IN DISSOLVED FRACTIONS OF HUMAN ENAMEL

    NARCIS (Netherlands)

    HAFSTROMBJORKMAN, U; SUNDSTROM, F; TENBOSCH, JJ

    Fluorescence induced by laser light is useful in early detection of enamel caries. The present work studied the fluorescence emission pattern in dissolved human enamel and in different molecular weight fractions obtained after gel chromatography or dialysis followed by ultrafiltration. For

  14. Control of excitation in the fluorescence microscope.

    Science.gov (United States)

    Lea, D J; Ward, D J

    1979-01-01

    In fluorescence microscopy image brightness and contrast and the rate of fading depend upon the intensity of illumination of the specimen. An iris diaphragm or neutral density filters may be used to reduce fluorescence excitation. Also the excitation bandwidth may be varied by using a broad band exciter filter with a set of interchangeable yellow glass filters at the lamphouse.

  15. Bridging fluorescence microscopy and electron microscopy

    NARCIS (Netherlands)

    Giepmans, Ben N. G.

    Development of new fluorescent probes and fluorescence microscopes has led to new ways to study cell biology. With the emergence of specialized microscopy units at most universities and research centers, the use of these techniques is well within reach for a broad research community. A major

  16. Bridging fluorescence microscopy and electron microscopy

    NARCIS (Netherlands)

    Giepmans, Ben N. G.

    2008-01-01

    Development of new fluorescent probes and fluorescence microscopes has led to new ways to study cell biology. With the emergence of specialized microscopy units at most universities and research centers, the use of these techniques is well within reach for a broad research community. A major breakth

  17. A sensitive fluorescent sensor of lanthanide ions

    CERN Document Server

    Bekiari, V; Lianos, P

    2003-01-01

    A fluorescent probe bearing a diazostilbene chromophore and a benzo-15-crown-5 ether moiety is a very efficient sensor of lanthanide ions. The ligand emits strong fluorescence only in the presence of specific ions, namely lanthanide ions, while the emission wavelength is associated with a particular ion providing high sensitivity and resolution.

  18. Hydrophilic fluorescent nanogel thermometer for intracellular thermometry.

    Science.gov (United States)

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

    2009-03-01

    The first methodology to measure intracellular temperature is described. A highly hydrophilic fluorescent nanogel thermometer developed for this purpose stays in the cytoplasm and emits stronger fluorescence at a higher temperature. Thus, intracellular temperature variations associated with biological processes can be monitored by this novel thermometer with a temperature resolution of better than 0.5 degrees C.

  19. 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.

  20. Holograms preparation using commercial fluorescent benzyl

    Energy Technology Data Exchange (ETDEWEB)

    Dorantes-GarcIa, V; Olivares-Perez, A; Ordonez-Padilla, M J; Mejias-Brizuela, N Y, E-mail: valdoga@Hotmail.com, E-mail: olivares@inaoep.mx [Instituto Nacional de Astrofisica, Optica y Electronica (INAOE), Coordinacion de Optica, Calle Luis Enrique Erro N0 1, Santa Maria Tonantzintla, Puebla (Mexico)

    2011-01-01

    We have been able to make holograms with substances such as fluorescence thought of light blue laser to make transmissions holograms, using ammonium dichromate as photo-sensitizer and polyvinyl alcohol (PVA) as matrix. Ammonium dichromate inhibit the fluorescence properties of inks, both mixed in a (PVA) matrix, but we avoid this chemical reaction and we show the results to use the method of painting hologram with fluorescents ink and we describe how the diffraction efficiency parameter changes as a function of the ink absorbed by the emulsion recorded with the gratings, we got good results, making holographic gratings with a blue light from laser diode 470 nm. And we later were painting with fluorescent ink, integrating fluorescence characteristics to the hologram.

  1. Magnetically modulated fluorescent probes in turbid media

    CERN Document Server

    Yang,; Chen, Hongyu; Anker, Jeffrey N

    2010-01-01

    Magnetically modulated optical nanoprobes (MagMOONs) were used to detect and distinguish probe fluorescence from autofluorescent backgrounds in turbid media. MagMOONs are micro/nano-sized particles with magnetically controlled orientation and orientation-dependent fluorescence. These probes blink when they rotate in response to rotating external magnetic fields. This blinking signal can be separated from backgrounds enabling spectrochemical sensing in media with strong autofluorescence. We explore the effect of scattering on MagMOON fluorescence. Turbid media reduce the modulated MagMOON signal due to a combination of attenuation of fluorescence signal and reduction in contrast between "On" and "Off" states. The blinking MagMOON fluorescence spectrum can be detected in turbid non-dairy creamer solution with extinction 2.0, and through 9 mm of chicken breast tissue, suggesting that whole mouse imaging is feasible by using this strategy.

  2. Fluorescence microscopy: A tool to study autophagy

    Science.gov (United States)

    Rai, Shashank; Manjithaya, Ravi

    2015-08-01

    Autophagy is a cellular recycling process through which a cell degrades old and damaged cellular components such as organelles and proteins and the degradation products are reused to provide energy and building blocks. Dysfunctional autophagy is reported in several pathological situations. Hence, autophagy plays an important role in both cellular homeostasis and diseased conditions. Autophagy can be studied through various techniques including fluorescence based microscopy. With the advancements of newer technologies in fluorescence microscopy, several novel processes of autophagy have been discovered which makes it an essential tool for autophagy research. Moreover, ability to tag fluorescent proteins with sub cellular targets has enabled us to evaluate autophagy processes in real time under fluorescent microscope. In this article, we demonstrate different aspects of autophagy in two different model organisms i.e. yeast and mammalian cells, with the help of fluorescence microscopy.

  3. Intense fluorescence of Au 20

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Chongqi; Harbich, Wolfgang; Sementa, Luca; Ghiringhelli, Luca; Aprá, Edoardo; Stener, Mauro; Fortunelli, Alessandro; Brune, Harald

    2017-08-21

    Ligand-protected Au clusters are non-bleaching fluorescence markers in bio- and medical applications. We show that their fluorescence is an intrinsic property of the Au cluster itself. We find a very intense and sharp fluorescence peak located at λ =739.2 nm (1.68 eV) for Au20 clusters in a Ne matrix held at 6 K. The fluorescence reflects the HOMO-LUMO diabatic bandgap of the cluster. The cluster shows a very rich absorption fine structure reminiscent of well defined molecule-like quantum levels. These levels are resolved since Au20 has only one stable isomer (tetrahedral), therefore our sample is mono-disperse in cluster size and conformation. Density-functional theory (DFT) and time-dependent DFT calculations clarify the nature of optical absorptionand predict both main absorption peaks and intrinsic fluorescence in good agreement with experiment.

  4. Fiber optical assembly for fluorescence spectrometry

    Science.gov (United States)

    Carpenter, II, Robert W.; Rubenstein, Richard; Piltch, Martin; Gray, Perry

    2010-12-07

    A system for analyzing a sample for the presence of an analyte in a sample. The system includes a sample holder for containing the sample; an excitation source, such as a laser, and at least one linear array radially disposed about the sample holder. Radiation from the excitation source is directed to the sample, and the radiation induces fluorescent light in the sample. Each linear array includes a plurality of fused silica optical fibers that receive the fluorescent light and transmits a fluorescent light signal from the first end to an optical end port of the linear array. An end port assembly having a photo-detector is optically coupled to the optical end port. The photo-detector detects the fluorescent light signal and converts the fluorescent light signal into an electrical signal.

  5. Radioactivity-synchronized fluorescence enhancement using a radionuclide fluorescence-quenched dye.

    Science.gov (United States)

    Berezin, Mikhail Y; Guo, Kevin; Teng, Bao; Edwards, W Barry; Anderson, Carolyn J; Vasalatiy, Olga; Gandjbakhche, Amir; Griffiths, Gary L; Achilefu, Samuel

    2009-07-08

    We demonstrate the first evidence of radioactivity-synchronized fluorescence quenching of a near-infrared light-emitting dye by a radionuclide, (64)Cu, and subsequent fluorescence enhancement upon (64)Cu decay to the daughter isotopes (64)Ni and (64)Zn. The dynamic switch from high radioactivity and low fluorescence to low radioactivity and high fluorescence is potentially useful for developing complementary multimodal imaging and detection platforms for chemical, environmental, and biomedical applications as well as for unraveling the mechanisms of metal-induced dynamic fluorescence changes.

  6. Fluorescent Approaches to High Throughput Crystallography

    Science.gov (United States)

    Pusey, Marc L.; Forsythe, Elizabeth; Achari, Aniruddha

    2006-01-01

    We have shown that by covalently modifying a subpopulation, less than or equal to 1%, of a macromolecule with a fluorescent probe, the labeled material will add to a growing crystal as a microheterogeneous growth unit. Labeling procedures can be readily incorporated into the final stages of purification, and the presence of the probe at low concentrations does not affect the X-ray data quality or the crystallization behavior. The presence of the trace fluorescent label gives a number of advantages when used with high throughput crystallizations. The covalently attached probe will concentrate in the crystal relative to the solution, and under fluorescent illumination crystals show up as bright objects against a dark background. Non-protein structures, such as salt crystals, will not incorporate the probe and will not show up under fluorescent illumination. Brightly fluorescent crystals are readily found against less bright precipitated phases, which under white light illumination may obscure the crystals. Automated image analysis to find crystals should be greatly facilitated, without having to first define crystallization drop boundaries as the protein or protein structures is all that shows up. Fluorescence intensity is a faster search parameter, whether visually or by automated methods, than looking for crystalline features. We are now testing the use of high fluorescence intensity regions, in the absence of clear crystalline features or "hits", as a means for determining potential lead conditions. A working hypothesis is that kinetics leading to non-structured phases may overwhelm and trap more slowly formed ordered assemblies, which subsequently show up as regions of brighter fluorescence intensity. Preliminary experiments with test proteins have resulted in the extraction of a number of crystallization conditions from screening outcomes based solely on the presence of bright fluorescent regions. Subsequent experiments will test this approach using a wider

  7. Fluorescent Probes and Fluorescence (Microscopy Techniques — Illuminating Biological and Biomedical Research

    Directory of Open Access Journals (Sweden)

    Gregor P. C. Drummen

    2012-11-01

    Full Text Available Fluorescence, the absorption and re-emission of photons with longer wavelengths, is one of those amazing phenomena of Nature. Its discovery and utilization had, and still has, a major impact on biological and biomedical research, since it enables researchers not just to visualize normal physiological processes with high temporal and spatial resolution, to detect multiple signals concomitantly, to track single molecules in vivo, to replace radioactive assays when possible, but also to shed light on many pathobiological processes underpinning disease states, which would otherwise not be possible. Compounds that exhibit fluorescence are commonly called fluorochromes or fluorophores and one of these fluorescent molecules in particular has significantly enabled life science research to gain new insights in virtually all its sub-disciplines: Green Fluorescent Protein. Because fluorescent proteins are synthesized in vivo, integration of fluorescent detection methods into the biological system via genetic techniques now became feasible. Currently fluorescent proteins are available that virtually span the whole electromagnetic spectrum. Concomitantly, fluorescence imaging techniques were developed, and often progress in one field fueled innovation in the other. Impressively, the properties of fluorescence were utilized to develop new assays and imaging modalities, ranging from energy transfer to image molecular interactions to imaging beyond the diffraction limit with super-resolution microscopy. Here, an overview is provided of recent developments in both fluorescence imaging and fluorochrome engineering, which together constitute the “fluorescence toolbox” in life science research.

  8. Further insights into metal-DOM interaction: consideration of both fluorescent and non-fluorescent substances.

    Directory of Open Access Journals (Sweden)

    Huacheng Xu

    Full Text Available Information on metal binding with fluorescent substances has been widely studied. By contrast, information on metal binding with non-fluorescent substances remains lacking despite the dominance of these substances in aquatic systems. In this study, the metal binding properties of both fluorescent and non-fluorescent substances were investigated by using metal titration combined with two-dimensional correlation spectroscopy (2D-COS analysis. The organic matters in the eutrophic algae-rich lake, including natural organic matters (NOM and algae-induced extracellular polymeric substances (EPS, both contained fluorescent and non-fluorescent substances. The peaks in the one-dimensional spectra strongly overlapped, while 2D-COS can decompose the overlapped peaks and thus enhanced the spectral resolution. Moreover, 2D FTIR COS demonstrated that the binding susceptibility of organic ligands in both NOM and algal EPS matrices followed the order: 3400>1380>1650 cm-1, indicative the significant contribution of non-fluorescent ligands in metal binding. The modified Stern-Volmer equation also revealed a substantial metal binding potential for the non-fluorescent substances (logKM: 3.57∼4.92. As for the effects of organic ligands on metal binding, EPS was characterized with higher binding ability than NOM for both fluorescent and non-fluorescent ligands. Algae-induced EPS and the non-fluorescent substances in eutrophic algae-rich lakes should not be overlooked because of their high metal binding potential.

  9. 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

  10. Expanding the potential of standard flow cytometry by extracting fluorescence lifetimes from cytometric pulse shifts

    National Research Council Canada - National Science Library

    Cao, Ruofan; Naivar, Mark A; Wilder, Mark; Houston, Jessica P

    2014-01-01

    Fluorescence lifetime measurements provide information about the fluorescence relaxation, or intensity decay, of organic fluorophores, fluorescent proteins, and other inorganic molecules that fluoresce...

  11. Practical use of corrected fluorescence excitation and emission spectra of fluorescent proteins in Förster Resonance Energy Transfer (FRET) studies

    NARCIS (Netherlands)

    Hink, M.A.; Visser, N.V.; Borst, J.W.; Hoek, van A.; Visser, A.J.W.G.

    2003-01-01

    Corrected fluorescence excitation and emission spectra have been obtained from several enhanced variants of the green fluorescent protein (EGFP) isolated from the jellyfish Aequorea victoria, blue fluorescence protein (EBFP), cyan fluorescent protein (ECFP), EGFP and yellow fluorescent protein (EYFP

  12. Practical use of corrected fluorescence excitation and emission spectra of fluorescent proteins in Förster Resonance Energy Transfer (FRET) studies

    NARCIS (Netherlands)

    Hink, M.A.; Visser, N.V.; Borst, J.W.; Hoek, van A.; Visser, A.J.W.G.

    2003-01-01

    Corrected fluorescence excitation and emission spectra have been obtained from several enhanced variants of the green fluorescent protein (EGFP) isolated from the jellyfish Aequorea victoria, blue fluorescence protein (EBFP), cyan fluorescent protein (ECFP), EGFP and yellow fluorescent protein

  13. Ultrafast fluorescence of photosynthetic crystals and light-harvesting complexes

    NARCIS (Netherlands)

    Oort, van B.F.

    2008-01-01

    This thesis focuses on the study of photosynthetic pigment protein complexes using time resolved fluorescence techniques. Fluorescence spectroscopy often requires attaching fluorescent labels to the proteins under investigation. With photosynthetic proteins this is not necessary, because these

  14. Ultrafast fluorescence of photosynthetic crystals and light-harvesting complexes

    NARCIS (Netherlands)

    Oort, van B.F.

    2008-01-01

    This thesis focuses on the study of photosynthetic pigment protein complexes using time resolved fluorescence techniques. Fluorescence spectroscopy often requires attaching fluorescent labels to the proteins under investigation. With photosynthetic proteins this is not necessary, because these prote

  15. Solid-State Camera System for Fluorescence Lifetime Microscopy

    NARCIS (Netherlands)

    Zhao, Q.

    2014-01-01

    Fluorescence microscopy is a well-established platform for biology and biomedical research (Chapter 2). Based on this platform, fluorescence lifetime imaging microscopy (FLIM) has been developed to measure fluorescence lifetimes, which are independent of fluorophore concentration and excitation inte

  16. Ultrafast fluorescence of photosynthetic crystals and light-harvesting complexes

    NARCIS (Netherlands)

    Oort, van B.F.

    2008-01-01

    This thesis focuses on the study of photosynthetic pigment protein complexes using time resolved fluorescence techniques. Fluorescence spectroscopy often requires attaching fluorescent labels to the proteins under investigation. With photosynthetic proteins this is not necessary, because these prote

  17. Caries diagnosis using laser fluorescence

    Science.gov (United States)

    Zanin, Fatima A. A.; Pinheiro, Antonio L. B.; Souza-Campos, Dilma H.; Brugnera, Aldo, Jr.; Pecora, Jesus D.

    2000-03-01

    Caries prevention is a goal to be achieved by dentist in order to promote health. There are several methods used to detect dental caries each one presenting advantages and disadvantages, especially regarding hidden occlusal caries. The improvement of laser technology has permitted the use of laser fluorescence for early diagnosis of hidden occlusal caries. The aim of this study was to assess the efficacy of the use of 655 nm laser light on the detection of hidden occlusal caries. Forty molar teeth from patients of both sexes which ages ranging from 10 - 18 years old were used on this study. Following manufacture's instructions regarding the use of the equipment, the teeth had their occlusal surface examined with the DIAGNOdent. Twenty six of 40 teeth had hidden occlusal caries detected by the DIAGNOdent. However only 17 of these 26 teeth showed radiographic signs of caries the other 9 teeth showed no radiological signs of the lesion. Radiographic examination was able to identify 34,61% of false negative cases. This means that many caries would be left untreated due to the lack of diagnosis using both visual and radiographic examination. The use of the DIAGNOdent was effective in successfully detecting hidden occlusal caries.

  18. Fluorescence spectroscopic behaviour of folic acid

    Energy Technology Data Exchange (ETDEWEB)

    Tyagi, A. [Institut II - Experimentelle und Angewandte Physik, Universitaet Regensburg, Universitaetstrasse 31, D-93053 Regensburg (Germany); Penzkofer, A., E-mail: alfons.penzkofer@physik.uni-regensburg.de [Institut II - Experimentelle und Angewandte Physik, Universitaet Regensburg, Universitaetstrasse 31, D-93053 Regensburg (Germany)

    2010-02-08

    The fluorescence spectroscopic behaviour of folic acid (FA) in 4 M HCl (dominant bi-cationic form), 0.1 M HCl (bi-cationic and cationic form), citric acid-NaOH pH 6 buffer (neutral form), 0.1 M and 4 M KOH (anionic form), and trifluoroacetic acid is studied. The thermal stability is investigated. Absolute absorption cross-section spectra are determined and compared with fluorescence excitation spectra. Intrinsic fluorescence quantum distributions and fluorescence quantum yields are extracted from fluorescence spectra measurements. The temporal fluorescence decay after picosecond pulse excitation is studied. The fluorescence quenching mechanisms for the different ionic forms of FA are discussed: excited-state proton release for bi-cationic FA, photo-physical non-radiative relaxation for cationic FA, and photo-induced intra-molecular electron transfer for neutral and anionic FA. Aerobic FA in 4 M KOH at elevated temperature dehydrated to 9,10-dehydro-folic acid. Its photo-dynamics was governed by twisted intra-molecular charge transfer and photo-isomerisation.

  19. Mean fluorescence lifetime and its error

    Energy Technology Data Exchange (ETDEWEB)

    Fiserova, Eva [Department of Mathematical Analysis and Applications of Mathematics, Faculty of Science, Palacky University in Olomouc, tr. 17. listopadu 12, CZE-77146 Olomouc (Czech Republic); Kubala, Martin, E-mail: mkubala@prfnw.upol.cz [Department of Biophysics, Faculty of Science, Palacky University in Olomouc, tr. 17. listopadu 12, CZE-77146 Olomouc (Czech Republic)

    2012-08-15

    Mean excited-state lifetime is one of the fundamental fluorescence characteristics and enters as an important parameter into numerous calculations characterizing molecular interactions, such as e.g. FRET or fluorescence quenching. Our experiments demonstrated that the intensity-weighted mean fluorescence lifetime is very robust characteristic, in contrast to the amplitude-weighted one, which value is dependent on the data quality and particularly on the used fitting model. For the first time, we also report the procedure for the error estimation for both the intensity- and amplitude-weighted mean fluorescence lifetimes. Furthermore, we present a method for estimation of the mean fluorescence lifetime directly from the fluorescence-decay curve recorded by TCSPC (Time-Correlated Single-Photon Counting) method. For its simplicity and low computational demands, it could be a useful tool in the high-throughput applications, such as FACS, FLIM-FRET or HPLC detectors. - Highlights: Black-Right-Pointing-Pointer Intensity-weighted mean fluorescence lifetime is very robust characteristic. Black-Right-Pointing-Pointer The amplitude-weighted mean lifetime depends on the selection of fitting model. Black-Right-Pointing-Pointer Rigorous procedure for estimation of confidence intervals for mean lifetime. Black-Right-Pointing-Pointer The mean lifetime can be estimated directly from the TCSPC histogram.

  20. Fluorescence of fullerene derivatives at room temperature

    Energy Technology Data Exchange (ETDEWEB)

    Lin, S.K.; Shiu, L.L.; Chien, K.M.; Luh, T.Y.; Lin, T.I. (National Taiwan Univ., Taipei (Taiwan, Province of China))

    1995-01-05

    The absorption and fluorescence spectral properties of fullerene (C[sub 60]) and its derivatives C[sub 60]C[sub 4]H[sub 6], C[sub 60]C[sub 5]H[sub 6], C[sub 60]CHCO[sub 2]Et, and C[sub 60]NCO[sub 2]Et at room temperature were investigated. Breaking the structural symmetry of C[sub 60] results in enhancing the fluorescence quantum yield 2-3-fold in some derivatives. Thus, the room temperature fluorescence of fullerene compounds could be detected more rapidly. New absorption bands and altered fluorescence spectra were observed in the derivatives. The Stokes' shifts of the derivatives are small, about 4-5 nm, compared to 68 nm for the parent compound. The time-resolved fluorescence decay study indicates that all four fullerene derivatives have a single fluorescence lifetime of ca. 1.2-1.4 as, which is about the same as that for C[sub 60] (ca. 1.3 ns). Aliphatic solvents have little influence on the absorption or fluorescence spectral profile except on the extinction coefficient whereas aromatic and polar solvents strongly interact with the fullerene derivatives, causing a peak broadening effect. 31 refs., 7 figs., 3 tabs.

  1. 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

  2. Fluorescence characterization of clinically-important bacteria.

    Directory of Open Access Journals (Sweden)

    Lewis R Dartnell

    Full Text Available Healthcare-associated infections (HCAI/HAI represent a substantial threat to patient health during hospitalization and incur billions of dollars additional cost for subsequent treatment. One promising method for the detection of bacterial contamination in a clinical setting before an HAI outbreak occurs is to exploit native fluorescence of cellular molecules for a hand-held, rapid-sweep surveillance instrument. Previous studies have shown fluorescence-based detection to be sensitive and effective for food-borne and environmental microorganisms, and even to be able to distinguish between cell types, but this powerful technique has not yet been deployed on the macroscale for the primary surveillance of contamination in healthcare facilities to prevent HAI. Here we report experimental data for the specification and design of such a fluorescence-based detection instrument. We have characterized the complete fluorescence response of eleven clinically-relevant bacteria by generating excitation-emission matrices (EEMs over broad wavelength ranges. Furthermore, a number of surfaces and items of equipment commonly present on a ward, and potentially responsible for pathogen transfer, have been analyzed for potential issues of background fluorescence masking the signal from contaminant bacteria. These include bedside handrails, nurse call button, blood pressure cuff and ward computer keyboard, as well as disinfectant cleaning products and microfiber cloth. All examined bacterial strains exhibited a distinctive double-peak fluorescence feature associated with tryptophan with no other cellular fluorophore detected. Thus, this fluorescence survey found that an emission peak of 340nm, from an excitation source at 280nm, was the cellular fluorescence signal to target for detection of bacterial contamination. The majority of materials analysed offer a spectral window through which bacterial contamination could indeed be detected. A few instances were found of

  3. Fluorescent Sensing of Fluoride in Cellular System

    Science.gov (United States)

    Jiao, Yang; Zhu, Baocun; Chen, Jihua; Duan, Xiaohong

    2015-01-01

    Fluoride ions have the important roles in a lot of physiological activities related with biological and medical system, such as water fluoridation, caries treatment, and bone disease treatment. Great efforts have been made to develop new methods and strategies for F- detection in the past decades. Traditional methods for the detection of F- including ion chromatography, ion-selective electrodes, and spectroscopic techniques have the limitations in the biomedicine research. The fluorescent probes for F- are very promising that overcome some drawbacks of traditional fluoride detection methods. These probes exhibit high selectivity, high sensitivity as well as quick response to the detection of fluoride anions. The review commences with a brief description of photophysical mechanisms for fluorescent probes for fluoride, including photo induced electron transfer (PET), intramolecular charge transfer (ICT), fluorescence resonance energy transfer (FRET), and excited-state intramolecular proton transfer (ESIPT). Followed by a discussion about common dyes for fluorescent fluoride probes, such as anthracene, naphalimide, pyrene, BODIPY, fluorescein, rhodamine, resorufin, coumarin, cyanine, and near-infrared (NIR) dyes. We divide the fluorescent probes for fluoride in cellular application systems into nine groups, for example, type of hydrogen bonds, type of cleavage of Si-O bonds, type of Si-O bond cleavage and cylization reactions, etc. We also review the recent reported carriers in the delivery of fluorescent fluoride probes. Seventy-four typical fluorescent fluoride probes are listed and compared in detail, including quantum yield, reaction medium, excitation and emission wavelengths, linear detection range, selectivity for F-, mechanism, and analytical applications. Finally, we discuss the future challenges of the application of fluorescent fluoride probes in cellular system and in vivo. We wish that more and more excellent fluorescent fluoride probes will be developed

  4. 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.

  5. Angle-Resolved Spectroscopy of Parametric Fluorescence

    CERN Document Server

    Hsu, Feng-kuo

    2013-01-01

    The parametric fluorescence from a nonlinear crystal forms a conical radiation pattern. We measure the angular and spectral distributions of parametric fluorescence in a beta-barium borate crystal pumped by a 405-nm diode laser employing angle-resolved imaging spectroscopy. The experimental angle-resolved spectra and the generation efficiency of parametric down conversion are compared with a plane-wave theoretical analysis. The parametric fluorescence is used as a broadband light source for the calibration of the instrument spectral response function in the wavelength range from 450 to 1000 nm.

  6. Fluorescent integrating sphere for the vacuum ultraviolet.

    Science.gov (United States)

    Brandenberg, W M

    1970-02-01

    An integrating sphere for absolute, hemispherical reflectance measurements on imperfectly diffuse surfaces in the wavelength range between 1250 A and 3500 A has been built. The sphere uses a double layer coating consisting of a sodium salicylate film on top of a diffuse white paint. The phosphor coating, under uv irradiation, emits fluorescent radiation in the blue, and the underlying paint layer serves as a diffuser of the fluorescent radiation. The usual problem, encountered in ordinary integrating spheres where direct irradiation of the detector by the sample can lead to erroneous signals, is easily eliminated in the fluorescent integrating sphere by proper filtering of the detector.

  7. Peptide-stabilized, fluorescent silver nanoclusters

    DEFF Research Database (Denmark)

    Gregersen, Simon; Vosch, Tom André Jos; Jensen, Knud Jørgen

    2016-01-01

    . Herein, we demonstrate how solid-phase methods can increase throughput dramatically in peptide ligand screening and in initial evaluation of fluorescence intensity and chemical stability of peptide-stabilized AgNCs (P-AgNCs). 9-Fluorenylmethyloxycarbonyl (Fmoc) solid-phase peptide synthesis......Few-atom silver nanoclusters (AgNCs) can exhibit strong fluorescence; however, they require ligands to prevent aggregation into larger nanoparticles. Fluorescent AgNCs in biopolymer scaffolds have so far mainly been synthesized in solution, and peptides have only found limited use compared to DNA...

  8. Containerless Atomic-Fluorescence Property Measurements

    Science.gov (United States)

    Nordine, P.; Schiffman, R.; Walker, C.

    1987-01-01

    Report describes studies conducted to establish and verify use of laser-induced fluorescence in monitoring and controlling high-temperature containerless processes. Specimens levitated by gas jets or electromagnetic fields and heated by laser beams or electromagnetic induction while being irradiated and detected by fluorescence technique. Makes quantitative and qualitative comparisons among three new methods of temperature measurement; all rely on laser-induced fluorescence. One method gas-density thermometry with seed gas. Other two methods involve measurements of velocities of evaporating atoms or of population ratios of different electronic states.

  9. Laser Excited Fluorescence For Forensic Diagnostics

    Science.gov (United States)

    McKinney, Robert E.

    1986-07-01

    The application of laser excited fluorescence to the detection and identification of latent fingerprints was first accomplished ten years ago. The development of the technology has progressed rapidly with the introduction of commercial equipment by several manufacturers. Systems based on Argon-ion, Copper-vapor, and frequency-doubled Nd:YAG lasers are compared. The theoretical basis of detection by fluorescence is discussed along with the more useful techniques of dye staining. Other applications of the laser excited fluorescence in forensic investigation include gunshot residue analysis, serology, collection of trace evidence, and document examination.

  10. 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.

  11. Welcome to Methods and Applications in Fluorescence

    Science.gov (United States)

    Birch, David; Mély, Yves; Wolfbeis, Otto S.

    2013-03-01

    On behalf of the Editorial Board of Methods and Applications in Fluorescence and IOP Publishing we are delighted to invite you to read the first articles in our new journal. Methods and Applications in Fluorescence is forged out of the renowned MAF conference series of the same name and we fully expect the natural synergy between the two to provide the ideal platform for moving the field of fluorescence forward. Our aim is for this new journal to reflect the truly global and diverse impact fluorescence is having across many disciplines and help fluorescence achieve its full potential. Just as MAF is the leading conference in fluorescence we are confident of the high impact of this new journal. Methods and Applications in Fluorescence has a distinguished Editorial Board that is drawn from the MAF conference Permanent Steering Committee. Together with the Editorial Board and the rest of the community, the journal will closely track the very latest developments in fluorescence while delivering a fair and constructive review process. We are very pleased that this journal is backed by the Institute of Physics, one of the world's premier learned societies. IOP Publishing has a wealth of experience in science publishing that dates back to 1874. It is a not-for-profit organization that publishes over 60 journals, many on multidisciplinary topics and many including seminal contributions from Nobel Laureates. Any funding surplus generated by IOP Publishing goes directly back into science through the Institute of Physics, thus helping to nurture science for future generations. We invite submissions as regular articles, review articles and technical notes within the scope of the journal, which includes all the major aspects of fluorescence. This covers both theory and experiment across spectroscopy, imaging, materials, labels, probes and sensors. The applications of fluorescence to emerging areas in bionanotechnology, nanotechnology and medicine are very much part of the

  12. Technical principles and neurosurgical applications of fluorescein fluorescence using a microscope-integrated fluorescence module.

    Science.gov (United States)

    Rey-Dios, Roberto; Cohen-Gadol, Aaron A

    2013-04-01

    Fluorescent technology has recently become a valuable tool in the surgical management of neoplastic and vascular lesions. The availability of microscope-integrated fluorescent modules has facilitated incorporation of this technology within the microsurgical workflow. The currently available microscope integrated modules use 5-aminolevulinic acid (5-ALA) and indocyanine green (ICG) as fluorophores. Fluorescein sodium is a fluorescent molecule that has been used specifically in ophthalmology for the treatment of retinal angiography. A new microscope-integrated fluorescent module has been recently developed for fluorescein. We employed this technology to maximize resection of tumors and perform intraoperative angiography to guide microsurgical management of aneurysms and arteriovenous malformations. Fluorescein fluorescence allows the surgeon to appreciate fluorescent structures through the oculars while visualizing non-fluorescent tissues in near natural colors. Therefore, the operator can proceed with microsurgery under the fluorescent mode. We present three representative cases in which the use of fluorescein fluorescence was found useful in the surgeon's decision making during surgery. The applications of this new microscope-integrated fluorescent module are multiple, and include vascular and oncologic neurosurgery. Further clinical investigations with large patient cohorts are needed to fully establish the role of this new technology.

  13. Multivariate analysis of endometrial tissue fluorescence spectra

    Science.gov (United States)

    Vaitkuviene, Aurelija; Auksorius, E.; Fuchs, D.; Gavriushin, V.

    2002-10-01

    Background and Objective: The detailed multivariate analysis of endometrial tissue fluorescence spectra was done. Spectra underlying features and classification algorithm were analyzed. An effort has been made to determine the importance of neopterin component in endometrial premalignization. Study Design/Materials and Methods: Biomedical tissue fluorescence was measured by excitation with the Nd YAG laser third harmonic. Multivariate analysis techniques were used to analyze fluorescence spectra. Biomedical optics group at Vilnius University analyzed the neopterin substance supplied by the Institute of Medical Chemistry and Biochemistry of Innsbruck University. Results: Seven statistically significant spectral compounds were found. The classification algorithm classifying samples to histopathological categories was developed and resulted in sensitivity of 80% and specificity 93% for malignant vs. hyperplastic and normal. Conclusions: Fluorescence spectra could be classified with high accuracy. Spectral variation underlying features can be extracted. Neopterin component might play an important role in endometrial hyperplasia development.

  14. Smart Phone Fluorescent Chem8 Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Ionu Biosystems will develop a fluorescent smart phone blood analyzer that can measure important physiological concentrations from a drop of blood. The approach will...

  15. Fluorescence assessment of Lactococcus lactis viability

    NARCIS (Netherlands)

    Bunthof, C.J.; Braak, van den S.; Breeuwer, P.; Rombouts, F.M.; Abee, T.

    2000-01-01

    The reproduction and activity of lactic acid bacteria (LAB) are essential in their applications in the dairy industry and other fermentations. Traditionally used methods like plate counting and acidification tests require long incubation times and provide limited information. Fluorescence techniques

  16. 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.

  17. Use of fluorescent screens for isotope radiography

    Energy Technology Data Exchange (ETDEWEB)

    Hubbard, S. K.

    1979-01-01

    Radiographic examination can be performed on items beyond the limitation of conventional isotope radiography without a great loss of resolution. With proper film and screen selection and scatter radiation control, fluorescent screens can be a valuable additional tool for radiography.

  18. Remote UV Fluorescence Lifetime Spectrometer Project

    Data.gov (United States)

    National Aeronautics and Space Administration — The goal of this project is to develop, demonstrate, and deliver to NASA an innovative, portable, and power efficient Remote UV Fluorescence Lifetime Spectrometer...

  19. Compact fluorescence spectroscopic tool for cancer detection

    Science.gov (United States)

    Nadeau, Valerie; Hamdan, Khaled; Hewett, Jacqueline; Makaryceva, Juljia; Tait, Iain; Cuschieri, Alfred; Padgett, Miles J.

    2002-05-01

    We describe a compact fluorescence spectroscopic tool for in vivo point monitoring of aminolaevulinic acid (ALA)-induced protoporphyrin IX (PpIX) fluorescence and autofluorescence, as a non-invasive method of differentiating normal and cancerous tissue. This instrument incorporates a 405nm diode laser with a shutter to prevent exposure of tissue to harmful light doses and reduce photobleaching, a bifurcated optical fibre to allow illumination of tissue and collection of fluorescence with a single fibre, a compact grating spectrometer for collection of spectra and a PC for system control. We present spectra obtained using this system both during routine gastro-intestinal (GI) endoscopy for cancer detection and during photodynamic therapy (PDT) of anal intraepithelial neoplasia (AIN) for monitoring of treatment progress. These results illustrate the potential of the system to be used for fluorescence monitoring in a variety of clinical applications.

  20. Fluorescence of berberine in microheterogeneous systems

    Energy Technology Data Exchange (ETDEWEB)

    Colina, Ariel N.; Díaz, Marta S.; Gutiérrez, María Isela, E-mail: isela@unpata.edu.ar

    2013-12-15

    Spectral properties of the alkaloid berberine were studied in micellar solution and microemulsions based on anionic sodium dodecyl sulfate, cationic cetyltrimethylammonium bromide and nonionic Triton X-100 surfactants. Absorption and fluorescence emission spectra were determined. For screening the influence of type and concentration of micelles on the fluorescence of berberine a 3{sup 2} full factorial design was used. Higher responses were obtained when berberine was dissolved in sodium dodecyl sulfate micelles 0.01 M. Comparative results of fluorescence quantum yields (Φ{sub f}) reveal that the highest values (Φ{sub f}≥0.01) were observed in microemulsions. In the microheterogeneous systems investigated the most probable location of berberine is the micellar interfacial region. -- Highlights: • Spectroscopic propereies of berberine in microheterogeneous media were investigated. • Berberine shows enhanced fluorescence in SDS micelles as compared to water • Berberine is probably located in the interface of the microheterogeneous systems.

  1. Depth-resolved fluorescence of biological tissue

    Science.gov (United States)

    Wu, Yicong; Xi, Peng; Cheung, Tak-Hong; Yim, So Fan; Yu, Mei-Yung; Qu, Jianan Y.

    2005-06-01

    The depth-resolved autofluorescence ofrabbit oral tissue, normal and dysplastic human ectocervical tissue within l20μm depth were investigated utilizing a confocal fluorescence spectroscopy with the excitations at 355nm and 457nm. From the topmost keratinizing layer of oral and ectocervical tissue, strong keratin fluorescence with the spectral characteristics similar to collagen was observed. The fluorescence signal from epithelial tissue between the keratinizing layer and stroma can be well resolved. Furthermore, NADH and FADfluorescence measured from the underlying non-keratinizing epithelial layer were strongly correlated to the tissue pathology. This study demonstrates that the depth-resolved fluorescence spectroscopy can reveal fine structural information on epithelial tissue and potentially provide more accurate diagnostic information for determining tissue pathology.

  2. Conjugation of fluorescent proteins with DNA oligonucleotides.

    Science.gov (United States)

    Lapiene, Vidmantas; Kukolka, Florian; Kiko, Kathrin; Arndt, Andreas; Niemeyer, Christof M

    2010-05-19

    This work describes the synthesis of covalent ssDNA conjugates of six fluorescent proteins, ECFP, EGFP, E(2)GFP, mDsRed, Dronpa, and mCherry, which were cloned with an accessible C-terminal cystein residue to enable site-selective coupling using a heterobispecific cross-linker. The resulting conjugates revealed similar fluorescence emission intensity to the unconjugated proteins, and the functionality of the tethered oligonucleotide was proven by specific Watson-Crick base pairing to cDNA-modified gold nanoparticles. Fluorescence spectroscopy analysis indicated that the fluorescence of the FP is quenched by the gold particle, and the extent of quenching varied with the intrinsic spectroscopic properties of FP as well as with the configuration of surface attachment. Since this study demonstrates that biological fluorophores can be selectively incorporated into and optically coupled with nanoparticle-based devices, applications in DNA-based nanofabrication can be foreseen.

  3. Fluorescence enhancement of photoswitchable metal ion sensors

    Science.gov (United States)

    Sylvia, Georgina; Heng, Sabrina; Abell, Andrew D.

    2016-12-01

    Spiropyran-based fluorescence sensors are an ideal target for intracellular metal ion sensing, due to their biocompatibility, red emission frequency and photo-controlled reversible analyte binding for continuous signal monitoring. However, increasing the brightness of spiropyran-based sensors would extend their sensing capability for live-cell imaging. In this work we look to enhance the fluorescence of spiropyran-based sensors, by incorporating an additional fluorophore into the sensor design. We report a 5-membered monoazacrown bearing spiropyran with metal ion specificity, modified to incorporate the pyrene fluorophore. The effect of N-indole pyrene modification on the behavior of the spiropyran molecule is explored, with absorbance and fluorescence emission characterization. This first generation sensor provides an insight into fluorescence-enhancement of spiropyran molecules.

  4. Second-Generation Fluorescent Quadracyclic Adenine Analogues

    DEFF Research Database (Denmark)

    Dumat, Blaise; Bood, Mattias; Wranne, Moa S.;

    2015-01-01

    Fluorescent base analogues comprise a group of increasingly important molecules for the investigation of nucleic acid structure, dynamics, and interactions with other molecules. Herein, we report on the quantum chemical calculation aided design, synthesis, and characterization of four new putativ...

  5. Do large fluorescent particles enhance the modulation efficiency of ultrasound-modulated fluorescence?

    Science.gov (United States)

    Liu, Yuan; Yuan, Baohong; Vignola, Joseph

    2011-03-01

    The question of whether particle size affects modulation efficiency, defined as the ratio of ultrasound-modulated fluorescence (UMF) signal to DC (direct current) signal, of the fluorescence emission from four different sized fluorescent particles was investigated experimentally. The four particles are streptavidin-conjugated Alexa Fluo 647 ({5 nm in diameter) and three carboxylate-modified fluorescent microspheres (FM) with different diameters of 0.02, 0.2, and 1.0 μm. Modulation efficiency was evaluated as a function of the fluorophore size and fluorophore concentration. The modulation efficiency was improved about two times when the size of the fluorescent particles is increased from 5 nm to 1 μm. This result implies that using large fluorescence particles can slightly improve the modulation efficiency but the improvement is limited.

  6. Fluorescence labeling of carbon nanotubes and visualization of a nanotube-protein hybrid under fluorescence microscope.

    Science.gov (United States)

    Yoshimura, Shige H; Khan, Shahbaz; Maruyama, Hiroyuki; Nakayama, Yoshikazu; Takeyasu, Kunio

    2011-04-11

    Biological applications of carbon nanotubes have been hampered by the inability to visualize them using conventional optical microscope, which is the most common tool for the observation and measurement of biological processes. Recently, a number of fluorescence labeling methods for biomolecules and various fluorescence probes have been developed and widely utilized in biological fields. Therefore, labeling carbon nanotubes with such fluorophores under physiological conditions will be highly useful in their biological applications. In this Article, we present a method to fluorescently label nanotubes by combining a detergent and a fluorophore commonly used in biological experiments. Fluorophores carrying an amino group (Texas Red hydrazide or BODIPY FL-hydrazide) were covalently attached to the hydroxyl groups of Tween 20 using carbonyldiimidazole. Fluorescence microscopy demonstrated that nanotubes were efficiently solubilized and labeled by this fluorescently labeled detergent. By using this technique, we also demonstrated multicolor fluorescence imaging of a nanotube-protein hybrid.

  7. Optimal Fluorescence Waveband Determination for Detecting Defective Cherry Tomatoes Using a Fluorescence Excitation-Emission Matrix

    Directory of Open Access Journals (Sweden)

    In-Suck Baek

    2014-11-01

    Full Text Available A multi-spectral fluorescence imaging technique was used to detect defective cherry tomatoes. The fluorescence excitation and emission matrix was used to measure for defects, sound surface and stem areas to determine the optimal fluorescence excitation and emission wavelengths for discrimination. Two-way ANOVA revealed the optimal excitation wavelength for detecting defect areas was 410 nm. Principal component analysis (PCA was applied to the fluorescence emission spectra of all regions at 410 nm excitation to determine the emission wavelengths for defect detection. The major emission wavelengths were 688 nm and 506 nm for the detection. Fluorescence images combined with the determined emission wavebands demonstrated the feasibility of detecting defective cherry tomatoes with >98% accuracy. Multi-spectral fluorescence imaging has potential utility in non-destructive quality sorting of cherry tomatoes.

  8. Fluorescence correlation spectroscopy in laser gradient field

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Fluorescence correlation spectroscopy (FCS) is capable of probing dynamic processes in living biological systems. From photon fluctuation of fluorescing particles which diffuse through a small detection volume, FCS reveals information on the concentration and the structure of the particles, as well as information on microscopic environment.In this note, we study the radiation forces experienced by Rayleigh particles in a laser field in details, and analyze the effects of gradient field on FCS measurements.

  9. Amide-based Fluorescent Macrocyclic Anion Receptors

    Institute of Scientific and Technical Information of China (English)

    ZENG, Zhen-Ya(曾振亚); XU, Kuo-Xi(徐括喜); HE, Yong-Bing(何永炳); LIU, Shun-Ying(刘顺英); WU, Jin-Long(吴进龙); WEI, Lan-Hua(隗兰华); MENG, Ling-Zhi(孟令芝)

    2004-01-01

    Two fluorescent anion receptors (1 and 2) based on amide macrocycle were synthesized and corresponding fluorescence quenching induced by anion complexation was observed in different degree. Receptors form 1: 1 complexes with anions by hydrogen bonding interactions. Receptor 1 bound anions in the order of F->Cl->H2PO4->CH3COO->>Br-, I- and receptor 2 showed high selectivity to F- over other anions.

  10. Chemical Address Tags of Fluorescent Bioimaging Probes

    OpenAIRE

    Shedden, Kerby; Rosania, Gus R.

    2010-01-01

    Chemical address tags can be defined as specific structural features shared by a set of bioimaging probes having a predictable influence on cell-associated visual signals obtained from these probes. Here, using a large image dataset acquired with a high content screening instrument, machine vision and cheminformatics analysis have been applied to reveal chemical address tags. With a combinatorial library of fluorescent molecules, fluorescence signal intensity, spectral, and spatial features c...

  11. Fluorescence-lifetime-based sensors for anions

    Science.gov (United States)

    Teichmann, Maria; Draxler, Sonja; Kieslinger, Dietmar; Lippitsch, Max E.

    1997-05-01

    Sensing of anions has been investigated using the fluorescence decaytime as the information carrier. The sensing mechanism is based on the coextraction of an anion and a proton, and the presence of a fluorophore with a rather long fluorescence decaytime inside the membrane to act as a pH indicator. The relevant theory is discussed shortly. As an example a sensor for nitrate is shown, and the influence of ionic additives on the working function has been investigated.

  12. A new fluorescent assay for sialyltransferase.

    Science.gov (United States)

    Kajihara, Y; Kamitani, T; Sakakibara, T

    2001-04-23

    A new fluorescent assay for the sialyltransferase reaction was established. After incubation of the sialyltransferase reaction, the sialyloligosaccharide obtained was treated by acid hydrolysis, and then the NeuAc that was released was labeled with 1,2-diamino-4,5-methylenedioxibenzene. The fluorescent-labeled NeuAc could be estimated by HPLC (excitation: 373 nm; emission: 448 nm) and a Lineweaver-Burk plot could be plotted with the data from this analysis.

  13. 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.

  14. Polymeric micellar drug carriers with fluorescent properties

    OpenAIRE

    Abreu, Ana Sofia Lemos Machado; Sá, Arsénio Vasconcelos; Oliveira, Manuel; Moura, I; Machado, A.V.

    2015-01-01

    Self-assembling polymeric surfactants, based on amphiphilic block copolymers into nanosized aggregates in aqueous solution, are of great interest in the biomedical fields as one class of promising carrier systems, for drug delivery, gene therapy and diagnostic biosensors.[1] The incorporation of fluorescent probes into polymeric micelles has been fulfilled either by physically encapsulation or chemically attachment of fluorophores. [2] These micelle-based fluorescent probes not only facili...

  15. Fluorescence derivatization of clarithromycin for high performance liquid chromatographic determination with fluorescence detection

    OpenAIRE

    Boonleang, J.

    2007-01-01

    Clarithromycin (CAM) is a semisynthetic macrolide antibiotic whose chemical structure has no suitable chromophore for highly sensitive and accurate direct determination. The aim of this study was toderivatize CAM with fluorescence-labeling compounds capable of enhancing the sensitivity of CAM determination. Two fluorescence-labeling compounds were used in this study, 9-fluorenylmethyloxycarbonylchloride (FMOC-Cl) and 1-naphthylisocyanate (NIC), both of which gave the fluorescent derivatives o...

  16. 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.

  17. Fluorescent Protein Based FRET Pairs with Improved Dynamic Range for Fluorescence Lifetime Measurements.

    Directory of Open Access Journals (Sweden)

    Bobin George Abraham

    Full Text Available Fluorescence Resonance Energy Transfer (FRET using fluorescent protein variants is widely used to study biochemical processes in living cells. FRET detection by fluorescence lifetime measurements is the most direct and robust method to measure FRET. The traditional cyan-yellow fluorescent protein based FRET pairs are getting replaced by green-red fluorescent protein variants. The green-red pair enables excitation at a longer wavelength which reduces cellular autofluorescence and phototoxicity while monitoring FRET. Despite the advances in FRET based sensors, the low FRET efficiency and dynamic range still complicates their use in cell biology and high throughput screening. In this paper, we utilized the higher lifetime of NowGFP and screened red fluorescent protein variants to develop FRET pairs with high dynamic range and FRET efficiency. The FRET variations were analyzed by proteolytic activity and detected by steady-state and time-resolved measurements. Based on the results, NowGFP-tdTomato and NowGFP-mRuby2 have shown high potentials as FRET pairs with large fluorescence lifetime dynamic range. The in vitro measurements revealed that the NowGFP-tdTomato has the highest Förster radius for any fluorescent protein based FRET pairs yet used in biological studies. The developed FRET pairs will be useful for designing FRET based sensors and studies employing Fluorescence Lifetime Imaging Microscopy (FLIM.

  18. Intracellular distribution of fluorescent copper and zinc bis(thiosemicarbazonato) complexes measured with fluorescence lifetime spectroscopy.

    Science.gov (United States)

    Hickey, James L; James, Janine L; Henderson, Clare A; Price, Katherine A; Mot, Alexandra I; Buncic, Gojko; Crouch, Peter J; White, Jonathan M; White, Anthony R; Smith, Trevor A; Donnelly, Paul S

    2015-10-05

    The intracellular distribution of fluorescently labeled copper and zinc bis(thiosemicarbazonato) complexes was investigated in M17 neuroblastoma cells and primary cortical neurons with a view to providing insights into the neuroprotective activity of a copper bis(thiosemicarbazonato) complex known as Cu(II)(atsm). Time-resolved fluorescence measurements allowed the identification of the Cu(II) and Zn(II) complexes as well as the free ligand inside the cells by virtue of the distinct fluorescence lifetime of each species. Confocal fluorescent microscopy of cells treated with the fluorescent copper(II)bis(thiosemicarbazonato) complex revealed significant fluorescence associated with cytoplasmic puncta that were identified to be lysosomes in primary cortical neurons and both lipid droplets and lysosomes in M17 neuroblastoma cells. Fluorescence lifetime imaging microscopy confirmed that the fluorescence signal emanating from the lipid droplets could be attributed to the copper(II) complex but also that some degree of loss of the metal ion led to diffuse cytosolic fluorescence that could be attributed to the metal-free ligand. The accumulation of the copper(II) complex in lipid droplets could be relevant to the neuroprotective activity of Cu(II)(atsm) in models of amyotrophic lateral sclerosis and Parkinson's disease.

  19. Fluorescent halite from Bochnia salt mine, Poland

    Science.gov (United States)

    Waluś, Edyta; Głąbińska, Dobrochna; Puławska, Aleksandra; Flasza, Michał; Manecki, Maciej

    2016-04-01

    The photoluminescence of selected halite crystals from Bochnia Salt Mine (Bochnia, Poland) were discovered in 2014. This is a result of contemporary precipitation from percolating waters. In most cases the fluorescence is observed in whole crystals or in zones of crystals. Only clear parts of transparent crystals are orange-red fluorescent in short UV light (320 nm). Chemical microanalysis by scanning electron microscopy/energy dispersive spectroscopy SEM/EDS indicates that this is activated by Mn and Pb. The concentration of Mn is similar in fluorescent and inactive salt and equals to 0.13 - 0.27 wt.%. The concentration of Pb, however, averages to 3.8 wt.% in fluorescent parts reaching only 1.9 wt.% elsewhere. There is no difference in the unit cell parameters determined by powder X-ray diffraction. The percolating waters contain some Mn (ca. 3.9 ppm) but the concentration of Pb is below the detection limits. The experiments of precipitation of halite from the solutions containing various concentrations of Mn and Pb were performed to simulate this fenomenon using solutions containing: 1 mg Pb/L and 80 mg Mn/L; 1 mg Pb/L and 0.8 mg Mn/L; 1 mg Pb/L and 0.6 mg Mn/L; and 0 mg Pb/L and 80 mg Mn/L. The results indicate that fluorescence is apparent when halite forms from solutions containing more than 0.8 mg Mn/L and more than 1 mg Pb/L. The presence of lead as co-activator is necessary requirement: Mn alone does not activate the fluorescence of halite. This is in accordance with the results of previous work (Murata et al., 1946; Sidike et al., 2002). Rock salt in the mine does not show fluorescence at all. Fluorescence of contemporary salt in Bochnia salt mine is a result of mining activity and slight, sporadic contamination with traces of Mn and Pb. This work is partially funded by AGH research grant no 11.11.140.319. Murata K. J., Smith R. L., 1946. Manganese and lead as coactivators of red fluorescence in halite, American Mineralogist, Volume 31, pages 527

  20. Biological applications of confocal fluorescence polarization microscopy

    Science.gov (United States)

    Bigelow, Chad E.

    Fluorescence polarization microscopy is a powerful modality capable of sensing changes in the physical properties and local environment of fluorophores. In this thesis we present new applications for the technique in cancer diagnosis and treatment and explore the limits of the modality in scattering media. We describe modifications to our custom-built confocal fluorescence microscope that enable dual-color imaging, optical fiber-based confocal spectroscopy and fluorescence polarization imaging. Experiments are presented that indicate the performance of the instrument for all three modalities. The limits of confocal fluorescence polarization imaging in scattering media are explored and the microscope parameters necessary for accurate polarization images in this regime are determined. A Monte Carlo routine is developed to model the effect of scattering on images. Included in it are routines to track the polarization state of light using the Mueller-Stokes formalism and a model for fluorescence generation that includes sampling the excitation light polarization ellipse, Brownian motion of excited-state fluorophores in solution, and dipole fluorophore emission. Results from this model are compared to experiments performed on a fluorophore-embedded polymer rod in a turbid medium consisting of polystyrene microspheres in aqueous suspension. We demonstrate the utility of the fluorescence polarization imaging technique for removal of contaminating autofluorescence and for imaging photodynamic therapy drugs in cell monolayers. Images of cells expressing green fluorescent protein are extracted from contaminating fluorescein emission. The distribution of meta-tetrahydroxypheny1chlorin in an EMT6 cell monolayer is also presented. A new technique for imaging enzyme activity is presented that is based on observing changes in the anisotropy of fluorescently-labeled substrates. Proof-of-principle studies are performed in a model system consisting of fluorescently labeled bovine

  1. Ratiometric fluorescent pH nanoprobes based on in situ assembling of fluorescence resonance energy transfer between fluorescent proteins.

    Science.gov (United States)

    Yu, Haijun; Chen, Chao; Cao, Xiaodan; Liu, Yueling; Zhou, Shengmin; Wang, Ping

    2017-08-01

    pH-dependent protein adsorption on mesoporous silica nanoparticle (MSN) was examined as a unique means for pH monitoring. Assuming that the degree of protein adsorption determines the distance separating protein molecules, we examined the feasibility of nanoscale pH probes based on fluorescence resonance energy transfer (FRET) between two fluorescent proteins (mTurquoise2 and mNeonGreen, as donor and acceptor, respectively). Since protein adsorption on MSN is pH-sensitive, both fluorescent proteins were modified to make their isoelectric points (pIs) identical, thus achieving comparable adsorption between the proteins and enhancing FRET signals. The adsorption behaviors of such modified fluorescent proteins were examined along with ratiometric FRET signal generation. Results demonstrated that the pH probes could be manipulated to show feasible sensitivity and selectivity for pH changes in hosting solutions, with a good linearity observed in the pH range of 5.5-8.0. In a demonstration test, the pH probes were successfully applied to monitor progress of enzymatic reactions. Such an "in situ-assembling" pH sensor demonstrates a promising strategy in developing nanoscale fluorescent protein probes. Graphical abstract Working principle of the developed pH sensor TNS; and FRET Ratio (I528/I460) as a function of pH under different protein feed ratios (mNeonGreen to mTurquoise2).

  2. Interconversion of Anthozoa GFP-like fluorescent and non-fluorescent proteins by mutagenesis

    Directory of Open Access Journals (Sweden)

    Mudrik Nikolay N

    2002-04-01

    Full Text Available Abstract Background Within the family of green fluorescent protein (GFP homologs, one can mark two main groups, specifically, fluorescent proteins (FPs and non-fluorescent or chromoproteins (CPs. Structural background of differences between FPs and CPs are poorly understood to date. Results Here, we applied site-directed and random mutagenesis in order to to transform CP into FP and vice versa. A purple chromoprotein asCP (asFP595 from Anemonia sulcata and a red fluorescent protein DsRed from Discosoma sp. were selected as representatives of CPs and FPs, respectively. For asCP, some substitutions at positions 148 and 165 (numbering in accordance to GFP were found to dramatically increase quantum yield of red fluorescence. For DsRed, substitutions at positions 148, 165, 167, and 203 significantly decreased fluorescence intensity, so that the spectral characteristics of these mutants became more close to those of CPs. Finally, a practically non-fluorescent mutant DsRed-NF was generated. This mutant carried four amino acid substitutions, specifically, S148C, I165N, K167M, and S203A. DsRed-NF possessed a high extinction coefficient and an extremely low quantum yield ( Conclusions We located a novel point in asCP sequence (position 165 mutations at which can result in red fluorescence appearance. Probably, this finding could be applied onto other CPs to generate red and far-red fluorescent mutants. A possibility to transform an FP into CP was demonstrated. Key role of residues adjacent to chromophore's phenolic ring in fluorescent/non-fluorescent states determination was revealed.

  3. Fluorescent Nanoparticle Uptake for Brain Tumor Visualization

    Directory of Open Access Journals (Sweden)

    Rachel Tréhin

    2006-04-01

    Full Text Available Accurate delineation of tumor margins is vital to the successful surgical resection of brain tumors. We have previously developed a multimodal nanoparticle CLIO-Cy5.5, which is detectable by both magnetic resonance imaging and fluorescence, to assist in intraoperatively visualizing tumor boundaries. Here we examined the accuracy of tumor margin determination of orthotopic tumors implanted in hosts with differing immune responses to the tumor. Using a nonuser-based signal intensity method applied to fluorescent micrographs of 9L gliosarcoma green fluorescent protein (GFP tumors, mean overestimations of 2 and 24 µm were obtained using Cy5.5 fluorescence, compared to the true tumor margin determined by GFP fluorescence, in nude mice and rats, respectively. To resolve which cells internalized the nanoparticle and to quantitate degree of uptake, tumors were disaggregated and cells were analyzed by flow cytometry and fluorescence microscopy. Nanoparticle uptake was seen in both CD11b+ cells (representing activated microglia and macrophages and tumor cells in both animal models by both methods. CD11b+ cells were predominantly found at the tumor margin in both hosts, but were more pronounced at the margin in the rat model. Additional metastatic (CT26 colon and primary (Gli36 glioma brain tumor models likewise demonstrated that the nanoparticle was internalized both by tumor cells and by host cells. Together, these observations suggest that fluorescent nanoparticles provide an accurate method of tumor margin estimation based on a combination of tumor cell and host cell uptake for primary and metastatic tumors in animal model systems and offer potential for clinical translation.

  4. Recent Progress on Plasmon-Enhanced Fluorescence

    Directory of Open Access Journals (Sweden)

    Dong Jun

    2015-12-01

    Full Text Available The optically generated collective electron density waves on metal–dielectric boundaries known as surface plasmons have been of great scientific interest since their discovery. Being electromagnetic waves on gold or silver nanoparticle’s surface, localised surface plasmons (LSP can strongly enhance the electromagnetic field. These strong electromagnetic fields near the metal surfaces have been used in various applications like surface enhanced spectroscopy (SES, plasmonic lithography, plasmonic trapping of particles, and plasmonic catalysis. Resonant coupling of LSPs to fluorophore can strongly enhance the emission intensity, the angular distribution, and the polarisation of the emitted radiation and even the speed of radiative decay, which is so-called plasmon enhanced fluorescence (PEF. As a result, more and more reports on surface-enhanced fluorescence have appeared, such as SPASER-s, plasmon assisted lasing, single molecule fluorescence measurements, surface plasmoncoupled emission (SPCE in biological sensing, optical orbit designs etc. In this review, we focus on recent advanced reports on plasmon-enhanced fluorescence (PEF. First, the mechanism of PEF and early results of enhanced fluorescence observed by metal nanostructure will be introduced. Then, the enhanced substrates, including periodical and nonperiodical nanostructure, will be discussed and the most important factor of the spacer between molecule and surface and wavelength dependence on PEF is demonstrated. Finally, the recent progress of tipenhanced fluorescence and PEF from the rare-earth doped up-conversion (UC and down-conversion (DC nanoparticles (NPs are also commented upon. This review provides an introduction to fundamentals of PEF, illustrates the current progress in the design of metallic nanostructures for efficient fluorescence signal amplification that utilises propagating and localised surface plasmons.

  5. Recent Progress on Plasmon-Enhanced Fluorescence

    Science.gov (United States)

    Dong, Jun; Zhang, Zhenglong; Zheng, Hairong; Sun, Mentao

    2015-12-01

    The optically generated collective electron density waves on metal-dielectric boundaries known as surface plasmons have been of great scientific interest since their discovery. Being electromagnetic waves on gold or silver nanoparticle's surface, localised surface plasmons (LSP) can strongly enhance the electromagnetic field. These strong electromagnetic fields near the metal surfaces have been used in various applications like surface enhanced spectroscopy (SES), plasmonic lithography, plasmonic trapping of particles, and plasmonic catalysis. Resonant coupling of LSPs to fluorophore can strongly enhance the emission intensity, the angular distribution, and the polarisation of the emitted radiation and even the speed of radiative decay, which is so-called plasmon enhanced fluorescence (PEF). As a result, more and more reports on surface-enhanced fluorescence have appeared, such as SPASER-s, plasmon assisted lasing, single molecule fluorescence measurements, surface plasmoncoupled emission (SPCE) in biological sensing, optical orbit designs etc. In this review, we focus on recent advanced reports on plasmon-enhanced fluorescence (PEF). First, the mechanism of PEF and early results of enhanced fluorescence observed by metal nanostructure will be introduced. Then, the enhanced substrates, including periodical and nonperiodical nanostructure, will be discussed and the most important factor of the spacer between molecule and surface and wavelength dependence on PEF is demonstrated. Finally, the recent progress of tipenhanced fluorescence and PEF from the rare-earth doped up-conversion (UC) and down-conversion (DC) nanoparticles (NPs) are also commented upon. This review provides an introduction to fundamentals of PEF, illustrates the current progress in the design of metallic nanostructures for efficient fluorescence signal amplification that utilises propagating and localised surface plasmons.

  6. 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.

  7. A highly selective and sensitive fluorescent chemosensor for Zn2+

    Institute of Scientific and Technical Information of China (English)

    Xiu Ying Zhang; Zuo Hui Wang; Lin Yang

    2008-01-01

    A new selective Zn2+ fluorescent chemosensor,o-vanillin-4-ethoxybenzoylhydrazone(1),was designed and prepared.Free 1 mainly displayed very weak fluorescence at 480 nm upone xcitation at 403 nln.It displayed high selectivity for Zn2+ and had a 518-fold fluorescent enhancement upon binding of Zn2+.while the other cation ions had only little influence on the fluorescence of 1.Mechanism of enhancement of 1's fluorescence by Zn2+ was briefly discussed.

  8. Visualizing Fluorescence: Using a Homemade Fluorescence "Microscope" to View Latent Fingerprints on Paper

    Science.gov (United States)

    LaFratta, Christopher N.; Huh, Sun Phill; Mallillin, Allistair C.; Riviello, Peter J.; Walt, David R.

    2010-01-01

    We describe an inexpensive hand-held fluorescence imager (low-magnification microscope), constructed from poly(vinyl chloride) pipe and other inexpensive components for use as a teaching tool to understand the principles of fluorescence detection. Optical filters are used to select the excitation and emission wavelengths and can be easily…

  9. 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

  10. Sensitive turn-on fluorescent detection of tartrazine based on fluorescence resonance energy transfer.

    Science.gov (United States)

    Huang, Sheng Tian; Shi, Yan; Li, Nian Bing; Luo, Hong Qun

    2012-01-18

    We introduce a sensitive, rapid, label-free and general fluorescent method for the determination of tartrazine by competitive binding to reduced graphene oxide (rGO) against fluorescein, and the fluorescence recovery upon fluorescein desorption from rGO provides a quantitative readout for tartrazine, giving a detection limit of 0.53 ng mL(-1).

  11. 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

  12. Is the flower fluorescence relevant in biocommunication?

    Science.gov (United States)

    Iriel, Analía; Lagorio, María Gabriela

    2010-10-01

    Flower fluorescence has been previously proposed as a potential visual signal to attract pollinators. In this work, this point was addressed by quantitatively measuring the fluorescence quantum yield ( Φ f) for flowers of Bellis perennis (white, yellow, pink, and purple), Ornithogalum thyrsoides (petals and ovaries), Limonium sinuatum (white and yellow), Lampranthus productus (yellow), Petunia nyctaginiflora (white), Bougainvillea spectabilis (white and yellow), Antirrhinum majus (white and yellow), Eustoma grandiflorum (white and blue), Citrus aurantium (petals and stigma), and Portulaca grandiflora (yellow). The highest values were obtained for the ovaries of O. thyrsoides ( Φ f = 0.030) and for Citrus aurantium petals ( Φ f = 0.014) and stigma ( Φ f = 0.013). Emitted photons as fluorescence were compared with reflected photons. It was concluded that the fluorescence emission is negligible compared to the reflected light, even for the most fluorescent samples, and it may not be considered as an optical signal in biocommunication. The work was complemented with the calculation of quantum catches for each studied flower species to describe the visual sensitization of eye photoreceptors.

  13. Fabrication of fluorescent chitosan-containing microcapsules

    Directory of Open Access Journals (Sweden)

    Zhang R.

    2013-08-01

    Full Text Available Intense emission peaks of Eu(DBM3Phen (DBM and Phen are dibenzoylmethane and 1,10-phenanthroline, respectively in the microcapsules containing molecules of quaternary ammonium chitosan (QACS and sodium alginate are observed. The microcapsules are assembled by using CaCO3 particles as template cores by the layer-by-layer (LbL technique. Observation of microcapsules by the fluorescence mode and the transmission mode in the confocal laser scanning microscopy shows that the microcapsules are intact after core decomposition. Fluorescence under ultraviolet irradiation comes directly from the Eu(DBM3Phen. Homogeneous assembly of Eu(DBM3Phen can be deduced due to the homogeneous fluorescence of the microcapsules in the fluorescence micrographs. The microcapsules show adherence to solid substrates due to large quantities of hydroxyl groups of QACS. AFM measurements of dried hollow microcapsules with only 4 bilayers of (CS/SA fabricated with Eu(DBM3Phen show the intact shell with a thickness of 3.0 nm. Regarding the biocompatible natural polysaccharides and the intense fluorescence emission, the microcapsules in this work might be of great importance in potential application in drug delivery and bioassay.

  14. Preclinical fluorescent mouse models of pancreatic cancer

    Science.gov (United States)

    Bouvet, Michael; Hoffman, Robert M.

    2007-02-01

    Here we describe our cumulative experience with the development and preclinical application of several highly fluorescent, clinically-relevant, metastatic orthotopic mouse models of pancreatic cancer. These models utilize the human pancreatic cancer cell lines which have been genetically engineered to selectively express high levels of the bioluminescent green fluorescent (GFP) or red fluorescent protein (RFP). Fluorescent tumors are established subcutaneously in nude mice, and tumor fragments are then surgically transplanted onto the pancreas. Locoregional tumor growth and distant metastasis of these orthotopic implants occurs spontaneously and rapidly throughout the abdomen in a manner consistent with clinical human disease. Highly specific, high-resolution, real-time visualization of tumor growth and metastasis may be achieved in vivo without the need for contrast agents, invasive techniques, or expensive imaging equipment. We have shown a high correlation between florescent optical imaging and magnetic resonance imaging in these models. Alternatively, transplantation of RFP-expressing tumor fragments onto the pancreas of GFP-expressing transgenic mice may be used to facilitate visualization of tumor-host interaction between the pancreatic tumor fragments and host-derived stroma and vasculature. Such in vivo models have enabled us to serially visualize and acquire images of the progression of pancreatic cancer in the live animal, and to demonstrate the real-time antitumor and antimetastatic effects of several novel therapeutic strategies on pancreatic malignancy. These fluorescent models are therefore powerful and reliable tools with which to investigate human pancreatic cancer and therapeutic strategies directed against it.

  15. Hitherto Unrecognized Fluorescence Properties of Coniferyl Alcohol

    Directory of Open Access Journals (Sweden)

    Anup Kumar Singh

    2010-03-01

    Full Text Available We instituted a quasi-quality assurance program for demonstrating coniferyl alcohol’s fluorescence and fluorescence diminishment following enzymatic oxidation. The magnitude of diminishment was a measure of catalysis. High throughput screening was performed in pseudo-kinetic and endpoint modes by measuring the fluorescence at 416 nm following excitation at 290, 310 or 340 nm. Dose-response tracings were linear between two and three orders of magnitude with average limits of detection and quantitation of 1.8 and 6.9 mM coniferyl alcohol, respectively. Oxidation was evident with 0.025 mg/mL laccase or 0.003 mg/mL peroxidase or inside 5 min using 0.5 mg/mL laccase or 5 mM substrate. Sodium chloride inhibited (IC50, 25 mM laccase oxidation of coniferyl alcohol. Fluorescence from 10 concentrations (1 to 1000 mM of coniferyl alcohol was stable for 24 hours over 14 excitation/emission cycles at 3 different combinations of excitation and emission wavelengths. In conclusion, coniferyl alcohol absorption and fluorescence assays should facilitate biomass lignin analyses and improve delignification.

  16. Use of astronomy filters in fluorescence microscopy.

    Science.gov (United States)

    Piper, Jörg

    2012-02-01

    Monochrome astronomy filters are well suited for use as excitation or suppression filters in fluorescence microscopy. Because of their particular optical design, such filters can be combined with standard halogen light sources for excitation in many fluorescent probes. In this "low energy excitation," photobleaching (fading) or other irritations of native specimens are avoided. Photomicrographs can be taken from living motile fluorescent specimens also with a flash so that fluorescence images can be created free from indistinctness caused by movement. Special filter cubes or dichroic mirrors are not needed for our method. By use of suitable astronomy filters, fluorescence microscopy can be carried out with standard laboratory microscopes equipped with condensers for bright-field (BF) and dark-field (DF) illumination in transmitted light. In BF excitation, the background brightness can be modulated in tiny steps up to dark or black. Moreover, standard industry microscopes fitted with a vertical illuminator for examinations of opaque probes in DF or BF illumination based on incident light (wafer inspections, for instance) can also be used for excitation in epi-illumination when adequate astronomy filters are inserted as excitatory and suppression filters in the illuminating and imaging light path. In all variants, transmission bands can be modulated by transmission shift.

  17. S - and N-alkylating agents diminish the fluorescence of fluorescent dye-stained DNA.

    Science.gov (United States)

    Giesche, Robert; John, Harald; Kehe, Kai; Schmidt, Annette; Popp, Tanja; Balzuweit, Frank; Thiermann, Horst; Gudermann, Thomas; Steinritz, Dirk

    2017-01-25

    Sulfur mustard (SM), a chemical warfare agent, causes DNA alkylation, which is believed to be the main cause of its toxicity. SM DNA adducts are commonly used to verify exposure to this vesicant. However, the required analytical state-of-the-art mass-spectrometry methods are complex, use delicate instruments, are not mobile, and require laboratory infrastructure that is most likely not available in conflict zones. Attempts have thus been made to develop rapid detection methods that can be used in the field. The analysis of SM DNA adducts (HETE-G) by immunodetection is a convenient and suitable method. For a diagnostic assessment, HETE-G levels must be determined in relation to the total DNA in the sample. Total DNA can be easily visualized by the use of fluorescent DNA dyes. This study examines whether SM and related compounds affect total DNA staining, an issue that has not been investigated before. After pure DNA was extracted from human keratinocytes (HaCaT cells), DNA was exposed to different S- and N-alkylating agents. Our experiments revealed a significant, dose-dependent decrease in the fluorescence signal of fluorescent dye-stained DNA after exposure to alkylating agents. After mass spectrometry and additional fluorescence measurements ruled out covalent modifications of ethidium bromide (EthBr) by SM, we assumed that DNA crosslinks caused DNA condensation and thereby impaired access of the fluorescent dyes to the DNA. DNA digestion by restriction enzymes restored fluorescence, a fact that strengthened our hypothesis. However, monofunctional agents, which are unable to crosslink DNA, also decreased the fluorescence signal. In subsequent experiments, we demonstrated that protons produced during DNA alkylation caused a pH decrease that was found responsible for the reduction in fluorescence. The use of an appropriate buffer system eliminated the adverse effect of alkylating agents on DNA staining with fluorescent dyes. An appropriate buffer system is thus

  18. Fluorescent multiplex cell flow systems and methods

    KAUST Repository

    Merzaban, Jasmeen

    2017-06-01

    Systems and methods are provided for simultaneously assaying cell adhesion or cell rolling for multiple cell specimens. One embodiment provides a system for assaying adhesion or cell rolling of multiple cell specimens that includes a confocal imaging system containing a parallel plate flow chamber, a pump in fluid communication with the parallel plate flow chamber via a flow chamber inlet line and a cell suspension in fluid communication with the parallel plate flow chamber via a flow chamber outlet line. The system also includes a laser scanning system in electronic communication with the confocal imaging system, and a computer in communication with the confocal imaging system and laser scanning system. In certain embodiments, the laser scanning system emits multiple electromagnetic wavelengths simultaneously it cause multiple fluorescent labels having different excitation wavelength maximums to fluoresce. The system can simultaneously capture real-time fluorescence images from at least seven cell specimens in the parallel plate flow chamber.

  19. 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

  20. Fluorescence energy transfer enhancement in aluminum nanoapertures

    CERN Document Server

    de Torres, Juan; Moparthi, Satish Babu; Grigoriev, Victor; Wenger, Jérome

    2015-01-01

    Zero-mode waveguides (ZMWs) are confining light into attoliter volumes, enabling single molecule fluorescence experiments at physiological micromolar concentrations. Among the fluorescence spectroscopy techniques that can be enhanced by ZMWs, F\\"{o}rster resonance energy transfer (FRET) is one of the most widely used in life sciences. Combining zero-mode waveguides with FRET provides new opportunities to investigate biochemical structures or follow interaction dynamics at micromolar concentration with single molecule resolution. However, prior to any quantitative FRET analysis on biological samples, it is crucial to establish first the influence of the ZMW on the FRET process. Here, we quantify the FRET rates and efficiencies between individual donor-acceptor fluorophore pairs diffusing in aluminum zero-mode waveguides. Aluminum ZMWs are important structures thanks to their commercial availability and the large literature describing their use for single molecule fluorescence spectroscopy. We also compare the ...

  1. 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.

  2. Oligothiophenes as Fluorescent Markers for Biological Applications

    Directory of Open Access Journals (Sweden)

    Antonio Manetto

    2012-01-01

    Full Text Available This paper summarizes some of our results on the application of oligothiophenes as fluorescent markers for biological studies. The oligomers of thiophene, widely known for their semiconductor properties in organic electronics, are also fluorescent compounds characterized by chemical and optical stability, high absorbance and quantum yield. Their fluorescent emission can be easily modulated via organic synthesis by changing the number of thiophene rings and the nature of side-chains. This review shows how oligothiophenes can be derivatized with active groups such as phosphoramidite, N-hydroxysuccinimidyl and 4-sulfotetrafluorophenyl esters, isothiocyanate and azide by which the (biomolecules of interest can be covalently bound. This paper also describes how molecules such as oligonucleotides, proteins and even nanoparticles, tagged with oligothiophenes, can be used in experiments ranging from hybridization studies to imaging of fixed and living cells. Finally, a few multilabeling experiments are described.

  3. Coupled external cavity photonic crystal enhanced fluorescence.

    Science.gov (United States)

    Pokhriyal, Anusha; Lu, Meng; Ge, Chun; Cunningham, Brian T

    2014-05-01

    We report a fundamentally new approach to enhance fluorescence in which surface adsorbed fluorophore-tagged biomolecules are excited on a photonic crystal surface that functions as a narrow bandwidth and tunable mirror of an external cavity laser. This scheme leads to ∼10× increase in the electromagnetic enhancement factor compared to ordinary photonic crystal enhanced fluorescence. In our experiments, the cavity automatically tunes its lasing wavelength to the resonance wavelength of the photonic crystal, ensuring optimal on-resonance coupling even in the presence of variable device parameters and variations in the density of surface-adsorbed capture molecules. We achieve ∼10(5) × improvement in the limit of detection of a fluorophore-tagged protein compared to its detection on an unpatterned glass substrate. The enhanced fluorescence signal and easy optical alignment make cavity-coupled photonic crystals a viable approach for further reducing detection limits of optically-excited light emitters that are used in biological assays.

  4. Tailored nanoporous gold for ultrahigh fluorescence enhancement.

    Science.gov (United States)

    Lang, X Y; Guan, P F; Fujita, T; Chen, M W

    2011-03-07

    We report molecular fluorescence enhancement of free-standing nanoporous gold in which the nanoporosity can be arbitrarily tailored by the combination of dealloying and electroless gold plating. The nanoporous gold fabricated by this facile method possesses unique porous structures with large gold ligaments and very small pores, and exhibits significant improvements in surface enhanced fluorescence as well as structure rigidity. It demonstrates that the confluence effect of improved quantum yield and excitation of fluorophores is responsible for the large fluorescence enhancement due to the near-field enhancement of nanoporous gold, which arises from the strong electromagnetic coupling between neighboring ligaments and the weakening of plasmon damping of the large ligaments because of the small pore size and large ligament size, respectively.

  5. Adaptive Optics for Fluorescence Correlation Spectroscopy

    CERN Document Server

    Leroux, Charles Edouard; Derouard, Jacques; Delon, Antoine

    2011-01-01

    Fluorescence Correlation Spectroscopy (FCS) yields measurement parameters (number of molecules, diffusion time) that characterize the concentration and kinetics of fluorescent molecules within a supposedly known observation volume. Absolute derivation of concentrations and diffusion constants therefore requires preliminary calibrations of the confocal Point Spread Function with phantom solutions under perfectly controlled environmental conditions. In this paper, we quantify the influence of optical aberrations on single photon FCS and demonstrate a simple Adaptive Optics system for aberration correction. Optical aberrations are gradually introduced by focussing the excitation laser beam at increasing depths in fluorescent solutions with various refractive indices, which leads to drastic depth-dependent bias in the estimated FCS parameters. Aberration correction with a Deformable Mirror stabilizes these parameters within a range of several tens of \\mum into the solution. We also demonstrate, both theoretically...

  6. Fluorescent protein integrated white LEDs for displays

    Science.gov (United States)

    Press, Daniel Aaron; Melikov, Rustamzhon; Conkar, Deniz; Nur Firat-Karalar, Elif; Nizamoglu, Sedat

    2016-11-01

    The usage time of displays (e.g., TVs, mobile phones, etc) is in general shorter than their functional life time, which worsens the electronic waste (e-waste) problem around the world. The integration of biomaterials into electronics can help to reduce the e-waste problem. In this study, we demonstrate fluorescent protein integrated white LEDs to use as a backlight source for liquid crystal (LC) displays for the first time. We express and purify enhanced green fluorescent protein (eGFP) and monomeric Cherry protein (mCherry), and afterward we integrate these proteins as a wavelength-converter on a blue LED chip. The protein-integrated backlight exhibits a high luminous efficacy of 248 lm/Wopt and the area of the gamut covers 80% of the NTSC color gamut. The resultant colors and objects in the image on the display can be well observed and distinguished. Therefore, fluorescent proteins show promise for display applications.

  7. 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.

  8. Plasmonically amplified fluorescence bioassay with microarray format

    Science.gov (United States)

    Gogalic, S.; Hageneder, S.; Ctortecka, C.; Bauch, M.; Khan, I.; Preininger, Claudia; Sauer, U.; Dostalek, J.

    2015-05-01

    Plasmonic amplification of fluorescence signal in bioassays with microarray detection format is reported. A crossed relief diffraction grating was designed to couple an excitation laser beam to surface plasmons at the wavelength overlapping with the absorption and emission bands of fluorophore Dy647 that was used as a label. The surface of periodically corrugated sensor chip was coated with surface plasmon-supporting gold layer and a thin SU8 polymer film carrying epoxy groups. These groups were employed for the covalent immobilization of capture antibodies at arrays of spots. The plasmonic amplification of fluorescence signal on the developed microarray chip was tested by using interleukin 8 sandwich immunoassay. The readout was performed ex situ after drying the chip by using a commercial scanner with high numerical aperture collecting lens. Obtained results reveal the enhancement of fluorescence signal by a factor of 5 when compared to a regular glass chip.

  9. UV fluorescence lidar detection of bioaerosols

    Science.gov (United States)

    Christesen, Steven D.; Merrow, Clifton N.; Desha, Michael S.; Wong, Anna; Wilson, Mark W.; Butler, John C.

    1994-06-01

    A UV fluorescence lidar system for the remote detection of bioaerosols has been built and tested. At the heart of the UV- LIDAR Fluorosensor system are a 200 mJ quadrupled Nd:YAG laser at 266 nm and a 16-inch Cassagrain telescope. Operating on three data collection channels, the UV lidar is capable of real time monitoring of 266 nm elastic backscatter, the total fluorescence between 300 and 400 nm, and the dispersed fluorescence spectrum (using a small spectrograph and gated intensified CCD array). Our goal in this effort was to assess the capabilities of biofluorescence for quantitative detection and discrimination of bioaerosols. To this end, the UV-LIDAR Fluorosensor system was tested against the aerosolized bacterial spore Bacillus subtilus var. niger sp. globiggi (BG) and several likely interferences at several ranges from approximately 600 to 3000 m. Our tests with BG indicate a detection limit of approximately 500 mg/cubic meter at a range of 3000 m.

  10. 荧光原位杂交技术在膀胱尿路上皮癌诊断中的应用价值%Utility of fluorescence in situ hybridization for the diagnosis of bladder urothelial carcinoma

    Institute of Scientific and Technical Information of China (English)

    黄建文; 安瑞华; 李运伟; 穆家贵; 甘秀国; 于雷; 原劲杨

    2012-01-01

    目的 探讨荧光原位杂交技术(fluorescence in situ hybridization,FISH)在膀胱尿路上皮癌诊断中的应用价值. 方法 收集81例膀胱尿路上皮癌患者的新鲜尿液标本,分别行FISH和尿细胞学检测,并行膀胱镜检查;病理诊断泌尿系良性疾病12例合并8例临床诊断泌尿系良性疾病患者的尿液标本作对照.统计学分析FISH和尿细胞学检测诊断的特征值. 结果 81例均经病理检查确诊膀胱尿路上皮癌.非肌层浸润性膀胱尿路上皮癌34例,肌层浸润性癌14例;低级别42例,高级别24例;由于送检组织不完整,无法准确分期、分级者分别为33例和15例.FISH、尿细胞学和膀胱镜检查诊断膀胱尿路上皮癌的敏感性分别为72.8% (59/81) 、27.2% (22/81)和97.5% (79/81),FISH诊断敏感性高于尿细胞学检测(P<0.05),但低于膀胱镜检查(P <0.05);FISH和尿细胞学检测诊断特异性分别为85.0%(17/20)、100.0% (20/20),二者比较差异无统计学意义(P>0.05).FISH和尿细胞学检测诊断符合率分别为75.2% (76/101)、41.6%(42/101),二者比较差异有统计学意义(P<0.05). 结论 FISH检测诊断膀胱尿路上皮癌敏感性高、特异性强、无创,临床应用价值高.%Objective To evaluate the clinical value of fluorescence in situ hybridization (FISH) method for diagnosis of bladder urothelial carcinoma.Methods Urine samples from 81 patients suspected of having bladder urothelial carcinoma were collected for immediate urine cytology and FISH analysis.All patients underwent cystoscopy for identification of bladder lesions.Urine samples from 8 patients with benign disease of urinary system werealso analyzed by means of FISH and cytology.The sensitivity and specificity of FISH were compared with cytology.Results 81 subjects of bladder urothelial carcinoma were verified with pathology,which included 34 non-muscle invasive carcinoma,14 cases with muscle invasive carcinoma,42 cases with high

  11. Diversity and evolution of coral fluorescent proteins.

    Directory of Open Access Journals (Sweden)

    Naila O Alieva

    Full Text Available GFP-like fluorescent proteins (FPs are the key color determinants in reef-building corals (class Anthozoa, order Scleractinia and are of considerable interest as potential genetically encoded fluorescent labels. Here we report 40 additional members of the GFP family from corals. There are three major paralogous lineages of coral FPs. One of them is retained in all sampled coral families and is responsible for the non-fluorescent purple-blue color, while each of the other two evolved a full complement of typical coral fluorescent colors (cyan, green, and red and underwent sorting between coral groups. Among the newly cloned proteins are a "chromo-red" color type from Echinopora forskaliana (family Faviidae and pink chromoprotein from Stylophora pistillata (Pocilloporidae, both evolving independently from the rest of coral chromoproteins. There are several cyan FPs that possess a novel kind of excitation spectrum indicating a neutral chromophore ground state, for which the residue E167 is responsible (numeration according to GFP from A. victoria. The chromoprotein from Acropora millepora is an unusual blue instead of purple, which is due to two mutations: S64C and S183T. We applied a novel probabilistic sampling approach to recreate the common ancestor of all coral FPs as well as the more derived common ancestor of three main fluorescent colors of the Faviina suborder. Both proteins were green such as found elsewhere outside class Anthozoa. Interestingly, a substantial fraction of the all-coral ancestral protein had a chromohore apparently locked in a non-fluorescent neutral state, which may reflect the transitional stage that enabled rapid color diversification early in the history of coral FPs. Our results highlight the extent of convergent or parallel evolution of the color diversity in corals, provide the foundation for experimental studies of evolutionary processes that led to color diversification, and enable a comparative analysis of

  12. Fluorescent DNA Stabilized Silver Nanoclusters as Biosensors

    Directory of Open Access Journals (Sweden)

    Alfonso Latorre

    2013-01-01

    Full Text Available DNA stabilized fluorescent silver nanoclusters are promising materials, of which fluorescent properties can be exploited to develop sensors. Particularly, the presence of a DNA strand in the structure has promoted the development of gene sensors where one part of the sensor is able to recognize the target gene sequence. Moreover, since oligonucleotides can be designed to have binding properties (aptamers a variety of sensors for proteins and cells have been developed using silver nanoclusters. In this review the applications of this material as sensors of different biomolecules are summarized.

  13. Materials for incandescent and fluorescent lamps

    DEFF Research Database (Denmark)

    Thorsen, Knud Aage

    1996-01-01

    The article gives an overview of the materials systems used for incandescent lamps as well as a brief introduction to the systems used for fluorescent lamps. The materials used for incandescent lamps are doped tungsten used for the filaments, metals and alloys used for terminal and support posts......, lead wires and internal reflectors and screens as well as glasses for the envelope. The physics of bulbs and changes in bulbs during use are elucidated. The cost and energy savings and environmental benefits by replacement of incandescent lamps by fluorescent lamps are presented....

  14. Robust, directed assembly of fluorescent nanodiamonds.

    Science.gov (United States)

    Kianinia, Mehran; Shimoni, Olga; Bendavid, Avi; Schell, Andreas W; Randolph, Steven J; Toth, Milos; Aharonovich, Igor; Lobo, Charlene J

    2016-10-27

    Arrays of fluorescent nanoparticles are highly sought after for applications in sensing, nanophotonics and quantum communications. Here we present a simple and robust method of assembling fluorescent nanodiamonds into macroscopic arrays. Remarkably, the yield of this directed assembly process is greater than 90% and the assembled patterns withstand ultra-sonication for more than three hours. The assembly process is based on covalent bonding of carboxyl to amine functional carbon seeds and is applicable to any material, and to non-planar surfaces. Our results pave the way to directed assembly of sensors and nanophotonics devices.

  15. Robust, directed assembly of fluorescent nanodiamonds

    CERN Document Server

    Kianinia, Mehran; Shimoni, Olga; Randolph, Steven J; Toth, Milos; Aharonovich, Igor; Lobo, Charlene J

    2016-01-01

    Arrays of fluorescent nanoparticles are highly sought after for applications in sensing and nanophotonics. Here we present a simple and robust method of assembling fluorescent nanodiamonds into macroscopic arrays. Remarkably, the yield of this directed assembly process is greater than 90% and the assembled patterns withstand ultra-sonication for more than three hours. The assembly process is based on covalent bonding of carboxyl to amine functional carbon seeds and is applicable to any material, and to non-planar surfaces. Our results pave the way to directed assembly of sensing and nanophotonics devices.

  16. A highly selective fluorescent sensor for glucosamine.

    Science.gov (United States)

    Tran, Tam Minh; Alan, Yuksel; Glass, Timothy Edward

    2015-05-07

    A new fluorescent chemical sensor for glucosamine is reported. The sensor is based on a boronic acid-containing coumarin aldehyde and shows excellent selectivity for glucosamine by forming a boronic ester with the sugar diol as well as an iminium ion with the amine group of glucosamine. The sensor successfully discriminates glucosamine over other similar biomolecules in terms of both fluorescence intensity and binding affinity. This method provides a new concept for the design and synthesis of very selective turn-on optical sensors for selective detection of multi-functional biomolecules.

  17. Chlorophyll a fluorescence analysis in forests

    OpenAIRE

    M. Pollastrini; Holland, V; Brüggemann, W.; F. Bussotti

    2016-01-01

    A European-wide assessment of chlorophyll a fluorescence (ChlF, prompt fluorescence on dark-adapted samples) parameters in forest ecosystems was carried out in the years 2012-2013, within the 7FP FunDivEUROPE project. A total of 1596 trees growing in 209 stands distributed in six countries, from Mediterranean to boreal sites, were sampled. This paper shows the applicability of the ChlF in forest ecology surveys, the protocols adopted for leaf sampling and ChlF measurements, the variability of...

  18. CHLOROPHYLL a FLUORESCENCE ANALYSIS IN FORESTS

    OpenAIRE

    M. Pollastrini; Holland, V; Brüggemann, W.; F. Bussotti

    2016-01-01

    A European-wide assessment of chlorophyll a fluorescence (ChlF, prompt fluorescence on dark-adapted samples) parameters in forest ecosystems was carried out in the years 2012-2013, within the 7FP FunDivEUROPE project. A total of 1596 trees growing in 209 stands distributed in six countries, from Mediterranean to boreal sites, were sampled. This paper shows the applicability of the ChlF in forest ecology surveys, the protocols adopted for leaf sampling and ChlF measurements, the variability of...

  19. Analysis of Cholesterol Trafficking with Fluorescent Probes

    DEFF Research Database (Denmark)

    Maxfield, Frederick R.; Wustner, Daniel

    2012-01-01

    Cholesterol plays an important role in determining the biophysical properties of biological membranes, and its concentration is tightly controlled by homeostatic processes. The intracellular transport of cholesterol among organelles is a key part of the homeostatic mechanism, but sterol transport...... that can bind to cholesterol to reveal its distribution in cells. We also discuss the use of intrinsically fluorescent sterols that closely mimic cholesterol, as well as some minimally modified fluorophore-labeled sterols. Methods for imaging these sterols by conventional fluorescence microscopy...... and by multiphoton microscopy are described. Some label-free methods for imaging cholesterol itself are also discussed briefly....

  20. Aggregate Formed by a Cationic Fluorescence Probe

    Institute of Scientific and Technical Information of China (English)

    TIAN, Juan; SANG, Da-Yong; JI, Guo-Zhen

    2007-01-01

    The aggregation behavior of a cationic fluorescence probe 10-(4,7,10,13,16-pentaoxa-1-azacyclooctadecyl-methyl)anthracen-9-ylmethyl dodecanoate (1) was observed and studied by a fluorescence methodology in acidic and neutral conditions. By using the Py scale, differences between simple aggregates and micelles have been discussed. The stability of simple aggregates was discussed in terms of hydrophobic interaction and electrostatic repulsion. The absence of excimer emission of the anthrancene moiety of probe 1 in neutral condition was attributed to the photoinduced electron transfer mechanism instead of photodimerization.

  1. 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.

  2. Novel Terbium Chelate Doped Fluorescent Silica Nanoparticles

    Institute of Scientific and Technical Information of China (English)

    Ning Qiaoyu; Meng Jianxin; Wang Haiming; Liu Yingliang; Man Shiqing

    2006-01-01

    Novel terbium chelate doped silica fluorescent nanoparticles were prepared and characterized.The preparation was carried out in water-in-oil (W/O) microemulsion containing monomer precursor (pAB-DTPAA-APTEOS), Triton X-100, n-hexanol, and cyclohexane by controlling copolymerization of tetraethyl orthosilicate and 3-aminopropyl-triethyloxysilane.The nanoparticles are spherical and uniform in size, about 30 nm in diameter, strongly fluorescent, and highly stable.The amino groups directly introduced to the surface of the nanoparticles using APTEOS during preparation made the surface modification and bioconjugation of the nanoparticles easier.The nanoparticles are expected as an efficient time-resolved luminescence biological label.

  3. UroVysion fluorescence in situ hybridization for monitoring the recurrence of bladder cancer%UroVysion荧光原位杂交监测膀胱癌复发的研究

    Institute of Scientific and Technical Information of China (English)

    李连印; 姜君仪; 刘俊江; 李守宾; 高双友

    2011-01-01

    Objective UroVysion fluorescence in situ hybridization ( FISH) is already applied abroad to the early detection of bladder cancee. This study was to assess the value of UroVysion in the diagnosis of bladder cancer. Methods A total of 58 bladder cancer patients treated by transurethral resection of the bladder were included in this study. UroVysion FlSH, urine cytology and cystoscopy were used to monitor the recurrence of the disease, and the results were compared. Results Recurrence was found in 41 of the patients. The results for the multi-target DNA FISH assay were compared with independent analyses of urine cytology. Both FISH and urine cytolugy achieved a 100% specificity in the detection of the recurrence of bladder cancer, and the sensitivity of FISH for bladder cancer was significant higher than that of urine cytology ( 62. 1% vs 31% ) . In the FISH-positive cases , 17 were G3 tumor , 12 were G2 and 7 were G1, as compared with 15 cases of G3 and 3 cases of G2 in the urine cytology test. Conclusion UroVysion FISH, superior to cytology and cystoscopy in monitoring the recurrence of bladder cancer, has much prospect of clinical application.%目的 荧光原位杂交技术目前在国外已用于膀胱癌的早期诊断,但在国内尚未应用于临床,同时缺乏膀胱癌早期诊断价值的相关资料.文中评估UroVysion荧光原位杂交在膀胱癌复发检测中的价值.方法 收集接受经尿道膀胱肿瘤电切术的膀胱尿路上皮癌患者58例,分别采用膀胱镜、UroVysion荧光原位杂交和尿脱落细胞学方法检测膀胱癌复发,并进行独立对比分析.结果 对患者的检测结果发现,41例患者确诊为膀胱癌术后复发.荧光原位杂交和尿脱落细胞学检查对诊断膀胱癌复发特异性均为100%.荧光原位杂交检测敏感性为62.1%,明显高于尿脱落细胞学检测敏感性的31.0%,P<0.05.在荧光原位杂交检测阳性的病例中,17例为G3级肿瘤,12例为G2级肿瘤,7例为G1级

  4. A fluorescence switch based on a controllable photochromic naphthopyran group

    Energy Technology Data Exchange (ETDEWEB)

    Chen Lizhen [Faculty of Chemistry, Northeast Normal University, Changchun 130024 (China); Wang Guang, E-mail: wangg923@nenu.edu.cn [Faculty of Chemistry, Northeast Normal University, Changchun 130024 (China); Zhao Xiancai [Faculty of Chemistry, Northeast Normal University, Changchun 130024 (China)

    2011-08-15

    A fluorescence switch based on photoisomerization of naphthopyran (NP) has been designed by employing 2-(pyridin-2-yl)-benzimidazole (BPI) and the naphthopyran containing two pyran rings (NP) as fluorescent dye and photochromic compound, respectively. The fluorescence switch of benzimidazole derivative can be modulated either by controlling the irradiation time of UV light or by adjusting the amount ratio of fluorescent benzimidazole derivative to photochromic naphthopyran in both solution and polymethyl methacrylate (PMMA) film. The experimental results indicated that the decrease of fluorescence intensity of benzimidazole derivative is attributed to the interaction of benzimidazole with naphthopyran. - Highlights: > Naphthopyran was first used to fabricate fluorescence switch with benzimidazole derivative. > Fluorescence intensity can be modulated by controlling the UV irradiation time. > Fluorescence intensity can be adjusted by changing the ratio of benzimidazole derivative to naphthopyran. > Decrease of fluorescence intensity is attributed to the interaction of benzimidazole derivative and naphthopyran.

  5. A fluorescent chromatophore changes the level of fluorescence in a reef fish.

    Directory of Open Access Journals (Sweden)

    Matthias F Wucherer

    Full Text Available Body coloration plays a major role in fish ecology and is predominantly generated using two principles: a absorbance combined with reflection of the incoming light in pigment colors and b scatter, refraction, diffraction and interference in structural colors. Poikilotherms, and especially fishes possess several cell types, so-called chromatophores, which employ either of these principles. Together, they generate the dynamic, multi-color patterns used in communication and camouflage. Several chromatophore types possess motile organelles, which enable rapid changes in coloration. Recently, we described red fluorescence in a number of marine fish and argued that it may be used for private communication in an environment devoid of red. Here, we describe the discovery of a chromatophore in fishes that regulates the distribution of fluorescent pigments in parts of the skin. These cells have a dendritic shape and contain motile fluorescent particles. We show experimentally that the fluorescent particles can be aggregated or dispersed through hormonal and nervous control. This is the first description of a stable and natural cytoskeleton-related fluorescence control mechanism in vertebrate cells. Its nervous control supports suggestions that fluorescence could act as a context-dependent signal in some marine fish species and encourages further research in this field. The fluorescent substance is stable under different chemical conditions and shows no discernible bleaching under strong, constant illumination.

  6. Effect of Fluorescent Particle Size on the Modulation Efficiency of Ultrasound-Modulated Fluorescence

    Directory of Open Access Journals (Sweden)

    Yuan Liu

    2012-01-01

    Full Text Available To investigate whether the size of fluorescent particles affects the modulation efficiency of ultrasound-modulated fluorescence (UMF, we measured UMF and DC (direct current signals of the fluorescence emission from four different-sized fluorescent particles: (1 three carboxylate-modified fluorescent microspheres (FM with diameters of 20 nm, 200 nm, and 1.0 μm and (2 streptavidin-conjugated Alexa Fluor 647 with a diameter of approximately 5 nm. The UMF and DC signals were simultaneously measured using a broadband lock-in amplifier and a narrowband amplifier, respectively. The ratio of the UMF strength to the DC signal strength is defined as the modulation efficiency. This modulation efficiency was then used to evaluate the effects of fluorophore size and concentration. Results show that the modulation efficiency was improved by approximately a factor of two when the size of the fluorescent particles is increased from 5 nm to 1 μm. In addition, the linear relationship between the UMF strength and ultrasound pressure (observed in our previous study was maintained regardless of the fluorescent particle sizes.

  7. Let's Exploit Available Knowledge on Vegetation Fluorescence

    Science.gov (United States)

    Magnani, Federico; Raddi, Sabrina; Mohammed, Gina; Middleton, Elizabeth M.

    2014-01-01

    The potential to measure vegetation fluorescence from space (1) and to derive from it direct information on the gross primary productivity (GPP) of terrestrial ecosystems is probably the most thrilling development in remote sensing and global ecology of recent years, as it moves Earth observation techniques from the detection of canopy biophysics (e.g., fraction of absorbed radiation) and biochemistry (chlorophyll and nitrogen content) to the realm of ecosystem function. The existence of a functional relationship between fluorescence and photosynthesis has been elucidated over the last decade by several laboratories, notably as part of the preliminary studies of the European Space Agency Fluorescence Explorer (FLEX) Earth Explorer Mission. The empirical observation presented by Guanter et al. (2) of a linear relationship between fluorescence radiance and GPP, however, provides the first experimental confirmation of the feasibility of the approach— already thoroughly tested at leaf level—at the desired scale, despite the confounding effects associated with the satellite detection of such a faint signal. A word of clarification is needed here. The use of fluorescence as a probe of leaf photochemistry has been a staple of plant ecophysiology for decades, rooted in a sound understanding of photosynthetic energy dissipation. However, most past studies had to rely for the interpretation of results on active (pulse-saturated) techniques, making them unsuitable for remote-sensing applications. Over recent years, however, novel process based models have been developed for the interpretation of steady-state, solar-induced fluorescence at the leaf to canopy scale (3). We are therefore in a position to move beyond the mere empirical observation of an association between GPP and fluorescence radiance. In particular, Guanter et al. (2) base their analysis on the assumption of a constant ratio between photosynthetic and fluorescence light use efficiencies (equation 3 in ref

  8. Origin of tryptophan fluorescence lifetimes. Part 2: fluorescence lifetimes origin of tryptophan in proteins.

    Science.gov (United States)

    Albani, J R

    2014-01-01

    Fluorescence intensity decays of L-tryptophan in proteins dissolved in pH 7 buffer, in ethanol and in 6 M guanidine pH 7.8 and in lyophilized proteins were measured. In all protein conditions, three lifetimes were obtained along the emission spectrum (310-410 nm). The two shortest lifetimes are in the same range of those obtained for L-Trp in water or in ethanol. Thus, these two lifetimes originate from specific two sub-structures existing in the excited state and are inherent to the tryptophan structure independently of the surrounding environment (amino acids residues, solvent, etc.) In proteins, the third lifetime originates from the interactions that are occurring between tryptophan residues and neighboring amino acids. Populations of these lifetimes are independent of the excitation wavelength and thus originate from pre-defined sub structures existing in the excited state and put into evidence after tryptophan excitation. Fluorescence decay studies of different tripeptides having a tryptophan residue in second position show that the best analysis is obtained with two fluorescence lifetimes. Consequently, this result seems to exclude the possibility that peptide bond induces the third fluorescence lifetimes. Indole dissolved in water and/or in ethanol emits with two fluorescence lifetimes that are completely different from those observed for L-Trp. Absence of the third lifetime in ethanol demonstrates that indole behaves differently when compared to tryptophan. Thus, it seems not adequate to attribute fluorescence lifetime or fluorescence properties of tryptophan to indole ring and to compare tryptophan fluorescence properties in proteins to molecules having close structures such as NATA which fluoresces with one lifetime.

  9. One- and two-photon excited fluorescence lifetimes and anisotropy decays of green fluorescent proteins.

    OpenAIRE

    2000-01-01

    We have used one- (OPE) and two-photon (TPE) excitation with time-correlated single-photon counting techniques to determine time-resolved fluorescence intensity and anisotropy decays of the wild-type Green Fluorescent Protein (GFP) and two red-shifted mutants, S65T-GFP and RSGFP. WT-GFP and S65T-GFP exhibited a predominant approximately 3 ns monoexponential fluorescence decay, whereas for RSGFP the main lifetimes were approximately 1.1 ns (main component) and approximately 3.3 ns. The anisotr...

  10. UV fluorescence enhancement from nanostructured aluminum materials

    Science.gov (United States)

    Montanari, Danielle E.; Dean, Nathan; Poston, Pete E.; Blair, Steve; Harris, Joel M.

    2016-09-01

    Interest in label-free detection of biomolecules has given rise to the need for UV plasmonic materials. DNA bases and amino acid residues have electronic resonances in the UV which allow for sensitive detection of these species by surface-enhanced UV fluorescence spectroscopy. Electrochemical roughening has been used extensively to generate plasmonically-active metal surfaces that produce localized enhancement of excitation and emission of electromagnetic radiation from surface-bound molecules. Electrochemically roughened gold and silver surfaces produce enhancement in the visible and near-IR regions, but to the best of our knowledge, application of this technique for producing UV-enhancing substrates has not been reported. Using electropolishing of aluminum, we are able to generate nanostructured surfaces that produce enhanced spectroscopic detection of molecules in the UV. Aluminum is a natural choice for substrate composition as it exhibits a relatively large quality factor in the UV. We have fabricated electropolished aluminum films with nanometer scale roughness and have studied UV-excited fluorescence enhancement from submonolayer coverage of tryptophan on these substrates using a UV-laser based spectrometer. Quantitative dosing by dip-coating was used to deposit known surface concentrations of the aromatic amino acid tryptophan, so that fluorescence enhancement could be evaluated. Compared to a dielectric substrate (surface-oxidized silicon), we observe a 180-fold enhancement in the total fluorescence emitted by tryptophan on electropolished aluminum under photobleaching conditions, allowing detection of sub-monolayer coverages of molecules essential for development of biosensor technologies.

  11. Induced systemic resistance by fluorescent Pseudomonas spp.

    NARCIS (Netherlands)

    Bakker, P.A.H.M.; Pieterse, C.M.J.; Loon, L.C. van

    2007-01-01

    Fluorescent Pseudomonas spp. have been studied for decades for their plant growth-promoting effects through effective suppression of soilborne plant diseases. The modes of action that play a role in disease suppression by these bacteria include siderophore-mediated competition for iron, antibiosis,

  12. Fluorescence emission of pyrene in surfactant solutions.

    Science.gov (United States)

    Piñeiro, Lucas; Novo, Mercedes; Al-Soufi, Wajih

    2015-01-01

    The systematic description of the complex photophysical behaviour of pyrene in surfactant solutions in combination with a quantitative model for the surfactant concentrations reproduces with high accuracy the steady-state and the time resolved fluorescence intensity of pyrene in surfactant solutions near the cmc, both in the monomer and in the excimer emission bands. We present concise model equations that can be used for the analysis of the pyrene fluorescence intensity in order to estimate fundamental parameters of the pyrene-surfactant system, such as the binding equilibrium constant K of pyrene to a given surfactant micelle, the rate constant of excimer formation in micelles, and the equilibrium constant of pyrene-surfactant quenching. The values of the binding equilibrium constant K(TX100)=3300·10³ M⁻¹ and K(SDS)=190·10³ M⁻¹ for Triton X-100 (TX100) and SDS micelles, respectively, show that the partition of pyrene between bulk water and micelles cannot be ignored, even at relatively high surfactant concentrations above the cmc. We apply the model to the determination of the cmc from the pyrene fluorescence intensity, especially from the intensity ratio at two vibronic bands in the monomer emission or from the ratio of excimer to monomer emission intensity. We relate the finite width of the transition region below and above the cmc with the observed changes in the pyrene fluorescence in this region.

  13. Computational Modeling of Fluorescence Loss in Photobleaching

    DEFF Research Database (Denmark)

    Hansen, Christian Valdemar; Schroll, Achim; Wüstner, Daniel

    2015-01-01

    Fluorescence loss in photobleaching (FLIP) is a modern microscopy method for visualization of transport processes in living cells. Although FLIP is widespread, an automated reliable analysis of image data is still lacking. This paper presents a framework for modeling and simulation of FLIP...

  14. Novel Fluorescent Dyes Based on Coumarin System

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Seven novel fluorescent coumarin derivatives were synthesized from 7-diethylamino-4-chloro-3-formyl coumarin. The spectra of absorption, excitation and emission were dependent not only on the structures and also on the concentration of dyes. The PPP-MO predictions can only be consistent with the spectra in dilute solutions.

  15. Fluorescent nanodiamonds embedded in biocompatible translucent shells.

    Science.gov (United States)

    Rehor, Ivan; Slegerova, Jitka; Kucka, Jan; Proks, Vladimir; Petrakova, Vladimira; Adam, Marie-Pierre; Treussart, François; Turner, Stuart; Bals, Sara; Sacha, Pavel; Ledvina, Miroslav; Wen, Amy M; Steinmetz, Nicole F; Cigler, Petr

    2014-03-26

    High pressure high temperature (HPHT) nanodiamonds (NDs) represent extremely promising materials for construction of fluorescent nanoprobes and nanosensors. However, some properties of bare NDs limit their direct use in these applications: they precipitate in biological solutions, only a limited set of bio-orthogonal conjugation techniques is available and the accessible material is greatly polydisperse in shape. In this work, we encapsulate bright 30-nm fluorescent nanodiamonds (FNDs) in 10-20-nm thick translucent (i.e., not altering FND fluorescence) silica shells, yielding monodisperse near-spherical particles of mean diameter 66 nm. High yield modification of the shells with PEG chains stabilizes the particles in ionic solutions, making them applicable in biological environments. We further modify the opposite ends of PEG chains with fluorescent dyes or vectoring peptide using click chemistry. High conversion of this bio-orthogonal coupling yielded circa 2000 dye or peptide molecules on a single FND. We demonstrate the superior properties of these particles by in vitro interaction with human prostate cancer cells: while bare nanodiamonds strongly aggregate in the buffer and adsorb onto the cell membrane, the shell encapsulated NDs do not adsorb nonspecifically and they penetrate inside the cells.

  16. Laser-fluorescence measurement of marine algae

    Science.gov (United States)

    Browell, E. V.

    1980-01-01

    Progress in remote sensing of algae by laser-induced fluorescence is subject of comprehensive report. Existing single-wavelength and four-wavelength systems are reviewed, and new expression for power received by airborne sensor is derived. Result differs by as much as factor of 10 from those previously reported. Detailed error analysis evluates factors affecting accuracy of laser-fluorosensor systems.

  17. Inositol phosphates induce DAPI fluorescence shift.

    Science.gov (United States)

    Kolozsvari, Bernadett; Parisi, Federica; Saiardi, Adolfo

    2014-06-15

    The polymer inorganic polyP (polyphosphate) and inositol phosphates, such as IP6 (inositol hexakisphosphate; also known as phytic acid), share many biophysical features. These similarities must be attributed to the phosphate groups present in these molecules. Given the ability of polyP to modify the excitation-emission spectra of DAPI we decided to investigate whether inositol phosphates possess the same property. We discovered that DAPI-IP6 complexes emit at approximately 550 nm when excited with light of wavelength 410-420 nm. IP5 (inositol pentakisphosphate) is also able to induce a similar shift in DAPI fluorescence. Conversely, IP3 (inositol trisphosphate) and IP4 (inositol tetrakisphosphate) are unable to shift DAPI fluorescence. We have employed this newly discovered feature of DAPI to study the enzymatic activity of the inositol polyphosphate multikinase and to monitor phytase phosphatase reactions. Finally, we used DAPI-IP6 fluorescence to determine the amount of IP6 in plant seeds. Using an IP6 standard curve this straight-forward analysis revealed that among the samples tested, borlotti beans possess the highest level of IP6 (9.4 mg/g of dry mass), whereas the Indian urad bean the lowest (3.2 mg/g of dry mass). The newly identified fluorescence properties of the DAPI-IP5 and DAPI-IP6 complexes allow the levels and enzymatic conversion of these two important messengers to be rapidly and reliably monitored.

  18. Phytoplankton productivity quantified from chlorophyll fluorescence

    DEFF Research Database (Denmark)

    Hancke, Kasper; Dalsgaard, Tage; Sejr, Mikael Kristian

    Phytoplankton are the main food source for marine life, and accurate uantification of its productivity is essential for understanding how marine food webs function. As a novel non-invasive technology, chlorophyll fluorescence can be used to assess in situ primary production in phytoplankton...

  19. Laser-stimulated fluorescence in paleontology.

    Science.gov (United States)

    Kaye, Thomas G; Falk, Amanda R; Pittman, Michael; Sereno, Paul C; Martin, Larry D; Burnham, David A; Gong, Enpu; Xu, Xing; Wang, Yinan

    2015-01-01

    Fluorescence using ultraviolet (UV) light has seen increased use as a tool in paleontology over the last decade. Laser-stimulated fluorescence (LSF) is a next generation technique that is emerging as a way to fluoresce paleontological specimens that remain dark under typical UV. A laser's ability to concentrate very high flux rates both at the macroscopic and microscopic levels results in specimens fluorescing in ways a standard UV bulb cannot induce. Presented here are five paleontological case histories that illustrate the technique across a broad range of specimens and scales. Novel uses such as back-lighting opaque specimens to reveal detail and detection of specimens completely obscured by matrix are highlighted in these examples. The recent cost reductions in medium-power short wavelength lasers and use of standard photographic filters has now made this technique widely accessible to researchers. This technology has the potential to automate multiple aspects of paleontology, including preparation and sorting of microfossils. This represents a highly cost-effective way to address paleontology's preparatory bottleneck.

  20. Fluorescence of gaseous tetraenes and pentaenes

    Energy Technology Data Exchange (ETDEWEB)

    Bouwman, W.G.; Jones, A.C.; Phillips, D. (Imperial College of Science, London (England)); Thibodeau, P.; Friel, C.; Christensen, R.L. (Bowdoin College, Brunswick, ME (USA))

    1990-09-20

    Fluorescence, fluorescence excitation, and absorption spectra of room temperature vapors of 1,3,5,7-octatetraene, 1,3,5,7-nonatetraene, 2,4,6,8-decatetraene, and 2,4,6,8,10-dodecapentaene have been obtained. All four polyenes show dual (S{sub 1} {yields} S{sub 0} and S{sub 2}{yields}S{sub 0}) fluorescences with the ratios of the two emissions (2{sup 1}A{sub g} {yields} 1{sup 1}A{sub g} and 1{sup 1}B{sub u} {yields} 1{sup 1}A{sub g}) being sensitive to the extent of substitution. For example, the (S{sub 1} {yields} S{sub 0})/(S{sub 2} {yields} S{sub 0}) fluorescence yield ratio increases from 0.06 to 0.7 when terminal methyl groups are added to octatetraene. The gas-phase S{sub 2}-S{sub 1} energy gaps are almost identical for these two molecules (6,600 cm{sup {minus}1} for octatetraene versus 6,300 cm{sup {minus}1} for decatetraene), suggesting that the apparent difference in internal conversion efficiencies is due to the larger density of vibronic states in the methyl-substituted compounds. These results are discussed in relation to the previous gas-phase spectroscopy of octatetraene and shorter polyenes.

  1. Global analysis of fluorescence fluctuation data

    NARCIS (Netherlands)

    Skakun, V.V.; Hink, M.A.; Digris, A.V.; Engel, R.; Novikov, E.G.; Apanasovich, V.V.; Visser, A.J.W.G.

    2005-01-01

    Over the last decade the number of applications of fluorescence correlation spectroscopy (FCS) has grown rapidly. Here we describe the development and application of a software package, FCS Data Processor, to analyse the acquired correlation curves. The algorithms combine strong analytical power

  2. Flow cytometry, fluorescent probes, and flashing bacteria

    NARCIS (Netherlands)

    Bunthof, C.J.

    2002-01-01

     


    Key words: fluorescent probes, flow cytometry, CSLM, viability, survival, microbial physiology, lactic acid bacteria, Lactococcus lactis , Lactobacillus plantarum , cheese, milk, probiotic In food industry there is a perceived need for rapid methods for detection and viability a

  3. Developing Fluorescent Hyaluronan Analogs for Hyaluronan Studies

    Directory of Open Access Journals (Sweden)

    Shi Ke

    2012-02-01

    Full Text Available Two kinds of fluorescent hyaluronan (HA analogs, one serving as normal imaging agent and the other used as a biosensitive contrast agent, were developed for the investigation of HA uptake and degradation. Our approach of developing HA imaging agents depends on labeling HA with varying molar percentages of a near-infrared (NIR dye. At low labeling ratios, the hyaluronan uptake can be directly imaged while at high labeling ratios, the fluorescent signal is quenched and signal generation occurs only after degradation. It is found that the conjugate containing 1%–2% NIR dye can be used as a normal optical imaging agent, while bioactivable imaging agents are formed at 6% to 17% dye loading. It was determined that the conjugation of dye to HA with different loading percentages does not impact HA biodegradation by hyaluronidase (Hyal. The feasibility of using these two NIR fluorescent hyaluronan analogs for HA investigation was evaluated in vivo with optical imaging. The data demonstrates that the 1% dye loaded fluorescent HA can be used to monitor the behavior of HA and its fragments, whereas bioactivatable HA imaging agent (17% dye in HA is more suitable for detecting HA fragments.

  4. Fluorescent hybridization probes for nucleic acid detection.

    Science.gov (United States)

    Guo, Jia; Ju, Jingyue; Turro, Nicholas J

    2012-04-01

    Due to their high sensitivity and selectivity, minimum interference with living biological systems, and ease of design and synthesis, fluorescent hybridization probes have been widely used to detect nucleic acids both in vivo and in vitro. Molecular beacons (MBs) and binary probes (BPs) are two very important hybridization probes that are designed based on well-established photophysical principles. These probes have shown particular applicability in a variety of studies, such as mRNA tracking, single nucleotide polymorphism (SNP) detection, polymerase chain reaction (PCR) monitoring, and microorganism identification. Molecular beacons are hairpin oligonucleotide probes that present distinctive fluorescent signatures in the presence and absence of their target. Binary probes consist of two fluorescently labeled oligonucleotide strands that can hybridize to adjacent regions of their target and generate distinctive fluorescence signals. These probes have been extensively studied and modified for different applications by modulating their structures or using various combinations of fluorophores, excimer-forming molecules, and metal complexes. This review describes the applicability and advantages of various hybridization probes that utilize novel and creative design to enhance their target detection sensitivity and specificity.

  5. Fluorescence Talbot microscope using incoherent source

    Science.gov (United States)

    Sun, Yangyang; Pang, Shuo

    2016-08-01

    Fluorescence Talbot microscope is a scalable field-of-view (FOV) imaging platform, which takes advantage of the phase sensitivity of the self-image of a periodic structure. Such a system can maintain the microscopic resolution and extend the FOV for the whole slide (15 mm×15 mm) scanning. Previously reported Talbot fluorescence systems, tabletop and on-chip device alike, rely on the coherence of the illumination source, limiting their potential applications in low-resource setting environment. A more cost-effective setup using a light-emitting diode, which has an area of 4 mm2 and a full width at half maximum of 16 nm in wavelength, is demonstrated. Compared to the illumination that is spatially filtered by a single pinhole, our system has achieved an illumination intensity that is 357 times higher. The reconstructed image quality is comparable to that of a 10× microscope objective. Various samples, such as fluorescent beads, green fluorescence protein-labeled HeLa cells, and a mouse kidney slide, were reconstructed by the system.

  6. A Fluorescent Broad-Spectrum Proteasome Inhibitor

    NARCIS (Netherlands)

    Verdoes, Martijn; Florea, Bogdan I.; Menendez-Benito, Victoria; Maynard, Christa J.; Witte, Martin D.; Linden, Wouter A. van der; Nieuwendijk, Adrianus M.C.H. van den; Hofmann, Tanja; Berkers, Celia R.; Leeuwen, Fijs W.B. van; Groothuis, Tom A.; Leeuwenburgh, Michiel A.; Ovaa, Huib; Neefjes, Jacques J.; Filippov, Dmitri V.; Marel, Gijs A. van der; Dantuma, Nico P.; Overkleeft, Herman S.

    2006-01-01

    The proteasome is an essential evolutionary conserved protease involved in many regulatory systems. Here, we describe the synthesis and characterization of the activity-based, fluorescent, and cell-permeable inhibitor Bodipy TMR-Ahx3L3VS (MV151), which specifically targets all active subunits of the

  7. Laser-stimulated fluorescence in paleontology.

    Directory of Open Access Journals (Sweden)

    Thomas G Kaye

    Full Text Available Fluorescence using ultraviolet (UV light has seen increased use as a tool in paleontology over the last decade. Laser-stimulated fluorescence (LSF is a next generation technique that is emerging as a way to fluoresce paleontological specimens that remain dark under typical UV. A laser's ability to concentrate very high flux rates both at the macroscopic and microscopic levels results in specimens fluorescing in ways a standard UV bulb cannot induce. Presented here are five paleontological case histories that illustrate the technique across a broad range of specimens and scales. Novel uses such as back-lighting opaque specimens to reveal detail and detection of specimens completely obscured by matrix are highlighted in these examples. The recent cost reductions in medium-power short wavelength lasers and use of standard photographic filters has now made this technique widely accessible to researchers. This technology has the potential to automate multiple aspects of paleontology, including preparation and sorting of microfossils. This represents a highly cost-effective way to address paleontology's preparatory bottleneck.

  8. Fluorescent compounds for plastic scintillation applications

    Energy Technology Data Exchange (ETDEWEB)

    Pla-Dalmau, A.; Bross, A.D.

    1994-04-01

    Several 2-(2{prime}-hydroxyphenyl)benzothiazole, -benzoxazole, and -benzimidazole derivatives have been prepared. Transmittance, fluorescence, light yield, and decay time characteristics of these compounds have been studied in a polystyrene matrix and evaluated for use in plastic scintillation detectors. Radiation damage studies utilizing a {sup 60}C source have also been performed.

  9. Dental resin cure monitoring by inherent fluorescence

    Science.gov (United States)

    Li, Qun; Zhou, Jack X.; Li, Qingxiong; Wang, Sean X.

    2008-02-01

    It is demonstrated that the inherent fluorescence of a dental composite resin can be utilized to monitor the curing status, i.e. degree of conversion of the resin. The method does not require any sample preparation and is potentially very fast for real time cure monitoring. The method is verified by Raman spectroscopy analysis.

  10. Refractometric sensing with fluorescent-core microcapillaries.

    Science.gov (United States)

    Manchee, C P K; Zamora, V; Silverstone, J W; Veinot, J G C; Meldrum, A

    2011-10-24

    Capillaries present a promising structure for microfluidic refractive index sensors. We demonstrate a capillary-type fluorescent core microcavity sensor based on whispering gallery mode (WGM) resonances. The device consists of a microcapillary having a layer of fluorescent silicon quantum dots (QDs) coated on the channel surface. The high effective index of the QD layer confines the electric field near the capillary channel and causes the development of WGM resonances in the fluorescence spectrum. Solutions consisting of sucrose dissolved in water were pumped through the capillary while the fluorescence WGMs were measured with a spectrometer. The device showed a refractometric sensitivity of 9.8 nm/RIU (up to 13.8 nm/RIU for higher solution refractive index) and a maximum detection limit of ~7.2 x 10(-3) RIU. Modeling the field inside the capillary structure, which is analogous to a layered hollow ring resonator, shows that sensitivities as high as 100 nm/RIU and detection limits as low as ~10(-5) RIU may be achievable by optimizing the QD film thickness.

  11. Recent Advances in Fluorescence in situ Hybridization

    OpenAIRE

    吉田, 廸弘; Michihiro C., Yoshida; 北海道大学理学部附属動物染色体研究施設; Chromosome Research Unit, Faculty of Science, Hokkaido University

    1992-01-01

    Fluorescence in situ hybridization (FISH) procedures that directly couple molecular and cytological information allow precise visualization of DNA sequences on metaphase chromosomes and interphase nuclei. These techniques can be used to identify chromosomes, detect chromosomal aberrations, and analyze linear and spetial genome organization. FISH procedures are also used to clinical fields for diagnosis of disease-related chromosome changes and tumor biology.

  12. Improving the Stability of Fluorescent Silver Nanoclusters

    Science.gov (United States)

    Swanson, Nicholas; Stanko, Danielle; Campbell, Ian; Wittmershaus, Bruce

    The quantum mechanical nature of noble metal nanoparticles results in them having optical properties much different from the bulk metal. Silver nanoclusters (AgNC), groups of 4 to 20 atoms, are characterized by strong optical transitions in the visible part of the spectrum giving them an appearance like fluorescent organic dyes. These nanoclusters can also have fluorescence quantum yields over 90%. Following the analysis of published results of DNA templated nanoclusters, we created a procedure for synthesizing AgNC. The AgNC have a high fluorescence quantum yield but degrade with a lifetime of only a few days when in solution at room temperature. Our goal in this study was to increase the stability of the AgNC towards improving their value as a fluorescent material in various applications, such as luminescent solar concentrators. To increase their stability, we've chosen to modify our procedure by removing oxygen from the solution after the sample has reacted. Oxygen removal caused a significant increase in the stability of the clusters over a given period of time. This material is based upon work supported by the National Science Foundation under Grant Number NSF-ECCS-1306157.

  13. Microgels for multiplex and direct fluorescence detection

    Science.gov (United States)

    Causa, Filippo; Aliberti, Anna; Cusano, Angela M.; Battista, Edmondo; Netti, Paolo A.

    2015-05-01

    Blood borne oligonucleotides fragments contain useful clinical information whose detection and monitoring represent the new frontier in liquid biopsy as they can transform the current diagnosis procedure. For instance, recent studies have identified a new class of circulating biomarkers such as s miRNAs, and demonstrated that changes in their concentration are closely associated with the development of cancer and other pathologies. However, direct detection of miRNAs in body fluids is particularly challenging and demands high sensitivity -concentration range between atto to femtomolarspecificity, and multiplexing Here we report on engineered multifunctional microgels and innovative probe design for a direct and multiplex detection of relevant clinical miRNAs in fluorescence by single particle assay. Polyethyleneglycol-based microgels have a coreshell architecture with two spectrally encoded fluorescent dyes for multiplex analyses and are endowed with fluorescent probes for miRNA detection. Encoding and detection fluorescence signals are distinguishable by not overlapping emission spectra. Tuneable fluorescence probe conjugation and corresponding emission confinement on single microgel allows for enhanced target detection. Such suspension array has indeed high selectivity and sensitivity with a detection limit of 10-15 M and a dynamic range from 10-9 to 10-15 M. We believe that sensitivity in the fM concentration range, signal background minimization, multiplexed capability and direct measurement of such microgels will translate into diagnostic benefits opening up new roots toward liquid biopsy in the context of point-of-care testing through an easy and fast detection of sensitive diagnostic biomarkers directly in serum.

  14. Blood Compatibility Evaluations of Fluorescent Carbon Dots.

    Science.gov (United States)

    Li, Sha; Guo, Zhong; Zhang, Yi; Xue, Wei; Liu, Zonghua

    2015-09-02

    Because of their unique advantages, fluorescent carbon dots are gaining popularity in various biomedical applications. For these applications, good biosafety is a prerequisite for their use in vivo. Studies have reported the preliminary biocompatibility evaluations of fluorescent carbon dots (mainly cytotoxicity); however, to date, little information is available about their hemocompatibility, which could impede their development from laboratory to bedside. In this work, we evaluated the hemocompatibility of fluorescent carbon dots, which we prepared by hydrothermal carbonization of α-cyclodextrin. The effects of the carbon dots on the structure and function of key blood components were investigated at cellular and molecular levels. In particular, we considered the morphology and lysis of human red blood cells, the structure and conformation of the plasma protein fibrinogen, the complement activation, platelet activation, and in vitro and in vivo blood coagulation. We found that the carbon dots have obvious concentration-dependent effects on the blood components. Overall, concentrations of the fluorescent carbon dots at ≤0.1 mg/mL had few adverse effects on the blood components, but at higher doses, the carbon dots impair the structure and function of the blood components, causing morphological disruptions and lysis of red blood cells, interference in the local microenvironments of fibrinogen, activation of the complement system, and disturbances in the plasma and whole blood coagulation function in vitro. However, the carbon dots tend to activate platelets only at low concentrations. Intravenous administration of the carbon dots at doses up to 50 mg/kg did not impair the blood coagulation function. These results provide valuable information for the clinical application of fluorescent carbon dots.

  15. Diagnosis of Dentin Caries – Ultraviolet Fluorescence

    Directory of Open Access Journals (Sweden)

    Uzunov Ts.

    2014-12-01

    Full Text Available The technology advance in recent years determines the need of construction of modern appliances for early diagnosis of dental caries, which are categorized by great precision, non-invasiveness, easy usage and wide availability. Such non-invasive and accurate tool for diagnostics of caries is Caries Detector (LED, Bulgarian product by “Optica Laser”. The detector emits a specific wavelength of near ultraviolet light, which causes fluorescence to porphyrins - metabolic products of the life cycle of caries-inducing bacteria. The purpose of the study is piloting a new diagnostic tool for detection and monitoring of caries excavation based on fluorescence - LED UV caries detector of company “Optica laser”. Subjected to examination by caries indicator dye and UV caries detector were sixty permanent teeth with deep dentine caries. Two methods were used to assess the dentin caries - UV fluorescence detector of “Optica Laser” and staining with caries indicator - dye (Sable ™ Seek®. It was found that among all sixty teeth, the fields, closed by margins of carious process overlap. Fifty-four of tested teeth has shown bigger field of images with staining method and six - smaller in comparison to the fluorescent method. Ultraviolet fLuorescence caries detector of “Optica Laser” company is affordable and easy applicable method for controlled excavation of dentine caries. The detector can be used in daily dental practice equally with other methods. The unit has a number of advantages - non-invasiveness, lack of interaction with tooth structures, speed, reliability, efficiency, predictability and repeatability of results.

  16. Mechanism of fluorescent cocoon sex identification for silkworms Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    By using silkworms,Bombyx mori, fluorescent cocoon sex identification (FCSI) as an experimental material, direct fluorescence spectrometry of the cocoon surface indicates that the fluorescent color of silkworm cocoons is made up of two peaks of yellow and blue-purple fluorescence emission. The fluorescent difference between male and female cocoons is attributed to the differential absorption of yellow fluorescent substances by the midgut tissue of 5th instar female silkworms. Thin layer chromatography (TLC) and fluorescent spectra indicate that blue-purple fluorescent substances are composed of at least five blue-purple fluorescent pigments, and yellow fluorescent substances are made up of at least three. UV spectra and AlCl3 color reaction show that the three fluorescent yellow pigments are flavonoids or their glycosides. Silkworm FCSI is due to selective absorption or accumulation of the yellow fluorescent pigments by the posterior midgut cells of female 5th instar larvae. The cells of the FCSI silkworm midgut, especially the cylinder intestinal cells of the posterior midgut have a component which is a yellow fluorescent pigment-specific binding protein that may be vigorously expressed in the 5th instar larvae.

  17. Highly Selective Fluorescent Sensing of Proteins Based on a Fluorescent Molecularly Imprinted Nanosensor

    Directory of Open Access Journals (Sweden)

    Shuo Wang

    2013-09-01

    Full Text Available A fluorescent molecularly imprinted nanosensor was obtained by grafting imprinted polymer onto the surface of multi-wall carbon nanotubes and post-imprinting treatment with fluorescein isothiocyanate (FITC. The fluorescence of lysozyme-imprinted polymer (Lys-MIP was quenched more strongly by Lys than that of nonimprinted polymer (NIP, which indicated that the Lys-MIP could recognize Lys. The resulted imprinted material has the ability to selectively sense a target protein, and an imprinting factor of 3.34 was achieved. The Lys-MIP also showed selective detection for Lys among other proteins such as cytochrome C (Cyt C, hemoglobin (HB and bovine serum albumin (BSA due to the imprinted sites in the Lys-MIP. This approach combines the high selectivity of surface molecular imprinting technology and fluorescence, and converts binding events into detectable signals by monitoring fluorescence spectra. Therefore, it will have further applications for Lys sensing.

  18. Fluorescence Emission Centres and the Corresponding Infrared Fluorescence Saturation in a Bismuth-Doped Silica Fibre

    Institute of Scientific and Technical Information of China (English)

    QIU Yan-Qing; SHEN Yong-Hang

    2008-01-01

    We investigate the fluorescence characteristics of bismuth doped silica fibres with and without Al co-dopant which are fabricated by means of modified chemical vapour deposition (MCVD) technique, and find that the fluorescences in the red region (centred around 750nm) and in the infrared region (centred around 1100nm) may originate from different emission sites in the fibre. Strong upconversion phenomena are observed in both Al-codoped and non Al codoped bismuth fibres when the fibres are excited by an acoustic-optic Q-switched Nd: YVO4 laser. Both the aspects indicate that the upper energy level absorption reported in the work of the bismuth doped silica fibre lasers may result from the fluorescence emission sites that are not responsible for the infrared emission. It is thus expected that optimizing the compositions and the fabrication conditions of the fibre and then transferring more fluorescence emission centres are helpful for the infrared emission.

  19. 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.

  20. Synthesis and fluorescence enhancement behavior of a novel fluorescent aqueous polyurethane emulsion DDAQ-TDI-PU

    Institute of Scientific and Technical Information of China (English)

    Xian Hai Hu; Xing Yuan Zhang; Jia Bing Dai

    2012-01-01

    A novel fluorescent aqueous polyurethane emulsion DDAQ-TDI-PU was synthesized by blocking the anthraquinone moiety of 1,4-diamino-2,3-diphenoxyanthraquinone (DDAQ) into polyurethane chain using 2,4-tolylene diisocyanate (TDI),poly(propylene glycol) and 2,2-dimethylol propionic acid.The chain structure of DDAQ-TDI-PU was confirmed by means of Fourier transform infrared spectroscopy and UV-vis analysis.Comparing to the UV-vis spectrum of DDAQ,DDAQ-TDI-PU showed a hypsochromic shift from the absorption maxima of 518,558,609 nm to 510,548,586 nm,respectively.It was found that the fluorescence intensity of DDAQ-TDI-PU emulsion was enhanced greatly comparing with that of DDAQ.The fluorescence of DDAQ-TDI-PU was very stable not only for the long term storage but also for the fluorescence quencher.

  1. Fluorescence resonance energy transfer between fluorescent proteins as powerful toolkits for in vivo studies

    Science.gov (United States)

    Rusanov, A. L.; Savitsky, A. P.

    2011-02-01

    To expand the field of research in biological systems development of extra-sensitive analytical methods is highly desirable. In this review, the latest advances in technologies relying on the fluorescence resonance energy transfer between fluorescent proteins (FP's) to visualize numerous molecular processes in living cells are discussed. Variety of FP's as well as of novel experimental techniques allows one to choose the most appropriate tools to attack concrete problems.

  2. STUDY ON ELIMINATING FLUORESCENCE IN COTTON PULP WITH PERACETIC ACID

    Institute of Scientific and Technical Information of China (English)

    LixinXu; BaoguoSun

    2004-01-01

    This paper has studied processing conditions and the influence of every variable to the pulp when the fluorescence in cotton linter pulp is eliminated with peracetic acid. The suitable variables of the elimination of fluorescence, are found.

  3. STUDY ON ELIMINATING FLUORESCENCE IN COTTON PULP WITH PERACETIC ACID

    Institute of Scientific and Technical Information of China (English)

    Lixin Xu; Baoguo Sun

    2004-01-01

    This paper has studied processing conditions and the influence of every variable to the pulp when the fluorescence in cotton linter pulp is eliminated with peracetic acid. The suitable variables of the elimination of fluorescence. are found.

  4. Generally Applicable Transformation Protocols for Fluorescent Nanodiamond Internalization into Cells

    NARCIS (Netherlands)

    Hemelaar, Simon R; van der Laan, Kiran J; Hinterding, Sophie R; Koot, Manon V; Ellermann, Else; Perona-Martinez, Felipe P; Roig, David; Hommelet, Severin; Novarina, Daniele; Takahashi, Hiroki; Chang, Michael; Schirhagl, Romana

    2017-01-01

    Fluorescent nanodiamonds (FNDs) are promising nanoprobes, owing to their stable and magnetosensitive fluorescence. Therefore they can probe properties as magnetic resonances, pressure, temperature or strain. The unprecedented sensitivity of diamond defects can detect the faint magnetic resonance of

  5. Highly fluorescent benzofuran derivatives of the GFP chromophore

    DEFF Research Database (Denmark)

    Christensen, Mikkel Andreas; Jennum, Karsten Stein; Abrahamsen, Peter Bæch

    2012-01-01

    Intramolecular cyclization reactions of Green Fluorescent Protein chromophores (GFPc) containing an arylethynyl ortho-substituent at the phenol ring provide new aryl-substituted benzofuran derivatives of the GFPc. Some of these heteroaromatic compounds exhibit significantly enhanced fluorescence...

  6. Synthesis and characterization of colloidal fluorescent silver nanoclusters.

    Science.gov (United States)

    Huang, Sherry; Pfeiffer, Christian; Hollmann, Jana; Friede, Sebastian; Chen, Justin Jin-Ching; Beyer, Andreas; Haas, Benedikt; Volz, Kerstin; Heimbrodt, Wolfram; Montenegro Martos, Jose Maria; Chang, Walter; Parak, Wolfgang J

    2012-06-19

    Ultrasmall water-soluble silver nanoclusters are synthesized, and their properties are investigated. The silver nanoclusters have high colloidal stability and show fluorescence in the red. This demonstrates that like gold nanoclusters also silver nanoclusters can be fluorescent.

  7. Non-radiographic intraoperative fluorescent cholangiography is feasible

    DEFF Research Database (Denmark)

    Larsen, Søren Schytt; Schulze, Svend; Bisgaard, Thue

    2014-01-01

    INTRODUCTION: Intraoperative fluorescent cholangiography (IFC) with concomitant fluorescent angiography was recently developed for non-invasive identification of the anatomy during laparoscopic cholecystectomy. The objective of this study was to assess the time required for routine-use of IFC...

  8. New fluorescence parameters for monitoring photosynthesis in plants

    NARCIS (Netherlands)

    Force, L.; Critchley, Ch.; Rensen, van J.J.S.

    2003-01-01

    Chlorophyll fluorescence measurements have a wide range of applications from basic understanding of photosynthesis functioning to plant environmental stress responses and direct assessments of plant health. The measured signal is the fluorescence intensity (expressed in relative units) and the most

  9. 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...

  10. The Lantibiotic Nisin Induces Transmembrane Movement of a Fluorescent Phospholipid

    NARCIS (Netherlands)

    Moll, Gert N.; Konings, Wil N.; Driessen, Arnold J.M.

    1998-01-01

    Nisin is a pore-forming antimicrobial peptide. The capacity of nisin to induce transmembrane movement of a fluorescent phospholipid in lipid vesicles was investigated. Unilamellar phospholipid vesicles that contained a fluorescent phospholipid

  11. Application of fluorescence in situ hybridization (FISH) to the ...

    African Journals Online (AJOL)

    Application of fluorescence in situ hybridization (FISH) to the analysis of ... In this study, fluorescent in situ hybridization (FISH) as a culture-independent molecular ... a high percentage and took place in an oily biological system under aerobic ...

  12. Fluorescent molecularly imprinted polymer film binds glucose with a concomitant changes in fluorescence.

    Science.gov (United States)

    Manju, S; Hari, P R; Sreenivasan, K

    2010-10-15

    A fluorescent molecularly imprinted polymeric formulation capable of picking up glucose from aqueous media is reported. The fluorescence intensity of the polymer film was found to reduce proportionally with the concentration of glucose facilitating its use as a glucose sensing element. We used commercially available tear fluid to demonstrate the ability of the film to recognize glucose among other sugar molecules. Fluorescence was measured after equilibrating the film in tear fluid in the presence of a mixture of different sugars. We observed a reduction in fluorescence intensity due to the nonspecific binding of the sugars. The intensity remains the same even if we added additional quantities of the sugars. Interestingly, the fluorescence intensity of the film was found to decrease proportionally when varied concentrations of glucose was added indicating the ability of the film to recognize and bind glucose from a mixture of other sugars. Detectable changes in fluorescence intensity were observed with a concentration of 10 μg/mL of glucose. The results show that the polymer film could be used for detecting glucose in aqueous fluids such as tear. Copyright © 2010 Elsevier B.V. All rights reserved.

  13. Forensic applications: Fluorescence properties of tooth-coloured restorative materials using a fluorescence DSLR camera.

    Science.gov (United States)

    Kiran, Ramya; Walsh, Laurence J; Forrest, Alexander; Tennant, Marc; Chapman, James

    2017-02-03

    The objective of this study was to compare the fluorescence properties of dry and wet samples of contemporary tooth-coloured restorative materials using a fluorescence based DSLR camera and a variety of LEDs emitting different wavelengths of visible light as excitation sources. The materials examined included resin composites; ceramics and hybrid restorative materials such as ormocers, Vita Enamic™ and resin reinforced glass-ionomer cements. The levels of fluorescence for each sample under different combinations of incident light wavelengths and filters was analysed by using histogram data for colour channels from Adobe Photoshop software. Fluorescence patterns were influenced by water sorption of the materials. UV-A/Violet light (405±nm) produced the greatest range of luminosity values (10-204) amongst the tooth-coloured restorative materials, and showed the greatest differences between restorations and tooth structure. The best filter combinations with violet light were orange or yellow filters. Under ultraviolet excitation, Fuji VIII A2 exhibited a unique bright pink fluorescence emission, while VitaEnamic™, ormocer and glass-ionomer cements emitted bluish-pink fluorescence emissions. In conclusion, restorative materials exhibited varied emission pattern under UV-A (405nm) light, which enables their detection and differentiation from natural tooth structure.

  14. Measuring and sorting cell populations expressing isospectral fluorescent proteins with different fluorescence lifetimes.

    Directory of Open Access Journals (Sweden)

    Bryan Sands

    Full Text Available Study of signal transduction in live cells benefits from the ability to visualize and quantify light emitted by fluorescent proteins (XFPs fused to different signaling proteins. However, because cell signaling proteins are often present in small numbers, and because the XFPs themselves are poor fluorophores, the amount of emitted light, and the observable signal in these studies, is often small. An XFP's fluorescence lifetime contains additional information about the immediate environment of the fluorophore that can augment the information from its weak light signal. Here, we constructed and expressed in Saccharomyces cerevisiae variants of Teal Fluorescent Protein (TFP and Citrine that were isospectral but had shorter fluorescence lifetimes, ∼ 1.5 ns vs ∼ 3 ns. We modified microscopic and flow cytometric instruments to measure fluorescence lifetimes in live cells. We developed digital hardware and a measure of lifetime called a "pseudophasor" that we could compute quickly enough to permit sorting by lifetime in flow. We used these abilities to sort mixtures of cells expressing TFP and the short-lifetime TFP variant into subpopulations that were respectively 97% and 94% pure. This work demonstrates the feasibility of using information about fluorescence lifetime to help quantify cell signaling in living cells at the high throughput provided by flow cytometry. Moreover, it demonstrates the feasibility of isolating and recovering subpopulations of cells with different XFP lifetimes for subsequent experimentation.

  15. Optimized Time-Gated Fluorescence Spectroscopy for the Classification and Recycling of Fluorescently Labeled Plastics.

    Science.gov (United States)

    Fomin, Petr; Zhelondz, Dmitry; Kargel, Christian

    2016-08-29

    For the production of high-quality parts from recycled plastics, a very high purity of the plastic waste to be recycled is mandatory. The incorporation of fluorescent tracers ("markers") into plastics during the manufacturing process helps overcome typical problems of non-tracer based optical classification methods. Despite the unique emission spectra of fluorescent markers, the classification becomes difficult when the host plastics exhibit (strong) autofluorescence that spectrally overlaps the marker fluorescence. Increasing the marker concentration is not an option from an economic perspective and might also adversely affect the properties of the plastics. A measurement approach that suppresses the autofluorescence in the acquired signal is time-gated fluorescence spectroscopy (TGFS). Unfortunately, TGFS is associated with a lower signal-to-noise (S/N) ratio, which results in larger classification errors. In order to optimize the S/N ratio we investigate and validate the best TGFS parameters-derived from a model for the fluorescence signal-for plastics labeled with four specifically designed fluorescent markers. In this study we also demonstrate the implementation of TGFS on a measurement and classification prototype system and determine its performance. Mean values for a sensitivity of [Formula: see text] = 99.93% and precision [Formula: see text] = 99.80% were achieved, proving that a highly reliable classification of plastics can be achieved in practice.

  16. Effect of refractive index on the fluorescence lifetime of green fluorescent protein.

    Science.gov (United States)

    Tregidgo, Carolyn; Levitt, James A; Suhling, Klaus

    2008-01-01

    The average fluorescence lifetime of the green fluorescent protein (GFP) in solution is a function of the refractive index of its environment. We report that this is also the case for GFP-tagged proteins in cells. Using time-correlated single-photon counting (TCSPC)-based fluorescence lifetime imaging (FLIM) with a confocal scanning microscope, images of GFP-tagged proteins in cells suspended in different refractive index media are obtained. It is found that the average fluorescence lifetime of GFP decreases on addition of glycerol or sucrose to the media in which the fixed cells are suspended. The inverse GFP lifetime is proportional to the refractive index squared. This is the case for GFP-tagged major histocompatibility complex (MHC) proteins with the GFP located inside the cytoplasm, and also for GPI-anchored GFP that is located outside the cell membrane. The implications of these findings are discussed with regard to total internal reflection fluorescence (TIRF) techniques where the change in refractive index is crucial in producing an evanescent wave to excite fluorophores near a glass interface. Our findings show that the GFP fluorescence lifetime is shortened in TIRF microscopy in comparison to confocal microscopy.

  17. Fluorescent fingerprints of edible oils and biodiesel by means total synchronous fluorescence and Tucker3 modeling

    Science.gov (United States)

    Insausti, Matías; de Araújo Gomes, Adriano; Camiña, José Manuel; de Araújo, Mario Cesar Ugulino; Band, Beatriz Susana Fernández

    2017-03-01

    The present work proposes the use of total synchronous fluorescence spectroscopy (TSFS) as a discrimination methodology for fluorescent compounds in edible oils, which are preserved after the transesterification processes in the biodiesel production. In the same way, a similar study is presented to identify fluorophores that do not change in expired vegetal oils, to associate physicochemical parameters to fluorescent measures, as contribution to a fingerprint for increasing the chemical knowledge of these products. The fluorescent fingerprints were obtained by Tucker3 decomposition of a three-way array of the total synchronous fluorescence matrices. This chemometric method presents the ability for modeling non-bilinear data, as Total Synchronous Fluorescence Spectra data, and consists in the decomposition of the three way data arrays (samples × Δλ × λ excitation), into four new data matrices: A (scores), B (profile in Δλ mode), C (profile in spectra mode) and G (relationships between A, B and C). In this study, 50 samples of oil from soybean, corn and sunflower seeds before and after its expiration time, as well as 50 biodiesel samples obtained by transesterification of the same oils were measured by TSFS. This study represents an immediate application of chemical fingerprint for the discrimination of non-expired and expired edible oils and biodiesel. This method does not require the use of reagents or laborious procedures for the chemical characterization of samples.

  18. 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.

  19. 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.

  20. Fluorescence Microscopy of Nanoscale Silver Oxide Thin Films

    Institute of Scientific and Technical Information of China (English)

    PAN Xin-Yu; JIANG Hong-Bing; LIU Chun-Ling; GONG Qi-Huang; ZHANG Xi-Yao; ZHANG Qi-Feng; XU Bei-Xue; WU Jin-Lei

    2003-01-01

    The experimental conditions for photoactivated intermittent fluorescence from nanoscale silver oxide were studied with fluorescence microscopy. Strong fluorescence was observed from the Ag?O particles with size of 10-20nm excited with both blue and green light. We observed the saturation of photoexcitation with blue light and explained the experimental results using the model of agglomeration of silver atoms to form small clusters and the fluorescence of Ag2 and Ags clusters.

  1. Using Fluorescent Viruses for Detecting Bacteria in Water

    Science.gov (United States)

    Tabacco, Mary Beth; Qian, Xiaohua; Russo, Jaimie A.

    2009-01-01

    A method of detecting water-borne pathogenic bacteria is based partly on established molecular-recognition and fluorescent-labeling concepts, according to which bacteria of a species of interest are labeled with fluorescent reporter molecules and the bacteria can then be detected by fluorescence spectroscopy. The novelty of the present method lies in the use of bacteriophages (viruses that infect bacteria) to deliver the fluorescent reporter molecules to the bacteria of the species of interest.

  2. Coumarinyl(thienyl)thiazoles: novel photochromes with modulated fluorescence.

    Science.gov (United States)

    Traven, Valery F; Bochkov, Andrei Y; Krayushkin, Mikhail M; Yarovenko, Vladimir N; Nabatov, Boris V; Dolotov, Sergei M; Barachevsky, Valery A; Beletskaya, Irina P

    2008-03-20

    Novel photochromic 5-(3'-coumarinyl)-4-(3''-thienyl)thiazoles have been synthesized. These compounds display intensive fluorescence emission in the open form A, which is modulated by light. Fluorescence intensity decreases significantly upon irradiation of A with UV-light (lambdaB. Irradiation of B with visible light (lambda>470 nm) promotes its opening and the recovering of fluorescence. Novel dihetarylethenes undergo photochromic modulation of fluorescence both in solution and in polymeric matrices.

  3. Determination of the Residual Anthracene Concentration in Cultures of Haloalkalitolerant Actinomycetes by Excitation Fluorescence, Emission Fluorescence, and Synchronous Fluorescence: Comparative Study

    Directory of Open Access Journals (Sweden)

    Reyna del Carmen Lara-Severino

    2016-01-01

    Full Text Available Polycyclic aromatic hydrocarbons (PAHs are compounds that can be quantified by fluorescence due to their high quantum yield. Haloalkalitolerant bacteria tolerate wide concentration ranges of NaCl and pH. They are potentially useful in the PAHs bioremediation of saline environments. However, it is known that salinity of the sample affects fluorescence signal regardless of the method. The objective of this work was to carry out a comparative study based on the sensitivity, linearity, and detection limits of the excitation, emission, and synchronous fluorescence methods, during the quantification of the residual anthracene concentration from the following haloalkalitolerant actinomycetes cultures Kocuria rosea, Kocuria palustris, Microbacterium testaceum, and 4 strains of Nocardia farcinica, in order to establish the proper fluorescence method to study the PAHs biodegrading capacity of haloalkalitolerant actinobacteria. The study demonstrated statistical differences among the strains and among the fluorescence methods regarding the anthracene residual concentration. The results showed that excitation and emission fluorescence methods performed very similarly but sensitivity in excitation fluorescence is slightly higher. Synchronous fluorescence using Δλ=150 nm is not the most convenient method. Therefore we propose the excitation fluorescence as the fluorescence method to be used in the study of the PAHs biodegrading capacity of haloalkalitolerant actinomycetes.

  4. Fluorescent method for monitoring cheese starter permeabilization and lysis

    NARCIS (Netherlands)

    Bunthof, C.J.; Schalkwijk, van S.; Meijer, W.; Abee, T.; Hugenholtz, J.

    2001-01-01

    A fluorescence method to monitor lysis of cheese starter bacteria using dual staining with the LIVE/DEAD BacLight bacterial viability kit is described. This kit combines membrane-permeant green fluorescent nucleic acid dye SYTO 9 and membrane-impermeant red fluorescent nucleic acid dye propidium iod

  5. Fluorescence Emission from Small Molecules Containing Amino Group

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    After the treatment of oxygen gas, the small molecules containing amine group could emit fluorescence. Oxidation was believed to play an important role in the formation of fluorescence centers. Compared to previous results, both small molecules and macromolecules might have the same fluorescence centers.

  6. Carboxyfluorescein Diacetate Succinimidyl Ester Fluorescent Dye for Cell Labeling

    Institute of Scientific and Technical Information of China (English)

    Xiao-Qi WANG; Xiu-Mei DUAN; Li-Hua LIU; Yan-Qiu FANG; Yan TAN

    2005-01-01

    Our objective was to study the properties of the carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and the methodology of cell labeling using CFDA-SE fluorescent dye. First, we analyzed the kinetics of CFDA-SE fluorescent dye intensity over time. Second, we determined the optimal concentration of CFDA-SE fluorescent dye for cell labeling. Third, we tested the toxicity of CFDA-SE fluorescent dye on labeled cells. Finally, we determined the optimal staining time of CFDA-SE fluorescent dye for cell labeling.The results show that the optimal concentration of CFDA-SE fluorescent dye for cell labeling varies according to different cell types. CFDA-SE fluorescent dye is non-toxic to cells as the cell death rate caused by CFDASE labeling is below 5%. The optimal cell labeling time was determined to be 8 min of incubation with CFDA-SE fluorescent dye. We concluded that the advantages of using CFDA-SE fluorescent dye for cell labeling are as follows: (1) the binding of CFDA-SE fluorescent dye to cells is stable; (2) CFDA-SE fluorescent dye is not toxic and does not modify the viability of labeled cells; and (3) CFDA-SE fluorescent dye is a suitable fluorochrome for cell labeling.

  7. 7 CFR 201.61 - Fluorescence percentages in ryegrasses.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Fluorescence percentages in ryegrasses. 201.61 Section... ACT FEDERAL SEED ACT REGULATIONS Tolerances § 201.61 Fluorescence percentages in ryegrasses. Tolerances for 400-seed fluorescence tests shall be those set forth in the following table plus one-half...

  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. Chlorophyll a fluorescence and herbicide efficacy, metabolism and selectivity

    DEFF Research Database (Denmark)

    Abbas Poor, Majid

    Analysis of chlorophyll fluorescence induction curve (Kautsky curve) parameters was used for the study of the efficacy, metabolism and selectivity of ACCase, PSII and EPSPS inhibitors. Fv/Fm, Fvj and area above Kautsky curve and maximum fluorescence were selected among numerous fluorescence...

  10. 21 CFR 872.1745 - Laser fluorescence caries detection device.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Laser fluorescence caries detection device. 872... SERVICES (CONTINUED) MEDICAL DEVICES DENTAL DEVICES Diagnostic Devices § 872.1745 Laser fluorescence caries detection device. (a) Identification. A laser fluorescence caries detection device is a laser,...

  11. DETERMINATION OF AMINOGLYCOSIDES IN FOOD BY FLUORESCENCE POLARIZATION IMMUNOASSAY

    OpenAIRE

    FARAFONOVA O.V.; Eremin, S. A.; ERMOLAEVA T.N.; VASILIEV S.V.

    2015-01-01

    The methodic for quantitative determination of aminoglycoside antibiotics (gentamicin, kanamycin, streptomycin, amikacin, neomycin) in food by polarization fluorescent immunoassay (FPIA) is developed. The size and structure influence of a fluorescent molecule on a fluorescence polarization degree is analyzed. Affinity constants of antibodies to compounds and tracers were estimated, optimized working concentration of tracers and antibodies that provide the maximum value of analytical signal. M...

  12. Syntheses of Some Organic Fluorescent Dyes for Security Tickers

    Institute of Scientific and Technical Information of China (English)

    LI Jun-fen; BAI Guan; LIN Pei-hua; TIAN Mei-lin; DONG Chuan; LI Du-xin

    2004-01-01

    Five organic fluorescence dyes were synthesized by two- or three-step reactions. These synthetic methods have an advantage of the simple processes, low costs and high yields. The compositions of the five compounds are characterized by IR, 1H NMR, elemental analyses and fluorescence spectroscopies. The quantum yields of fluorescence were measured.

  13. 荧光原位杂交技术在膀胱尿路上皮癌术后复发的早期检测中的临床应用价值%Clinical value of fluorescence in situ hybridization in early detection of postoperative recurrence of bladder urothelial cancer

    Institute of Scientific and Technical Information of China (English)

    闫伟; 康雯婷; 陈山; 刘跃新

    2014-01-01

    目的:应用荧光原位杂交技术(fluorescence in situ hybridization,FISH)对膀胱尿路上皮肿瘤患者术后复发进行早期评估预测。方法2009年1月至2011年11月,选取首都医科大学附属北京同仁医院40例临床诊断的高危膀胱尿路上皮癌患者,术后第3,6,9,12,18个月行尿细胞学检查、膀胱镜检查,并行尿液 FISH 检查,记录膀胱肿瘤复发的时间。观察 FISH 对于膀胱尿路上皮肿瘤患者术后复发的早期预测价值。结果60%(12/20)的 FISH 阳性者膀胱肿瘤复发,FISH 阴性者无1例出现膀胱肿瘤复发。 FISH 阳性者膀胱尿路上皮癌复发率高于 FISH 阴性者(60% vs 0%,P<0.05)。6例(50%)肿瘤复发患者 FISH 阳性先于膀胱镜检查及尿细胞学检查。结论 FISH 对膀胱肿瘤检测的阳性率高于细胞学检测,FISH 对膀胱肿瘤的诊断早于膀胱镜和细胞学。 FISH 技术可作为膀胱尿路上皮肿瘤术后复发的监测指标之一。%Objective To early diagnose postoperative recurrence of bladder urothelial cancer with fluorescence in situ hybridization (FISH). Methods Forty patients with bladder urothelial cancer were chosen between January, 2009 and November, 2011. Urine cytologic examination, cystoscopy, and FISH were performed in these patients at 3, 6, 9, 12, and 18 months post-operation, and tumor recurrence was recorded. Results Among the 40 patients tested, 12 of 20 FISH-positive patients developed tumor recurrence within the follow-up period, while none of the FISH-negative patients developed recurrence during the same period. The recurrence rate in FISH-positive patients(60% ) was significantly higher than that of FISH-negative patients(P<0. 05). The sensitivity to detect tumor recurrence by FISH analysis was higher than that by cystoscopy and urine cytologic examination. Conclusion FISH analysis is effective in early detection of postoperative recurrence of bladder urothelial cancer.

  14. Thermophoresis of DNA determined by microfluidic fluorescence.

    Science.gov (United States)

    Duhr, S; Arduini, S; Braun, D

    2004-11-01

    We describe a microfluidic all-optical technique to measure the thermophoresis of molecules. Within micrometer-thick chambers, we heat aqueous solutions with a micrometer-sized focus of infrared light. The temperature increase of about 1 K is monitored with temperature-sensitive fluorescent dyes. We test the approach in measuring the thermophoresis of DNA. We image the concentration of DNA in a second fluorescence-color channel. DNA is depleted away from the heated spot. The profile of depletion is fitted by the thermophoretic theory to reveal the Soret coefficient. We evaluate the method with numerical 3D calculations of temperature profiles, drift, convection and thermophoretic depletion using finite element methods. The approach opens new ways to monitor thermophoresis at the single molecule level, near boundaries and in complex mixtures. The flexible microfluidic setting is a good step towards microfluidic applications of thermophoresis in biotechnology.

  15. Filtered fluorescer x-ray detector

    Energy Technology Data Exchange (ETDEWEB)

    Bruns, H.C.; Emig, J.A.; Thoe, R.S.; Springer, P.T.; Hernandez, J.A.

    1995-04-01

    Recently, an instrument capable of measuring x-rays between 8 and 90 keV was conceived to help understand conditions pertaining to pulsed power research. This resulted in the development of a versatile device that would incrementally detect x-rays emitted at predetermined energy bands over this range. To accomplish this, an array of well characterized filter-fluorescer combinations were produced which would allow fluoresced x-rays to be observed by time resolved electro-optical devices. As many as sixteen channels could be utilized with each channel having a corresponding background channel. Upon completion of the device, a three week series of experiments was then successfully carried out.

  16. Portable fluorescence meter for medical applications

    Science.gov (United States)

    Kornilin, Dmitriy V.; Grishanov, Vladimir N.

    2016-04-01

    Recently, there are great deals of skin fluorescence studies for diagnostic purposes in medicine. Measurement of the intensity of autofluorescence (AF) is suitable method for diagnostic, because it does not require traumatic procedures. Skin AF is widely used by doctors in order to assess the concentration of advanced glycation endproduct (AGE). There are no in vivo fluorescence meters made in Russia, which are affordable, portable, easy-to-use and easily replicable. This paper is devoted to study of the fluorimeter and its mathematical model of spectral characteristics that were developed by authors. Fluorimeter and its software are fully operational and they were given to doctors for testing in the real clinic conditions in order to get a set of AF statistics for patients.

  17. Introduction to fluorescence probing of biological membranes.

    Science.gov (United States)

    Demchenko, Alexander P; Duportail, Guy; Oncul, Sule; Klymchenko, Andrey S; Mély, Yves

    2015-01-01

    Fluorescence is one of the most powerful and commonly used tools in biophysical studies of biomembrane structure and dynamics that can be applied on different levels, from lipid monolayers and bilayers to living cells, tissues, and whole animals. Successful application of this method relies on proper design of fluorescence probes with optimized photophysical properties. These probes are efficient for studying the microscopic analogs of viscosity, polarity, and hydration, as well as the molecular order, environment relaxation, and electrostatic potentials at the sites of their location. Being smaller than the membrane width they can sense the gradients of these parameters across the membrane. We present examples of novel dyes that achieve increased spatial resolution and information content of the probe responses. In this respect, multiparametric environment-sensitive probes feature considerable promise.

  18. Fluorescent silver nanoparticles via exploding wire technique

    Indian Academy of Sciences (India)

    Alqudami Abdullah; S Annapoorni

    2005-11-01

    Aqueous solution containing spherical silver nanoparticles of 20–80 nm size have been generated using a newly developed novel electro-exploding wire (EEW) technique where thin silver wires have been exploded in double distilled water. Structural properties of the resulted nanoparticles have been studied by means of X-ray diffractometer (XRD) and transmission electron microscopy (TEM). The absorption spectrum of the aqueous solution of silver nanoparticles showed the appearance of a broad surface plasmon resonance (SPR) peak centered at a wavelength of 390 nm. The theoretically generated SPR peak seems to be in good agreement with the experimental one. Strong green fluorescence emission was observed from the water-suspended silver nanoparticles excited with light of wavelengths 340, 360 and 390 nm. The fluorescence of silver nanoparticles could be due to the excitation of the surface plasmon coherent electronic motion with the small size effect and the surface effect considerations.

  19. Laser induced fluorescence of dental caries

    Science.gov (United States)

    Albin, S.; Byvik, C. E.; Buoncristiani, A. M.

    1988-01-01

    Significant differences between the optical spectra taken from sound regions of teeth and carious regions have been observed. These differences appear both in absorption and in laser induced fluorescence spectra. Excitation by the 488 nm line of an argon ion laser beam showed a peak in the emission intensity around 553 nm for the sound dental material while the emission peak from the carious region was red-shifted by approximately 40 nm. The relative absorption of carious region was significantly higher at 488 nm; however its fluorescence intensity peak was lower by an order of magnitude compared to the sound tooth. Implications of these results for a safe, reliable and early detection of dental caries are discussed.

  20. Laser induced fluorescence of dental caries

    Science.gov (United States)

    Albin, S.; Byvik, C. E.; Buoncristiani, A. M.

    1988-01-01

    Significant differences between the optical spectra taken from sound regions of teeth and carious regions have been observed. These differences appear both in absorption and in laser induced fluorescence spectra. Excitation by the 488 nm line of an argon ion laser beam showed a peak in the emission intensity around 553 nm for the sound dental material while the emission peak from the carious region was red-shifted by approximately 40 nm. The relative absorption of carious region was significantly higher at 488 nm; however its fluorescence intensity peak was lower by an order of magnitude compared to the sound tooth. Implications of these results for a safe, reliable and early detection of dental caries are discussed.

  1. Fluorescence bronchoscope for lung tumor localization

    Energy Technology Data Exchange (ETDEWEB)

    Profio, A.E. (Univ. of California, Santa Barbara); Doiron, D.R.; Huth, G.C.

    1977-02-01

    A fluorescence bronchoscope has been developed for localization of small bronchogenic tumors at the carcinoma in situ stage. Injected hematoporphyrin-derivative is preferentially taken up or retained by a malignant tumor, and very small amounts can be detected by the red fluorescence under excitation by violet light. The target lesion is 80 ..mu..m thick, with a mass of 250 ..mu..g, containing 250 pg of hematoporphyrin-derivative. A fiberoptic bronchoscope system with a 200W high pressure mercury arc lamp, primary filter passing 405 nm light, special violet transmitting light conductor, coherent imaging bundle, red secondary filter, and three-stage electrostatic focus image intensifier tube was designed for this application. Tumors have been visualized in animals and preparations for clinical use are underway.

  2. Fluorescent Bioactive Corrole Grafted-Chitosan Films.

    Science.gov (United States)

    Barata, Joana F B; Pinto, Ricardo J B; Vaz Serra, Vanda I R C; Silvestre, Armando J D; Trindade, Tito; Neves, Maria Graça P M S; Cavaleiro, José A S; Daina, Sara; Sadocco, Patrizia; Freire, Carmen S R

    2016-04-11

    Transparent corrole grafted-chitosan films were prepared by chemical modification of chitosan with a corrole macrocycle, namely, 5,10,15-tris(pentafluorophenyl)corrole (TPFC), followed by solvent casting. The obtained films were characterized in terms of absorption spectra (UV-vis), FLIM (fluorescence lifetime imaging microscopy), structure (FTIR, XPS), thermal stability (TGA), thermomechanical properties (DMA), and antibacterial activity. The results showed that the chemical grafting of chitosan with corrole units did not affect its film-forming ability and that the grafting yield increased with the reaction time. The obtained transparent films presented fluorescence which increases with the amount of grafted corrole units. Additionally, all films showed bacteriostatic effect against S. aureus, as well as good thermomechanical properties and thermal stability. Considering these features, promising applications may be envisaged for these corrole-chitosan films, such as biosensors, bioimaging agents, and bioactive optical devices.

  3. Fluorescence analysis of iodinated acetophenone derivatives.

    Science.gov (United States)

    Crivelaro, F; Oliveira, M R S; Lima, S M; Andrade, L H C; Casagrande, G A; Raminelli, C; Caires, A R L

    2015-03-15

    In the present paper the synthesis and optical characterization of iodinated acetophenone, 4-hydroxy-3-iodoacetophenone and 4-hydroxy-3,5-diiodoacetophenone obtained from 4-hydroxyacetophenone, were carried out. The optical features of iodinated molecules were determined by performing the UV-Vis absorption, fluorescence and thermal lens spectroscopies. The results showed that the optical properties of the 4-hydroxyacetophenone is altered when the iodine atom is inserted, as substituent, in the aromatic ring. Although it was determined that the optical feature was changed when one iodine atom was inserted in the aromatic ring (4-hydroxy-3-iodoacetophenone), the results revealed that emission behavior was strongly altered when two iodine atoms (4-hydroxy-3,5-diiodoacetophenone) were acting as substituents: the fluorescence quantum efficiency increases approximately 60%.

  4. Fluorescence Spectroscopic Studies on Ovis Lactoperoxidase

    Directory of Open Access Journals (Sweden)

    P. V. Joseph

    2004-01-01

    Full Text Available Ovis lactoperoxidase (sLP, on excitation at 280 nm shows fluorescence emission of a single broad maximum at 332 nm. The conformational stability was measured by unfolding studies in urea and guanidine hydrochloride. The fluorescence intensity gradually decreased with increase in urea concentrations. The decline might have been caused by partial unfolding, affecting some of the tryptophan residues. In 5 M GuHCl concentrations, a red shift in emission maximum to 356 nm was observed. It indicates that tryptophan is buried in the interior of the hydrophobic environment in native folded state and inaccessible to solvent water but on unfolding all get exposed to aqueous environment. Acrylamide is an efficient quencher and the quenching process is essentially homogenous with all tryptophan being accessible. A little quenching is observed for KI is interpreted as sLP has tryptophan residues that are buried inside the core of the protein.

  5. Highly Fluorescent Noble Metal Quantum Dots

    Science.gov (United States)

    Zheng, Jie; Nicovich, Philip R.; Dickson, Robert M.

    2009-01-01

    Highly fluorescent, water-soluble, few-atom noble metal quantum dots have been created that behave as multi-electron artificial atoms with discrete, size-tunable electronic transitions throughout the visible and near IR. These “molecular metals” exhibit highly polarizable transitions and scale in size according to the simple relation, Efermi/N1/3, predicted by the free electron model of metallic behavior. This simple scaling indicates that fluorescence arises from intraband transitions of free electrons and that these conduction electron transitions are the low number limit of the plasmon – the collective dipole oscillations occurring when a continuous density of states is reached. Providing the “missing link” between atomic and nanoparticle behavior in noble metals, these emissive, water-soluble Au nanoclusters open new opportunities for biological labels, energy transfer pairs, and light emitting sources in nanoscale optoelectronics. PMID:17105412

  6. X-ray microtome by fluorescence tomography

    CERN Document Server

    Simionovici, A S; Guenzler, F; Schrör, C; Snigirev, A; Snigireva, I; Tümmler, J; Weitkamp, T

    2001-01-01

    The X-ray fluorescence microtomography method is presented, which is capable of virtually slicing samples to obtain cross-sections of their inner structure. High precision experimental results of fluo-tomography in 'pencil-beam' geometry with up to 1.2 mu m resolution are described. Image reconstructions are based on either a simplified algebraic reconstruction method (ART) or the filtered back-projection method (FBP). Phantoms of inhomogeneous test objects as well as biological samples are successfully analyzed.

  7. Estimating phytoplankton photosynthesis by active fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Falkowski, P.G.; Kolber, Z.

    1992-01-01

    Photosynthesis can be described by target theory, At low photon flux densities, photosynthesis is a linear function of irradiance (I), The number of reaction centers (n), their effective absorption capture cross section {sigma}, and a quantum yield {phi}. As photosynthesis becomes increasingly light saturated, an increased fraction of reaction centers close. At light saturation the maximum photosynthetic rate is given as the product of the number of reaction centers (n) and their maximum electron transport rate (I/{tau}). Using active fluorometry it is possible to measure non-destructively and in real time the fraction of open or closed reaction centers under ambient irradiance conditions in situ, as well as {sigma} and {phi} {tau} can be readily, calculated from knowledge of the light saturation parameter, I{sub k} (which can be deduced by in situ by active fluorescence measurements) and {sigma}. We built a pump and probe fluorometer, which is interfaced with a CTD. The instrument measures the fluorescence yield of a weak probe flash preceding (f{sub 0}) and succeeding (f{sub 0}) a saturating pump flash. Profiles of the these fluorescence yields are used to derive the instantaneous rate of gross photosynthesis in natural phytoplankton communities without any incubation. Correlations with short-term simulated in situ radiocarbon measurements are extremely high. The average slope between photosynthesis derived from fluorescence and that measured by radiocarbon is 1.15 and corresponds to the average photosynthetic quotient. The intercept is about 15% of the maximum radiocarbon uptake and corresponds to the average net community respiration. Profiles of photosynthesis and sections showing the variability in its composite parameters reveal a significant effect of nutrient availability on biomass specific rates of photosynthesis in the ocean.

  8. Estimating phytoplankton photosynthesis by active fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Falkowski, P.G.; Kolber, Z.

    1992-10-01

    Photosynthesis can be described by target theory, At low photon flux densities, photosynthesis is a linear function of irradiance (I), The number of reaction centers (n), their effective absorption capture cross section {sigma}, and a quantum yield {phi}. As photosynthesis becomes increasingly light saturated, an increased fraction of reaction centers close. At light saturation the maximum photosynthetic rate is given as the product of the number of reaction centers (n) and their maximum electron transport rate (I/{tau}). Using active fluorometry it is possible to measure non-destructively and in real time the fraction of open or closed reaction centers under ambient irradiance conditions in situ, as well as {sigma} and {phi} {tau} can be readily, calculated from knowledge of the light saturation parameter, I{sub k} (which can be deduced by in situ by active fluorescence measurements) and {sigma}. We built a pump and probe fluorometer, which is interfaced with a CTD. The instrument measures the fluorescence yield of a weak probe flash preceding (f{sub 0}) and succeeding (f{sub 0}) a saturating pump flash. Profiles of the these fluorescence yields are used to derive the instantaneous rate of gross photosynthesis in natural phytoplankton communities without any incubation. Correlations with short-term simulated in situ radiocarbon measurements are extremely high. The average slope between photosynthesis derived from fluorescence and that measured by radiocarbon is 1.15 and corresponds to the average photosynthetic quotient. The intercept is about 15% of the maximum radiocarbon uptake and corresponds to the average net community respiration. Profiles of photosynthesis and sections showing the variability in its composite parameters reveal a significant effect of nutrient availability on biomass specific rates of photosynthesis in the ocean.

  9. APD detectors for biological fluorescence spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Mazeres, S. [Institut de Pharmacologie et de Biologie Structurale, IPBS-CNRS, 205 route de Narbonne 31077 Toulouse Cedex 4 (France)]. E-mail: serge.mazeres@ipbs.fr; Borrel, V. [GIATHE/CESR, 9 avenue du Colonel Roche BP 4346, 31029 Toulouse Cedex (France); Magenc, C. [GIATHE/CESR, 9 avenue du Colonel Roche BP 4346, 31029 Toulouse Cedex (France); Courrech, J.L. [GIATHE/CESR, 9 avenue du Colonel Roche BP 4346, 31029 Toulouse Cedex (France); Bazer-Bachi, R. [GIATHE/CESR, 9 avenue du Colonel Roche BP 4346, 31029 Toulouse Cedex (France)

    2006-11-01

    Fluorescence spectroscopy is a very convenient and widely used method for studying the molecular background of biological processes [L. Salome, J.L. Cazeil, A. Lopez, J.F. Tocanne, Eur. Biophys. J. 27 (1998) 391-402]. Chromophores are included in the structure under study and a flash of laser light induces fluorescence (Fluorescence Recovery After Photo-bleaching), the decay of which yields information on the polarity, the speed of rotation, and the speed of diffusion as well as on the temporal and spatial evolution of interactions between molecular species. The method can even be used to study living cells [J.F. Tocanne, L. Cezanne, A. Lopez, Prog. Lipid Res. 33 (1994) 203-237, L. Cezanne, A. Lopez, F. Loste, G. Parnaud, O. Saurel, P. Demange, J.F. Tocanne, Biochemistry 38 (1999) 2779-2786]. This is classically performed with a PM-based system. For biological reasons a decrease of the excitation of the cells is highly desirable. Because the fluorescence response then becomes fainter a significant improvement in detector capability would be welcome. We present here results obtained with an Avalanche Photo Diode (APD)-based system. The small sensitive area of detection allows a very significant improvement in signal/noise ratio, improvement in gain, and the opening-up of a new parameter space. With these new detectors we can begin the study of information transmission between cells through morphine receptors. This work involves both electronics engineers and biophysicists, so results and techniques in both fields will be presented here.

  10. Fluorescent beeswax for surface flow velocity observations

    Science.gov (United States)

    Grimaldi, S.; Tauro, F.; Petroselli, A.; Mocio, G.; Capocci, I.; Rapiti, E.; Rapiti, R.; Cipollari, G.; Porfiri, M.

    2012-12-01

    Watershed surface processes control downstream runoff phenomena, waste and pollutant diffusion, erosion mechanics, and sediment transport. A quantitative understanding of the flow physics is currently limited by the lack of effective tracing techniques suitable for basin-scale observations. More specifically, field experiments require environmentally resilient, non-invasive, and low cost measurement systems that can potentially operate in remotely-controlled or unmanned conditions. Traditional tracing methodologies are largely not capable to cope with extreme in-situ conditions, including practical logistic challenges as well as inherent flow complexity. Specifically, most of available technologies need physical sampling to estimate the tracer concentration and do not allow for continuous-time measurements. In addition, commonly used tracers, such as isotopes, dyes, and chemicals, are not directly applicable to monitor surface hillslope processes and large-scale microchannel networks due to elaborate detection processes and dispersion issues. In this context, the feasibility of using buoyant fluorescent microspheres as particle tracers in natural water flows is investigated. Specifically, a novel fabrication methodology is designed to manufacture particles from natural beeswax and a highly diluted solution of a nontoxic fluorescent red dye. The fabrication procedure allows for adjusting the size of the particles from tens of microns up to a few millimeters and their density from positively to negatively-buoyant with respect to water. An array of experimental techniques is employed to conduct a thorough characterization of the fluorescence and morphology of the tracers. In addition, ad-hoc experiments are designed to assess the fluorescence response due to Ultra Violet (UV) exposure and thermal processes. Proof-of-concept laboratory analysis are conducted to illustrate the integration of the novel particle tracers in existing tracing methods for surface flow

  11. Photophysics of thermally activated delayed fluorescence molecules

    Science.gov (United States)

    Dias, Fernando B.; Penfold, Thomas J.; Monkman, Andrew P.

    2017-03-01

    Thermally activated delayed fluorescence (TADF) has recently emerged as one of the most attractive methods for harvesting triplet states in metal-free organic materials for application in organic light emitting diodes (OLEDs). A large number of TADF molecules have been reported in the literature with the purpose of enhancing the efficiency of OLEDs by converting non-emissive triplet states into emissive singlet states. TADF emitters are able to harvest both singlets and triplet states through fluorescence (prompt and delayed), the latter due to the thermally activated reverse intersystem crossing mechanism that allows up-conversion of low energy triplet states to the emissive singlet level. This allows otherwise pure fluorescent OLEDs to overcome their intrinsic limit of 25% internal quantum efficiency (IQE), which is imposed by the 1:3 singlet-triplet ratio arising from the recombination of charges (electrons and holes). TADF based OLEDS with IQEs close to 100% are now routinely fabricated in the green spectral region. There is also significant progress for blue emitters. However, red emitters still show relatively low efficiencies. Despite the significant progress that has been made in recent years, still significant challenges persist to achieve full understanding of the TADF mechanism and improve the stability of these materials. These questions need to be solved in order to fully implement TADF in OLEDs and expand their application to other areas. To date, TADF has been exploited mainly in the field of OLEDs, but applications in other areas, such as sensing and fluorescence microscopies, are envisaged. In this review, the photophysics of TADF molecules is discussed, summarising current methods to characterise these materials and the current understanding of the TADF mechanism in various molecular systems.

  12. Fluorescent prey traps in carnivorous plants.

    Science.gov (United States)

    Kurup, R; Johnson, A J; Sankar, S; Hussain, A A; Sathish Kumar, C; Sabulal, B

    2013-05-01

    Carnivorous plants acquire most of their nutrients by capturing ants, insects and other arthropods through their leaf-evolved biological traps. So far, the best-known attractants in carnivorous prey traps are nectar, colour and olfactory cues. Here, fresh prey traps of 14 Nepenthes, five Sarracenia, five Drosera, two Pinguicula species/hybrids, Dionaea muscipula and Utricularia stellaris were scanned at UV 366 nm. Fluorescence emissions of major isolates of fresh Nepenthes khasiana pitcher peristomes were recorded at an excitation wavelength of 366 nm. N. khasiana field pitcher peristomes were masked by its slippery zone extract, and prey capture rates were compared with control pitchers. We found the existence of distinct blue fluorescence emissions at the capture spots of Nepenthes, Sarracenia and Dionaea prey traps at UV 366 nm. These alluring blue emissions gradually developed with the growth of the prey traps and diminished towards their death. On excitation at 366 nm, N. khasiana peristome 3:1 CHCl3–MeOH extract and its two major blue bands showed strong fluorescence emissions at 430–480 nm. Masking of blue emissions on peristomes drastically reduced prey capture in N. khasiana pitchers. We propose these molecular emissions as a critical factor attracting arthropods and other visitors to these carnivorous traps. Drosera, Pinguicula and Utricularia prey traps showed only red chlorophyll emissions at 366 nm.

  13. Metal-enhanced fluorescence of carbon nanotubes.

    Science.gov (United States)

    Hong, Guosong; Tabakman, Scott M; Welsher, Kevin; Wang, Hailiang; Wang, Xinran; Dai, Hongjie

    2010-11-17

    The photoluminescence (PL) quantum yield of single-walled carbon nanotubes (SWNTs) is relatively low, with various quenching effects by metallic species reported in the literature. Here, we report the first case of metal enhanced fluorescence (MEF) of surfactant-coated carbon nanotubes on nanostructured gold substrates. The photoluminescence quantum yield of SWNTs is observed to be enhanced more than 10-fold. The dependence of fluorescence enhancement on metal-nanotube distance and on the surface plasmon resonance (SPR) of the gold substrate for various SWNT chiralities is measured to reveal the mechanism of enhancement. Surfactant-coated SWNTs in direct contact with metal exhibit strong MEF without quenching, suggesting a small quenching distance for SWNTs on the order of the van der Waals distance, beyond which the intrinsically fast nonradiative decay rate in nanotubes is little enhanced by metal. The metal enhanced fluorescence of SWNTs is attributed to radiative lifetime shortening through resonance coupling of SWNT emission to the reradiating dipolar plasmonic modes in the metal.

  14. Fluorescent lighting with aluminum nitride phosphors

    Energy Technology Data Exchange (ETDEWEB)

    Cherepy, Nerine J.; Payne, Stephen A.; Seeley, Zachary M.; Srivastava, Alok M.

    2016-05-10

    A fluorescent lamp includes a glass envelope; at least two electrodes connected to the glass envelope; mercury vapor and an inert gas within the glass envelope; and a phosphor within the glass envelope, wherein the phosphor blend includes aluminum nitride. The phosphor may be a wurtzite (hexagonal) crystalline structure Al.sub.(1-x)M.sub.xN phosphor, where M may be drawn from beryllium, magnesium, calcium, strontium, barium, zinc, scandium, yttrium, lanthanum, cerium, praseodymium, europium, gadolinium, terbium, ytterbium, bismuth, manganese, silicon, germanium, tin, boron, or gallium is synthesized to include dopants to control its luminescence under ultraviolet excitation. The disclosed Al.sub.(1-x)M.sub.xN:Mn phosphor provides bright orange-red emission, comparable in efficiency and spectrum to that of the standard orange-red phosphor used in fluorescent lighting, Y.sub.2O.sub.3:Eu. Furthermore, it offers excellent lumen maintenance in a fluorescent lamp, and does not utilize "critical rare earths," minimizing sensitivity to fluctuating market prices for the rare earth elements.

  15. Molecular cytogenetics using fluorescence in situ hybridization

    Energy Technology Data Exchange (ETDEWEB)

    Gray, J.W.; Kuo, Wen-Lin; Lucas, J.; Pinkel, D.; Weier, H-U.; Yu, Loh-Chung.

    1990-12-07

    Fluorescence in situ hybridization (FISH) with chromosome-specific probes enables several new areas of cytogenetic investigation by allowing visual determination of the presence and normality of specific genetic sequences in single metaphase or interphase cells. in this approach, termed molecular cytogenetics, the genetic loci to be analyzed are made microscopically visible in single cells using in situ hybridization with nucleic acid probes specific to these loci. To accomplish this, the DNA in the target cells is made single stranded by thermal denaturation and incubated with single-stranded, chemically modified probe under conditions where the probe will anneal only with DNA sequences to which it has high DNA sequence homology. The bound probe is then made visible by treatment with a fluorescent reagent such as fluorescein that binds to the chemical modification carried by the probe. The DNA to which the probe does not bind is made visible by staining with a dye such as propidium iodide that fluoresces at a wavelength different from that of the reagent used for probe visualization. We show in this report that probes are now available that make this technique useful for biological dosimetry, prenatal diagnosis and cancer biology. 31 refs., 3 figs.

  16. NANODIAMONDS FOR FLUORESCENT CELL AND SENSOR NANOTECHNOLOGIES

    Directory of Open Access Journals (Sweden)

    V. I. Nazarenko

    2013-10-01

    Full Text Available This review addresses the analysis of properties and applications of fluorescent nanodiamonds. They are carbon nanostructures with atomic arrangement of a diamond and carry all its properties, including record — high density, rigidity and refraction index. They are of almost spherical shape, and their small size (~4–10 nm creates substantial surface area that can be used for absorption of different compounds including drugs. Their surface is formed by different chemical groups (hydroxyls, carboxyls, etc. exhibits also chemical reactivity that allows different types of modifications. This opens innumerable possibilities for constructing different functional nanomaterials. The technologies have been developed for making these nanodiamonds fluorescent. Particularly, these properties are achieved by radioactive treatment with the formation of N–V impurities. These particles absorb and emit light in convenient for observation visible range of spectrum. They do not photobleach, which is very attractive for fluorescent microscopy of the cell. And, finally, these nanoparticles do not display toxicity on the cellular or whole — body level, and because of their biocompatibility they can be used in vivo as contrast agents and drug carriers. It is expected that future biotechnological applications of these nanoparticles will be connected with the creation of nanocomposites that combine multiple useful functions.

  17. Nonlinear inversion schemes for fluorescence optical tomography.

    Science.gov (United States)

    Freiberger, Manuel; Egger, Herbert; Scharfetter, Hermann

    2010-11-01

    Fluorescence optical tomography is a non-invasive imaging modality that employs the absorption and re-emission of light by fluorescent dyes. The aim is to reconstruct the fluorophore distribution in a body from measurements of light intensities at the boundary. Due to the diffusive nature of light propagation in tissue, fluorescence tomography is a nonlinear and severely ill-posed problem, and some sort of regularization is required for a stable solution. In this paper we investigate reconstruction methods based on Tikhonov regularization with nonlinear penalty terms, namely total-variation regularization and a levelset-type method using a nonlinear parameterization of the unknown function. Moreover, we use the full threedimensional nonlinear forward model, which arises from the governing system of partial differential equations. We discuss the numerical realization of the regularization schemes by Newtontype iterations, present some details of the discretization by finite element methods, and outline the efficient implementation of sensitivity systems via adjoint methods. As we will demonstrate in numerical tests, the proposed nonlinear methods provide better reconstructions than standard methods based on linearized forward models and linear penalty terms. We will additionally illustrate, that the careful discretization of the methods derived on the continuous level allows to obtain reliable, mesh independent reconstruction algorithms.

  18. Fluorescent Polystyrene Sulfonate for Polyelectrolyte Studies

    Science.gov (United States)

    Huberty, Wayne; Tong, Xiaowei; Balamurugan, Sreelatha; Zhang, Donghui; Russo, Paul

    2012-02-01

    The slow-mode decay found by dynamic light scattering for polyelectrolytes in low-salt conditions has perplexed investigators since its first observation. Many characterization methods have suggested temporary or transient aggregation, although there is still no consensus on the cause. Many different polyelectrolytes demonstrate the slow-mode decay, but the sodium salt of polystyrene sulfonate (NaPSS) is the most popular choice for study. Commercially available NaPSS may have hydrophobic patches due to incomplete sulfonation leading to associations apart from any putative ionic mechanisms. Therefore, essentially full sulfonation, or ``patchless'', NaPSS should be synthesized. To facilitate fluorescence measurements, which can provide new insights to the slow-mode phenomenon, the material must be rendered fluorescent (F-NaPSS). Several approaches to F-NaPSS have appeared; some labeled a previously synthesized NaPSS without concern for its hydrophobic patches. Other strategies include a free radical copolymerization of styrene sulfonate and a vinyl amine to provide side chains viable for labeling. This method is successful, but yields only small amounts of nearly monodisperse polymer after fractionation. In this presentation, a high-yield synthesis of fully sulfonated, low-polydispersity, fluorescently tagged polymer will be discussed.

  19. Fluorescence spectroscopic studies of DNA dynamics

    Energy Technology Data Exchange (ETDEWEB)

    Scalettar, B.A.

    1987-04-01

    Random solvent induced motions of DNA are manifest as nanosecond torsional oscillations of the helix backbone, nanosecond through millisecond bending deformations and overall rotational and translational diffusion of the polymer. Fluorescence spectroscopy is used to study this spectrum of DNA motions while ethidium monoazide was covalently bounded. The steady state fluorescence depolarization data indicate that the covalent monoazide/DNA complex exhibits internal motions characterized by an average angular amplitude of 26 degrees confirming reports of fast torsional oscillations in noncovalent ethidium bromide/DNA systems. Data obtained by use of a new polarized photobleaching recovery technique (FPR) reflect both the rotational dynamics of the polymer and the reversible photochemistry of the dye. To isolate the reorientational motion of the DNA, the FPR experiments were ran in two modes that differ only in the polarization of the bleaching light. A quotient function constructed from the data obtained in these two modes monitors only the rotational component of the FPR recovery. In specific applications those bending deformations of long DNA molecules that have characteristic relaxation times on the order of 100 microseconds have been resolved. A fluorescence correlation technique that relates fluctuations in particle number to center-of-mass motion was used to measure translational diffusion on coefficients of the plasmid PBR322 and a short oligomeric DNA. A theory that describes angular correlation in systems exhibiting cyclic, biologically directed reorientation and random Brownian rotation is developed.

  20. Gonadoblastoma and Y-chromosome fluorescence.

    Science.gov (United States)

    Lukusa, T; Fryns, J P; van den Berghe, H

    1986-04-01

    In this report we summarize our experience in 4 patients with 45,X/46,XY, one patient with 45,X/47,XYY mosaicism, and one patient with 46,XY karyotype and ambiguous external genitalia. In the 3 patients with a fluorescent Y-chromosome, the development of one or two gonadoblastomas was found, independent of the age of the patients at the time of examination. In the 3 patients with 45,X/46,XYnf mosaicism no gonadoblastoma was detected. This finding prompted us to review the data on patients reported with 45,X/46,XYnf mosaicism. Up to now, no patient with well documented 45,X/46,XYnf mosaicism and convincing evidence of development of gonadoblastoma has been reported. These data seem to confirm that alterations of the characteristic distal fluorescence of Yq may protect the dysgenetic gonad against tumoral degeneration in patients with 45,X/46,XY mosaicism. Possible mechanisms responsible for these changes in the oncogenic potential of Yq in relation with the Y chromosome fluorescence are discussed.

  1. UV fluorescence lidar detection of bioaerosols

    Energy Technology Data Exchange (ETDEWEB)

    Christesen, S.D.; DeSha, M.S.; Wong, A. [Army Edgewood Research, Development and Engineering Center, Aberdeen Proving Ground, MD (United States); Merrow, C.N.; Wilson, M.W.; Butler, J. [Science and Technology Corp., Hampton, VA (United States)

    1994-12-31

    Biological agents (e.g. bacterial spores, viruses, toxins) pose a serious threat to military forces on the modern battlefield. Remote detection of these agents is crucial to providing early warning of an attack and to allow for the avoidance of contaminated areas. Here, a UV fluorescence lidar system for the remote detection of bioaerosols has been built and tested. At the heart of the UV-LIDAR Fluorosensor system are a 200mJ quadrupled ND:YAG laser at 266nm and a 16 inch cassagrain telescope. Operating on three data collection channels, the UV lidar is capable of real time monitoring of 266nm elastic backscatter, the total fluorescence between 300 and 400nm, and the dispersed fluorescence spectrum (using a small spectrograph and gated intensified CCD array). The goal in this effort was to assess the capabilities of biofluorescence for quantitative detection and discrimination of bioaerosols. To this end, the UV-LIDAR Fluorosensor system was tested against the aerosolized bacterial spore Bacillus subtilus var. niger sp. globiggi (BG) and several likely interferences at several ranges from approximately 600 to 3000 meters. The tests with BG indicate a detection limit of approximately 500 mg/cubic meter at a range of 3000m.

  2. Fluorescence quantum yield measurements of fluorescent proteins: a laboratory experiment for a biochemistry or molecular biophysics laboratory course.

    Science.gov (United States)

    Wall, Kathryn P; Dillon, Rebecca; Knowles, Michelle K

    2015-01-01

    Fluorescent proteins are commonly used in cell biology to assess where proteins are within a cell as a function of time and provide insight into intracellular protein function. However, the usefulness of a fluorescent protein depends directly on the quantum yield. The quantum yield relates the efficiency at which a fluorescent molecule converts absorbed photons into emitted photons and it is necessary to know for assessing what fluorescent protein is the most appropriate for a particular application. In this work, we have designed an upper-level, biochemistry laboratory experiment where students measure the fluorescence quantum yields of fluorescent proteins relative to a standard organic dye. Four fluorescent protein variants, enhanced cyan fluorescent protein (ECFP), enhanced green fluorescent protein (EGFP), mCitrine, and mCherry, were used, however the methods described are useful for the characterization of any fluorescent protein or could be expanded to fluorescent quantum yield measurements of organic dye molecules. The laboratory is designed as a guided inquiry project and takes two, 4 hr laboratory periods. During the first day students design the experiment by selecting the excitation wavelength, choosing the standard, and determining the concentration needed for the quantum yield experiment that takes place in the second laboratory period. Overall, this laboratory provides students with a guided inquiry learning experience and introduces concepts of fluorescence biophysics into a biochemistry laboratory curriculum.

  3. Fluorescence Quantum Yield Measurements of Fluorescent Proteins: A Laboratory Experiment for a Biochemistry or Molecular Biophysics Laboratory Course

    Science.gov (United States)

    Wall, Kathryn P.; Dillon, Rebecca; Knowles, Michelle K.

    2015-01-01

    Fluorescent proteins are commonly used in cell biology to assess where proteins are within a cell as a function of time and provide insight into intracellular protein function. However, the usefulness of a fluorescent protein depends directly on the quantum yield. The quantum yield relates the efficiency at which a fluorescent molecule converts…

  4. 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.

  5. Chemical approaches for mimicking logic functions within fluorescent MPT dyes

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    The progress of the design, synthesis, fluorescence properties and application of a new family of fluorescent molecular switches towards information processing at the molecular level was reviewed. On the basis of the high fluorescence quantum yields and surroundings-sensitive fluorescent properties of the 5-methoxy-2-(2-pyridyl)-thiazole (2-MPT, 1) and a series of its derivatives as prepared, multiple binary logic and arithmetic functionalities were realized through encoding the controllable fluorescence switching properties with binary digit. Combined with the microfluidic platform, the fabrication of the molecular logic devices was attempted.

  6. Fluorescence spectroscopy and multi-way techniques. PARAFAC

    DEFF Research Database (Denmark)

    Murphy, Kathleen R.; Stedmon, Colin A.; Graeber, Daniel;

    2013-01-01

    PARAllel FACtor analysis (PARAFAC) is increasingly used to decompose fluorescence excitation emission matrices (EEMs) into their underlying chemical components. In the ideal case where fluorescence conforms to Beers Law, this process can lead to the mathematical identification and quantification...... of independently varying fluorophores. However, many practical and analytical hurdles stand between EEM datasets and their chemical interpretation. This article provides a tutorial in the practical application of PARAFAC to fluorescence datasets, demonstrated using a dissolved organic matter (DOM) fluorescence...... dataset. A new toolbox for MATLAB is presented to support improved visualisation and sensitivity analyses of PARAFAC models in fluorescence spectroscopy. © 2013 The Royal Society of Chemistry....

  7. Quantum Efficiency of Fluorescent Dyes and Color Matching

    Institute of Scientific and Technical Information of China (English)

    LI Rong; CHEN Dong-hui

    2002-01-01

    Because of the special optical characters, the color matching of fluorescent dyes is quite complicated. In order to find the algorithm of the color matching of fluorescent dyes, some experiments and measurements of one kind of fluorescent dye were carried out. An elementary probe into the method of color matching of fluorescent dyes has been made through the expression deduced by James S. Bonham and standard KubelkaMunk theory. The results prove that the method has a great applicability for the color matching of fabric dyed with only one kind of fluorescent dye.

  8. On the fluorescence of C60 at room temperature

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    The fluorescence properties of C 60 in different organic solvents have been investigated at room temperature. Three fluorescence emission centers are discovered and ascribed to different aggregations of C 60 in solvent. A series of blue fluorescence peaks centered at 440 nm derive from C 60 nanoparticles; a distinctive yellow-green fluorescence band in 575 nm region arises from the aggregates of C 60 nanoparticles; a more informative salmon fluorescence band around 700 nm originates from C 60 microcrystals. And the distinct configurations of C 60 aggregations are closely associated with the characteristic interaction between C 60 and solvent molecules.

  9. Chemical approaches for mimicking logic functions within fluorescent MPT dyes

    Institute of Scientific and Technical Information of China (English)

    XU ChunHu; SUN Wei; ZHANG Chao; BAI YanChun; FANG ChenJie; LI WenTao; HUANG YanYi; YAN ChunHua

    2009-01-01

    The progress of the design, synthesis, fluorescence properties and application of a new family of fluorescent molecular switches towards information processing at the molecular level was reviewed. On the basis of the high fluorescence quantum yields and surroundings-sensitive fluorescent proper-ties of the 5-methoxy-2-(2-pyridyl)-thiazole (2-MPT, 1) and a series of its derivatives as prepared, multi-ple binary logic and arithmetic functionalities were realized through encoding the controllable fluo-rescence switching properties with binary digit. Combined with the microfluidic platform, the fabrica-tion of the molecular logic devices was attempted.

  10. Fluorescence amplification by electrochemically deposited silver nanowires with fractal architecture.

    Science.gov (United States)

    Goldys, Ewa M; Drozdowicz-Tomsia, Krystyna; Xie, Fang; Shtoyko, Tanya; Matveeva, Eva; Gryczynski, Ignacy; Gryczynski, Zygmunt

    2007-10-10

    Electrochemically deposited silver structures with nanowires 50-100 nm in diameter show high fluorescence amplification and strongly reduced fluorescence lifetimes. Both quantities depend on the structure thickness. With increasing thickness the fluorescence amplification proportionally increases and the fluorescence lifetime decreases. This thickness dependence is caused by fluorophore interaction with a system of plasmon excitations in coupled nanowires extending over micrometer size regions. Thus the amplification is attributed to a combination of extended structure area and strong plasmonic coupling between nanowires which also help to radiatively scatter the fluorescence emission.

  11. [Fluorescent carbon dots and the application in biomedicine].

    Science.gov (United States)

    Zhang, Shuang; Gao, Hui-Le; Shen, Shun; Wang, Wei-Liang; Qian, Jun

    2014-09-01

    As a new type of carbon nanomaterials, fluorescent carbon dots (fluorescent CDs) have many advantages when compared with the traditional fluorescent probes. They are photoluminescence stable and resistance to photo bleaching. Moreover, they are excellent in biocompatibility, low-toxic and easy to modify. All these above make them a promising optical image material as a probe in optical image. This article reviews structure, the common carbon sources, the preparation methods, and the light-emitting principles of the carbon dots. We also introduce the research progress of fluorescent carbon dots in biomedicine, and the problems need to be resolved in the study of fluorescent CDs.

  12. Fluorescent nanodiamonds as highly stable biomarker for endotoxin verification

    Science.gov (United States)

    Bergmann, Thorsten; Burg, Jan Michael; Lilholt, Maria; Maeder, Ulf; Beer, Sebastian; Salzig, Denise; Ebrahimi, Mehrdad; Czermak, Peter; Fiebich, Martin

    2012-03-01

    Fluorescent nanodiamonds (ND) provide advantageous properties as a fluorescent biomarker for in vitro and in vivo studies. The maximum fluorescence occurs around 700 nm, they do not show photobleaching or blinking and seem to be nontoxic. After a pretreatment with strong acid fluorescent ND can be functionalized and coupled to endotoxin. Endotoxin is a decay product of bacteria and causes strong immune reactions. Therefore endotoxin has to be removed for most applications. An effective removal procedure is membrane filtration. The endotoxin, coupled to fluorescent ND can be visualized by using confocal microscopy which allows the investigation of the separation mechanisms of the filtration process within the membranes.

  13. Accurate fluorescent polymeric thermometers containing an ionic component.

    Science.gov (United States)

    Gota, Chie; Uchiyama, Seiichi; Ohwada, Tomohiko

    2007-02-01

    Fluorescent polymeric thermometers consisting of only N-alkylacrylamide and fluorescent components show rather low temperature resolution in their functional ranges (ca. 15-50 degrees C) because of the occurrence of intermolecular aggregation, which causes hysteresis in their fluorescence response to changes in temperature. By adding an ionic component to prevent such intermolecular aggregation, we obtained four fluorescent polymeric thermometers that offer high temperature resolution (<0.2 degrees C). Each new fluorescent polymeric thermometer covered the temperature range, 9-33 degrees C, 30-51 degrees C, 49-66 degrees C or 4-38 degrees C.

  14. Phasor approaches simplify the analysis of tryptophan fluorescence data in protein denaturation studies

    NARCIS (Netherlands)

    Bader, A.N.; Visser, N.V.; Amerongen, van H.; Visser, A.J.W.G.

    2014-01-01

    The intrinsic fluorescence of tryptophan is frequently used to investigate the structure of proteins. The analysis of tryptophan fluorescence data is challenging: fluorescence (anisotropy) decays typically have multiple lifetime (correlation time) components and fluorescence spectra are broad and ex

  15. Time-resolved spectroscopy of the fluorescence quenching of a donor — acceptor pair by halothane

    Science.gov (United States)

    Sharma, A.; Draxler, S.; Lippitsch, M. E.

    1992-04-01

    Donor (anthracene) sensitized acceptor (perylene) fluorescence is quenched more efficiently by halothane than is intrinsic perylene fluorescence. The underlying process of dynamic fluorescence quenching is investigated by time-resolved fluorescence spectroscopy.

  16. Fluorescence detection of natural RNA using rationally designed "clickable" oligonucleotide probes

    DEFF Research Database (Denmark)

    Okholm, Anders; Kjems, Jørgen; Astakhova, Kira

    2014-01-01

    Herein a reliable approach to the design of effective fluorescent probes for RNA detection is described. The fluorescence signalling of hybridization by internally positioned polyaromatic hydrocarbons and rhodamine dyes was achieved with a low fluorescence background signal, high fluorescence...

  17. Highly sensitive turn-on fluorescence detection of thrombomodulin based on fluorescence resonance energy transfer

    Science.gov (United States)

    Kong, Liyan; Zhu, Jiaming; Wang, Wen; Jin, Lehe; Fu, Yanjiao; Duan, Bohui; Tan, Liang

    2017-02-01

    As an integral glycoprotein on the surface of endothelial cells, thrombomodulin (TM) has very high affinity for thrombin. TM has been regarded to be a marker of endothelial damage since it can be released during endothelial cell injury. In this work, a highly sensitive fluorescence method for the quantitative detection of TM was developed. TM antibody (Ab) and bovine serum albumin (BSA) were bound on gold nanoparticles (AuNPs) to construct BSA-AuNPs-Ab nanocomposites and they were characterized by transmission electron microscope and UV-vis spectrophotometry. The fluorescence of acridine orange (AO) was quenched by the prepared gold nanocomposites based on fluorescence resonance energy transfer (FRET). In the presence of TM, the fluorescence was turned on due to the effective separation of AO from the surface of gold nanocomposites. Under optimum conditions, the enhanced fluorescence intensity displayed a linear relationship with the logarithm of the TM concentration from 0.1 pg mL- 1 to 5 ng mL- 1 with a low detection limit of 12 fg mL- 1. The release of soluble thrombomodulin (sTM) by the injured HUVEC-C cells in the presence of H2O2 was investigated using the proposed method. The released sTM content in the growth medium was found to be increased with the enhancement of contact time of the cells with H2O2.

  18. Fluorescence derivatization of clarithromycin for high performance liquid chromatographic determination with fluorescence detection

    Directory of Open Access Journals (Sweden)

    Boonleang, J.

    2007-03-01

    Full Text Available Clarithromycin (CAM is a semisynthetic macrolide antibiotic whose chemical structure has no suitable chromophore for highly sensitive and accurate direct determination. The aim of this study was toderivatize CAM with fluorescence-labeling compounds capable of enhancing the sensitivity of CAM determination. Two fluorescence-labeling compounds were used in this study, 9-fluorenylmethyloxycarbonylchloride (FMOC-Cl and 1-naphthylisocyanate (NIC, both of which gave the fluorescent derivatives of CAM with approximately the same fluorescence intensity. The derivatization reactions in the concentration rangestudied (0.1-2.4 μg/ml were reproducible with coefficient of variation of less than 6.01% and the fluorescence responses were linearly proportional to CAM concentration with r2 of greater than 0.99. The reaction of CAM with FMOC-Cl optimally occurred in a solvent mixture of acetonitrile and phosphate buffer pH 7.5(4:1 volume ratio at 40oC for 40 min. The optimum derivatization reaction of CAM with NIC took place in acetonitrile with triethylamine as catalyst at 30oC for 60 min. It was mild and quantitative giving CAM-NICfluorescent derivative, which is more stable at room temperature than CAM-FMOC derivative. This derivatization should, therefore, be more applicable for highly sensitive CAM determination especially for thestudy involving the analysis of several samples.

  19. Solid-State Carbon Dots with Red Fluorescence and Efficient Construction of Dual-Fluorescence Morphologies.

    Science.gov (United States)

    He, Jiangling; He, Youling; Chen, Yonghao; Lei, Bingfu; Zhuang, Jianle; Xiao, Yong; Liang, Yeru; Zheng, Mingtao; Zhang, Haoran; Liu, Yingliang

    2017-07-01

    Stable solid-state red fluorescence from organosilane-functionalized carbon dots (CDs) with sizes around 3 nm is reported for the first time. Meanwhile, a novel method is also first reported for the efficient construction of dual-fluorescence morphologies. The quantum yield of these solid-state CDs and their aqueous solution is 9.60 and 50.7%, respectively. The fluorescence lifetime is 4.82 ns for solid-state CDs, and 15.57 ns for their aqueous solution. These CDs are detailedly studied how they can exhibit obvious photoluminescence overcoming the self-quenching in solid state. Luminescent materials are constructed with dual fluorescence based on as-prepared single emissive CDs (red emission) and nonfluorescence media (starch, Al2 O3 , and RnOCH3 COONa), with the characteristic peaks located at nearly 440 and 600 nm. Tunable photoluminescence can be successfully achieved by tuning the mass ratio of CDs to solid matrix (such as starch). These constructed dual-fluorescence CDs/starch composites can also be applied in white light-emitting diodes with UV chips (395 nm), and oxygen sensing. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Toward structurally defined carbon dots as ultracompact fluorescent probes.

    Science.gov (United States)

    LeCroy, Gregory Ethan; Sonkar, Sumit Kumar; Yang, Fan; Veca, L Monica; Wang, Ping; Tackett, Kenneth N; Yu, Jing-Jiang; Vasile, Eugeniu; Qian, Haijun; Liu, Yamin; Luo, Pengju George; Sun, Ya-Ping

    2014-05-27

    There has been much discussion on the need to develop fluorescent quantum dots (QDs) as ultracompact probes, with overall size profiles comparable to those of the genetically encoded fluorescent tags. In the use of conventional semiconductor QDs for such a purpose, the beautifully displayed dependence of fluorescence color on the particle diameter becomes a limitation. More recently, carbon dots have emerged as a new platform of QD-like fluorescent nanomaterials. The optical absorption and fluorescence emissions in carbon dots are not bandgap in origin, different from those in conventional semiconductor QDs. The absence of any theoretically defined fluorescence color-dot size relationships in carbon dots may actually be exploited as a unique advantage in the size reduction toward having carbon dots serve as ultracompact QD-like fluorescence probes. Here we report on carbon dots of less than 5 nm in the overall dot diameter with the use of 2,2'-(ethylenedioxy)bis(ethylamine) (EDA) molecules for the carbon particle surface passivation. The EDA-carbon dots were found to be brightly fluorescent, especially over the spectral range of green fluorescent protein. These aqueous soluble smaller carbon dots also enabled more quantitative characterizations, including the use of solution-phase NMR techniques, and the results suggested that the dot structures were relatively simple and better-defined. The potential for these smaller carbon dots to serve as fluorescence probes of overall sizes comparable to those of fluorescent proteins is discussed.

  1. Photoreaction of indole-containing mycotoxins to fluorescent products.

    Science.gov (United States)

    Maragos, C M

    2009-06-01

    Photochemical reaction of the non-fluorescent mycotoxin cyclopiazonic acid (CPA) to fluorescent products was recently reported. Because CPA contains an indole moiety, believed to contribute to the fluorescence, it was of interest to determine whether the effect might be more generally applicable to indole-containing mycotoxins. Three indole-containing tremorgens (penitrem A, paxilline, verruculogen) that have not previously been reported to be fluorescent were rendered fluorescent by exposure to ultraviolet light in a photoreactor. Naturally fluorescent ergot alkaloids, which also contain an indole-moiety, exhibited a diminished response after exposure. This suggests that the phenomenon may be most useful for detection of indole-containing tremorgens that are non-fluorescent, rather than for the enhancement of materials that are already fluorescent, such as the ergot alkaloids. The extent to which fluorescence enhancement was seen was strongly influenced by the reaction environment, in particular the solvent used and whether cyclodextrins were present. In an HPLC format, placement of the photoreactor post-column allowed for the fluorescence detection of penitrem A, paxilline, and verruculogen. The ability to photoreact indole-containing tremorgens and detect them by fluorescence may open up new avenues for detection of these mycotoxins alone or in combination.

  2. 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.

  3. Fluorescent-magnetic Janus particles prepared via seed emulsion polymerization.

    Science.gov (United States)

    Kaewsaneha, Chariya; Bitar, Ahmad; Tangboriboonrat, Pramuan; Polpanich, Duangporn; Elaissari, Abdelhamid

    2014-06-15

    Anisotropic polymeric colloidal or Janus particles possessing simultaneous magnetic and fluorescent properties were successfully prepared via the swelling-diffusion or the in situ emulsion polymerization method. In the swelling-diffusion process, magnetic emulsions (an organic ferrofluid dispersed in aqueous medium) were synthesized and used for seeds of submicron magnetic Janus particles. After swelling the anisotropic particles obtained by 1-pyrene-carboxaldehyde fluorescent dye dissolved in tetrahydrofuran, well-defined fluorescent-magnetic Janus particles were produced. In the in situ emulsion polymerization, styrene monomer mixed with fluorescent dye monomers, i.e., 1-pyrenylmethyl methacrylate (PyMMA) or fluorescein dimethacrylate (FDMA), and an oil-soluble initiator (2,2'-azobis(2-isobutyronitrile)) were emulsified in the presence of magnetic seed emulsions. The confocal microscopic images showed the fluorescent-magnetic Janus particles with high fluorescent intensity when a fluorescent crosslinker monomer FDMA was employed.

  4. Use of laser fluorescence in dental caries diagnosis: a fluorescence x biomolecular vibrational spectroscopic comparative study.

    Science.gov (United States)

    Carvalho, Fabíola Bastos de; Barbosa, Artur Felipe Santos; Zanin, Fátima Antonia Aparecida; Brugnera Júnior, Aldo; Silveira Júnior, Landulfo; Pinheiro, Antonio Luiz Barbosa

    2013-01-01

    The aim of this work was to verify the existence of correlation between Raman spectroscopy readings of phosphate apatite (~960 cm-1), fluoridated apatite (~575 cm-1) and organic matrix (~1450 cm-1) levels and Diagnodent® readings at different stages of dental caries in extracted human teeth. The mean peak value of fluorescence in the carious area was recorded and teeth were divided in enamel caries, dentin caries and sound dental structure. After fluorescence readings, Raman spectroscopy was carried out on the same sites. The results showed significant difference (ANOVA, pcaries. There was significant negative correlation (pcaries. It may be concluded that the higher the fluorescence detected by Diagnodent the lower the peaks of phosphate apatite and fluoridated apatite. As the early diagnosis of caries is directly related to the identification of changes in the inorganic tooth components, Raman spectroscopy was more sensitive to variations of these components than Diagnodent.

  5. 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.

  6. On the purported "backbone fluorescence" in protein three-dimensional fluorescence spectra

    DEFF Research Database (Denmark)

    Bortolotti, Annalisa; Wong, Yin How; Korsholm, Stine S.

    2016-01-01

    as any traditional protein emission spectrum. The many papers in reputable journals erroneously reporting this peak assignment, contradicting 5 decades of prior knowledge, have led to the creation of a new dogma, where many authors and reviewers now take the purported backbone fluorescence......In this study, several proteins (albumin, lysozyme, insulin) and model compounds (Trp, Tyr, homopolypeptides) were used to demonstrate the origin of the fluorescence observed upon their excitation at 220-230 nm. In the last 10 years we have observed a worrying increase in the number of articles...... claiming that this fluorescence originates from the protein backbone, contrary to the established knowledge that UV protein emission is due to aromatic amino acids only. Overall, our data clearly demonstrate that the observed emission upon excitation at 220-230 nm is due to the excitation of Tyr and/or Trp...

  7. 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.

  8. Global analysis of fluorescence decays to probe the internal dynamics of fluorescently labeled macromolecules.

    Science.gov (United States)

    Duhamel, Jean

    2014-03-11

    The aim of this review is to introduce the reader first to the mathematical complexity associated with the analysis of fluorescence decays acquired with solutions of macromolecules labeled with a fluorophore and its quencher that are capable of interacting with each other via photophysical processes within the macromolecular volume, second to the experimental and mathematical approaches that have been proposed over the years to handle this mathematical complexity, and third to the information that one can expect to retrieve with respect to the internal dynamics of such fluorescently labeled macromolecules. In my view, the ideal fluorophore-quencher pair to use in studying the internal dynamics of fluorescently labeled macromolecules would involve a long-lived fluorophore, a fluorophore and a quencher that do not undergo energy migration, and a photophysical process that results in a change in fluorophore emission upon contact between the excited fluorophore and quencher. Pyrene, with its ability to form an excimer on contact between excited-state and ground-state species, happens to possess all of these properties. Although the concepts described in this review apply to any fluorophore and quencher pair sharing pyrene's exceptional photophysical properties, this review focuses on the study of pyrene-labeled macromolecules that have been characterized in great detail over the past 40 years and presents the main models that are being used today to analyze the fluorescence decays of pyrene-labeled macromolecules reliably. These models are based on Birks' scheme, the DMD model, the fluorescence blob model, and the model free analysis. The review also provides a step-by-step protocol that should enable the noneducated user to achieve a successful decay analysis exempt of artifacts. Finally, some examples of studies of pyrene-labeled macromolecules are also presented to illustrate the different types of information that can be retrieved from these fluorescence decay

  9. Ultrafast excited state dynamics of the green fluorescent protein chromophore and its kindling fluorescent protein analogue.

    Science.gov (United States)

    Addison, Kiri; Heisler, Ismael A; Conyard, Jamie; Dixon, Tara; Page, Philip C Bulman; Meech, Stephen R

    2013-01-01

    Fluorescent proteins exhibit a very diverse range of photochemical behaviour, from efficient fluorescence through photochromism to photochemical reactivity. Remarkably this diverse behaviour arises from chromophores which have very similar structures. Here we describe measurements and modelling of the excited state dynamics in the chromophores of GFP (HBDI) and the kindling fluorescent protein, KFP (FHBMI). The methods are ultrafast fluorescence spectroscopy with sub 50 fs time resolution and the modelling is based on the Smoluchowski equation. The excited state decays of both chromophores are very fast, longer for their anions than for the neutral form and independent of wavelength. Detailed studies show the mean fluorescence wavelength to be independent of time. The excited state decay times are also observed to be a very weak function of solvent polarity and viscosity. These results are modelled utilising recently calculated potential energy surfaces for the ground and excited states as a function of the twist coordinates about the two bridging bonds of the chromophore. For FHBMI and the scarce data on the neutral HBDI the calculations are not successful suggesting the need for refinement of these potential energy surfaces. For HBDI in methanol the simulation is successful provided a strong dependence of the radiationless decay rate on the coordinate is assumed. Such dependence should be included in future calculations of excited state dynamics. When the simulations are extended to more viscous solvents they fail to reproduce the observed weak viscosity dependence. The implications of these results for the nature of the coordinate leading to radiationless decay in the chromophore and for the photodynamics of fluorescent proteins are discussed.

  10. Waveguide evanescent field fluorescence microscopy: Thin film fluorescence intensities and its application in cell biology

    Science.gov (United States)

    Hassanzadeh, Abdollah; Nitsche, Michael; Mittler, Silvia; Armstrong, Souzan; Dixon, Jeff; Langbein, Uwe

    2008-06-01

    We demonstrate an inexpensive alternative to total internal reflection fluorescence microscopy. A method for imaging ultrathin films and living cells located on waveguides—illuminated with their evanescent fields—is introduced. An extensive analysis of ion-exchanged waveguides focusing on their application as microscopy substrates for studying interfacial phenomena is presented. Experimental results are in excellent agreement with the simulations. As an application osteoblasts (bone matrix forming cells) and ultrathin Langmuir-Blodgett films were imaged. The fluorescence intensity has been used to determine the cell attachment.

  11. Surface-Enhanced X-Ray Fluorescence

    Science.gov (United States)

    Anderson, Mark

    2010-01-01

    Surface-enhanced x-ray fluorescence (SEn-XRF) spectroscopy is a form of surface- enhanced spectroscopy that was conceived as a means of obtaining greater sensitivity in x-ray fluorescence (XRF) spectroscopy. As such, SEn-XRF spectroscopy joins the ranks of such other, longer-wavelength surface-enhanced spectroscopies as those based on surface-enhanced Raman scattering (SERS), surface-enhanced resonance Raman scattering (SERRS), and surfaceenhanced infrared Raman absorption (SEIRA), which have been described in previous NASA Tech Briefs articles. XRF spectroscopy has been used in analytical chemistry for determining the elemental compositions of small samples. XRF spectroscopy is rapid and quantitative and has been applied to a variety of metal and mineralogical samples. The main drawback of XRF spectroscopy as practiced heretofore is that sensitivity has not been as high as required for some applications. In SEn-XRF as in the other surface-enhanced spectroscopies, one exploits several interacting near-field phenomena, occurring on nanotextured surfaces, that give rise to local concentrations of incident far-field illumination. In this case, the far-field illumination comes from an x-ray source. Depending on the chemical composition and the geometry of a given nanotextured surface, these phenomena could include the lightning-rod effect (concentration of electric fields at the sharpest points on needlelike surface features), surface plasmon resonances, and grazing incidence geometric effects. In the far field, the observable effect of these phenomena is an increase in the intensity of the spectrum of interest - in this case, the x-ray fluorescence spectrum of chemical elements of interest that may be present within a surface layer at distances no more than a few nanometers from the surface.

  12. Salt stress change chlorophyll fluorescence in mango

    Directory of Open Access Journals (Sweden)

    Cicero Cartaxo de Lucena

    2012-12-01

    Full Text Available This study evaluated the tolerance of mango cultivars 'Haden', 'Palmer', 'Tommy Atkins' and 'Uba' grafted on rootstock 'Imbú' to salt stress using chlorophyll fluorescence. Plants were grown in modified Hoagland solution containing 0, 15, 30, and 45 mmol L-1 NaCl. At 97 days the parameters of the chlorophyll fluorescence (F0, Fm, Fv, F0/Fm, Fv/Fm, Fv'/Fm', ΦPSII = [(Fm'-Fs/(Fm'], D = (1- Fv'/Fm' and ETR = (ΦPSII×PPF×0,84×0,5 were determined. At 100 days, the leaf emission and leaf area, toxicity and leaf abscission indexes were determined. In all cultivars evaluated, in different degree, there were decreases in photochemical efficiency of photosystem II, enhanced concentrations from 15 mmol L-1 NaCl. The decreases in the potential quantum yield of photosystem II (Fv/Fm were 27.9, 18.7, 20.5, and 27.4%, for cultivars 'Haden', 'Palmer', 'Tommy Atkins', and 'Uba', respectively, when grown in 45 mmol L-1 NaCl. It was found decreases in leaf emission and mean leaf area in all cultivars from 15 mmol L-1 NaCl. There were increases in leaf toxicity of 33.0, 67.5, 41.6 and 80.8% and in leaf abscission of 71.8, 29.2, 32.5, and 67.9% for the cultivars 'Haden', 'Palmer', 'Tommy Atkins', and 'Uba' respectively, when grown in 45 mmol L-1 NaCl. Leaf toxicity and leaf abscission were not observed in 15 mmol L-1 NaCl. The decrease in Fv/Fm ratio were accompanied by decreasing in leaf emission and increased leaf toxicity index, showing, therefore, the potential of chlorophyll fluorescence in the early detection of salt stress in mango tree.

  13. Reductive fluorescence quenching of DMP with aniline

    Energy Technology Data Exchange (ETDEWEB)

    Asha Jhonsi, M. [B.S. Abdur Rahman University, Vandalur, Chennai 600048, Tamil Nadu (India); Kathiravan, A., E-mail: akathir23@hotmail.com [National Centre for Ultrafast Processes, University of Madras, Taramani Campus, Chennai 600113, Tamil Nadu (India)

    2014-01-15

    The photoinduced electron transfer (PET) between 8-(4-methoxyphenyl)-3,5-di[(E)-1-(4-methoxyphenyl)methylidene]-1,2,3,5,6, 7-hexahydrodicyclopenta[b,e]pyridine (DMP) and aniline is studied in acetonitrile medium by using steady state and time resolved absorption and fluorescence spectroscopic methods. Bimolecular quenching rate constants (k{sub q}) were calculated from the obtained linear Stern–Volmer plots from both steady state and time resolved measurements. The rate constant (k{sub q}) for PET between DMP and aniline is 1.4×10{sup 10} M{sup −1} s{sup −1}, which is in diffusion control limit. The free energy change (ΔG{sup 0}) has been evaluated by using Rehm–Weller equation for the evidence of electron transfer from aniline to DMP. Direct evidence for the electron transfer reaction in the present system has been obtained by characterizing the aniline cation radical using nanosecond time resolved absorption measurements in the visible region. Further, this quenching mechanism is attributed to be reductive in nature i.e. electron transfer occurs from ground state aniline to excited DMP. This is the first example of reductive fluorescence quenching of DMP with aniline in acetonitrile ever known. -- Highlights: • Photoinduced electron transfer between DMP and aniline using time resolved absorption and fluorescence spectroscopy has been investigated. • Reductive quenching behavior was observed. • Direct evidence for the ET reaction in the present system has been obtained by characterizing the aniline cation radical.

  14. 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.

  15. Red fluorescent biofilm: the thick, the old, and the cariogenic

    Directory of Open Access Journals (Sweden)

    Catherine M.C. Volgenant

    2016-04-01

    Full Text Available Background: Some dental plaque fluoresces red. The factors involved in this fluorescence are yet unknown. Objective: The aim of this study was to assess systematically the effect of age, thickness, and cariogenicity on the extent of red fluorescence produced by in vitro microcosm biofilms. Design: The effects of biofilm age and thickness on red fluorescence were tested in a constant depth film fermentor (CDFF by growing biofilms of variable thicknesses that received a constant supply of defined mucin medium (DMM and eight pulses of sucrose/day. The influence of cariogenicity on red fluorescence was tested by growing biofilm on dentin disks receiving DMM, supplemented with three or eight pulses of sucrose/day. The biofilms were analyzed at different time points after inoculation, up to 24 days. Emission spectra were measured using a fluorescence spectrophotometer (λexc405 nm and the biofilms were photographed with a fluorescence camera. The composition of the biofilms was assessed using 454-pyrosequecing of the 16S rDNA gene. Results: From day 7 onward, the biofilms emitted increasing intensities of red fluorescence as evidenced by the combined red fluorescence peaks. The red fluorescence intensity correlated with biofilm thickness but not in a linear way. Biofilm fluorescence also correlated with the imposed cariogenicity, evidenced by the induced dentin mineral loss. Increasing the biofilm age or increasing the sucrose pulsing frequency led to a shift in the microbial composition. These shifts in composition were accompanied by an increase in red fluorescence. Conclusions: The current study shows that a thicker, older, or more cariogenic biofilm results in a higher intensity of red fluorescence.

  16. Red fluorescent biofilm: the thick, the old, and the cariogenic

    Science.gov (United States)

    Volgenant, Catherine M.C.; Hoogenkamp, Michel A.; Buijs, Mark J.; Zaura, Egija; ten Cate, Jacob (Bob) M.; van der Veen, Monique H.

    2016-01-01

    Background Some dental plaque fluoresces red. The factors involved in this fluorescence are yet unknown. Objective The aim of this study was to assess systematically the effect of age, thickness, and cariogenicity on the extent of red fluorescence produced by in vitro microcosm biofilms. Design The effects of biofilm age and thickness on red fluorescence were tested in a constant depth film fermentor (CDFF) by growing biofilms of variable thicknesses that received a constant supply of defined mucin medium (DMM) and eight pulses of sucrose/day. The influence of cariogenicity on red fluorescence was tested by growing biofilm on dentin disks receiving DMM, supplemented with three or eight pulses of sucrose/day. The biofilms were analyzed at different time points after inoculation, up to 24 days. Emission spectra were measured using a fluorescence spectrophotometer (λexc405 nm) and the biofilms were photographed with a fluorescence camera. The composition of the biofilms was assessed using 454-pyrosequecing of the 16S rDNA gene. Results From day 7 onward, the biofilms emitted increasing intensities of red fluorescence as evidenced by the combined red fluorescence peaks. The red fluorescence intensity correlated with biofilm thickness but not in a linear way. Biofilm fluorescence also correlated with the imposed cariogenicity, evidenced by the induced dentin mineral loss. Increasing the biofilm age or increasing the sucrose pulsing frequency led to a shift in the microbial composition. These shifts in composition were accompanied by an increase in red fluorescence. Conclusions The current study shows that a thicker, older, or more cariogenic biofilm results in a higher intensity of red fluorescence. PMID:27060056

  17. Picosecond kinetic measurements of the metalloporphyrin fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Aaviskoq, Y.Y.; Freiburg, A.M.; Savikhin, S.F.; Stel' makh, G.F.

    1986-08-01

    The authors attempt to directly measure the deactivation kinetics of the short-lived excited S/sub 2/ and S/sub 1/ states of metalloporphyrins and compare the results with those obtained by other (either direct or indirect) methods. The studies were carried out on diamagnetic metallocomplexes of tetrabenzoporphyrin (MeTBP) exhibiting measurable fluorescence from the S/sub 1/ and S/sub 2/ states. The complexes with Lu, Cd, and Zn in dilute solutions were studied at room temperature. The results of direct kinetic experiments confirm the previously obtained data on picosecond deactivation processes in photoexcited metalloporphyrins.

  18. Absolute calibration of the Auger fluorescence detectors

    Energy Technology Data Exchange (ETDEWEB)

    Bauleo, P.; Brack, J.; Garrard, L.; Harton, J.; Knapik, R.; Meyhandan, R.; Rovero, A.C.; /Buenos Aires, IAFE; Tamashiro, A.; Warner, D.

    2005-07-01

    Absolute calibration of the Pierre Auger Observatory fluorescence detectors uses a light source at the telescope aperture. The technique accounts for the combined effects of all detector components in a single measurement. The calibrated 2.5 m diameter light source fills the aperture, providing uniform illumination to each pixel. The known flux from the light source and the response of the acquisition system give the required calibration for each pixel. In the lab, light source uniformity is studied using CCD images and the intensity is measured relative to NIST-calibrated photodiodes. Overall uncertainties are presently 12%, and are dominated by systematics.

  19. Validation of fluorescence transition probability calculations

    CERN Document Server

    Pia, M G; Sudhaka, Manju

    2009-01-01

    A systematic and quantitative validation of the K and L shell X-ray transition probability calculations according to different theoretical methods has been performed against experimental data. This study is relevant to the optimization of data libraries used by software systems, namely Monte Carlo codes, dealing with X-ray fluorescence. The results support the adoption of transition probabilities calculated according to the Hartree-Fock approach, which manifest better agreement with experimental measurements than calculations based on the Hartree-Slater method.

  20. RNA fluorescence with light-up aptamers

    Science.gov (United States)

    Ouellet, Jonathan

    2016-06-01

    Seeing is not only believing; it also includes understanding. Cellular imaging with GFP in live cells has been transformative in many research fields. Modulation of cellular regulation is tightly regulated and innovative imaging technologies contribute to further understand cellular signaling and physiology. New types of genetically encoded biosensors have been developed over the last decade. They are RNA aptamers that bind with their cognate fluorogen ligands and activate their fluorescence. The emergence and the evolution of these RNA aptamers as well as their conversion into a wide spectrum of applications are examined in a global way.

  1. Novel Nanophosphors for High Efficiency Fluorescent Lamps

    Energy Technology Data Exchange (ETDEWEB)

    Alok Srivatava

    2007-03-31

    This is the Final Report of the Novel Nanophosphors for High Efficiency Fluorescent Lamps, Department of Energy (DOE). The overall goal of this three-year program is to develop novel hybrid phosphors by coating commercially available lamp phosphors with highly stable wide band-gap nanocrystalline phosphors (NCP). The prime technical approach is the development of NCP quantum-splitting phosphor (QSP) and ultra-violet (UV) emitting phosphors with quantum efficiencies exceeding that of the conventional phosphors at 185 nm. The novel hybrid phosphors will increase the efficiency of the fluorescent lamps by up to 32%, enabling total energy savings of 0.26 quads, the reduction in the U.S. energy bill by $6.5 billion and the reduction of the annual carbon emission by 4.1 billion kilogram. Our work started by investigating through modeling calculations the requirement for the particle size of the NCP. Our work to develop suitable nanocrystalline phosphors started with the known oxide quantum splitting and UV emitting phosphors. We demonstrated several synthesis techniques for the production of high quality nanocrystalline materials that crystallizes in the desired phase and with the desired particle size. In collaboration with our subcontractor we demonstrated the feasibility for the manufacture of NC phosphors. We also demonstrated novel techniques of coating the NCP on the surface of micron sized phosphors. Our chief achievement pertains to the successful testing of the coated hybrid phosphor systems in linear fluorescent lamps. In linear fluorescent lamp tests, we have demonstrated up to 7% increase in the efficacy of hybrid phosphors over the conventional (uncoated) phosphors. We have also demonstrated the improvement in the lumen maintenance of the coated phosphors. A hybrid phosphor system based on the commercial red emitting phosphor, Y{sub 2}O{sub 3}:Eu{sup 3+} did not show the anticipated improvement in lamp efficacy. We explored the reasons for this observation

  2. Fluorescence Resonance Energy Transfer (FRET) sensor

    CERN Document Server

    Hussain, Syed Arshad; Chakraborty, Sekhar; Saha, Jaba; Roy, Arpan Datta; Chakraborty, Santanu; Debnath, Pintu; Bhattacharjee, D

    2014-01-01

    The applications of Fluorescence resonance energy transfer (FRET) have expanded tremendously in the last 25 years, and the technique has become a staple technique in many biological and biophysical fields. FRET can be used as spectroscopic ruler in various areas such as structural elucidation of biological molecules and their interactions, in vitro assays, in vivo monitoring in cellular research, nucleic acid analysis, signal transduction, light harvesting, and metallic nanomaterials etc. Based on the mechanism of FRET a variety of novel chemical sensors and Biosensors have been developed. This review highlights the recent applications of sensitive and selective ratiometric FRET based sensors.

  3. Generation of red fluorescent protein transgenic dogs.

    Science.gov (United States)

    Hong, So Gun; Kim, Min Kyu; Jang, Goo; Oh, Hyun Ju; Park, Jung Eun; Kang, Jung Taek; Koo, Ok Jae; Kim, Teoan; Kwon, Mo Sun; Koo, Bon Chul; Ra, Jeong Chan; Kim, Dae Yong; Ko, CheMyong; Lee, Byeong Chun

    2009-05-01

    Dogs (Canis familiaris) share many common genetic diseases with humans and development of disease models using a transgenic approach has long been awaited. However, due to the technical difficulty in obtaining fertilizable eggs and the unavailability of embryonic stem cells, no transgenic dog has been generated. Canine fetal fibroblasts were stably transfected with a red fluorescent protein (RFP) gene-expressing construct using retrovirus gene delivery method. Somatic cell nuclear transfer was then employed to replace the nucleus of an oocyte with the nucleus of the RFP-fibroblasts. Using this approach, we produced the first generation of transgenic dogs with four female and two male expressing RFP.

  4. High Fluorescent Porphyrin-PAMAM-Fluorene Dendrimers.

    Science.gov (United States)

    Garfias-Gonzalez, Karla I; Organista-Mateos, Ulises; Borja-Miranda, Andrés; Gomez-Vidales, Virginia; Hernandez-Ortega, Simon; Cortez-Maya, Sandra; Martínez-García, Marcos

    2015-05-13

    Two new classes of dendrimers bearing 8 and 32 fluorene donor groups have been synthesized. The first and second generations of these porphyrin-PAMAM-fluorene dendrimers were characterized by 1H-NMR, 13C-NMR, FTIR, UV-vis spectroscopy, elemental analyses and MALDI-TOF mass spectrometry. The UV-vis spectra showed that the individual properties of donor and acceptor moieties were preserved, indicating that the new dendrimers could be used as photosynthetic antennae. Furthermore, for fluorescent spectroscopy, these dendrimers showed good energy transfer.

  5. Laser induced fluorescence technique for environmental applications

    Science.gov (United States)

    Utkin, Andrei B.; Felizardo, Rui; Gameiro, Carla; Matos, Ana R.; Cartaxana, Paulo

    2014-08-01

    We discuss the development of laser induced fluorescence sensors and their application in the evaluation of water pollution and physiological status of higher plants and algae. The sensors were built on the basis of reliable and robust solid-state Nd:YAG lasers. They demonstrated good efficiency in: i) detecting and characterizing oil spills and dissolved organic matter; ii) evaluating the impact of stress on higher plants (cork oak, maritime pine, and genetically modified Arabidopsis); iii) tracking biomass changes in intertidal microphytobenthos; and iv) mapping macroalgal communities in the Tagus Estuary.

  6. 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

  7. Confocal microscopy via multimode fibers: fluorescence bandwidth

    Science.gov (United States)

    Loterie, Damien; Psaltis, Demetri; Moser, Christophe

    2016-03-01

    We recently described a method for confocal reflection imaging through fibers, as a way to increase contrast when imaging unstained biological specimens. Using a transmission matrix, focused spots can be created at the distal end of a fiber. The backscattered field coming back from the sample can be filtered using optical correlation to obtain spatial selectivity in the detection. In this proceedings article, we briefly review the working principle of this method, and we discuss how the scheme could be adapted to confocal fluorescence imaging. In particular, we show simulations of the achievable detection bandwidth when using step-index multimode fibers as imaging devices.

  8. 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.

  9. Investigation of fluorescence spectra disturbances influencing the classification performance of fluorescently labeled plastic flakes

    Science.gov (United States)

    Fomin, Petr; Brunner, Siegfried; Kargel, Christian

    2013-04-01

    The recycling of plastic products becomes increasingly attractive not only from an environmental point of view, but also economically. For recycled (engineering) plastic products with the highest possible quality, plastic sorting technologies must provide clean and virtually mono-fractional compositions from a mixture of many different types of (shredded) plastics. In order to put this high quality sorting into practice, the labeling of virgin plastics with specific fluorescent markers at very low concentrations (ppm level or less) during their manufacturing process is proposed. The emitted fluorescence spectra represent "optical fingerprints" - each being unique for a particular plastic - which we use for plastic identification and classification purposes. In this study we quantify the classification performance using our prototype measurement system and 15 different plastic types when various influence factors most relevant in practice cause disturbances of the fluorescence spectra emitted from the labeled plastics. The results of these investigations help optimize the development and incorporation of appropriate fluorescent markers as well as the classification algorithms and overall measurement system in order to achieve the lowest possible classification error rates.

  10. 5-Aminolevulinic Acid Fluorescence in High Grade Glioma Surgery: Surgical Outcome, Intraoperative Findings, and Fluorescence Patterns

    Directory of Open Access Journals (Sweden)

    Alessandro Della Puppa

    2014-01-01

    Full Text Available Background. 5-Aminolevulinic acid (5-ALA fluorescence is a validated technique for resection of high grade gliomas (HGG; the aim of this study was to evaluate the surgical outcome and the intraoperative findings in a consecutive series of patients. Methods. Clinical and surgical data from patients affected by HGG who underwent surgery guided by 5-ALA fluorescence at our Department between June 2011 and February 2014 were retrospectively evaluated. Surgical outcome was evaluated by assessing the resection rate as gross total resection (GTR>98% and GTR>90%. We finally stratified data for recurrent surgery, tumor location, tumor size, and tumor grade (IV versus III grade sec. WHO. Results. 94 patients were finally enrolled. Overall GTR>98% and GTR>90% was achieved in 93% and 100% of patients. Extent of resection (GTR>98% was dependent on tumor location, tumor grade (P<0.05, and tumor size (P<0.05. In 43% of patients the boundaries of fluorescent tissue exceeded those of tumoral tissue detected by neuronavigation, more frequently in larger (57% (P<0.01 and recurrent (60% tumors. Conclusions. 5-ALA fluorescence in HGG surgery enables a GTR in 100% of cases even if selection of patients remains a main bias. Recurrent surgery, and location, size, and tumor grade can predict both the surgical outcome and the intraoperative findings.

  11. Fluorescence kinetics of Trp-Trp dipeptide and its derivatives in water via ultrafast fluorescence spectroscopy.

    Science.gov (United States)

    Jia, Menghui; Yi, Hua; Chang, Mengfang; Cao, Xiaodan; Li, Lei; Zhou, Zhongneng; Pan, Haifeng; Chen, Yan; Zhang, Sanjun; Xu, Jianhua

    2015-08-01

    Ultrafast fluorescence dynamics of Tryptophan-Tryptophan (Trp-Trp/Trp2) dipeptide and its derivatives in water have been investigated using a picosecond resolved time correlated single photon counting (TCSPC) apparatus together with a femtosecond resolved upconversion spectrophotofluorometer. The fluorescence decay profiles at multiple wavelengths were fitted by a global analysis technique. Nanosecond fluorescence kinetics of Trp2, N-tert-butyl carbonyl oxygen-N'-aldehyde group-l-tryptophan-l-tryptophan (NBTrp2), l-tryptophan-l-tryptophan methyl ester (Trp2Me), and N-acetyl-l-tryptophan-l-tryptophan methyl ester (NATrp2Me) exhibit multi-exponential decays with the average lifetimes of 1.99, 3.04, 0.72 and 1.22ns, respectively. Due to the intramolecular interaction between two Trp residues, the "water relaxation" lifetime was observed around 4ps, and it is noticed that Trp2 and its derivatives also exhibit a new decay with a lifetime of ∼100ps, while single-Trp fluorescence decay in dipeptides/proteins shows 20-30ps. The intramolecular interaction lifetime constants of Trp2, NBTrp2, Trp2Me and NATrp2Me were then calculated to be 3.64, 0.93, 11.52 and 2.40ns, respectively. Candidate mechanisms (including heterogeneity, solvent relaxation, quasi static self-quenching or ET/PT quenching) have been discussed.

  12. Folding and unfolding of a non-fluorescent mutant of green fluorescent protein

    Energy Technology Data Exchange (ETDEWEB)

    Wielgus-Kutrowska, Beata [Department of Biophysics, Institute of Experimental Physics, University of Warsaw, Zwirki and Wigury 93, 02-089 (Poland); Narczyk, Marta [Department of Biophysics, Institute of Experimental Physics, University of Warsaw, Zwirki and Wigury 93, 02-089 (Poland); Buszko, Anna [Department of Biophysics, Institute of Experimental Physics, University of Warsaw, Zwirki and Wigury 93, 02-089 (Poland); Bzowska, Agnieszka [Department of Biophysics, Institute of Experimental Physics, University of Warsaw, Zwirki and Wigury 93, 02-089 (Poland); Clark, Patricia L [Department of Chemistry and Biochemistry, University of Notre Dame, 251 Nieuwland Science Hall, Notre Dame, IN 46556-5670 (United States)

    2007-07-18

    Green fluorescent protein (GFP), from the Pacific jellyfish A. victoria, has numerous uses in biotechnology and cell and molecular biology as a protein marker because of its specific chromophore, which is spontaneously created after proper protein folding. After formation, the chromophore is very stable and it remains intact during protein unfolding, meaning that the GFP unfolding process is not the reverse of the original folding reaction; i.e., the principles of microscopic reversibility do not apply. We have generated the mutant S65T/G67A-GFP, which is unable to efficiently form the cyclic chromophore, with the goal of investigating the folding, unfolding and competing aggregation of GFP under fully reversible conditions. Our studies have been performed in the presence of guanidinium hydrochloride (GdnHCl). The GFP conformation was monitored using intrinsic tryptophan fluorescence, and fluorescence of 1,1'-bis(4-anilino-5-naphthalenesulphonic acid) (bis-ANS). Light scattering was used to follow GFP aggregation. We conclude from these fluorescence measurements that S65T/G67A-GFP folding is largely reversible. During equilibrium folding, the first step is the formation of a molten globule, prone to aggregation.

  13. Steady state and time-resolved fluorescence spectroscopy of quinine sulfate dication bound to sodium dodecylsulfate micelles: Fluorescent complex formation

    Energy Technology Data Exchange (ETDEWEB)

    Joshi, Sunita; Pant, Debi D., E-mail: ddpant@pilani.bits-pilani.ac.in

    2014-01-15

    Interaction of quinine sulfate dication (QSD) with anionic, sodium dodecylsulphate (SDS) surfactant has been studied at different premicellar, micellar and postmicellar concentrations in aqueous phase using steady state, time-resolved fluorescence and fluorescence anisotropy techniques. At premicellar concentrations of SDS, the decrease in absorbance, appearance of an extra fluorescence band at lower wavelengths and tri-exponential decay behavior of fluorescence, are attributed to complex formation between QSD molecules and surfactant monomers. At postmicellar concentrations the red shift in fluorescence spectrum, increase in quantum yield and increase in fluorescence lifetimes are attributed to incorporation of solute molecules to micelles. At lower concentrations of SDS, a large shift in fluorescence is observed on excitation at the red edge of absorption spectrum and this is explained in terms of distribution of ion pairs of different energies in the ground state and the observed fluorescence lifetime behavior corroborates with this model. The temporal fluorescence anisotropy decay of QSD in SDS micelles allowed determination of restriction on the motion of the fluorophore. All the different techniques used in this study reveal that the photophysics of QSD is very sensitive to the microenvironments of SDS micelles and QSD molecules reside at the water-micelle interface. -- Highlights: • Probe molecule is very sensitive to microenvironment of micelles. • Highly fluorescent ion-pair formation has been observed. • Modulated photophysics of probe molecule in micellar solutions has been observed. • Probe molecules strongly bind with micelles and reside at probe–micelle interface.

  14. Portable fluorescence meter with reference backscattering channel

    Science.gov (United States)

    Kornilin, Dmitriy V.; Grishanov, Vladimir N.; Zakharov, Valery P.; Burkov, Dmitriy S.

    2016-09-01

    Methods based on fluorescence and backscattering are intensively used for determination of the advanced glycation end products (AGE) concentration in the biological tissues. There are strong correlation between the AGE concentration and the severity of such diseases like diabetes, coronary heart disease and renal failure. This fact can be used for diagnostic purposes in medical applications. Only few investigations in this area can be useful for development of portable and affordable in vivo AGE meter because the most of them are oriented on using spectrometers. In this study we describe the design and the results of tests on volunteers of portable fluorescence meter based on two photodiodes. One channel of such fluorimeter is used for measurement of the autofluorescence (AF) intensity, another one - for the intensity of elastically scattered radiation, which can be used as a reference. This reference channel is proposed for normalization of the skin autofluorescence signal to the human skin photo type. The fluorimeter, that was developed is relatively compact and does not contain any expensive optical and electronic components. The experimental results prove that proposed tool can be used for the AGE estimation in human skin.

  15. Fluorescent peptide sensors for tyrosylprotein sulfotransferase activity.

    Science.gov (United States)

    Zhou, Wenbo; Duckworth, Benjamin P; Geraghty, Robert J

    2014-09-15

    Tyrosine sulfurylation is a post-translational modification important for protein-protein interactions in the extracellular space that are instrumental in cell adhesion, cell signaling, immune responses, and pathogen recognition of host cells. Tyrosine sulfurylation is catalyzed by the tyrosylprotein sulfotransferases (TPSTs), and in humans there are two isoforms: hTPST1 and hTPST2. The study of hTPST function and the development of small molecule probes to examine the role of hTPSTs in cell biology have been delayed by the absence of a continuous direct assay for hTPST activity. We have developed a fluorescent peptide-based assay to directly monitor tyrosine sulfurylation in real time. TPST-mediated tyrosine sulfurylation of the peptides disrupts fluorophore quenching and results in increased fluorescence emission. The assay can be used to study TPST enzymatic activity, and we show that recombinant hTPSTs are active in the absence of divalent metal ions and that optimal activity is at pH 6.0. We further show that the assay can also be used to identify inhibitors of tyrosine sulfurylation. A clear understanding of hTPST function in normal cell biology and in disease states will require the identification of small molecule inhibitors or probes to modulate enzymatic activity, and our results will facilitate that process.

  16. Cell tracking using a photoconvertible fluorescent protein.

    Science.gov (United States)

    Hatta, Kohei; Tsujii, Hitomi; Omura, Tomomi

    2006-01-01

    The tracking of cell fate, shape and migration is an essential component in the study of the development of multicellular organisms. Here we report a protocol that uses the protein Kaede, which is fluorescent green after synthesis but can be photoconverted red by violet or UV light. We have used Kaede along with confocal laser scanning microscopy to track labeled cells in a pattern of interest in zebrafish embryos. This technique allows the visualization of cell movements and the tracing of neuronal shapes. We provide illustrative examples of expression by mRNA injection, mosaic expression by DNA injection, and the creation of permanent transgenic fish with the UAS-Gal4 system to visualize morphogenetic processes such as neurulation, placode formation and navigation of early commissural axons in the hindbrain. The procedure can be adapted to other photoconvertible and reversible fluorescent molecules, including KikGR and Dronpa; these molecules can be used in combination with two-photon confocal microscopy to specifically highlight cells buried in tissues. The total time needed to carry out the protocol involving transient expression of Kaede by injection of mRNA or DNA, photoconversion and imaging is 2-8 d.

  17. Modulated CMOS camera for fluorescence lifetime microscopy.

    Science.gov (United States)

    Chen, Hongtao; Holst, Gerhard; Gratton, Enrico

    2015-12-01

    Widefield frequency-domain fluorescence lifetime imaging microscopy (FD-FLIM) is a fast and accurate method to measure the fluorescence lifetime of entire images. However, the complexity and high costs involved in construction of such a system limit the extensive use of this technique. PCO AG recently released the first luminescence lifetime imaging camera based on a high frequency modulated CMOS image sensor, QMFLIM2. Here we tested and provide operational procedures to calibrate the camera and to improve the accuracy using corrections necessary for image analysis. With its flexible input/output options, we are able to use a modulated laser diode or a 20 MHz pulsed white supercontinuum laser as the light source. The output of the camera consists of a stack of modulated images that can be analyzed by the SimFCS software using the phasor approach. The nonuniform system response across the image sensor must be calibrated at the pixel level. This pixel calibration is crucial and needed for every camera settings, e.g. modulation frequency and exposure time. A significant dependency of the modulation signal on the intensity was also observed and hence an additional calibration is needed for each pixel depending on the pixel intensity level. These corrections are important not only for the fundamental frequency, but also for the higher harmonics when using the pulsed supercontinuum laser. With these post data acquisition corrections, the PCO CMOS-FLIM camera can be used for various biomedical applications requiring a large frame and high speed acquisition.

  18. Comparative Two-Dimensional Fluorescence Gel Electrophoresis.

    Science.gov (United States)

    Ackermann, Doreen; König, Simone

    2018-01-01

    Two-dimensional comparative fluorescence gel electrophoresis (CoFGE) uses an internal standard to increase the reproducibility of coordinate assignment for protein spots visualized on 2D polyacrylamide gels. This is particularly important for samples, which need to be compared without the availability of replicates and thus cannot be studied using differential gel electrophoresis (DIGE). CoFGE corrects for gel-to-gel variability by co-running with the sample proteome a standardized marker grid of 80-100 nodes, which is formed by a set of purified proteins. Differentiation of reference and analyte is possible by the use of two fluorescent dyes. Variations in the y-dimension (molecular weight) are corrected by the marker grid. For the optional control of the x-dimension (pI), azo dyes can be used. Experiments are possible in both vertical and horizontal (h) electrophoresis devices, but hCoFGE is much easier to perform. For data analysis, commercial software capable of warping can be adapted.

  19. Fluorescence properties of Laurdan in cochleate phases.

    Science.gov (United States)

    Ramani, Karthik; Balasubramanian, Sathyamangalam V

    2003-12-03

    Cochleates are lipid-based delivery system that have found application in drug and gene delivery. They are precipitates, formed as a result of interaction between cations (e.g. Ca2+) and negatively charged phospholipids such as phosphatidylserine (PS). In the present study, we investigated the utility of fluorescent probe Laurdan (6-dodecanoyl-2-dimethylamino naphthalene) to monitor cochleate phase formation. Following addition of Ca2+ to Laurdan labeled lipid vesicles comprised of brain phosphatidylserine (BPS), a significant blue shift in the emission peak maximum of Laurdan was observed and the spectral features were distinct from those observed for the gel and liquid-crystalline (LC) phases. This is consistent with the formation of anhydrous cochleate cylinders that was further confirmed by electron microscopy studies. Due to dipolar relaxation, excitation and emission generalized polarization (GPEx and GPEm) indicate transition from a LC to a rigid and dehydrated (RD) cochleate phase. These spectral changes were utilized to monitor the influence of lipid composition, ionic strength and lamellarity on the formation of cochleate phase. The results indicated that the presence of phosphatidylcholine (PC) and bulk Na+ concentration influenced the formation of cochleate structures from small unilamellar vesicles (SUV) and multilamellar vesicles (MLV) composed of PS. The presence of PC and higher bulk Na+ concentration stabilized the PS vesicles against collapse and total loss of contents, intermediate molecular events in the formation of cochleate structures. From these studies, we conclude that Laurdan fluorescence is a sensitive and a rapid method to detect cochleate phase formation.

  20. DNA Duplex Engineering for Enantioselective Fluorescent Sensor.

    Science.gov (United States)

    Hu, Yuehua; Lin, Fan; Wu, Tao; Zhou, Yufeng; Li, Qiusha; Shao, Yong; Xu, Zhiai

    2017-02-21

    The rapid identification of biomacromolecule structure that has a specific association with chiral enantiomers especially from natural sources will be helpful in developing enantioselective sensor and in speeding up drug exploitation. Herein, owing to its existence also in living cells, apurinic/apyrimidinic site (AP site) was first engineered into ds-DNA duplex to explore its competence in enantiomer selectivity. An AP site-specific fluorophore was utilized as an enantioselective discrimination probe to develop a straightforward chiral sensor using natural tetrahydropalmatine (L- and D-THP) as enantiomer representatives. We found that only L-THP can efficiently replace the prebound fluorophore to cause a significant fluorescence increase due to its specific binding with the AP site (two orders magnitude higher in affinity than binding with D-THP). The AP site binding specificity of L-THP over D-THP was assessed via intrinsic fluorescence, isothermal titration calorimetry, and DNA stability. The enantioselective performance can be easily tuned by the sequences near the AP site and the number of AP sites. A single AP site provides a perfect binding pocket to differentiate the chiral atom-induced structure discrepancy. We expect that our work will inspire interest in engineering local structures into a ds-DNA duplex for developing novel enantioselective sensors.

  1. Non-radiative excitation fluorescence microscopy

    Science.gov (United States)

    Riachy, Lina; Vézy, Cyrille; Jaffiol, Rodolphe

    2016-03-01

    Non-radiative Excitation Fluorescence Microscopy (NEFM) constitutes a new way to observe biological samples beyond the diffraction limit. Non-radiative excitation of the samples is achieved by coating the substrate with donor species, such as quantum dots (QDs). Thus the dyes are not excited directly by the laser source, as in common fluorescence microscopy, but through a non-radiative energy transfer. To prevent dewetting of the donor film, we have recently implemented a silanization process to covalently bond the QDs on the substrate. An homogeneous monolayer of QDs was then deposited on only one side of the coverslips. Atomic force microscopy was then used to characterize the QD layer. We highlight the potential of our method through the study of Giant Unilamellar Vesicles (GUVs) labeled with DiD as acceptor, in interaction with surface functionalized with poly-L-lysine. In the presence of GUVs, we observed a quenching of QDs emission, together with an emission of DiD located in the membrane, which clearly indicated that non-radiative energy transfer from QDs to DiD occurs.

  2. The fluorescence microprobe at the LNLS

    Energy Technology Data Exchange (ETDEWEB)

    Radtke, Martin; Perez, Carlos A.; Tolentino, Helio; Vicentin, Flavio C.; Neuenschwander, Regis T.; Barg, Bill [Laboratorio Nacional de Luz Sincrotron (LNLS), Campinas, SP (Brazil); Sanchez, Hector J.; Rubio, Marcelo [Cordoba Univ. Nacional (Argentina). Facultad de Matematica, Astronomia y Fisica; Bueno, Maria Izabel Silveira; Raimundo, Ivo M.; Rohwedder, Jarbas J.R. [Universidade Estadual de Campinas, SP (Brazil). Inst. de Quimica

    1999-11-01

    At the Laboratorio Nacional de Luz Sincrotron (LNLS) the microfocus fluorescence beamline will be operational in the spring of 1998. X-ray fluorescence with synchrotron radiation is an analytical method for determining the concentration of elements at the ppm level. The use of synchrotron radiation instead of conventional X-ray tubes increases the sensitivity of the method substantially. This is due to the higher intensity, the spectral composition, beam focusing and the decrease of background as a result of the polarization of the exciting spectrum. The sample can be excited by the `white` spectrum or monoenergetic beam. The choice of the excitation conditions has a considerable influence on the detection limits. The quantification of the measurements is possible by different mathematical techniques, like Monte Carlo simulation, fundamental parameter programs or, most easily with the calibration by standards if suitable standards are available. The use of focusing capillaries or mirror optics offers the possibility to examine a sample with a spatial resolution of a few {mu}m. One important function is the use of the setup as a scanning microprobe. A two dimensional scan over an unopened fluid inclusion is shown a s example. (author) 10 refs., 7 figs., 1 tab.

  3. 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.

  4. Nitrogen fluorescence in air for observing extensive air showers

    CERN Document Server

    Keilhauer, B; Fraga, M; Matthews, J; Sakaki, N; Tameda, Y; Tsunesada, Y; Ulrich, A

    2012-01-01

    Extensive air showers initiate the fluorescence emissions from nitrogen molecules in air. The UV-light is emitted isotropically and can be used for observing the longitudinal development of extensive air showers in the atmosphere over tenth of kilometers. This measurement technique is well-established since it is exploited for many decades by several cosmic ray experiments. However, a fundamental aspect of the air shower analyses is the description of the fluorescence emission in dependence on varying atmospheric conditions. Different fluorescence yields affect directly the energy scaling of air shower reconstruction. In order to explore the various details of the nitrogen fluorescence emission in air, a few experimental groups have been performing dedicated measurements over the last decade. Most of the measurements are now finished. These experimental groups have been discussing their techniques and results in a series of \\emph{Air Fluorescence Workshops} commenced in 2002. At the 8$^{\\rm{th}}$ Air Fluoresc...

  5. Study on fluorescence spectra of thiamine, riboflavin and pyridoxine

    Science.gov (United States)

    Yang, Hui; Xiao, Xue; Zhao, Xuesong; Hu, Lan; Lv, Caofang; Yin, Zhangkun

    2016-01-01

    This paper presents the intrinsic fluorescence characteristics of vitamin B1, B2 and B6 measured with 3D fluorescence Spectrophotometer. Three strong fluorescence areas of vitamin B2 locate at λex/λem=270/525nm, 370/525nm and 450/525nm, one fluorescence areas of vitamin B1 locates at λex/λem=370/460nm, two fluorescence areas of vitamin B6 locate at λex/λem=250/370nm and 325/370nm were found. The influence of pH of solution to the fluorescence profile was also discussed. Using the PARAFAC algorithm, 10 vitamin B1, B2 and B6 mixed solutions were successfully decomposed, and the emission profiles, excitation profiles, central wavelengths and the concentration of the three components were retrieved precisely through about 5 iteration times.

  6. Superior optical nonlinearity of an exceptional fluorescent stilbene dye

    Energy Technology Data Exchange (ETDEWEB)

    He, Tingchao [College of Physics Science and Technology, Shenzhen University, Shenzhen 518060 (China); Division of Physics and Applied Physics, Centre for Disruptive Photonic Technologies (CDPT), School of Physical and Mathematical Sciences, Nanyang Technological University, 21 Nanyang Link, Singapore 637371 (Singapore); Sreejith, Sivaramapanicker; Zhao, Yanli [Division of Chemistry and Biological Chemistry, School of Physical and Mathematical Sciences, Nanyang Technological University, 21 Nanyang Link, Singapore 637371 (Singapore); Gao, Yang; Grimsdale, Andrew C. [School of Materials Science and Engineering, Nanyang Technological University, Singapore, Singapore 639798 (Singapore); Lin, Xiaodong, E-mail: linxd@szu.edu.cn, E-mail: hdsun@ntu.edu.sg [College of Physics Science and Technology, Shenzhen University, Shenzhen 518060 (China); Sun, Handong, E-mail: linxd@szu.edu.cn, E-mail: hdsun@ntu.edu.sg [Division of Physics and Applied Physics, Centre for Disruptive Photonic Technologies (CDPT), School of Physical and Mathematical Sciences, Nanyang Technological University, 21 Nanyang Link, Singapore 637371 (Singapore)

    2015-03-16

    Strong multiphoton absorption and harmonic generation in organic fluorescent chromophores are, respectively, significant in many fields of research. However, most of fluorescent chromophores fall short of the full potential due to the absence of the combination of such different nonlinear upconversion behaviors. Here, we demonstrate that an exceptional fluorescent stilbene dye could exhibit efficient two- and three-photon absorption under the excitation of femtosecond pulses in solution phase. Benefiting from its biocompatibility and strong excited state absorption behavior, in vitro two-photon bioimaging and superior optical limiting have been exploited, respectively. Simultaneously, the chromophore could generate efficient three-photon excited fluorescence and third-harmonic generation (THG) when dispersed into PMMA film, circumventing the limitations of classical fluorescent chromophores. Such chromophore may find application in the production of coherent light sources of higher photon energy. Moreover, the combination of three-photon excited fluorescence and THG can be used in tandem to provide complementary information in biomedical studies.

  7. Superior optical nonlinearity of an exceptional fluorescent stilbene dye

    Science.gov (United States)

    He, Tingchao; Sreejith, Sivaramapanicker; Gao, Yang; Grimsdale, Andrew C.; Zhao, Yanli; Lin, Xiaodong; Sun, Handong

    2015-03-01

    Strong multiphoton absorption and harmonic generation in organic fluorescent chromophores are, respectively, significant in many fields of research. However, most of fluorescent chromophores fall short of the full potential due to the absence of the combination of such different nonlinear upconversion behaviors. Here, we demonstrate that an exceptional fluorescent stilbene dye could exhibit efficient two- and three-photon absorption under the excitation of femtosecond pulses in solution phase. Benefiting from its biocompatibility and strong excited state absorption behavior, in vitro two-photon bioimaging and superior optical limiting have been exploited, respectively. Simultaneously, the chromophore could generate efficient three-photon excited fluorescence and third-harmonic generation (THG) when dispersed into PMMA film, circumventing the limitations of classical fluorescent chromophores. Such chromophore may find application in the production of coherent light sources of higher photon energy. Moreover, the combination of three-photon excited fluorescence and THG can be used in tandem to provide complementary information in biomedical studies.

  8. IR-stimulated visible fluorescence in pink and brown diamond.

    Science.gov (United States)

    Byrne, K S; Chapman, J G; Luiten, A N

    2014-03-19

    Irradiation of natural pink and brown diamond by middle-ultraviolet light (photon energy ϵ ≥ 4.1 eV ) is seen to induce anomalous fluorescence phenomena at N3 defect centres (structure N3-V). When diamonds primed in this fashion are subsequently exposed to infrared light (even with a delay of many hours), a transient burst of blue N3 fluorescence is observed. The dependence of this IR-triggered fluorescence on pump wavelength and intensity suggest that this fluorescence phenomena is intrinsically related to pink diamond photochromism. An energy transfer process between N3 defects and other defect species can account for both the UV-induced fluorescence intensity changes, and the apparent optical upconversion of IR light. From this standpoint, we consider the implications of this N3 fluorescence behaviour for the current understanding of pink diamond photochromism kinetics.

  9. Multiple stimulated emission fluorescence photoacoustic sensing and spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Li, Gaoming [School of Electrical and Electronic Engineering, Nanyang Technological University, Singapore 639798 (Singapore); Key Laboratory of OptoElectronic Science and Technology for Medicine, Ministry of Education, Fujian Normal University, Fuzhou 350007 (China); Gao, Fei; Feng, Xiaohua; Zheng, Yuanjin, E-mail: yjzheng@ntu.edu.sg [School of Electrical and Electronic Engineering, Nanyang Technological University, Singapore 639798 (Singapore); Qiu, Yishen [Key Laboratory of OptoElectronic Science and Technology for Medicine, Ministry of Education, Fujian Normal University, Fuzhou 350007 (China)

    2016-07-04

    Multiple stimulated emission fluorescence photoacoustic (MSEF-PA) phenomenon is demonstrated in this letter. Under simultaneous illumination of pumping light and stimulated emission light, the fluorescence emission process is speeded up by the stimulated emission effect. This leads to nonlinear enhancement of photoacoustic signal while the quantity of absorbed photons is more than that of fluorescent molecules illuminated by pumping light. The electronic states' specificity of fluorescent molecular can also be labelled by the MSEF-PA signals, which can potentially be used to obtain fluorescence excitation spectrum in deep scattering tissue with nonlinearly enhanced photoacoustic detection. In this preliminary study, the fluorescence excitation spectrum is reconstructed by MSEF-PA signals through sweeping the wavelength of exciting light, which confirms the theoretical derivation well.

  10. Some fluorescence properties of dimethylaminochalcone and its novel cyclic analogues

    Science.gov (United States)

    Tomečková, Vladimíra; Poškrobová, Martina; Štefanišinová, Miroslava; Perjési, Pál

    2009-12-01

    This paper demonstrates the basic character (polarity, solubility, colour, absorption and fluorescence quantum yield) of synthetic dimethylaminochalcone ( 1) and its cyclic analogues measured in toluene, chloroform, dimethylsulfoxide and ethanol, which have been studied by absorption and fluorescence spectroscopy. The biologically active dye 4'-dimethylaminochalcone ( 1b) and its less flexible analogues 4-dimethylaminoindanone ( 2b), -tetralone ( 3b), and -benzosuberone ( 4b) are lipophilic molecules that displayed the best solubility in toluene and chloroform. The highest fluorescence and quantum yields of compounds 1 and 2 have been obtained in DMSO and chloroform. Quenching effect of fluorescence compounds ( 1- 4) has been studied in the mixture of the most polar organic solvents DMSO and water. In the presence of water, fluorescence of compound 1 has been quenched the best from all studied chalcones and emission maxima of molecules 1- 4 have been shifted to the longer wavelengths. Quenching effect of fluorescence by water was in order 1 > 2 > 3 > 4.

  11. Synthesis of Fluorescent Gelators and Direct Observation of Gelation with a Fluorescence Microscope.

    Science.gov (United States)

    Hanabusa, Kenji; Ueda, Takuya; Takata, Shingo; Suzuki, Masahiro

    2016-11-14

    Fluorescein-, benzothiazole-, quinoline-, stilbene-, and carbazole-containing fluorescent gelators have been synthesized by connecting gelation-driving segments, including l-isoleucine, l-valine, l-phenylalanine, l-leucine residue, cyclo(l-asparaginyl-l-phenylalanyl), and trans-(1R,2R)-diaminocyclohexane. The emission behaviors of the gelators were investigated, and their gelation abilities studied against 15 solvents. The minimum gel concentration, variable-temperature spectroscopy, transmission electron microscopy, scanning electron microscopy, fluorescence microscopy (FM), and confocal laser scanning microscopy (CLSM) were used to characterize gelation. The intermolecular hydrogen bonding between the N-H and C=O of amide, van der Waals interactions and π-π stacking play important roles in gelation. The colors of emission are related to the fluorescence structures of gelators. Fibrous aggregates characterized by the color of their emission were observed by FM. 3D images are produced by the superposition of images captured by CLSM every 0.1 μm to a settled depth. The 3D images show that the large micrometer-sized aggregates spread out three dimensionally. FM observations of mixed gelators are studied. In the case of gelation, two structurally related gelators with the same gelation-driving segment lead to the gelators build up of the same aggregates through similar hydrogen-bonding patterns. When two gelators with structurally different gelation-driving segments induce gelation, the gelators build up each aggregate through individual hydrogen-bonding patterns. A fluorescent reagent that was incorporated into the aggregates of gels through van der Waals interactions was developed. The addition of this fluorescent reagent enables the successful observation of nonfluorescent gelators' aggregates by FM.

  12. Quantitative Fluorescence Studies in Living Cells: Extending Fluorescence Fluctuation Spectroscopy to Peripheral Membrane Proteins

    Science.gov (United States)

    Smith, Elizabeth Myhra

    The interactions of peripheral membrane proteins with both membrane lipids and proteins are vital for many cellular processes including membrane trafficking, cellular signaling, and cell growth/regulation. Building accurate biophysical models of these processes requires quantitative characterization of the behavior of peripheral membrane proteins, yet methods to quantify their interactions inside living cells are very limited. Because peripheral membrane proteins usually exist both in membrane-bound and cytoplasmic forms, the separation of these two populations is a key challenge. This thesis aims at addressing this challenge by extending fluorescence fluctuation spectroscopy (FFS) to simultaneously measure the oligomeric state of peripheral membrane proteins in the cytoplasm and at the plasma membrane. We developed a new method based on z-scan FFS that accounts for the fluorescence contributions from cytoplasmic and membrane layers by incorporating a fluorescence intensity z-scan through the cell. H-Ras-EGFP served as a model system to demonstrate the feasibility of the technique. The resolvability and stability of z-scanning was determined as well as the oligomeric state of H-Ras-EGFP at the plasma membrane and in the cytoplasm. Further, we successfully characterized the binding affinity of a variety of proteins to the plasma membrane by quantitative analysis of the z-scan fluorescence intensity profile. This analysis method, which we refer to as z-scan fluorescence profile deconvoution, was further used in combination with dual-color competition studies to determine the lipid specificity of protein binding. Finally, we applied z-scan FFS to provide insight into the early assembly steps of the HTLV-1 retrovirus.

  13. A never ending race for new and improved fluorescent proteins.

    Science.gov (United States)

    Jones, Alexander M; Ehrhardt, David W; Frommer, Wolf B

    2012-05-03

    Bioluminescent and fluorescent proteins are now used as tools for research in all organisms. There has been massive progress over the past 15 years in creating a palette of fluorescent proteins with a wide spectrum of specific properties. One of the big challenges is to decide which variant may be best for a certain application. A recent article by Mann et al. in BMC Biotechnology describes a new orange fluorescent protein in plants.

  14. Cutin fluorescence in early embryos of Pinus and Tsuga

    OpenAIRE

    Ewa Szczuka; Irena Gielwanowska

    2014-01-01

    Embryos of Pinus nigra Arnold and Tsuga canadensis Carr. (Pinaceae) at different stages of development were dissected from fresh, unfixed seeds and examined in a fluorescence microscope with 400 nm excitation light. The embryos of the investigated species showed cutin fluorescence after auramine 0 staining. At first the fluorescing cutin layer was formed on the apical part of the embryo with a well developed secondary suspensor, then it extended over the lateral surface of the embryo; the sus...

  15. DETERMINATION OF AMINOGLYCOSIDES IN FOOD BY FLUORESCENCE POLARIZATION IMMUNOASSAY

    Directory of Open Access Journals (Sweden)

    FARAFONOVA O.V.

    2015-01-01

    Full Text Available The methodic for quantitative determination of aminoglycoside antibiotics (gentamicin, kanamycin, streptomycin, amikacin, neomycin in food by polarization fluorescent immunoassay (FPIA is developed. The size and structure influence of a fluorescent molecule on a fluorescence polarization degree is analyzed. Affinity constants of antibodies to compounds and tracers were estimated, optimized working concentration of tracers and antibodies that provide the maximum value of analytical signal. Methods were tested in the antibiotics identification in milk, eggs and chicken.

  16. Energy Dependence of Air Fluorescence Yield measured by AIRFLY

    CERN Document Server

    Ave, M

    2007-01-01

    In the fluorescence detection of ultra high energy (> 10**18 eV) cosmic rays, the number of emitted fluorescence photons is assumed to be proportional to the energy deposited in air by shower particles. We have performed measurements of the fluorescence yield in atmospheric gases excited by electrons over energies ranging from keV to hundreds of MeV in several accelerators. We found that within the measured energy ranges the proportionality holds at the level of few %.

  17. DBD dyes as fluorescent probes for sensing lipophilic environments.

    Science.gov (United States)

    Wawrzinek, Robert; Wessig, Pablo; Möllnitz, Kristian; Nikolaus, Jörg; Schwarzer, Roland; Müller, Peter; Herrmann, Andreas

    2012-09-01

    Small fluorescent organic molecules based on [1,3]dioxolo[4,5-f][1,3]benzodioxole (DBD) could be used as probes for lipophillic microenvironments in aqueous solutions by indicating the critical micelles concentration of detergents and staining cell organelles. Their fluorescence lifetime decreases drastically by the amount of water in their direct environment. Therefore they are potential probes for fluorescence lifetime imaging microscopy (FLIM).

  18. High-Definition X-Ray Fluorescence: Applications

    OpenAIRE

    Gibson, Walter M.; Chen, Z W; Li, Danhong

    2008-01-01

    Energy dispersive X-ray fluorescence (EDXRF) is a well-established and powerful tool for nondestructive elemental analysis of virtually any material. It is widely used for environmental, industrial, pharmaceutical, forensic, and scientific research applications to measure the concentration of elemental constituents or contaminants. The fluorescing atoms can be excited by energetic electrons, ions, or photons. A particular EDXRF method, monochromatic microbeam X-ray fluorescence (MμEDXRF), has...

  19. High Definition X-Ray Fluorescence: Principles and Techniques

    OpenAIRE

    Chen, Z W; Gibson, Walter M.; Huang, Huapeng

    2008-01-01

    Energy dispersive X-ray fluorescence (EDXRF) is a well-established and powerful tool for nondestructive elemental analysis of virtually any material. It is widely used for environmental, industrial, pharmaceutical, forensic, and scientific research applications to measure the concentration of elemental constituents or contaminants. The fluorescing atoms can be excited by energetic electrons, ions, or photons. A particular EDXRF method, monochromatic microfocus X-ray fluorescence (MμEDXRF), ha...

  20. Solid State NMR and Fluorescence Studies of Conjugated Polymer Nanocomposties

    Institute of Scientific and Technical Information of China (English)

    Chao Jun JING; Liu Sheng CHEN; Yi SHI; Xi Gao JIN

    2005-01-01

    13C spin-lattice relaxation times (T1) of a conjugated polymer MEH-PPV in polymer/layered silicate nanocomposites together with the steady state fluorescence emission and transient fluorescence decay measurements have been investigated. The T1 values of the conjugated carbons decrease dramatically according to the reduction of polymer concentration in the nano composites, while the fluorescence life times (τ) show a linear prolonging tendency. The results are explained from the point of view of molecular dynamics.

  1. Fluorescence Analysis of E. coli Bacteria in Water

    OpenAIRE

    Bulycheva, Elizaveta Vladimirovna; Korotkova, Elena Ivanovna; Voronova, Olesya Aleksandrovna; Kustova, A. A.; Petrova, Ekaterina Viktorovna

    2014-01-01

    The fluorescence analysis of Escherichia coli (E. coli) bacteria was done. It has been established that a luminescent signal from the one of metabolites (reduction form of nicotinamide adenine dinucleotide, NADH) can be adopted as a vitality indicator of the bacteria. This signal was chosen as an analytical signal. It was determined that the nature of this signal is fluorescence. In order to eliminate influence of the light scattering on this fluorescence signal optimal conditions were chosen.

  2. Fluorescence spectroscopy and multi-way techniques. PARAFAC

    DEFF Research Database (Denmark)

    Murphy, Kathleen R.; Stedmon, Colin A.; Graeber, Daniel;

    2013-01-01

    of independently varying fluorophores. However, many practical and analytical hurdles stand between EEM datasets and their chemical interpretation. This article provides a tutorial in the practical application of PARAFAC to fluorescence datasets, demonstrated using a dissolved organic matter (DOM) fluorescence...... dataset. A new toolbox for MATLAB is presented to support improved visualisation and sensitivity analyses of PARAFAC models in fluorescence spectroscopy. © 2013 The Royal Society of Chemistry....

  3. 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.

  4. Synchronous fluorescence and excitation emission characteristics of transformer oil ageing.

    Science.gov (United States)

    Deepa, Subbiah; Sarathi, R; Mishra, Ashok K

    2006-11-15

    This paper describes the evaluation of synchronous fluorescence spectroscopy (SFS) and excitation emission matrix fluorescence (EEMF) spectroscopy as means of monitoring transformer oil degradation. When accelerated thermal ageing method is used, the onset of degradation of transformer oil on 17th day and transformer oil with polypropylene and cellulosic paper on 23rd and 27th days is sensitively reflected in the SFS and EEMF fluorescence spectral characteristics.

  5. Binding of several benzodiazepines to bovine serum albumin: Fluorescence study

    Science.gov (United States)

    Machicote, Roberta G.; Pacheco, María E.; Bruzzone, Liliana

    2010-10-01

    The interactions of lorazepam, oxazepam and bromazepam with bovine serum albumin (BSA) were studied by fluorescence spectrometry. The Stern-Volmer quenching constants and corresponding thermodynamic parameters Δ H, Δ G and Δ S were calculated. The binding constants and the number of binding sites were also investigated. The distances between the donor (BSA) and the acceptors (benzodiazepines) were obtained according to fluorescence resonance energy transfer and conformational changes of BSA were observed from synchronous fluorescence spectra.

  6. Application value of fluorescence in situ hybridization in patients with urothelial bladder cancer%荧光原位杂交技术在膀胱尿路上皮癌诊断中的应用价值

    Institute of Scientific and Technical Information of China (English)

    余义; 刘飞; 黄永辉; 陆文宝; 黄红卫

    2012-01-01

    Objective:To evaluate the application value of fluorescence in situ hybridization(FISH)technique in the diagnosis of urothelial bladder cancer. Method: Urine samples from 60 patients suspected of having bladder cancer were collected before cystoscopy for immediate urine cytology and FISH analysis. The criteria for FISH abnormality were determined by evaluating the urine specimens from 20 subjects without urogenital neoplasm. Result; The overall sensitivity of cytology and FISH was 42. 0% and 82. 2% ,and their specificity was 93. 3% and 86. 7% , respectively. The differences between FISH and cytology showed statistical significance in the sensitivity of low grade and non-muscle-invasive cancer(P<0. 05). Conclusion: FISH can improve remarkablely the detection rate of urothelial bladder cancer,especially for the low stage and low grade tumors,and will be a new noninvasive screening method for urothelial bladder cancer.%目的:评估荧光原位杂交技术(FISH)在膀胱尿路上皮癌诊断中的应用价值.方法:收集60例疑似膀胱癌的血尿患者的尿液标本,分别作尿细胞学检测和荧光原位杂交分析.20例正常人尿液标本,用于建立FISH阀值,作为阳性判断的标准.结果:细胞学和FISH的总敏感性分别为42.0%、82.2%,特异性分别为:93.3%、86.7%.细胞学和FISH在低级别及非肌层浸润性肿瘤等敏感性的均差异有统计学意义(P<0.05).结论:FISH技术能明显提高膀胱尿路上皮癌的检出率,尤其是早期和低级别病变,可以成为筛查膀胱尿路上皮癌的一种新的无创性检查方法.

  7. Genetically encoded biosensors based on engineered fluorescent proteins.

    Science.gov (United States)

    Frommer, Wolf B; Davidson, Michael W; Campbell, Robert E

    2009-10-01

    Fluorescent proteins have revolutionized cell biology by allowing researchers to non-invasively peer into the inner workings of cells and organisms. While the most common applications of fluorescent proteins are to image expression, localization, and dynamics of protein chimeras, there is a growing interest in using fluorescent proteins to create biosensors for minimally invasive imaging of concentrations of ions and small molecules, the activity of enzymes, and changes in the conformation of proteins in living cells. This tutorial review provides an overview of the progress made in the development of fluorescent protein-based biosensors to date.

  8. Fluorescein sodium fluorescence microscope-integrated lymphangiography for lymphatic supermicrosurgery.

    Science.gov (United States)

    Ayestaray, Benoit; Bekara, Farid

    2015-07-01

    Microscope-integrated lymphangiography is a useful method in the field of lymphatic supermicrosurgery. Fluorescence based on indocyanine green (ICG) is the most commonly used. Fluorescein sodium is a fluorescent tracer used for retinal and neurosurgical angiography but not yet for lymphatic supermicrosurgery. In this report, we present a case in which the fluorescein sodium fluorescence microscope-integrated lymphangiography was used for assessment of lymphatic drainage pathway and patency in a patient treated for secondary lymphedema by lymphaticovenular anastomoses. Fluorescein sodium fluorescence microscope-integrated lymphangiography was evaluated in a 67-year-old female presented for a Campisi clinical stage IV lymphedema of the upper limb. Transcutaneous guidance and vascular fluorescence were assessed. A comparison with ICG fluorescence was made intraoperatively. Two lymphaticovenular anastomoses were performed and their patency were checked by lymphangiography. Transcutaneous signal was found higher with fluorescein sodium fluorescence. Intraluminal visualization was possible with fluorescein sodium coloration during lymphaticovenular anastomoses. No adverse reaction occurred. The circumferential differential reduction rate of affected limb was 8.1% 3 months after lymphaticovenular anastomoses. The use of fluorescence microscope-integrated lymphangiography with fluorescein sodium may be superior to ICG fluorescence in assistance of lymphaticovenular anastomoses. © 2015 Wiley Periodicals, Inc.

  9. Fluorescence chemosensors with pyrene and their interaction with nucleotide phosphate

    Institute of Scientific and Technical Information of China (English)

    李华平; 汪鹏飞; 吴世康

    1999-01-01

    A group of fluorescence chemosensor with pyrene, compounds (Ⅰ), (Ⅱ) and (Ⅲ), were synthesized The fluorescence spectra and the lifetime of these compounds were carefully measured. The fluorescence quenching spec tra of pyrenyl butyric acid, compounds (Ⅰ), (Ⅱ) and (Ⅲ) by different nucleotide phosphates, AMP ADP, ATP dTTP, were also recorded and studied. The quenching and the stability constants were calculated by Stern-Volmer equa tion and eq. (2), respectively. The mechanism of interaction between fluorescence chemosensor and nucleotide phos phate was didscussed based on the comparison of the results obtained with the CPK model of free molecules of these com pounds in the ground state.

  10. Fluorescent nanoparticles based on AIE fluorogens for bioimaging

    Science.gov (United States)

    Yan, Lulin; Zhang, Yan; Xu, Bin; Tian, Wenjing

    2016-01-01

    Fluorescent nanoparticles (FNPs) have recently attracted increasing attention in the biomedical field because of their unique optical properties, easy fabrication and outstanding performance in imaging. Compared with conventional molecular probes including small organic dyes and fluorescent proteins, FNPs based on aggregation-induced emission (AIE) fluorogens have shown significant advantages in tunable emission and brightness, good biocompatibility, superb photo- and physical stability, potential biodegradability and facile surface functionalization. In this review, we summarize the latest advances in the development of fluorescent nanoparticles based on AIE fluorogens including polymer nanoparticles and silica nanoparticles over the past few years, and the various biomedical applications based on these fluorescent nanoparticles are also elaborated.

  11. Turn on Fluorescent Probes for Selective Targeting of Aldehydes

    Directory of Open Access Journals (Sweden)

    Ozlem Dilek

    2016-03-01

    Full Text Available Two different classes of fluorescent dyes were prepared as a turn off/on sensor system for aldehydes. Amino derivatives of a boron dipyrromethene (BDP fluorophore and a xanthene-derived fluorophore (rosamine were prepared. Model compounds of their product with an aldehyde were prepared using salicylaldehyde. Both amino boron dipyrromethene and rosamine derivatives are almost non-fluorescent in polar and apolar solvent. However, imine formation with salicylaldehyde on each fluorophore increases the fluorescence quantum yield by almost a factor of 10 (from 0.05 to 0.4. These fluorophores are therefore suitable candidates for development of fluorescence-based sensors for aldehydes.

  12. Development of Enantioselective Fluorescent Sensors for Chiral Recognition

    Institute of Scientific and Technical Information of China (English)

    Lin Pu

    2004-01-01

    Novel chiral compounds have been synthesized for the enantioselective fluorescent recognition of alpha-hydroxycarboxylic acids and amino acids. By introducing dendritic branches to the chiral receptor units, the fluorescence signals of the receptors are significantly amplified because of the light-harvesting effect of the dendritic structure. This has greatly increased the sensitivity of the sensors in the fluorescent recognition. Highly enantioselective fluorescent responses have also been achieved. These sensors are potentially useful for the high throughput screening of chiral catalysts for asymmetric synthesis.

  13. Quenching of fluorescence in membrane protein by hypocrellin B.

    Science.gov (United States)

    Yue, J; Pang, S

    1997-04-01

    The hypocrellin B (HB) was used as a fluorescence quencher to study the basic physical charactcristics of HB in membrane systems, including the diffusion speed of quencher from aqueous phase into membrane phase, the partition coefficient (P) of quenchtr between membrane and water, and the fluorescence quenching constant of protein (K(sv); K(q),). The experimental results show that the quenching of fluorescence in membrane protein by HB can be determined by the principle of dynamic quenching. The experimental process of fluorescence quenching was observed in detail by using the ESR technique. The signal of HB- was found to arise from an electron transfer from excited trytophan to HB.

  14. Deconvolution of calcium fluorescent indicator signal from AFM cantilever reflection.

    Science.gov (United States)

    Lopez-Ayon, G Monserratt; Oliver, David J; Grutter, Peter H; Komarova, Svetlana V

    2012-08-01

    Atomic force microscopy (AFM) can be combined with fluorescence microscopy to measure the changes in intracellular calcium levels (indicated by fluorescence of Ca²⁺ sensitive dye fluo-4) in response to mechanical stimulation performed by AFM. Mechanical stimulation using AFM is associated with cantilever movement, which may interfere with the fluorescence signal. The motion of the AFM cantilever with respect to the sample resulted in changes of the reflection of light back to the sample and a subsequent variation in the fluorescence intensity, which was not related to changes in intracellular Ca²⁺ levels. When global Ca²⁺ responses to a single stimulation were assessed, the interference of reflected light with the fluorescent signal was minimal. However, in experiments where local repetitive stimulations were performed, reflection artifacts, correlated with cantilever motion, represented a significant component of the fluorescent signal. We developed a protocol to correct the fluorescence traces for reflection artifacts, as well as photobleaching. An added benefit of our method is that the cantilever reflection in the fluorescence recordings can be used for precise temporal correlation of the AFM and fluorescence measurements.

  15. Quenching of fluorescence in membrane protein by hypocrellin B

    Institute of Scientific and Technical Information of China (English)

    乐加昌; 庞素珍

    1997-01-01

    The hypocrellin B (HB) was used as a fluorescence quencher to study the basic physical characteris-tics of HB in membrane systems, including the diffusion speed of quencher from aqueous phase into membrane phase, the partition coefficient (P) of quencher between membrane and water, and the fluorescence quenching constant of protein (Ksv; Kq). The experimental results show that the quenching of fluorescence in membrane protein by HB can be determined by the principle of dynamic quenching. The experimental process of fluorescence quenching was ob-served in detail by using the ESR technique. The signal of HB" was found to arise from an electron transfer from ex-cited trytophan to HB.

  16. 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.

  17. [Development of Zn(2+) selective fluorescent probes for biological applications].

    Science.gov (United States)

    Hagimori, Masayori

    2013-01-01

    Zn(2+) is an essential element for life and is known to play important roles in biological processes including gene expression, apoptosis, enzyme regulation, immune system and neurotransmission. To investigate physiological roles of free or chelatable Zn(2+) in living cells, Zn(2+)-selective fluorescent probes are valuable tools. A variety of fluorescent probes based on quinoline, BF2 chelated dipyrromethene, fluorescein, etc. has been developed recently. In principle, such tools can provide useful information about zinc biology. However, most of the fluorescent probes presented so far possess a fluorescent core and a separate part for binding to Zn(2+) within the molecule, so that the molecular weight is usually large and the molecules are hydrophobic. As a result, the applications of such molecules in biological systems often face difficulties. Therefore, we need to develop a new class of fluorescent probes for Zn(2+) with improved molecular characteristics. If the initial core structure is small enough, the fluorescent probes may still be molecular weight below 500 with desirable physico-chemical properties, even after the modifications. In this review, we described novel low-molecular-weight fluorescent probes for Zn(2+) based on pyridine-pyridone. Small modification of pyridine-pyridone core structure brought about a marked improvement such as aqueous solubility, affinity toward Zn(2+), and fluorescence ON/OFF switching. Fluorescence images of Zn(2+) in cells showed that the pyridine-pyridone probe can be used in biological applications.

  18. The fluorescence and dynamics properties in phenoxy-phthalocyanines liquid

    Science.gov (United States)

    Yao, Cheng-Bao; Yan, Xiao-Yan; Tan, Ming-Yue; Li, Jin; Sun, Wen-Jun; Yang, Shou-Bin

    2015-06-01

    We investigated the one/two-photon fluorescence and excited state dynamics properties of two synthesized phenoxy-phthalocyanines (Pc1 and Pc2) using mild reaction coordination method. The results show that the fast decay component in the time-resolved fluorescence technique dynamics comes from the intramolecular vibrational relaxation, the slower ones from the internal conversion. Furthermore, in comparison with one-photon fluorescence spectra, the red shift of two-photon fluorescence spectra can be explained by the reabsorption effect of molecules. The samples are expected to be a potential candidate for optical applications and photodynamic therapy.

  19. Fluorescent eco-particles for surface flow physics analysis

    Science.gov (United States)

    Tauro, F.; Porfiri, M.; Grimaldi, S.

    2013-03-01

    In this letter, we describe a novel methodology for fabricating inexpensive environmentally-friendly fluorescent microparticles for quantitative surface flow visualization. Particles are synthesized from natural white beeswax and a highly diluted solution of a nontoxic fluorescent red dye. Bead fluorescence exhibits a long lifetime in adverse conditions, such as exposure to weathering agents, and is enhanced by Ultra Violet radiation. The fluorescent eco-particles are integrated in a particle image velocimetry study of circular hydraulic jump to demonstrate their feasibility in tracing complex surface flows.

  20. Protein conformation in solution by three-dimensional fluorescence spectrometry

    Institute of Scientific and Technical Information of China (English)

    鄢远; 许金钩; 陈国珍

    1996-01-01

    The conformations of bovine serum albumin (USA) and egg albumin (EA) in solution and their conformation changes under different conditions were studied by using three-dimensional fluorescence spectrometry (TDFS) such as three-dimensional fluorescence (TDF) spectra and three-dimensional fluorescence polarization (TDFP) spectra with tryptophan residues in protein molecules as an intrinsic fluorescent probe. The results show that the microenvironment of tryptophan residues of protein molecules in various solutions can be directly indicated and TDFS is an effective tool for studying protein conformation in solution. Meantime, some valuable results were obtained.

  1. Stink Bug Feeding Induces Fluorescence in Developing Cotton Bolls

    Directory of Open Access Journals (Sweden)

    Toews Michael D

    2011-08-01

    Full Text Available Abstract Background Stink bugs (Hemiptera: Pentatomidae comprise a critically important insect pest complex affecting 12 major crops worldwide including cotton. In the US, stink bug damage to developing cotton bolls causes boll abscission, lint staining, reduced fiber quality, and reduced yields with estimated losses ranging from 10 to 60 million dollars annually. Unfortunately, scouting for stink bug damage in the field is laborious and excessively time consuming. To improve scouting accuracy and efficiency, we investigated fluorescence changes in cotton boll tissues as a result of stink bug feeding. Results Fluorescent imaging under long-wave ultraviolet light showed that stink bug-damaged lint, the inner carpal wall, and the outside of the boll emitted strong blue-green fluorescence in a circular region near the puncture wound, whereas undamaged tissue emissions occurred at different wavelengths; the much weaker emission of undamaged tissue was dominated by chlorophyll fluorescence. We further characterized the optimum emission and excitation spectra to distinguish between stink bug damaged bolls from undamaged bolls. Conclusions The observed characteristic fluorescence peaks associated with stink bug damage give rise to a fluorescence-based method to rapidly distinguish between undamaged and stink bug damaged cotton bolls. Based on the fluorescent fingerprint, we envision a fluorescence reflectance imaging or a fluorescence ratiometric device to assist pest management professionals with rapidly determining the extent of stink bug damage in a cotton field.

  2. 3D fluorescence spectral data interpolation by using IDW.

    Science.gov (United States)

    He, Qinghang; Zhang, Zhenxi; Yi, Chao

    2008-12-01

    Because measured precision of some spectral instruments such as fluorescence spectrometer HITACHI F-4500 cannot reach the requirement of spectral analytical technique, and measured data is finite, which causes some three-dimensional (3D) fluorescence spectral data missed. The fact of missing data can result in errors in interpretations of 3D fluorescence spectral data. This paper takes ethanol solution (volume percentage phi(beta)=0.400) 3D fluorescence spectral data for example, applies inverse distance weighting (IDW) to 3D fluorescence spectral data interpolation. The results prove that the more details of 3D fluorescence spectra are expressed well by using IDW in contrast to that of original 3D fluorescence spectra. To evaluate the effectiveness of interpolation by using IDW, this paper compares standard deviation, coefficient of variation, and the mean, median, maximum and minimum values of original ethanol solution (phi(beta)=0.400) 3D fluorescence spectral data and that of the interpolated, whose results suggest that the interpolation of the 3D fluorescence spectra data by using IDW is exact.

  3. Tomato seeds maturity detection system based on chlorophyll fluorescence

    Science.gov (United States)

    Li, Cuiling; Wang, Xiu; Meng, Zhijun

    2016-10-01

    Chlorophyll fluorescence intensity can be used as seed maturity and quality evaluation indicator. Chlorophyll fluorescence intensity of seed coats is tested to judge the level of chlorophyll content in seeds, and further to judge the maturity and quality of seeds. This research developed a detection system of tomato seeds maturity based on chlorophyll fluorescence spectrum technology, the system included an excitation light source unit, a fluorescent signal acquisition unit and a data processing unit. The excitation light source unit consisted of two high power LEDs, two radiators and two constant current power supplies, and it was designed to excite chlorophyll fluorescence of tomato seeds. The fluorescent signal acquisition unit was made up of a fluorescence spectrometer, an optical fiber, an optical fiber scaffolds and a narrowband filter. The data processing unit mainly included a computer. Tomato fruits of green ripe stage, discoloration stage, firm ripe stage and full ripe stage were harvested, and their seeds were collected directly. In this research, the developed tomato seeds maturity testing system was used to collect fluorescence spectrums of tomato seeds of different maturities. Principal component analysis (PCA) method was utilized to reduce the dimension of spectral data and extract principal components, and PCA was combined with linear discriminant analysis (LDA) to establish discriminant model of tomato seeds maturity, the discriminant accuracy was greater than 90%. Research results show that using chlorophyll fluorescence spectrum technology is feasible for seeds maturity detection, and the developed tomato seeds maturity testing system has high detection accuracy.

  4. Lymphocyte fluorescent profiles (LFP): a possible screening technique in neoplasia.

    Science.gov (United States)

    Love, L A; Metcalf, N F; Metcalf, W K; Sharp, J G

    1979-05-01

    A technique involving fluorescent protein staining and microfluorometry has been developed for measuring the lymphocyte fluorescent profile (LFP) of peripheral blood lymphocytes. In contrast to normal humans who display a regular bell-shaped curve, the profile from patients with cancer is irregular, showing a bimodal distribution of fluorescence, with a significant population of cells fluorescing at a higher relative intensity. It is suggested that this elevation in protein concentration is due to an immune response to the presence of a neoplasm, and thus this technique may prove to be a useful indicator of malignancy.

  5. Native Fluorescence Life Detection instrument for planetary surfaces.

    Science.gov (United States)

    Smith, Heather

    2016-07-01

    For this research project we designed an instrument to detect organisms (amino acids, metabolites) via biomolecular fluorescence. We proposed a novel technique for searching for direct evidence of life on planetary bodies. Results indicate the fluorescence of the biotic component of desert soils is approximately as strong as the fluorescence of the mineral component. Fluorescence laboratory measurements using the portable instrument reveal microbial concentration in the Mojave Desert soil is 107 bacteria per gram of soil. Soil microbial concentrations in the Mojave Desert, determined in situ via fluorescence, show that the number varies from 104 to 107 cells per gram of soil. Biomolecules and polycyclic aromatic hydrocarbons are highly fluorescent at wavelengths in the ultra violet (266 nm, 355 nm), but not as much in the visible 532 nm range. Preliminary results show minerals discovered, such as perchlorate, fluoresce highest when excited by 355 nm. Overall, we conclude the fluorescent instrument described is suitable to detect soil microbes, organics, biomolecules, and some minerals via fluorescence, offering a high scientific return for minimal cost with non-contact applications in extreme environments on Earth and on future planetary missions.

  6. Fluorescent eco-particles for surface flow physics analysis

    Directory of Open Access Journals (Sweden)

    F. Tauro

    2013-03-01

    Full Text Available In this letter, we describe a novel methodology for fabricating inexpensive environmentally-friendly fluorescent microparticles for quantitative surface flow visualization. Particles are synthesized from natural white beeswax and a highly diluted solution of a nontoxic fluorescent red dye. Bead fluorescence exhibits a long lifetime in adverse conditions, such as exposure to weathering agents, and is enhanced by Ultra Violet radiation. The fluorescent eco-particles are integrated in a particle image velocimetry study of circular hydraulic jump to demonstrate their feasibility in tracing complex surface flows.

  7. An overview of remote sensing of chlorophyll fluorescence

    Science.gov (United States)

    Xing, Xiao-Gang; Zhao, Dong-Zhi; Liu, Yu-Guang; Yang, Jian-Hong; Xiu, Peng; Wang, Lin

    2007-03-01

    Besides empirical algorithms with the blue-green ratio, the algorithms based on fluorescence are also important and valid methods for retrieving chlorophyll-a concentration in the ocean waters, especially for Case II waters and the sea with algal blooming. This study reviews the history of initial cognitions, investigations and detailed approaches towards chlorophyll fluorescence, and then introduces the biological mechanism of fluorescence remote sensing and main spectral characteristics such as the positive correlation between fluorescence and chlorophyll concentration, the red shift phenomena. Meanwhile, there exist many influence factors that increase complexity of fluorescence remote sensing, such as fluorescence quantum yield, physiological status of various algae, substances with related optical property in the ocean, atmospheric absorption etc. Based on these cognitions, scientists have found two ways to calculate the amount of fluorescence detected by ocean color sensors: fluorescence line height and reflectance ratio. These two ways are currently the foundation for retrieval of chlorophyl l - a concentration in the ocean. As the in-situ measurements and synchronous satellite data are continuously being accumulated, the fluorescence remote sensing of chlorophyll-a concentration in Case II waters should be recognized more thoroughly and new algorithms could be expected.

  8. Novel Instrument to Measure Aerosol Fluorescence, Absorption, and Scattering Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Picarro, Inc proposes to develop the first cavity ringdown spectroscopy (CRDS) system to measure fluorescence, absorption, and scattering properties of atmospheric...

  9. A novel fluorescent label based on biological fluores-cent nanoparticles and its application in cell recognition

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Uniform-sized fluorescent nanoparticles have been prepared by employing silica as the shell and a highly luminescent dye complex of ruthenium ion and bipyridyl, tris(2,2′-bipyridyl) dichlororuthenium(Ⅱ) hexahydrate as the core of the nanoparticles. A novel fluorescent label method is proposed, which is based on the biological fluorescent nanoparticles on the foundation of nanotechnology, biotechnology and fluorescent label technology. In comparison with the conventional fluorophores as fluorescent labels such as fluorescein isothiocyanate (FITC) label, this new label shows more superiority in photochemical stability, detection sensitivity and application scope for the biomedicine research. SmIgG+ B lymphocytes isolated from the circulating blood of human beings can be easily recognized by using this new fluorescent label.

  10. Max Tech and Beyond: Fluorescent Lamps

    Energy Technology Data Exchange (ETDEWEB)

    Scholand, Michael

    2012-04-01

    Fluorescent lamps are the most widely used artificial light source today, responsible for approximately 70% of the lumens delivered to our living spaces globally. The technology was originally commercialized in the 1930's, and manufacturers have been steadily improving the efficacy of these lamps over the years through modifications to the phosphors, cathodes, fill-gas, operating frequency, tube diameter and other design attributes. The most efficient commercially available fluorescent lamp is the 25 Watt T5 lamp. This lamp operates at 114-116 lumens per watt while also providing good color rendering and more than 20,000 hours of operating life. Industry experts interviewed indicated that while this lamp is the most efficient in the market today, there is still a further 10 to 14% of potential improvements that may be introduced to the market over the next 2 to 5 years. These improvements include further developments in phosphors, fill-gas, cathode coatings and ultraviolet (UV) reflective glass coatings. The commercialization of these technology improvements will combine to bring about efficacy improvements that will push the technology up to a maximum 125 to 130 lumens per watt. One critical issue raised by researchers that may present a barrier to the realization of these improvements is the fact that technology investment in fluorescent lamps is being reduced in order to prioritize research into light emitting diodes (LEDs) and ceramic metal halide high intensity discharge (HID) lamps. Thus, it is uncertain whether these potential efficacy improvements will be developed, patented and commercialized. The emphasis for premium efficacy will continue to focus on T5 lamps, which are expected to continue to be marketed along with the T8 lamp. Industry experts highlighted the fact that an advantage of the T5 lamp is the fact that it is 40% smaller and yet provides an equivalent lumen output to that of a T8 or T12 lamp. Due to its smaller form factor, the T5 lamp

  11. Fluorescence activated cell sorting of plant protoplasts.

    Science.gov (United States)

    Bargmann, Bastiaan O R; Birnbaum, Kenneth D

    2010-02-18

    High-resolution, cell type-specific analysis of gene expression greatly enhances understanding of developmental regulation and responses to environmental stimuli in any multicellular organism. In situ hybridization and reporter gene visualization can to a limited extent be used to this end but for high resolution quantitative RT-PCR or high-throughput transcriptome-wide analysis the isolation of RNA from particular cell types is requisite. Cellular dissociation of tissue expressing a fluorescent protein marker in a specific cell type and subsequent Fluorescence Activated Cell Sorting (FACS) makes it possible to collect sufficient amounts of material for RNA extraction, cDNA synthesis/amplification and microarray analysis. An extensive set of cell type-specific fluorescent reporter lines is available to the plant research community. In this case, two marker lines of the Arabidopsis thaliana root are used: P(SCR;)::GFP (endodermis and quiescent center) and P(WOX5;)::GFP (quiescent center). Large numbers (thousands) of seedlings are grown hydroponically or on agar plates and harvested to obtain enough root material for further analysis. Cellular dissociation of plant material is achieved by enzymatic digestion of the cell wall. This procedure makes use of high osmolarity-induced plasmolysis and commercially available cellulases, pectinases and hemicellulases to release protoplasts into solution. FACS of GFP-positive cells makes use of the visualization of the green versus the red emission spectra of protoplasts excited by a 488 nm laser. GFP-positive protoplasts can be distinguished by their increased ratio of green to red emission. Protoplasts are typically sorted directly into RNA extraction buffer and stored for further processing at a later time. This technique is revealed to be straightforward and practicable. Furthermore, it is shown that it can be used without difficulty to isolate sufficient numbers of cells for transcriptome analysis, even for very scarce

  12. Magnetic resonance tracking of fluorescent nanodiamond fabrication

    Science.gov (United States)

    Shames, A. I.; Osipov, V. Yu; Boudou, J. P.; Panich, A. M.; von Bardeleben, H. J.; Treussart, F.; Vul', A. Ya

    2015-04-01

    Magnetic resonance techniques (electron paramagnetic resonance (EPR) and nuclear magnetic resonance (NMR)) are used for tracking the multi-stage process of the fabrication of fluorescent nanodiamonds (NDs) produced by high-energy electron irradiation, annealing, and subsequent nano-milling. Pristine commercial high pressure and high temperature microdiamonds (MDs) with mean size 150 μm contain ~5  ×  1018 spins/g of singlet (S = 1/2) substitutional nitrogen defects P1, as well as sp3 C-C dangling bonds in the crystalline lattice. The half-field X-band EPR clearly shows (by the appearance of the intense ‘forbidden’ g = 4.26 line) that high-energy electron irradiation and annealing of MDs induce a large amount (~5  ×  1017 spins/g) of triplet (S = 1) magnetic centers, which are identified as negatively charged nitrogen vacancy defects (NV-). This is supported by EPR observations of the ‘allowed’ transitions between Zeeman sublevels of the triplet state. After progressive milling of the fluorescent MDs down to an ultrasubmicron scale (≤100 nm), the relative abundance of EPR active NV- defects in the resulting fluorescent NDs (FND) substantially decreases and, vice versa, the content of C-inherited singlet defects correlatively increases. In the fraction of the finest FNDs (mean particle size dried supernatant of ultracentrifuged aqueous dispersion of FNDs, the NV- content is found to be reduced by one order of magnitude whereas the singlet defects content increases up to ~2  ×  1019 spins/g. In addition, another triplet-type defect, which is characterized by the g = 4.00 ‘forbidden’ line, appears. On reduction of the particle size below the 20 nm limit, the ‘allowed’ EPR lines become practically unobservable, whereas the ‘forbidden’ lines remain as a reliable fingerprint of the presence of NV- centers in small ND systems. The same size reduction causes the disappearance of the characteristic hyperfine satellites in the

  13. Fluorescence behaviour and dural infiltration of meningioma analyzed by 5-ALA based fluorescence: Operating microscope versus mini-spectrometer.

    Science.gov (United States)

    Knipps, Johannes; Beseoglu, Kerim; Kamp, Marcel; Fischer, Igor; Felsberg, Joerg; Neumann, Lisa M; Steiger, Hans-Jakob; Cornelius, Jan F

    2017-08-30

    To compare fluorescence intensity of tumor specimens as measured by a FGS-microscope and a spectrometer, 2.) to evaluate tumor infiltration of dura mater around meningiomas with help of these two different 5-ALA based fluorescence tools and 3.) to correlate fluorescence intensity with histopathological data. In a clinical series menigiomas were resected by 5-ALA fluorescence guided surgery. Fluorescence intensity was semi-quantitatively rated by the surgeon at pre-defined points. Biopsies were harvested and fluorescence intensity measured by a spectrometer and histopathologically analysed. Sampling was realized at the level of the dura in a centrifugal direction. 104 biopsies (n= 13 tumors) were analysed. Specificity and sensitivity of the microscope were 0.96 and 0.53 and of the spectrometer 0.95 and 0.93, respectively. Fluorescence intensity as measured by the spectrometer was correlated to histologically confirmed tumor burden. In a centrifugal direction, tumor burden and fluorescence intensity continuously decreased (along the dural tail). Below a threshold value of 639 arbitrary units no tumor was histologically detectable. At the level of the dura the spectrometer was highly sensitive for detection of meningioma cells. The surgical microscope showed false negative results and missed residual tumor cells in more than half of the cases. The complementary use of both fluorescence tools may improve resection quality. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. Preparation of Micron-size Functional Fluorescent Microspheres

    Institute of Scientific and Technical Information of China (English)

    WANG Di-qiang; LIU Bai-ling; LI He; HU Jie

    2004-01-01

    As a kind of special functional microspheres, fluorescent polymer microspheres could be used in cell label and separation, blood flow assay, flow cytometer marking, chemical reaction assay,and in analyst of the transform and diffusion of particles in soil 1. However, one of the most important applications of fluorescent microspheres is in the high-throughput screening of drugs (HTS) 2. Through affinity interaction, radioactive ligands (latent drugs) are bound to fluorescent microspheres covered by receptor, and luminescence is produced by radioactivity, so ligands can be assayed and screened.In this study, we developed a technique for preparing micron-size fluorescent microspheres with different functional groups. The methods included the synthesis of micron-size polystyrene microspheres through the dispersion polymerization of styrene in different media such as ethanol,ethanol-water, and isopropanol; the functional polystyrene microspheres were prepared by introduction of functional monomers into the reaction system of styrene; the functional fluorescent microspheres were obtained by the way of dying functional microspheres in the fluorescent material's ethanol solvent.The average diameter of microspheres was in the range of 1~10 μm, and the distribution was normal distribution. The functional groups included -OH, -CHO, -COOH, -CONH2, and SO3H. The absorbing spectrum and exciting spectrum were tested, the results showed that the maximal absorbance of fluorescent microsphere was near 306.5 nm, and its maximal excitation was near 362 nm. The excitation spectrum of fluorescent material (DPO) and fluorescent microspheres were shown in figure 1, and it indicated that the developed fluorescent microspheres showed the same excitation behavior like DPO, which related to the fluorescent microspheres had stable luminescence property.

  15. Efficacy of fluorescence diagnosis for pleural tumors with alasens

    Directory of Open Access Journals (Sweden)

    O. V. Pikin

    2013-01-01

    Full Text Available The study of efficacy of thoracoscopy-assisted fluorescence diagnosis with Alasens is described in the article. The results of fluorescence diagnosis in 27 patients with suspicion on pleral tumor are represented. Before thoracoscopy-assisted fluorescence diagnosis in 21 patients according to radiological studies there was a fluid in pleural ca, in 19 patients of them tumor cells were found by cytological study of pleural fluid, in 10 patients differential diagnosis was performed between mesothelioma and adenogenic cancer. For fluorescence diagnosis fluorescence system by company Кarl Storz and xenon lamp with set of light filters was used: fluorescence study was performed by excitation at wavelength 380–460 nm. 3 h before investigation the patient received alasens per os in dose of 30 mg/kg body weight in 100 ml of water. For routine thoracoscopy tumor lesions were determined in 20 (87.0% patients, other 3 (13.0% patients had no tumors. In the group of patients with tumor lesions determined by routine thoracoscopy the fluorescence during fluorescence study was registered in all lesions determined in white light, besides this 24 additional foci of fluorescence were noticed, according to morphological study 21 of them had tumor nature, 3 lesions were inflammatory. In 1 of 3 patients with no lesion in white light there was one focus of fluorescence, morphological study proved the metastasis of adenocarcinoma in this area. According to morphological study of pleural biopsy specimens the true-positive results for fluorescence thoracoscopy accounted for 82, false-negative – 10, true-negative – 23, false-positive – 3. The sensitivity of the method was 89,1%, the specificity – 88,4%, the diagnostic accuracy – 88,9%. 

  16. Fluorescence self-quenching of tetraphenylporphyrin in liquid medium

    Energy Technology Data Exchange (ETDEWEB)

    Ghosh, Mihir [Integrated Science Education and Research Centre, Siksha-Bhavana, Visva-Bharati, Santiniketan 731235 (India); Nath, Sukhendu [Radiation and Photochemistry Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400085 (India); Hajra, Alakananda [Department of Chemistry, Siksha-Bhavana, Visva-Bharati, Santiniketan 731235 (India); Sinha, Subrata, E-mail: subratasinha67@rediffmail.com [Integrated Science Education and Research Centre, Siksha-Bhavana, Visva-Bharati, Santiniketan 731235 (India)

    2013-09-15

    Self-quenching of the fluorescence emission of tetraphenylporphyrin at high concentrations in toluene at the ambient temperature (300 K) is discussed in detail based on steady state and time-resolved fluorescence measurements. The fluorescence self-quenching is mainly attributed to re-absorption effect and the Förster type resonance energy transfer process (homotransfer). The re-absorption effect is found to deform the fluorescence emission spectra significantly in energy positions as well as relative intensities of different peaks at high concentrations. Nearly ideal fluorescence emission spectra are observed at a concentration ∼10{sup −7} mol/L. Moreover, there is an apparent enhancement of the fluorescence lifetime value of tetraphenylporphyrin in toluene at high concentrations, especially on the blue side of the fluorescence emission spectra. To the best knowledge of the authors, this is the first detail report on the fluorescence self-quenching of porphyrins in liquid medium. This finding carries great importance in view of the widespread research on porphyrins in the fields of solar light harvesting, artificial photosynthesis, photodynamic therapy, etc. -- Highlights: • The effect of concentration on the fluorescence emission spectra of tetraphenylporphyrin (TPhP) in toluene at 300 K is investigated by using steady state and time-resolved fluorescence techniques. • Both re-absorption effect and the Förster type resonance energy transfer are found to be responsible for the observed fluorescence self-quenching at high concentrations. • These investigations are extremely important in view of the extensive applications of porphyrins in the fabrication of molecular electronic devices, especially for solar and artificial photosynthetic devices, where highly concentrated porphyrins are often used for efficient light harvesting.

  17. DAPI fluorescence in nuclei isolated from tumors.

    Science.gov (United States)

    Krishan, Awtar; Dandekar, Payal D

    2005-08-01

    In DNA histograms of some human solid tumors stained with nuclear isolation medium--4,6-diamidino-2-phenylindole dihydrochloride (NIM-DAPI), the coefficient of variation (CV) of the G0/G1 peak was broad, and in nuclear volume vs DNA scattergrams, a prominent slope was seen. To determine the cause for this, nuclei from frozen breast tumors were stained with NIM-DAPI and analyzed after dilution or resuspension in PBS. In two-color (blue vs red) analysis, most of the slope and broad CV was due to red fluorescence of nuclei stained with NIM-DAPI, which was reduced on dilution or resuspension in PBS, resulting in elimination of the slope and tightening of the CV.

  18. Glucose Recognition in Vitro Using Fluorescent Spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Noronha, G; Heiss, A M; Reilly, J R; Vachon, Jr, D J; Cary, D R; Zaitseva, N P; Reibold, R A; Lane, S M; Peyser, T A; Satcher, J H

    2001-04-25

    Diabetes is a disease that affects over 16 million people in the USA at a cost of 100 billion dollars annually. The ability to regulate insulin delivery in people with Type 1 diabetes is imperative as is the need to manage glucose levels in all people with this disease. Our current method for monitoring glucose is a (FDA approved) minimally invasive enzymatic sensor that can measure glucose levels in vivo for three days. We are focused on developing a noninvasive implantable glucose sensor that will be interrogated by an external device. The material must be robust, easy to process, biocompatible and resistant to biofouling. In this Presentation we will discuss the development of a new polymeric matrix that can recognize physiological levels of glucose in vitro using fluorescent spectroscopy.

  19. Distance dependence of fluorescence resonance energy transfer

    Indian Academy of Sciences (India)

    R S Swathi; K L Sebastian

    2009-09-01

    Deviations from the usual -6 dependence of the rate of fluorescence resonance energy transfer (FRET) on the distance between the donor and the acceptor have been a common scenario in the recent times. In this paper, we present a critical analysis of the distance dependence of FRET, and try to illustrate the non--6 type behaviour of the rate for the case of transfer from a localized electronic excitation on the donor, a dye molecule to three different energy acceptors with delocalized electronic excitations namely, graphene, a two-dimensional semiconducting sheet and the case of such a semiconducting sheet rolled to obtain a nanotube. We use simple analytic models to understand the distance dependence in each case.

  20. Enhancing molecule fluorescence with asymmetrical plasmonic antennas.

    Science.gov (United States)

    Lu, Guowei; Liu, Jie; Zhang, Tianyue; Shen, Hongming; Perriat, Pascal; Martini, Matteo; Tillement, Olivier; Gu, Ying; He, Yingbo; Wang, Yuwei; Gong, Qihuang

    2013-07-21

    We propose and justify by the finite-difference time-domain method an efficient strategy to enhance the spontaneous emission of a fluorophore with a multi-resonance plasmonic antenna. The custom-designed asymmetrical antenna consists of two plasmonic nanoparticles with different sizes and is able to couple efficiently to free space light through multiple localized surface plasmon resonances. This design simultaneously permits a large near-field excitation near the antenna as well as a high quantum efficiency, which results in an unusual and significant enhancement of the fluorescence of a single emitter. Such an asymmetrical antenna presents intrinsic advantages over single particle or dimer based antennas made using two identical nanostructures. This promising concept can be exploited in the large domain of light-matter interaction processes involving multiple frequencies.