Palladium Nanoparticles-Based Fluorescence Resonance Energy Transfer Aptasensor for Highly Sensitive Detection of Aflatoxin M₁ in Milk.

Science.gov (United States)

Li, Hui; Yang, Daibin; Li, Peiwu; Zhang, Qi; Zhang, Wen; Ding, Xiaoxia; Mao, Jin; Wu, Jing

2017-10-13

A highly sensitive aptasensor for aflatoxin M₁ (AFM₁) detection was constructed based on fluorescence resonance energy transfer (FRET) between 5-carboxyfluorescein (FAM) and palladium nanoparticles (PdNPs). PdNPs (33 nm) were synthesized through a seed-mediated growth method and exhibited broad and strong absorption in the whole ultraviolet-visible (UV-Vis) range. The strong coordination interaction between nitrogen functional groups of the AFM₁ aptamer and PdNPs brought FAM and PdNPs in close proximity, which resulted in the fluorescence quenching of FAM to a maximum extent of 95%. The non-specific fluorescence quenching caused by PdNPs towards fluorescein was negligible. After the introduction of AFM₁ into the FAM-AFM₁ aptamer-PdNPs FRET system, the AFM₁ aptamer preferentially combined with AFM₁ accompanied by conformational change, which greatly weakened the coordination interaction between the AFM₁ aptamer and PdNPs. Thus, fluorescence recovery of FAM was observed and a linear relationship between the fluorescence recovery and the concentration of AFM₁ was obtained in the range of 5-150 pg/mL in aqueous buffer with the detection limit of 1.5 pg/mL. AFM₁ detection was also realized in milk samples with a linear detection range from 6 pg/mL to 150 pg/mL. The highly sensitive FRET aptasensor with simple configuration shows promising prospect in detecting a variety of food contaminants.

Highly Sensitive Ratiometric Fluorescent Sensor for Trinitrotoluene Based on the Inner Filter Effect between Gold Nanoparticles and Fluorescent Nanoparticles.

Science.gov (United States)

Lu, Hongzhi; Quan, Shuai; Xu, Shoufang

2017-11-08

In this work, we developed a simple and sensitive ratiometric fluorescent assay for sensing trinitrotoluene (TNT) based on the inner filter effect (IFE) between gold nanoparticles (AuNPs) and ratiometric fluorescent nanoparticles (RFNs), which was designed by hybridizing green emissive carbon dots (CDs) and red emissive quantum dots (QDs) into a silica sphere as a fluorophore pair. AuNPs in their dispersion state can be a powerful absorber to quench CDs, while the aggregated AuNPs can quench QDs in the IFE-based fluorescent assays as a result of complementary overlap between the absorption spectrum of AuNPs and emission spectrum of RFNs. As a result of the fact that TNT can induce the aggregation of AuNPs, with the addition of TNT, the fluorescent of QDs can be quenched, while the fluorescent of CDs would be recovered. Then, ratiometric fluorescent detection of TNT is feasible. The present IFE-based ratiometric fluorescent sensor can detect TNT ranging from 0.1 to 270 nM, with a detection limit of 0.029 nM. In addition, the developed method was successfully applied to investigate TNT in water and soil samples with satisfactory recoveries ranging from 95 to 103%, with precision below 4.5%. The simple sensing approach proposed here could improve the sensitivity of colorimetric analysis by changing the ultraviolet analysis to ratiometric fluorescent analysis and promote the development of a dual-mode detection system.

Screen-printed fluorescent sensors for rapid and sensitive anthrax biomarker detection

International Nuclear Information System (INIS)

Lee, Inkyu; Oh, Wan-Kyu; Jang, Jyongsik

2013-01-01

Highlights: •We fabricated flexible anthrax sensors with a simple screen-printing method. •The sensors selectively detected B. anthracis biomarker. •The sensors provide the visible alarm against anthrax attack. -- Abstract: Since the 2001 anthrax attacks, efforts have focused on the development of an anthrax detector with rapid response and high selectivity and sensitivity. Here, we demonstrate a fluorescence sensor for detecting anthrax biomarker with high sensitivity and selectivity using a screen-printing method. A lanthanide–ethylenediamine tetraacetic acid complex was printed on a flexible polyethersulfone film. Screen-printing deposition of fluorescent detecting moieties produced fluorescent patterns that acted as a visual alarm against anthrax

High-Resolution Ultrasound-Switchable Fluorescence Imaging in Centimeter-Deep Tissue Phantoms with High Signal-To-Noise Ratio and High Sensitivity via Novel Contrast Agents.

Science.gov (United States)

Cheng, Bingbing; Bandi, Venugopal; Wei, Ming-Yuan; Pei, Yanbo; D'Souza, Francis; Nguyen, Kytai T; Hong, Yi; Yuan, Baohong

2016-01-01

For many years, investigators have sought after high-resolution fluorescence imaging in centimeter-deep tissue because many interesting in vivo phenomena-such as the presence of immune system cells, tumor angiogenesis, and metastasis-may be located deep in tissue. Previously, we developed a new imaging technique to achieve high spatial resolution in sub-centimeter deep tissue phantoms named continuous-wave ultrasound-switchable fluorescence (CW-USF). The principle is to use a focused ultrasound wave to externally and locally switch on and off the fluorophore emission from a small volume (close to ultrasound focal volume). By making improvements in three aspects of this technique: excellent near-infrared USF contrast agents, a sensitive frequency-domain USF imaging system, and an effective signal processing algorithm, for the first time this study has achieved high spatial resolution (~ 900 μm) in 3-centimeter-deep tissue phantoms with high signal-to-noise ratio (SNR) and high sensitivity (3.4 picomoles of fluorophore in a volume of 68 nanoliters can be detected). We have achieved these results in both tissue-mimic phantoms and porcine muscle tissues. We have also demonstrated multi-color USF to image and distinguish two fluorophores with different wavelengths, which might be very useful for simultaneously imaging of multiple targets and observing their interactions in the future. This work has opened the door for future studies of high-resolution centimeter-deep tissue fluorescence imaging.

Enhancement of uranyl fluorescence using trimesic acid: Ligand sensitization and co-fluorescence

Energy Technology Data Exchange (ETDEWEB)

Maji, S. [Chemistry Group, Materials Chemistry Division, Indira Gandhi Centre for Atomic Research, Kalpakkam 603102 (India); Viswanathan, K.S., E-mail: vish@igcar.gov.in [Chemistry Group, Materials Chemistry Division, Indira Gandhi Centre for Atomic Research, Kalpakkam 603102 (India)

2011-09-15

Trimesic acid (TMA) was shown to sensitize and enhance uranyl fluorescence in aqueous medium, with the enhancement being a maximum at pH 5.0. Fluorescence spectra and lifetime data together suggest that TMA complexes with uranyl (UO{sub 2}{sup 2+}). The fluorescence of UO{sub 2}{sup 2+} in its acid complex is further enhanced by more than two orders of magnitude following the addition of Y{sup 3+}; a process referred to as co-fluorescence, leading to the possibility of detecting uranium at sub ng/mL level. The present study demonstrates, for the first time, fluorescence enhancement of the uranyl species due to co-fluorescence. - Highlights: > Trimesic acid was shown to sensitize and enhance the fluorescence of uranium in aqueous medium. > This ligand also exhibited co-fluorescence of uranium with Y{sup 3+}. > To the best of our knowledge this is the first report of co-fluorescence in uranium. > The enhancement of uranium fluorescence, resulted in detection limits in the ng/mL regime.

An extremely sensitive monoboronic acid based fluorescent sensor for glucose

International Nuclear Information System (INIS)

Sun Xiangying; Liu Bin; Jiang Yunbao

2004-01-01

An extremely sensitive monoboronic acid based fluorescent sensor for glucose was developed. This was carried out by assembling a fluorescent monoboronic acid, 3-aminophenylboronic acid (PBA) indirectly onto gold surface via its electrostatic interaction with cysteine (Cys) that was directly assembled on the gold surface. The formation of self-assembled bilayers (SAB) was confirmed and primarily characterized by cyclic voltammetry and X-ray photoelectron spectra (XPS). The SAB containing PBA was found fluorescent and its fluorescence showed an extremely high sensitivity to the presence of glucose and other monosaccharides such as galactose and fructose with quenching constants at 10 8 M -1 order of magnitude compared to those at 10 2 M -1 in bulk solutions. The quenching constants were found to vary in the order of D-glucose>D-galactose>D-fructose>D-mannose that is different from that in bulk solution which shows the highest binding affinity toward D-fructose and very low sensitivity toward glucose. The reported monoboronic acid based SAB fluorescent sensor showed the highest sensitivity towards glucose with the capacity of detecting saccharides of concentration down to nanomolar level. It was also demonstrated that the fluorescence from PBA/Cys/Au can be easily recovered after each measurement event and therefore also represents a new reusable method for immobilizing reagent in fabricating chemosensors

Graphitic Carbon Nitride Nanosheets-Based Ratiometric Fluorescent Probe for Highly Sensitive Detection of H2O2 and Glucose.

Science.gov (United States)

Liu, Jin-Wen; Luo, Ying; Wang, Yu-Min; Duan, Lu-Ying; Jiang, Jian-Hui; Yu, Ru-Qin

2016-12-14

Graphitic carbon nitride (g-C 3 N 4 ) nanosheets, an emerging graphene-like carbon-based nanomaterial with high fluorescence and large specific surface areas, hold great potential for biosensor applications. Current g-C 3 N 4 nanosheets based fluorescent biosensors majorly rely on single fluorescent intensity reading through fluorescence quenching interactions between the nanosheets and metal ions. Here we report for the first time the development of a novel g-C 3 N 4 nanosheets-based ratiometric fluorescence sensing strategy for highly sensitive detection of H 2 O 2 and glucose. With o-phenylenediamine (OPD) oxidized by H 2 O 2 in the presence of horseradish peroxidase (HRP), the oxidization product can assemble on the g-C 3 N 4 nanosheets through hydrogen bonding and π-π stacking, which effectively quenches the fluorescence of g-C 3 N 4 while delivering a new emission peak. The ratiometric signal variations enable robust and sensitive detection of H 2 O 2 . On the basis of the glucose converting into H 2 O 2 through the catalysis of glucose oxidase, the g-C 3 N 4 -based ratiometric fluorescence sensing platform is also exploited for glucose assay. The developed strategy is demonstrated to give a detection limit of 50 nM for H 2 O 2 and 0.4 μM for glucose, at the same time, it has been successfully used for glucose levels detection in human serum. This strategy may provide a cost-efficient, robust, and high-throughput platform for detecting various species involving H 2 O 2 -generation reactions for biomedical applications.

Fluorescent probes for "off-on" highly sensitive detection of Hg²⁺ and L-cysteine based on nitrogen-doped carbon dots.

Science.gov (United States)

Zhang, Yi; Cui, Peipei; Zhang, Feng; Feng, Xiaoting; Wang, Yaling; Yang, Yongzhen; Liu, Xuguang

2016-05-15

Fluorescent nitrogen-doped carbon dots (NCDs) were synthesized by a facile, and low-cost one-step hydrothermal strategy using citric acid as carbon source and ammonia solution as nitrogen source for the first time. The obtained NCDs show stable blue fluorescence with a high quantum yield of 35.4%, along with the fluorescence lifetime of ca. 6.75 ns. Most importantly, Hg(2+) can completely quench the fluorescence of NCDs as a result of the formation of a non-fluorescent stable NCDs-Hg(2+) complex. Static fluorescence quenching towards Hg(2+) is proved by the Stern-Volmer equation, ultraviolet-visible absorption spectra, temperature dependent quenching and fluorescence lifetime measurements. Subsequently, the fluorescence of the NCDs-Hg(2+) system is completely recovered with the addition L-cysteine (L-Cys) owing to the dissociation of NCDs-Hg(2+) complex to form a more stable Hg(2+)-L-Cys complex by Hg(2+)-S bonding. Therefore, such NCDs can be used as an effective fluorescent "turn-off" probe for rapid, rather highly selective and sensitive detection of Hg(2+), with a limit of detection (LOD) as low as 1.48 nM and a linear detection range of 0-10 μM. Interestingly, NCDs-Hg(2+) system can be conveniently employed as a fluorescent "turn-on" sensor for highly selective and sensitive detection of L-Cys with a low LOD of 0.79 nM and a wide linear detection range of 0-50 μM. Further, the sensitivity of NCDs to Hg(2+) is preserved in tap water with a LOD of 1.65 nM and a linear detection range of 0-10 μM. Copyright © 2016 Elsevier B.V. All rights reserved.

Aminoquinoline based highly sensitive fluorescent sensor for lead(II) and aluminum(III) and its application in live cell imaging

International Nuclear Information System (INIS)

Anand, Thangaraj; Sivaraman, Gandhi; Mahesh, Ayyavu; Chellappa, Duraisamy

2015-01-01

Highlights: • Aminoquinoline derivative was synthesized and used to recognize Pb 2+ /Al 3+ . • ANQ was high sensitive, selective and turn-on sensor for Pb 2+ /Al 3+ . • The Pb 2+ detection limit (2.08 × 10 −9 mol L −1 ) is reported. • This fluorescence change was further supported by DFT/TD-DFT calculations. • The probe is applied successfully for recognizing intracellular Pb 2+ /Al 3+ within living cells. - Abstract: We have synthesized a new probe 5-((anthracen-9-ylmethylene) amino)quinolin-10-ol (ANQ) based on anthracene platform. The probe was tested for its sensing behavior toward heavy metal ions Hg 2+ , Pb 2+ , light metal Al 3+ ion, alkali, alkaline earth, and transition metal ions by UV–visible and fluorescent techniques in ACN/H 2 O mixture buffered with HEPES (pH 7.4). It shows high selectivity toward sensing Pb 2+ /Al 3+ metal ions. Importantly, 10-fold and 5- fold fluorescence enhancement at 429 nm was observed for probe upon complexation with Pb 2+ and Al 3+ ions, respectively. This fluorescence enhancement is attributable to the prevention of photoinduced electron transfer. The photonic studies indicate that the probe can be adopted as a sensitive fluorescent chemosensor for Pb 2+ and Al 3+ ions

Improved Diffuse Fluorescence Flow Cytometer Prototype for High Sensitivity Detection of Rare Circulating Cells In Vivo

Science.gov (United States)

Pestana, Noah Benjamin

Accurate quantification of circulating cell populations is important in many areas of pre-clinical and clinical biomedical research, for example, in the study of cancer metastasis or the immune response following tissue and organ transplants. Normally this is done "ex-vivo" by drawing and purifying a small volume of blood and then analyzing it with flow cytometry, hemocytometry or microfludic devices, but the sensitivity of these techniques are poor and the process of handling samples has been shown to affect cell viability and behavior. More recently "in vivo flow cytometry" (IVFC) techniques have been developed where fluorescently-labeled cells flowing in a small blood vessel in the ear or retina are analyzed, but the sensitivity is generally poor due to the small sampling volume. To address this, our group recently developed a method known as "Diffuse Fluorescence Flow Cytometry" (DFFC) that allows detection and counting of rare circulating cells with diffuse photons, offering extremely high single cell counting sensitivity. In this thesis, an improved DFFC prototype was designed and validated. The chief improvements were three-fold, i) improved optical collection efficiency, ii) improved detection electronics, and iii) development of a method to mitigate motion artifacts during in vivo measurements. In combination, these improvements yielded an overall instrument detection sensitivity better than 1 cell/mL in vivo, which is the most sensitive IVFC system reported to date. Second, development and validation of a low-cost microfluidic device reader for analysis of ocular fluids is described. We demonstrate that this device has equivalent or better sensitivity and accuracy compared a fluorescence microscope, but at an order-of-magnitude reduced cost with simplified operation. Future improvements to both instruments are also discussed.

Fluorescent Gold Nanoprobes for the Sensitive and Selective Detection for Hg2+

Directory of Open Access Journals (Sweden)

Chai Fang

2010-01-01

Full Text Available Abstract A simple, cost-effective yet rapid and sensitive sensor for on-site and real-time Hg2+ detection based on bovine serum albumin functionalized fluorescent gold nanoparticles as novel and environmentally friendly fluorescent probes was developed. Using this probe, aqueous Hg2+ can be detected at 0.1 nM in a facile way based on fluorescence quenching. This probe was also applied to determine the Hg2+ in the lake samples, and the results demonstrate low interference and high sensitivity.

Ratiometric fluorescent pH-sensitive polymers for high-throughput monitoring of extracellular pH†

OpenAIRE

Zhang, Liqiang; Su, Fengyu; Kong, Xiangxing; Lee, Fred; Day, Kevin; Gao, Weimin; Vecera, Mary E.; Sohr, Jeremy M.; Buizer, Sean; Tian, Yanqing; Meldrum, Deirdre R

2016-01-01

Extracellular pH has a strong effect on cell metabolism and growth. Precisely detecting extracellular pH with high throughput is critical for cell metabolism research and fermentation applications. In this research, a series of ratiometric fluorescent pH sensitive polymers are developed and the ps-pH-neutral is characterized as the best one for exculsive detection of extracellular pH. Poly(N-(2-hydroxypropyl)methacrylamide) (PHPMA) is used as the host polymer to increase the water solubility ...

Fluorescent metal-organic framework MIL-53(Al) for highly selective and sensitive detection of Fe3+ in aqueous solution.

Science.gov (United States)

Yang, Cheng-Xiong; Ren, Hu-Bo; Yan, Xiu-Ping

2013-08-06

Fluorescent metal-organic frameworks (MOFs) have received great attention in sensing application. Here, we report the exploration of fluorescent MIL-53(Al) for highly selective and sensitive detection of Fe(3+) in aqueous solution. The cation exchange between Fe(3+) and the framework metal ion Al(3+) in MIL-53(Al) led to the quenching of the fluorescence of MIL-53(Al) due to the transformation of strong-fluorescent MIL-53(Al) to weak-fluorescent MIL-53(Fe), allowing highly selective and sensitive detection of Fe(3+) in aqueous solution with a linear range of 3-200 μM and a detection limit of 0.9 μM. No interferences from 0.8 M Na(+); 0.35 M K(+); 11 mM Cu(2+); 10 mM Ni(2+); 6 mM Ca(2+), Pb(2+), and Al(3+); 5.5 mM Mn(2+); 5 mM Co(2+) and Cr(3+); 4 mM Hg(2+), Cd(2+), Zn(2+), and Mg(2+); 3 mM Fe(2+); 0.8 M Cl(-); 60 mM NO2(-) and NO3(-); 10 mM HPO4(2-), H2PO4(-), SO3(2-), SO4(2-), and HCOO(-); 8 mM CO3(2-), HCO3(-), and C2O4(2-); and 5 mM CH3COO(-) were found for the detection of 150 μM Fe(3+). The possible mechanism for the quenching effect of Fe(3+) on the fluorescence of MIL-53(Al) was elucidated by inductively coupled plasma-mass spectrometry, X-ray diffraction spectrometry, and Fourier transform infrared spectrometry. The specific cation exchange behavior between Fe(3+) and the framework Al(3+) along with the excellent stability of MIL-53(Al) allows highly selective and sensitive detection of Fe(3+) in aqueous solution. The developed method was applied to the determination of Fe(3+) in human urine samples with the quantitative spike recoveries from 98.2% to 106.2%.

A highly sensitive, single selective, fluorescent sensor for Al3+ detection and its application in living cell imaging

International Nuclear Information System (INIS)

Ye, Xing-Pei; Sun, Shao-bo; Li, Ying-dong; Zhi, Li-hua; Wu, Wei-na; Wang, Yuan

2014-01-01

A new o-aminophenol-based fluorogenic chemosensor methyl 3,5-bis((E)-(2-hydroxyphenylimino)methyl)-4-hydroxybenzoate 1 have been synthesized by Schiff base condensation of methyl 3,5-diformyl-4-hydroxybenzoate with o-aminophenol, which exhibits high selectivity and sensitivity toward Al 3+ . Fluorescence titration studies of receptors 1 with different metal cations in CH 3 OH medium showed highly selective and sensitive towards Al 3+ ions even in the presence of other commonly coexisting metal ions. The detection limit of Al 3+ ions is at the parts per billion level. Interestingly, the Al(III) complex of 1 offered a large Stokes shift (>120 nm), which can miximize the selfquenching effect. In addition, possible utilization of this receptor as bio-imaging fluorescent probe to detect Al 3+ in human cervical HeLa cancer cell lines was also investigated by confocal fluorescence microscopy. - Highlights: • A new Schiff base chemosensor is reported. • The sensor for Al 3+ offers large Stokes shift. • The detection limit of Al 3+ in CH 3 OH solution is at the parts per billion level. • The utilization of sensor for the monitoring of Al 3+ levels in living cells was examined

Ultra-sensitive high performance liquid chromatography-laser-induced fluorescence based proteomics for clinical applications.

Science.gov (United States)

Patil, Ajeetkumar; Bhat, Sujatha; Pai, Keerthilatha M; Rai, Lavanya; Kartha, V B; Chidangil, Santhosh

2015-09-08

An ultra-sensitive high performance liquid chromatography-laser induced fluorescence (HPLC-LIF) based technique has been developed by our group at Manipal, for screening, early detection, and staging for various cancers, using protein profiling of clinical samples like, body fluids, cellular specimens, and biopsy-tissue. More than 300 protein profiles of different clinical samples (serum, saliva, cellular samples and tissue homogenates) from volunteers (normal, and different pre-malignant/malignant conditions) were recorded using this set-up. The protein profiles were analyzed using principal component analysis (PCA) to achieve objective detection and classification of malignant, premalignant and healthy conditions with high sensitivity and specificity. The HPLC-LIF protein profiling combined with PCA, as a routine method for screening, diagnosis, and staging of cervical cancer and oral cancer, is discussed in this paper. In recent years, proteomics techniques have advanced tremendously in life sciences and medical sciences for the detection and identification of proteins in body fluids, tissue homogenates and cellular samples to understand biochemical mechanisms leading to different diseases. Some of the methods include techniques like high performance liquid chromatography, 2D-gel electrophoresis, MALDI-TOF-MS, SELDI-TOF-MS, CE-MS and LC-MS techniques. We have developed an ultra-sensitive high performance liquid chromatography-laser induced fluorescence (HPLC-LIF) based technique, for screening, early detection, and staging for various cancers, using protein profiling of clinical samples like, body fluids, cellular specimens, and biopsy-tissue. More than 300 protein profiles of different clinical samples (serum, saliva, cellular samples and tissue homogenates) from healthy and volunteers with different malignant conditions were recorded by using this set-up. The protein profile data were analyzed using principal component analysis (PCA) for objective

Aminoquinoline based highly sensitive fluorescent sensor for lead(II) and aluminum(III) and its application in live cell imaging

Energy Technology Data Exchange (ETDEWEB)

Anand, Thangaraj; Sivaraman, Gandhi [School of Chemistry, Madurai Kamaraj University, Madurai 625021 (India); Mahesh, Ayyavu, E-mail: mahesh.a06@gmail.com [School of Biological Sciences, Madurai Kamaraj University, Madurai 625021 (India); Chellappa, Duraisamy, E-mail: dcmku123@gmail.com [School of Chemistry, Madurai Kamaraj University, Madurai 625021 (India)

2015-01-01

Highlights: • Aminoquinoline derivative was synthesized and used to recognize Pb{sup 2+}/Al{sup 3+}. • ANQ was high sensitive, selective and turn-on sensor for Pb{sup 2+}/Al{sup 3+}. • The Pb{sup 2+} detection limit (2.08 × 10{sup −9} mol L{sup −1}) is reported. • This fluorescence change was further supported by DFT/TD-DFT calculations. • The probe is applied successfully for recognizing intracellular Pb{sup 2+}/Al{sup 3+} within living cells. - Abstract: We have synthesized a new probe 5-((anthracen-9-ylmethylene) amino)quinolin-10-ol (ANQ) based on anthracene platform. The probe was tested for its sensing behavior toward heavy metal ions Hg{sup 2+}, Pb{sup 2+}, light metal Al{sup 3+} ion, alkali, alkaline earth, and transition metal ions by UV–visible and fluorescent techniques in ACN/H{sub 2}O mixture buffered with HEPES (pH 7.4). It shows high selectivity toward sensing Pb{sup 2+}/Al{sup 3+} metal ions. Importantly, 10-fold and 5- fold fluorescence enhancement at 429 nm was observed for probe upon complexation with Pb{sup 2+} and Al{sup 3+} ions, respectively. This fluorescence enhancement is attributable to the prevention of photoinduced electron transfer. The photonic studies indicate that the probe can be adopted as a sensitive fluorescent chemosensor for Pb{sup 2+} and Al{sup 3+} ions.

Synthesis of molecularly imprinted dye-silica nanocomposites with high selectivity and sensitivity: Fluorescent imprinted sensor for rapid and efficient detection of τ-fluvalinate in vodka.

Science.gov (United States)

Wang, Yunyun; Wang, Jixiang; Cheng, Rujia; Sun, Lin; Dai, Xiaohui; Yan, Yongsheng

2018-04-01

An imprinted fluorescent sensor was fabricated based on SiO 2 nanoparticles encapsulated with a molecularly imprinted polymer containing allyl fluorescein. High fluorine cypermethirin as template molecules, methyl methacrylate as functional monomer, and allyl fluorescein as optical materials synthesized a core-shell fluorescent molecular imprinted sensor, which showed a high and rapid sensitivity and selectivity for the detection of τ-fluvalinate. The sensor presented appreciable sensitivity with a limit of 13.251 nM, rapid detection that reached to equilibrium within 3 min, great linear relationship in the relevant concentration range from 0 to 150 nM, and excellent selectivity over structural analogues. In addition, the fluorescent sensor demonstrated desirable regeneration ability (eight cycling operations). The molecularly imprinted polymers ensured specificity, while the fluorescent dyes provided the stabile sensitivity. Finally, an effective application of the sensor was implemented by the detection of τ-fluvalinate in real samples from vodka. The molecularly imprinted fluorescent sensor showed a promising potential in environmental monitoring and food safety. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Environmentally Sensitive Fluorescent Sensors Based on Synthetic Peptides

Directory of Open Access Journals (Sweden)

Laurence Choulier

2010-03-01

Full Text Available Biosensors allow the direct detection of molecular analytes, by associating a biological receptor with a transducer able to convert the analyte-receptor recognition event into a measurable signal. We review recent work aimed at developing synthetic fluorescent molecular sensors for a variety of analytes, based on peptidic receptors labeled with environmentally sensitive fluorophores. Fluorescent indicators based on synthetic peptides are highly interesting alternatives to protein-based sensors, since they can be synthesized chemically, are stable, and can be easily modified in a site-specific manner for fluorophore coupling and for immobilization on solid supports.

Sensitive and selective tumor imaging with novel and highly activatable fluorescence probes

International Nuclear Information System (INIS)

Urano, Yasuteru

2008-01-01

Selective and sensitive tumor imaging in vivo is one of the most requested methodologies in medical sciences. Although several imaging modalities have been developed including positron emission tomography (PET) and magnetic resonance (MR) imaging for the detection of tumors, none of these modalities can activate the signals upon being accumulated or uptaken to tumor sites. Among these modalities, only optical fluorescence imaging has a marked advantage, that is, their signals can be dramatically increased upon detecting some biological features. In this short review, I will introduce some recent strategies for activatable optical fluorescence imaging of tumors, and discuss their advantages over other modalities. (author)

Design and synthesis of new fluorescent probe for rapid and highly sensitive detection of proteins via electrophoretic gel stain.

Science.gov (United States)

Suzuki, Yoshio; Takagi, Nobuyuki; Chimuro, Tomoyuki; Shinohara, Atsushi; Sakaguchi, Nao; Hiratsuka, Atsunori; Yokoyama, Kenji

2011-06-01

A new fluorescent molecular probe, 2,2'-(1E,1'E)-2,2'-(4-(dicyanomethylene)-4H-pyrane-2,6-diyl)bis(ethene-2,1-diyl)bis(sodium benzenesulfonate) salt (1), possessing the cyanopyranyl moieties and two benzene sulfonic acid groups was designed and synthesized to detect proteins in solution and for high-throughput SDS-PAGE. Compound 1 exhibited no fluorescence in the absence of proteins; however, it exhibited strong fluorescence on the addition of bovine serum albumin as a result of intramolecular charge transfer. Compared with the conventional protocols for in-gel protein staining, such as SYPRO Ruby and silver staining, 1 achieves higher sensitivity, even though it offers a simplified, higher throughput protocol. In fact, the total time required for protein staining was 60-90 min under optimum conditions much shorter than that required by the less-sensitive silver staining or SYPRO Ruby staining protocols. Moreover, 1 was successfully applied to protein identification by mass spectrometry via in-gel tryptic digestion, Western blotting, and native PAGE together with protein staining by 1, which is a modified protocol of blue native PAGE (BN-PAGE). Thus, 1 may facilitate high-sensitivity protein detection, and it may be widely applicable as a convenient tool in various scientific and medical fields. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A label-free fluorescence biosensor for highly sensitive detection of lectin based on carboxymethyl chitosan-quantum dots and gold nanoparticles

International Nuclear Information System (INIS)

Liu, Ziping; Liu, Hua; Wang, Lei; Su, Xingguang

2016-01-01

In this work, we report a novel label-free fluorescence “turn off-on” biosensor for lectin detection. The highly sensitive and selective sensing system is based on the integration of carboxymethyl chitosan (CM-CHIT), CuInS_2 quantum dots (QDs) and Au nanoparticles (NPs). Firstly, CuInS_2 QDs featuring carboxyl groups were directly synthesized via a hydrothermal synthesis method. Then, the carboxyl groups on the CuInS_2 QDs surface were interacted with the amino groups (−NH_2), carboxyl groups (−COOH) and hydroxyl groups (−OH) within CM-CHIT polymeric chains via electrostatic interactions and hydrogen bonding to form CM-CHIT-QDs assemblies. Introduction of Au NPs could quench the fluorescence of CM-CHIT-QDs through electron and energy transfer. In the presence of lectin, lectin could bind exclusively with CM-CHIT-QDs by means of specific multivalent carbohydrate-protein interaction. Thus, the electron and energy transfer process between CM-CHIT-QDs and Au NPs was inhibited, and as a result, the fluorescence of CM-CHIT-QDs was effectively “turned on”. Under the optimum conditions, there was a good linear relationship between the fluorescence intensity ratio I/I_0 (I and I_0 were the fluorescence intensity of CM-CHIT-QDs-Au NPs in the presence and absence of lectin, respectively) and lectin concentration in the range of 0.2–192.5 nmol L"−"1, And the detection limit could be down to 0.08 nmol L"−"1. Furthermore, the proposed biosensor was employed for the determination of lectin in fetal bovine serum samples with satisfactory results. - Graphical abstract: A label-free fluorescence biosensor for highly sensitive detection of lectin based on the integration of carboxymethyl chitosan, CuInS_2 quantum dots and gold nanoparticles. - Highlights: • A label-free near-infrared fluorescence “turn off-on” biosensor for detection of lectin was established. • The highly sensitive biosensor was based on the inner filter effect of Au NPs on CM

A label-free fluorescence biosensor for highly sensitive detection of lectin based on carboxymethyl chitosan-quantum dots and gold nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Liu, Ziping; Liu, Hua; Wang, Lei; Su, Xingguang, E-mail: suxg@jlu.edu.cn

2016-08-17

In this work, we report a novel label-free fluorescence “turn off-on” biosensor for lectin detection. The highly sensitive and selective sensing system is based on the integration of carboxymethyl chitosan (CM-CHIT), CuInS{sub 2} quantum dots (QDs) and Au nanoparticles (NPs). Firstly, CuInS{sub 2} QDs featuring carboxyl groups were directly synthesized via a hydrothermal synthesis method. Then, the carboxyl groups on the CuInS{sub 2} QDs surface were interacted with the amino groups (−NH{sub 2}), carboxyl groups (−COOH) and hydroxyl groups (−OH) within CM-CHIT polymeric chains via electrostatic interactions and hydrogen bonding to form CM-CHIT-QDs assemblies. Introduction of Au NPs could quench the fluorescence of CM-CHIT-QDs through electron and energy transfer. In the presence of lectin, lectin could bind exclusively with CM-CHIT-QDs by means of specific multivalent carbohydrate-protein interaction. Thus, the electron and energy transfer process between CM-CHIT-QDs and Au NPs was inhibited, and as a result, the fluorescence of CM-CHIT-QDs was effectively “turned on”. Under the optimum conditions, there was a good linear relationship between the fluorescence intensity ratio I/I{sub 0} (I and I{sub 0} were the fluorescence intensity of CM-CHIT-QDs-Au NPs in the presence and absence of lectin, respectively) and lectin concentration in the range of 0.2–192.5 nmol L{sup −1}, And the detection limit could be down to 0.08 nmol L{sup −1}. Furthermore, the proposed biosensor was employed for the determination of lectin in fetal bovine serum samples with satisfactory results. - Graphical abstract: A label-free fluorescence biosensor for highly sensitive detection of lectin based on the integration of carboxymethyl chitosan, CuInS{sub 2} quantum dots and gold nanoparticles. - Highlights: • A label-free near-infrared fluorescence “turn off-on” biosensor for detection of lectin was established. • The highly sensitive biosensor was based on the

Highly Sensitive Fluorescent Sensor for Cartap Based on Fluorescence Resonance Energy Transfer Between Gold Nanoparticles and Rhodamine B.

Science.gov (United States)

Dong, Liang; Hou, Changjun; Fa, Huanbao; Yang, Mei; Wu, Huixiang; Zhang, Liang; Huo, Danqun

2018-04-01

Cartap residue poses a great threat to human health and its derivatives would remain in soils, natural waters and other environmental domains for a long time. Herein, a simple, rapid and ultrasensitive analytical method for the determination of cartap based on fluorescence resonance energy transfer (FRET) between Au nanoparticles (AuNPs) and rhodamine B (RB) is first described. With the presence of citrate-stabilized AuNPs, the fluorescence of RB was remarkably quenched by AuNPs via FRET. The fluorescence of the AuNPs-RB system was recovered upon addition of cartap, cartap can be adsorbed on the surface of AuNPs due to its amino group that has good affinity with gold, which could induce the aggregation of AuNPs accompanying color change from red to blue. Thus, the FRET between AuNPs and RB was weakened and the PL intensity of RB was recovered accordingly. A good linear correlation for detection of RB was exhibited from 1 nM to 180 nM, and the detection limit reached 0.88 nM, which was much lower than the safety limit required by USA, UK and China. To the best of our knowledge, it has been the lowest detection ever without the aid of costly instrumentation. This method was successfully carried out for the assessment of cartap in real samples with satisfactory results, which revealed many advantages such as high sensitivity, low cost and non-time-consuming compared with traditional methods.

High-quality substrate for fluorescence enhancement using agarose-coated silica opal film.

Science.gov (United States)

Xu, Ming; Li, Juan; Sun, Liguo; Zhao, Yuanjin; Xie, Zhuoying; Lv, Linli; Zhao, Xiangwei; Xiao, Pengfeng; Hu, Jing; Lv, Mei; Gu, Zhongze

2010-08-01

To improve the sensitivity of fluorescence detection in biochip, a new kind of substrates was developed by agarose coating on silica opal film. In this study, silica opal film was fabricated on glass substrate using the vertical deposition technique. It can provide stronger fluorescence signals and thus improve the detection sensitivity. After coating with agarose, the hybrid film could provide a 3D support for immobilizing sample. Comparing with agarose-coated glass substrate, the agarose-coated opal substrates could selectively enhance particular fluorescence signals with high sensitivity when the stop band of the silica opal film in the agarose-coated opal substrate overlapped the fluorescence emission wavelength. A DNA hybridization experiment demonstrated that fluorescence intensity of special type of agarose-coated opal substrates was about four times that of agarose-coated glass substrate. These results indicate that the optimized agarose-coated opal substrate can be used for improving the sensitivity of fluorescence detection with high quality and selectivity.

The enhanced cyan fluorescent protein: a sensitive pH sensor for fluorescence lifetime imaging.

Science.gov (United States)

Poëa-Guyon, Sandrine; Pasquier, Hélène; Mérola, Fabienne; Morel, Nicolas; Erard, Marie

2013-05-01

pH is an important parameter that affects many functions of live cells, from protein structure or function to several crucial steps of their metabolism. Genetically encoded pH sensors based on pH-sensitive fluorescent proteins have been developed and used to monitor the pH of intracellular compartments. The quantitative analysis of pH variations can be performed either by ratiometric or fluorescence lifetime detection. However, most available genetically encoded pH sensors are based on green and yellow fluorescent proteins and are not compatible with multicolor approaches. Taking advantage of the strong pH sensitivity of enhanced cyan fluorescent protein (ECFP), we demonstrate here its suitability as a sensitive pH sensor using fluorescence lifetime imaging. The intracellular ECFP lifetime undergoes large changes (32 %) in the pH 5 to pH 7 range, which allows accurate pH measurements to better than 0.2 pH units. By fusion of ECFP with the granular chromogranin A, we successfully measured the pH in secretory granules of PC12 cells, and we performed a kinetic analysis of intragranular pH variations in living cells exposed to ammonium chloride.

A highly sensitive, single selective, fluorescent sensor for Al{sup 3+} detection and its application in living cell imaging

Energy Technology Data Exchange (ETDEWEB)

Ye, Xing-Pei [Department of Physics and Chemistry, Henan Polytechnic University, Jiaozuo 454000 (China); Sun, Shao-bo; Li, Ying-dong [Institute of Integrated Traditional and Western Medicine, Gansu University of Traditional Chinese Medicine, Lanzhou 730000 (China); Zhi, Li-hua [Department of Physics and Chemistry, Henan Polytechnic University, Jiaozuo 454000 (China); Wu, Wei-na, E-mail: wuwn08@hpu.edu.cn [Department of Physics and Chemistry, Henan Polytechnic University, Jiaozuo 454000 (China); Wang, Yuan, E-mail: wangyuan08@hpu.edu.cn [Department of Physics and Chemistry, Henan Polytechnic University, Jiaozuo 454000 (China)

2014-11-15

A new o-aminophenol-based fluorogenic chemosensor methyl 3,5-bis((E)-(2-hydroxyphenylimino)methyl)-4-hydroxybenzoate 1 have been synthesized by Schiff base condensation of methyl 3,5-diformyl-4-hydroxybenzoate with o-aminophenol, which exhibits high selectivity and sensitivity toward Al{sup 3+}. Fluorescence titration studies of receptors 1 with different metal cations in CH{sub 3}OH medium showed highly selective and sensitive towards Al{sup 3+} ions even in the presence of other commonly coexisting metal ions. The detection limit of Al{sup 3+} ions is at the parts per billion level. Interestingly, the Al(III) complex of 1 offered a large Stokes shift (>120 nm), which can miximize the selfquenching effect. In addition, possible utilization of this receptor as bio-imaging fluorescent probe to detect Al{sup 3+} in human cervical HeLa cancer cell lines was also investigated by confocal fluorescence microscopy. - Highlights: • A new Schiff base chemosensor is reported. • The sensor for Al{sup 3+} offers large Stokes shift. • The detection limit of Al{sup 3+} in CH{sub 3}OH solution is at the parts per billion level. • The utilization of sensor for the monitoring of Al{sup 3+} levels in living cells was examined.

A novel pH sensitive water soluble fluorescent nanomicellar sensor for potential biomedical applications.

Science.gov (United States)

Georgiev, Nikolai I; Bryaskova, Rayna; Tzoneva, Rumiana; Ugrinova, Iva; Detrembleur, Christophe; Miloshev, Stoyan; Asiri, Abdullah M; Qusti, Abdullah H; Bojinov, Vladimir B

2013-11-01

Herein we report on the synthesis and sensor activity of a novel pH sensitive probe designed as highly water-soluble fluorescent micelles by grafting of 1,8-naphthalimide-rhodamine bichromophoric FRET system (RNI) to the PMMA block of a well-defined amphiphilic diblock copolymer-poly(methyl methacrylate)-b-poly(methacrylic acid) (PMMA48-b-PMAA27). The RNI-PMMA48-b-PMAA27 adduct is capable of self-assembling into micelles with a hydrophobic PMMA core, containing the anchored fluorescent probe, and a hydrophilic shell composed of PMAA block. Novel fluorescent micelles are able to serve as a highly sensitive pH probe in water and to internalize successfully HeLa and HEK cells. Furthermore, they showed cell specificity and significantly higher photostability than that of a pure organic dye label such as BODIPY. The valuable properties of the newly prepared fluorescent micelles indicate the high potential of the probe for future biological and biomedical applications. Copyright © 2013 Elsevier Ltd. All rights reserved.

A sensitive fluorescence biosensor for alkaline phosphatase activity based on the Cu(II)-dependent DNAzyme

Energy Technology Data Exchange (ETDEWEB)

Zhao, Mengmeng; Guo, Yajuan; Wang, Lixu [College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116 (China); Luo, Fang, E-mail: luofang0812@163.com [College of Biological Science and Technology, Fuzhou University, Fuzhou, Fujian 350116 (China); Lin, Cuiying, E-mail: lcuiying@fzu.edu.cn [College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116 (China); Lin, Zhenyu; Chen, Guonan [College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116 (China)

2016-12-15

Alkaline phosphatase (ALP) plays an important role in phosphate metabolism processes; deviation from its normal level may indicate different kinds of diseases, so it is highly necessary to develop some simple and sensitive methods to monitor the ALP level. In this study, a simple, high selective, and sensitive fluorescent biosensor has been proposed for ALP activity determination. The Cu(II)-dependent DNAzyme (Cu-Enzyme) are divided into two parts: Cu-Enzyme 1 and Cu-Enzyme 2, and labelled with alkyne and azido groups, respectively. The Cu-substrate (Cu-Sub) is labelled with a FAM fluorophore (6-carboxyfluorescein) at the 3′-end and an additional quencher (BHQ1) at the 5′-end. The 5′-end of Cu-Enzyme 1 is labelled with BHQ1 as well. The hybridization of the Cu-Enzyme 1 and Cu-Enzyme 2 with Cu-Sub strand results in the low background fluorescence signal because the fluorescence from FAM is quenched. The addition of ALP can hydrolyze AA-P into AA, which can reduce Cu(II) into Cu(I) and in turn catalyze the cycloaddition of Cu-Enzyme 1 and Cu-Enzyme 2 to form a modified Cu-Enzyme. Then the modified Cu-Enzyme catalyzes the cleavage of the Cu-Sub strands into two pieces. One piece containing FAM fluorophore can easily diffuse into solution and give off a strong fluorescence signal. The enhanced fluorescent intensity has a linear relationship with the ALP concentration in the range of 0.36–54.55 U L{sup −1} with the detection limit of 0.14 U L{sup −1} (S/N = 3). The proposed biosensor has been successfully applied to detect ALP in serum samples with satisfied results. - Highlights: • We have proposed a simple, high selective and sensitive fluorescent biosensor for ALP. • The biosensor combines the high selectivity and the click reaction and the high sensitivity of the fluorescence detection. • The biosensor has been successfully applied to detect ALP in serum samples with satisfied results.

High sensitivity optical molecular imaging system

Science.gov (United States)

An, Yu; Yuan, Gao; Huang, Chao; Jiang, Shixin; Zhang, Peng; Wang, Kun; Tian, Jie

2018-02-01

Optical Molecular Imaging (OMI) has the advantages of high sensitivity, low cost and ease of use. By labeling the regions of interest with fluorescent or bioluminescence probes, OMI can noninvasively obtain the distribution of the probes in vivo, which play the key role in cancer research, pharmacokinetics and other biological studies. In preclinical and clinical application, the image depth, resolution and sensitivity are the key factors for researchers to use OMI. In this paper, we report a high sensitivity optical molecular imaging system developed by our group, which can improve the imaging depth in phantom to nearly 5cm, high resolution at 2cm depth, and high image sensitivity. To validate the performance of the system, special designed phantom experiments and weak light detection experiment were implemented. The results shows that cooperated with high performance electron-multiplying charge coupled device (EMCCD) camera, precision design of light path system and high efficient image techniques, our OMI system can simultaneously collect the light-emitted signals generated by fluorescence molecular imaging, bioluminescence imaging, Cherenkov luminance and other optical imaging modality, and observe the internal distribution of light-emitting agents fast and accurately.

Highly Sensitive Detection of Glucose by a "Turn-Off-On" Fluorescent Probe Using Gadolinium-Doped Carbon Dots and Carbon Microparticles.

Science.gov (United States)

Hu, Meixin; Qi, Jianrong; Ruan, Jing; Shen, Guangxia

2018-06-01

Carbon dots, as a potential substitute for semiconductor quantum dots, have drawn great interest in recent years. The preparation of fluorescent carbon dots has been made easy with many significant advances, but the complicated purifying processes, low quantum yield, and blue emission wavelength still limit its wider application in biosensors, biomedicine, and photonic devices. Here we report a strategy to synthesis Gd-doped carbon dots (Gd-Cdots) of super-high quantum yield with a microwave assisted hydrothermal method. The Gd-Cdots, with a diameter of 47∼8 nm, can be purified easily with conventional centrifugal techniques. Carbon microparticles (CMPs) have also been synthesized with a similar procedure. Meanwhile, we demonstrated a novel "turn-off-on" fluorescent biosensor, which has been developed for highly sensitive detection of glucose using Gd-doped carbon dots as probes. The proposed biosensor has exhibited low-cost and non-toxic properties, with high sensitivity and good specificity. In addition, the results in real blood samples further confirmed it as a promising application in diabetes diagnosis.

Fluorescent Metal-Organic Framework (MOF) as a Highly Sensitive and Quickly Responsive Chemical Sensor for the Detection of Antibiotics in Simulated Wastewater.

Science.gov (United States)

Zhu, Xian-Dong; Zhang, Kun; Wang, Yu; Long, Wei-Wei; Sa, Rong-Jian; Liu, Tian-Fu; Lü, Jian

2018-02-05

A Zn(II)-based fluorescent metal-organic framework (MOF) was synthesized and applied as a highly sensitive and quickly responsive chemical sensor for antibiotic detection in simulated wastewater. The fluorescent chemical sensor, denoted FCS-1, exhibited enhanced fluorescence derived from its highly ordered, 3D MOF structure as well as excellent water stability in the practical pH range of simulated antibiotic wastewater (pH = 3.0-9.0). Remarkably, FCS-1 was able to effectively detect a series of sulfonamide antibiotics via photoinduced electron transfer that caused detectable fluorescence quenching, with fairly low detection limits. Two influences impacting measurements related to wastewater treatment and water quality monitoring, the presence of heavy-metal ions and the pH of solutions, were studied in terms of fluorescence quenching, which was nearly unaffected in sulfonamide-antibiotic detection. Additionally, the effective detection of sulfonamide antibiotics was rationalized by the theoretical computation of the energy bands of sulfonamide antibiotics, which revealed a good match between the energy bands of FCS-1 and sulfonamide antibiotics, in connection with fluorescence quenching in this system.

Highly sensitive colorimetric and fluorescent sensor for cyanazine based on the inner filter effect of gold nanoparticles

Science.gov (United States)

Dong, Liang; Hou, Changjun; Yang, Mei; Fa, Huanbao; Wu, Huixiang; Shen, Caihong; Huo, Danqun

2016-06-01

Cyanazine residue poses a great threat to human health and its derivatives would remain in soils, natural waters, and other environmental domains for a long time. Herein, a simple, rapid, and ultra-sensitive analytical method for the determination of cyanazine (CZ) based on inner filter effect (IFE) of Au nanoparticles (AuNPs) on the fluorescence of CdTe quantum dots (QDs) is first described in this study. With the presence of citrate-stabilized AuNPs, the fluorescence of GSH-capped CdTe QDs was remarkably quenched by AuNPs via IFE. The fluorescence of the AuNP-CdTe QD system was recovered upon addition of CZ. CZ can adsorb on to the surface of AuNPs due to its cyano group that has good affinity with gold, which could induce the aggregation of AuNPs accompanying color change from red to blue. Thus, the IFE of AuNPs on CdTe QDs was weakened, and the fluorescence intensity of CdTe QDs was recovered accordingly. A good linear correlation for detection of CZ was exhibited from 0.05 to 9 μM, and the detection limit reached 0.1568 μM, which was much lower than the safety limit required by the USA, the UK, and China. In order to probe into the selectivity of AuNPs towards CZ over other pesticides, various frequently used pesticides were mixed with AuNPs. AuNP composite solution shows good selectivity towards CZ among other pesticides. This method was successfully carried out for the assessment of CZ in real samples with satisfactory results, which revealed many advantages such as high sensitivity, low cost, and non-time-consuming compared with traditional methods.

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).

Highly sensitive and selective fluorescent assay for guanine based on the Cu2 +/eosin Y system

Science.gov (United States)

Shi, Huimin; Cui, Yi; Gong, Yijun; Feng, Suling

2016-05-01

A fluorescent probe has been developed for the determination of guanine based on the quenched fluorescence signal of Cu2 +/eosin Y. Cu2 + interacted with eosin Y, resulting in fluorescence quenching. Subsequently, with the addition of guanine to the Cu2 +/eosin Y system, guanine reacted with Cu2 + to form 1:1 chelate cation, which further combined with eosin Y to form a 1:1 ternary ion-association complex by electrostatic attraction and hydrophobic interaction, resulting in significant decrease of the fluorescence. Hence, a fluorescent system was constructed for rapid, sensitive and selective detection of guanine with a detection limit as low as 1.5 nmol L- 1 and a linear range of 3.3-116 nmol L- 1. The method has been applied satisfactorily to the determination of guanine in DNA and urine samples with the recoveries from 98.7% to 105%. This study significantly expands the realm of application of ternary ion-association complex in fluorescence probe.

Highly selective and sensitive detection of Cu2+ with lysine enhancing bovine serum albumin modified-carbon dots fluorescent probe.

Science.gov (United States)

Liu, Jia-Ming; Lin, Li-ping; Wang, Xin-Xing; Lin, Shao-Qin; Cai, Wen-Lian; Zhang, Li-Hong; Zheng, Zhi-Yong

2012-06-07

Based on the ability of lysine (Lys) to enhance the fluorescence intensity of bovine serum albumin modified-carbon dots (CDs-BSA) to decrease surface defects and quench fluorescence of the CDs-BSA-Lys system in the presence of Cu(2+) under conditions of phosphate buffer (PBS, pH = 5.0) at 45 °C for 10 min, a sensitive Lys enhancing CDs-BSA fluorescent probe was designed. The environment-friendly, simple, rapid, selective and sensitive fluorescent probe has been utilized to detect Cu(2+) in hair and tap water samples and it achieved consistent results with those obtained by inductively coupled plasma mass spectroscopy (ICP-MS). The mechanism of the proposed assay for the detection of Cu(2+) is discussed.

Mobile Phone Ratiometric Imaging Enables Highly Sensitive Fluorescence Lateral Flow Immunoassays without External Optical Filters.

Science.gov (United States)

Shah, Kamal G; Singh, Vidhi; Kauffman, Peter C; Abe, Koji; Yager, Paul

2018-05-14

Paper-based diagnostic tests based on the lateral flow immunoassay concept promise low-cost, point-of-care detection of infectious diseases, but such assays suffer from poor limits of detection. One factor that contributes to poor analytical performance is a reliance on low-contrast chromophoric optical labels such as gold nanoparticles. Previous attempts to improve the sensitivity of paper-based diagnostics include replacing chromophoric labels with enzymes, fluorophores, or phosphors at the expense of increased fluidic complexity or the need for device readers with costly optoelectronics. Several groups, including our own, have proposed mobile phones as suitable point-of-care readers due to their low cost, ease of use, and ubiquity. However, extant mobile phone fluorescence readers require costly optical filters and were typically validated with only one camera sensor module, which is inappropriate for potential point-of-care use. In response, we propose to couple low-cost ultraviolet light-emitting diodes with long Stokes-shift quantum dots to enable ratiometric mobile phone fluorescence measurements without optical filters. Ratiometric imaging with unmodified smartphone cameras improves the contrast and attenuates the impact of excitation intensity variability by 15×. Practical application was shown with a lateral flow immunoassay for influenza A with nucleoproteins spiked into simulated nasal matrix. Limits of detection of 1.5 and 2.6 fmol were attained on two mobile phones, which are comparable to a gel imager (1.9 fmol), 10× better than imaging gold nanoparticles on a scanner (18 fmol), and >2 orders of magnitude better than gold nanoparticle-labeled assays imaged with mobile phones. Use of the proposed filter-free mobile phone imaging scheme is a first step toward enabling a new generation of highly sensitive, point-of-care fluorescence assays.

Ultra-sensitive and selective Hg{sup 2+} detection based on fluorescent carbon dots

Energy Technology Data Exchange (ETDEWEB)

Liu, Ruihua; Li, Haitao; Kong, Weiqian; Liu, Juan [Institute of Functional Nano and Soft Materials (FUNSOM) and Jiangsu Key Laboratory for Carbon-Based Functional Materials and Devices, Soochow University, Suzhou 215123 (China); Liu, Yang, E-mail: yangl@suda.edu.cn [Institute of Functional Nano and Soft Materials (FUNSOM) and Jiangsu Key Laboratory for Carbon-Based Functional Materials and Devices, Soochow University, Suzhou 215123 (China); Tong, Cuiyan, E-mail: tongcy959@nenu.edu.cn [Chemisty Department, Northeast Normal University, Changchun 130024 (China); Zhang, Xing [Institute of Functional Nano and Soft Materials (FUNSOM) and Jiangsu Key Laboratory for Carbon-Based Functional Materials and Devices, Soochow University, Suzhou 215123 (China); Kang, Zhenhui, E-mail: zhkang@suda.edu.cn [Institute of Functional Nano and Soft Materials (FUNSOM) and Jiangsu Key Laboratory for Carbon-Based Functional Materials and Devices, Soochow University, Suzhou 215123 (China)

2013-07-15

Graphical abstract: Fluorescent carbon dots were efficiently synthesized by one-step sodium hydroxide-assisted reflux method from PEG and demonstrated to show high selectivity toward Hg2+ ions detection. - Highlights: • FCDs were synthesized by one-step sodium hydroxide-assisted reflux method from PEG. • The FCDs emit blue photoluminescence and have upconversion fluorescent property. • The FCDs show ultra-sensitive detective ability for Hg{sup 2+} ions. - Abstract: Fluorescent carbon dots (FCDs) were efficiently synthesized by one-step sodium hydroxide-assisted reflux method from poly(ethylene glycol) (PEG). The obtained FCDs exhibit excellent water-solubility and high stability. Under the UV irradiation, the FCDs could emit bright blue photoluminescence, and also they were found to show excellent up-conversion fluorescence. It was further demonstrated that such FCDs can serve as effective fluorescent sensing platform for Hg{sup 2+} ions detection with ultra-sensitivity and selectivity. The sensing system achieved a limit of detection as low as 1 fM, which is much lower than all the previous reported sensing systems for Hg{sup 2+} ions detection. This FCDs sensing system has been successfully applied for the analysis of Hg{sup 2+} ions in water samples from river, lake, and tap water, showing good practical feasibility.

Fast and Sensitive Interferon-γ Assay Using Supercritical Angle Fluorescence

Directory of Open Access Journals (Sweden)

Stefan Seeger

2013-02-01

Full Text Available We present an immunoassay for Interferon-γ (IFN-γ with a limit of detection of 1.9 pM (30 pg/mL and a linear concentration range spanning three orders of magnitude. The developed one-step assay takes only 12 min and can replace the time-consuming and labor-intensive enzyme-linked immunosorbent assay (ELISA. The solid-phase sandwich assay is performed on a new measurement system comprising single-use test tubes and a compact fluorescence reader. The polymer tubes contain an optical configuration for the detection of supercritical angle fluorescence, allowing for highly sensitive real-time binding measurements.

Highly sensitive colorimetric and fluorescent sensor for cyanazine based on the inner filter effect of gold nanoparticles

International Nuclear Information System (INIS)

Dong, Liang; Hou, Changjun; Yang, Mei; Fa, Huanbao; Wu, Huixiang; Shen, Caihong; Huo, Danqun

2016-01-01

Cyanazine residue poses a great threat to human health and its derivatives would remain in soils, natural waters, and other environmental domains for a long time. Herein, a simple, rapid, and ultra-sensitive analytical method for the determination of cyanazine (CZ) based on inner filter effect (IFE) of Au nanoparticles (AuNPs) on the fluorescence of CdTe quantum dots (QDs) is first described in this study. With the presence of citrate-stabilized AuNPs, the fluorescence of GSH-capped CdTe QDs was remarkably quenched by AuNPs via IFE. The fluorescence of the AuNP–CdTe QD system was recovered upon addition of CZ. CZ can adsorb on to the surface of AuNPs due to its cyano group that has good affinity with gold, which could induce the aggregation of AuNPs accompanying color change from red to blue. Thus, the IFE of AuNPs on CdTe QDs was weakened, and the fluorescence intensity of CdTe QDs was recovered accordingly. A good linear correlation for detection of CZ was exhibited from 0.05 to 9 μM, and the detection limit reached 0.1568 μM, which was much lower than the safety limit required by the USA, the UK, and China. In order to probe into the selectivity of AuNPs towards CZ over other pesticides, various frequently used pesticides were mixed with AuNPs. AuNP composite solution shows good selectivity towards CZ among other pesticides. This method was successfully carried out for the assessment of CZ in real samples with satisfactory results, which revealed many advantages such as high sensitivity, low cost, and non-time-consuming compared with traditional methods.

Highly sensitive colorimetric and fluorescent sensor for cyanazine based on the inner filter effect of gold nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Dong, Liang; Hou, Changjun, E-mail: houcj@cqu.edu.cn; Yang, Mei [Chongqing University, Key Laboratory of Biorheology Science and Technology, Ministry of Education, College of Bioengineering (China); Fa, Huanbao [Chongqing University, College of Chemistry and Chemical Engineering (China); Wu, Huixiang [Chongqing University, Key Laboratory of Biorheology Science and Technology, Ministry of Education, College of Bioengineering (China); Shen, Caihong [Luzhou Laojiao Group Co.Ltd, National Engineering Research Center of Solid-State Brewing (China); Huo, Danqun, E-mail: huodq@cqu.edu.cn [Chongqing University, Key Laboratory of Biorheology Science and Technology, Ministry of Education, College of Bioengineering (China)

2016-06-15

Cyanazine residue poses a great threat to human health and its derivatives would remain in soils, natural waters, and other environmental domains for a long time. Herein, a simple, rapid, and ultra-sensitive analytical method for the determination of cyanazine (CZ) based on inner filter effect (IFE) of Au nanoparticles (AuNPs) on the fluorescence of CdTe quantum dots (QDs) is first described in this study. With the presence of citrate-stabilized AuNPs, the fluorescence of GSH-capped CdTe QDs was remarkably quenched by AuNPs via IFE. The fluorescence of the AuNP–CdTe QD system was recovered upon addition of CZ. CZ can adsorb on to the surface of AuNPs due to its cyano group that has good affinity with gold, which could induce the aggregation of AuNPs accompanying color change from red to blue. Thus, the IFE of AuNPs on CdTe QDs was weakened, and the fluorescence intensity of CdTe QDs was recovered accordingly. A good linear correlation for detection of CZ was exhibited from 0.05 to 9 μM, and the detection limit reached 0.1568 μM, which was much lower than the safety limit required by the USA, the UK, and China. In order to probe into the selectivity of AuNPs towards CZ over other pesticides, various frequently used pesticides were mixed with AuNPs. AuNP composite solution shows good selectivity towards CZ among other pesticides. This method was successfully carried out for the assessment of CZ in real samples with satisfactory results, which revealed many advantages such as high sensitivity, low cost, and non-time-consuming compared with traditional methods.

Sensitivity of in vivo X-ray fluorescence determination of skeletal lead stores

International Nuclear Information System (INIS)

Sokas, R.K.; Besarab, A.; McDiarmid, M.A.; Shapiro, I.M.; Bloch, P.

1990-01-01

Eighteen patients with known past occupational lead exposure underwent parenteral diagnostic chelation with ethylenediaminetetraacetic acid and x-ray fluorescent determination of in vivo skeletal lead stores at the distal styloid process of the ulna and at the temporal base bone using a cobalt 57 source and measuring lead Ka x-rays. X-ray fluorescent lead measurements in both locations correlated with results of diagnostic chelation. Using a post-chelation urinary excretion of greater than 600 micrograms lead/24 h as the definition of high-lead stores, sensitivity of x-ray fluorescence at the wrist and temple was 56% and 39%, respectively

Highly sensitive and selective fluorescent assay for guanine based on the Cu(2+)/eosin Y system.

Science.gov (United States)

Shi, Huimin; Cui, Yi; Gong, Yijun; Feng, Suling

2016-05-15

A fluorescent probe has been developed for the determination of guanine based on the quenched fluorescence signal of Cu(2+)/eosin Y. Cu(2+) interacted with eosin Y, resulting in fluorescence quenching. Subsequently, with the addition of guanine to the Cu(2+)/eosin Y system, guanine reacted with Cu(2+) to form 1:1 chelate cation, which further combined with eosin Y to form a 1:1 ternary ion-association complex by electrostatic attraction and hydrophobic interaction, resulting in significant decrease of the fluorescence. Hence, a fluorescent system was constructed for rapid, sensitive and selective detection of guanine with a detection limit as low as 1.5 nmol L(-1) and a linear range of 3.3-116 nmol L(-1). The method has been applied satisfactorily to the determination of guanine in DNA and urine samples with the recoveries from 98.7% to 105%. This study significantly expands the realm of application of ternary ion-association complex in fluorescence probe. Copyright © 2016 Elsevier B.V. All rights reserved.

A sensitive fluorescence quenching method for determination of bismuth with tiron

Energy Technology Data Exchange (ETDEWEB)

Taher, Mohammad Ali; Rahimi, Mina [Department of Chemistry, Shahid Bahonar University of Kerman, Kerman (Iran, Islamic Republic of); Fazelirad, Hamid, E-mail: hamidfazelirad@gmail.com [Department of Chemistry, Shahid Bahonar University of Kerman, Kerman (Iran, Islamic Republic of); Department of Chemistry, Science and Research Branch, Islamic Azad University, Yazd (Iran, Islamic Republic of); Young Researchers Society, Shahid Bahonar University of Kerman, P.O. Box 76175-133, Kerman (Iran, Islamic Republic of)

2014-01-15

We describe a fluorescence quenching method for determination of bismuth with tiron. The method is based on the reaction of tiron by bismuth(III) in acidic media. The influence of variables such as the pH, type of buffer, tiron concentration, reaction time and temperature were investigated. Under optimized conditions, the fluorescence quenching extent is proportional to the concentration of bismuth for Bi–tiron system at the range 0.13–2.09 μg mL{sup −1} and the detection limit is 0.05 μg mL{sup −1}. The proposed sensor presented good repeatability, evaluated in terms of relative standard deviation (R.S.D.=±0.498%) for 11 replicates. This sensitive, rapid and accurate method has been successfully applied to the determination of trace bismuth(III) in water and hair samples and certified reference materials. -- Highlights: • No previous paper report on use of fluorescence quenching for determination of Bi. • Fluorescence quenching of trion is a sensitive method for determination of Bi(III). • Under the optimum conditions the detection limit is very low (0.05 μg mL{sup −1}). • The procedure is simple and safe and has high tolerance limit to interferences.

Quaternary ammonium promoted ultra selective and sensitive fluorescence detection of fluoride ion in water and living cells.

Science.gov (United States)

Li, Long; Ji, Yuzhuo; Tang, Xinjing

2014-10-21

Highly selective and sensitive fluorescent probes with a quaternary ammonium moiety have been rationally designed and developed for fast and sensitive fluorescence detection of fluoride ion (F(-) from NaF, not TBAF) in aqueous solution and living cells. With the sequestration effect of quaternary ammonium, the detection time was less than 2 min and the detection limit of fluoride ion was as low as 0.57 ppm that is among the lowest detection limits in aqueous solutions of many fluoride fluorescence probes in the literature.

Amplified fluorescent aptasensor through catalytic recycling for highly sensitive detection of ochratoxin A.

Science.gov (United States)

Wei, Yin; Zhang, Ji; Wang, Xu; Duan, Yixiang

2015-03-15

This paper describes a novel approach utilizing nano-graphite-aptamer hybrid and DNase I for the amplified detection of ochratoxin A (OTA) for the first time. Nano-graphite can effectively quench the fluorescence of carboxyfluorescein (FAM) labeled OTA specific aptamer due to their strong π-π; stacking interactions; while upon OTA addition, it will bind with aptamer to fold into an OTA-aptamerG-quadruplex structure, which does not adsorb on the surface of nano-graphite and thus retains the dye fluorescence. Meanwhile, the G-quadruplex structure can be cleaved by DNase I, and in such case OTA is delivered from the complex. The released OTA then binds other FAM-labeled aptamers on the nano-graphite surface, and touches off another target recycling, resulting in the successive release of dye-labeled aptamers from the nano-graphite, which leads to significant amplification of the signal. Under the optimized conditions, the present amplified sensing system exhibits high sensitivity toward OTA with a limit of detection of 20nM (practical measurement), which is about 100-fold higher than that of traditional unamplified homogeneous assay. Our developed method also showed high selectivity against other interference molecules and can be applied for the detection of OTA in real red wine samples. The proposed assay is simple, cost-effective, and might open a door for the development of new assays for other biomolecules. This aptasensor is of great practical importance in food safety and could be widely extended to the detection of other toxins by replacing the sequence of the recognition aptamer. Copyright © 2014 Elsevier B.V. All rights reserved.

Sensitive detection of alkaline phosphatase by switching on gold nanoclusters fluorescence quenched by pyridoxal phosphate.

Science.gov (United States)

Halawa, Mohamed Ibrahim; Gao, Wenyue; Saqib, Muhammad; Kitte, Shimeles Addisu; Wu, Fengxia; Xu, Guobao

2017-09-15

In this work, we designed highly sensitive and selective luminescent detection method for alkaline phosphatase using bovine serum albumin functionalized gold nanoclusters (BSA-AuNCs) as the nanosensor probe and pyridoxal phosphate as the substrate of alkaline phosphatase. We found that pyridoxal phosphate can quench the fluorescence of BSA-AuNCs and pyridoxal has little effect on the fluorescence of BSA-AuNCs. The proposed mechanism of fluorescence quenching by PLP was explored on the basis of data obtained from high-resolution transmission electron microscopy (HRTEM), dynamic light scattering (DLS), UV-vis spectrophotometry, fluorescence spectroscopy, fluorescence decay time measurements and circular dichroism (CD) spectroscopy. Alkaline phosphatase catalyzes the hydrolysis of pyridoxal phosphate to generate pyridoxal, restoring the fluorescence of BSA-AuNCs. Therefore, a recovery type approach has been developed for the sensitive detection of alkaline phosphatase in the range of 1.0-200.0U/L (R 2 =0.995) with a detection limit of 0.05U/L. The proposed sensor exhibit excellent selectivity among various enzymes, such as glucose oxidase, lysozyme, trypsin, papain, and pepsin. The present switch-on fluorescence sensing strategy for alkaline phosphatase was successfully applied in human serum plasma with good recoveries (100.60-104.46%), revealing that this nanosensor probe is a promising tool for ALP detection. Copyright © 2017 Elsevier B.V. All rights reserved.

Time-Resolved Fluorescent Immunochromatography of Aflatoxin B1 in Soybean Sauce: A Rapid and Sensitive Quantitative Analysis.

Science.gov (United States)

Wang, Du; Zhang, Zhaowei; Li, Peiwu; Zhang, Qi; Zhang, Wen

2016-07-14

Rapid and quantitative sensing of aflatoxin B1 with high sensitivity and specificity has drawn increased attention of studies investigating soybean sauce. A sensitive and rapid quantitative immunochromatographic sensing method was developed for the detection of aflatoxin B1 based on time-resolved fluorescence. It combines the advantages of time-resolved fluorescent sensing and immunochromatography. The dynamic range of a competitive and portable immunoassay was 0.3-10.0 µg·kg(-1), with a limit of detection (LOD) of 0.1 µg·kg(-1) and recoveries of 87.2%-114.3%, within 10 min. The results showed good correlation (R² > 0.99) between time-resolved fluorescent immunochromatographic strip test and high performance liquid chromatography (HPLC). Soybean sauce samples analyzed using time-resolved fluorescent immunochromatographic strip test revealed that 64.2% of samples contained aflatoxin B1 at levels ranging from 0.31 to 12.5 µg·kg(-1). The strip test is a rapid, sensitive, quantitative, and cost-effective on-site screening technique in food safety analysis.

Graphene Oxide-terpyridine Conjugate: A Highly Selective Colorimetric and Sensitive Fluorescence Nano-chemosensor for Fe2+ in Aqueous Media

Directory of Open Access Journals (Sweden)

Bagher Eftekhari-Sis

2016-07-01

Full Text Available A graphene oxide-terpyridine conjugate (GOTC based colorimetric and fluorescent nano-chemosensor was synthesized. It showed high selectivity and sensitivity for Fe2+ and Fe3+ ions in neutral aqueous solution over other metal ions such as Li+, Na+, Ba2+, Ca2+, Al3+, Cd2+, Co2+, Cu2+, Hg2+, Mn2+, Ni2+, Pb2+, Zn2+, Cr3+ and Ag+. In absorption spectra, upon addition of Fe2+ or Fe3+, the sensor displayed a peak at 568 nm, by changing the color of the solution from light pink for GOTC to light magenta and deep magenta for Fe3+ and Fe2+, respectively. Also, the fluorescence studies revealed that, Fe2+, Fe3+ and Co2+ quench the emission of GOTC at 473 nm, while other metal ions do not quench the fluorescence of GOTC in solution. Colorimetric and fluorescence techniques could be used for detection of Fe2+ ion concentration at least about 6-10 μM in water solution. The sensing on test paper was also investigated for the naked-eye detection of Fe2+.

Fluorescent immunochromatography for rapid and sensitive typing of seasonal influenza viruses.

Directory of Open Access Journals (Sweden)

Akira Sakurai

Full Text Available Lateral flow tests also known as Immunochromatography (IC is an antigen-detection method conducted on a nitrocellulose membrane that can be completed in less than 20 min. IC has been used as an important rapid test for clinical diagnosis and surveillance of influenza viruses, but the IC sensitivity is relatively low (approximately 60% and the limit of detection (LOD is as low as 10³ pfu per reaction. Recently, we reported an improved IC assay using antibodies conjugated with fluorescent beads (fluorescent immunochromatography; FLIC for subtyping H5 influenza viruses (FLIC-H5. Although the FLIC strip must be scanned using a fluorescent reader, the sensitivity (LOD is significantly improved over that of conventional IC methods. In addition, the antibodies which are specific against the subtypes of influenza viruses cannot be available for the detection of other subtypes when the major antigenicity will be changed. In this study, we established the use of FLIC to type seasonal influenza A and B viruses (FLIC-AB. This method has improved sensitivity to 100-fold higher than that of conventional IC methods when we used several strains of influenza viruses. In addition, FLIC-AB demonstrated the ability to detect influenza type A and influenza type B viruses from clinical samples with high sensitivity and specificity (Type A: sensitivity 98.7% (74/75, specificity 100% (54/54, Type B: sensitivity 100% (90/90, specificity 98.2% (54/55 in nasal swab samples in comparison to the results of qRT-PCR. And furthermore, FLIC-AB performs better in the detection of early stage infection (under 13 h than other conventional IC methods. Our results provide new strategies to prevent the early-stage transmission of influenza viruses in humans during both seasonal outbreaks and pandemics.

High resolution and high sensitivity methods for oligosaccharide mapping and characterization by normal phase high performance liquid chromatography following derivatization with highly fluorescent anthranilic acid.

Science.gov (United States)

Anumula, K R; Dhume, S T

1998-07-01

Facile labeling of oligosaccharides (acidic and neutral) in a nonselective manner was achieved with highly fluorescent anthranilic acid (AA, 2-aminobenzoic acid) (more than twice the intensity of 2-aminobenzamide, AB) for specific detection at very high sensitivity. Quantitative labeling in acetate-borate buffered methanol (approximately pH 5.0) at 80 degreesC for 60 min resulted in negligible or no desialylation of the oligosaccharides. A high resolution high performance liquid chromatographic method was developed for quantitative oligosaccharide mapping on a polymeric-NH2bonded (Astec) column operating under normal phase and anion exchange (NP-HPAEC) conditions. For isolation of oligosaccharides from the map by simple evaporation, the chromatographic conditions developed use volatile acetic acid-triethylamine buffer (approximately pH 4.0) systems. The mapping and characterization technology was developed using well characterized standard glycoproteins. The fluorescent oligosaccharide maps were similar to the maps obtained by the high pH anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD), except that the fluorescent maps contained more defined peaks. In the map, the oligosaccharides separated into groups based on charge, size, linkage, and overall structure in a manner similar to HPAEC-PAD with contribution of -COOH function from the label, anthranilic acid. However, selectivity of the column for sialic acid linkages was different. A second dimension normal phase HPLC (NP-HPLC) method was developed on an amide column (TSK Gel amide-80) for separation of the AA labeled neutral complex type and isomeric structures of high mannose type oligosaccharides. The oligosaccharides labeled with AA are compatible with biochemical and biophysical techniques, and use of matrix assisted laser desorption mass spectrometry for rapid determination of oligosaccharide mass map of glycoproteins is demonstrated. High resolution of NP-HPAEC and NP-HPLC methods

Project Title: Radiochemical Analysis by High Sensitivity Dual-Optic Micro X-ray Fluorescence

International Nuclear Information System (INIS)

Havrilla, George J.; Gao, Ning

2002-01-01

A novel dual-optic micro X-ray fluorescence instrument will be developed to do radiochemical analysis of high-level radioactive wastes at DOE sites such as Savannah River Site and Hanford. This concept incorporates new X-ray optical elements such as monolithic polycapillaries and double bent crystals, which focus X-rays. The polycapillary optic can be used to focus X-rays emitted by the X-ray tube thereby increasing the X-ray flux on the sample over 1000 times. Polycapillaries will also be used to collect the X-rays from the excitation site and screen the radiation background from the radioactive species in the specimen. This dual-optic approach significantly reduces the background and increases the analyte signal thereby increasing the sensitivity of the analysis. A doubly bent crystal used as the focusing optic produces focused monochromatic X-ray excitation, which eliminates the bremsstrahlung background from the X-ray source. The coupling of the doubly bent crystal for monochromatic excitation with a polycapillary for signal collection can effectively eliminate the noise background and radiation background from the specimen. The integration of these X-ray optics increases the signal-to-noise and thereby increases the sensitivity of the analysis for low-level analytes. This work will address a key need for radiochemical analysis of high-level waste using a non-destructive, multi-element, and rapid method in a radiation environment. There is significant potential that this instrumentation could be capable of on-line analysis for process waste stream characterization at DOE sites

Highly sensitive ratiometric detection of heparin and its oversulfated chondroitin sulfate contaminant by fluorescent peptidyl probe.

Science.gov (United States)

Mehta, Pramod Kumar; Lee, Hyeri; Lee, Keun-Hyeung

2017-05-15

The selective and sensitive detection of heparin, an anticoagulant in clinics as well as its contaminant oversulfated chondroitin sulfate (OSCS) is of great importance. We first reported a ratiometric sensing method for heparin as well as OSCS contaminants in heparin using a fluorescent peptidyl probe (Pep1, pyrene-GSRKR) and heparin-digestive enzyme. Pep1 exhibited a highly sensitive ratiometric response to nanomolar concentration of heparin in aqueous solution over a wide pH range (2~11) and showed highly selective ratiometric response to heparin among biological competitors such as hyaluronic acid and chondroitin sulfate. Pep1 showed a linear ratiometric response to nanomolar concentrations of heparin in aqueous solutions and in human serum samples. The detection limit for heparin was calculated to be 2.46nM (R 2 =0.99) in aqueous solutions, 2.98nM (R 2 =0.98) in 1% serum samples, and 3.43nM (R 2 =0.99) in 5% serum samples. Pep1 was applied to detect the contaminated OSCS in heparin with heparinase I, II, and III, respectively. The ratiometric sensing method using Pep1 and heparinase II was highly sensitive, fast, and efficient for the detection of OSCS contaminant in heparin. Pep1 with heparinase II could detect as low as 0.0001% (w/w) of OSCS in heparin by a ratiometric response. Copyright © 2017 Elsevier B.V. All rights reserved.

Environment-sensitive behavior of fluorescent molecular rotors

Directory of Open Access Journals (Sweden)

Theodorakis Emmanuel A

2010-09-01

Full Text Available Abstract Molecular rotors are a group of fluorescent molecules that form twisted intramolecular charge transfer (TICT states upon photoexcitation. When intramolecular twisting occurs, the molecular rotor returns to the ground state either by emission of a red-shifted emission band or by nonradiative relaxation. The emission properties are strongly solvent-dependent, and the solvent viscosity is the primary determinant of the fluorescent quantum yield from the planar (non-twisted conformation. This viscosity-sensitive behavior gives rise to applications in, for example, fluid mechanics, polymer chemistry, cell physiology, and the food sciences. However, the relationship between bulk viscosity and the molecular-scale interaction of a molecular rotor with its environment are not fully understood. This review presents the pertinent theories of the rotor-solvent interaction on the molecular level and how this interaction leads to the viscosity-sensitive behavior. Furthermore, current applications of molecular rotors as microviscosity sensors are reviewed, and engineering aspects are presented on how measurement accuracy and precision can be improved.

Fluorescence ELISA for sensitive detection of ochratoxin A based on glucose oxidase-mediated fluorescence quenching of CdTe QDs

International Nuclear Information System (INIS)

Liang, Yi; Huang, Xiaolin; Yu, Ruijin; Zhou, Yaofeng; Xiong, Yonghua

2016-01-01

The present study described a novel fluorescence enzyme-linked immunosorbent assay (ELISA) used to detect ochratoxin A (OTA) by using the glucose oxidase (GOx)-mediated fluorescence quenching of mercaptopropionic acid-capped CdTe quantum dots (MPA-QDs), in which GOx was used as an alternative to horseradish peroxidase (HRP) for the oxidization of glucose into hydrogen peroxide (H_2O_2) and gluconic acid. The MPA-QDs were used as a fluorescent signal output, whose fluorescence variation was extremely sensitive to the presence of H_2O_2 or hydrogen ions in the solution. Under the optimized conditions, the proposed fluorescence ELISA demonstrated a good linear detection of OTA in corn extract from 2.4 pg mL"−"1 to 625 pg mL"−"1 with a limit of detection of 2.2 pg mL"−"1, which was approximately 15-fold lower than that of conventional HRP-based ELISA. Our developed fluorescence immunoassay was also similar to HRP-based ELISA in terms of selectivity, accuracy, and reproducibility. In summary, this study was the first to use the GOx-mediated fluorescence quenching of QDs in immunoassay to detect OTA, offering a new possibility for the analysis of other mycotoxins and biomolecules. - Highlights: • A novel fluorescence ELISA was first developed for the detection of OTA by using GOx-mediated fluorescence quenching of QDs. • The pH- and H_2O_2-sensitive MPA-capped CdTe QDs were used as a fluorescent signal output to improve the detection sensitivity. • This novel method open up a different vision to detect other mycotoxins and biomolecules.

Highly sensitive immunoassay of protein molecules based on single nanoparticle fluorescence detection in a nanowell

Science.gov (United States)

Han, Jin-Hee; Kim, Hee-Joo; Lakshmana, Sudheendra; Gee, Shirley J.; Hammock, Bruce D.; Kennedy, Ian M.

2011-03-01

A nanoarray based-single molecule detection system was developed for detecting proteins with extremely high sensitivity. The nanoarray was able to effectively trap nanoparticles conjugated with biological sample into nanowells by integrating with an electrophoretic particle entrapment system (EPES). The nanoarray/EPES is superior to other biosensor using immunoassays in terms of saving the amounts of biological solution and enhancing kinetics of antibody binding due to reduced steric hindrance from the neighboring biological molecules. The nanoarray patterned onto a layer of PMMA and LOL on conductive and transparent indium tin oxide (ITO)-glass slide by using e-beam lithography. The suspension of 500 nm-fluorescent (green emission)-carboxylated polystyrene (PS) particles coated with protein-A followed by BDE 47 polyclonal antibody was added to the chip that was connected to the positive voltage. The droplet was covered by another ITO-coated-glass slide and connected to a ground terminal. After trapping the particles into the nanowells, the solution of different concentrations of anti-rabbit- IgG labeled with Alexa 532 was added for an immunoassay. A single molecule detection system could quantify the anti-rabbit IgG down to atto-mole level by counting photons emitted from the fluorescent dye bound to a single nanoparticle in a nanowell.

Luminescent conjugated oligothiophenes for sensitive fluorescent assignment of protein inclusion bodies.

Science.gov (United States)

Klingstedt, Therése; Blechschmidt, Cristiane; Nogalska, Anna; Prokop, Stefan; Häggqvist, Bo; Danielsson, Olof; Engel, W King; Askanas, Valerie; Heppner, Frank L; Nilsson, K Peter R

2013-03-18

Small hydrophobic ligands identifying intracellular protein deposits are of great interest, as protein inclusion bodies are the pathological hallmark of several degenerative diseases. Here we report that fluorescent amyloid ligands, termed luminescent conjugated oligothiophenes (LCOs), rapidly and with high sensitivity detect protein inclusion bodies in skeletal muscle tissue from patients with sporadic inclusion body myositis (s-IBM). LCOs having a conjugated backbone of at least five thiophene units emitted strong fluorescence upon binding, and showed co-localization with proteins reported to accumulate in s-IBM protein inclusion bodies. Compared with conventional amyloid ligands, LCOs identified a larger fraction of immunopositive inclusion bodies. When the conjugated thiophene backbone was extended with terminal carboxyl groups, the LCO revealed striking spectral differences between distinct protein inclusion bodies. We conclude that 1) LCOs are sensitive, rapid and powerful tools for identifying protein inclusion bodies and 2) LCOs identify a wider range of protein inclusion bodies than conventional amyloid ligands. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Highly selective ratiometric fluorescent detection of Fe{sup 3+} with a polyphenyl derivative

Energy Technology Data Exchange (ETDEWEB)

Li, Zhan-Xian, E-mail: lizx@zzu.edu.cn [The College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou 450001 (China); Zhou, Wan; Zhang, Li-Feng; Yuan, Rui-Li; Liu, Xing-Jiang; Wei, Liu-He [The College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou 450001 (China); Yu, Ming-Ming, E-mail: yumm@zzu.edu.cn [The College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou 450001 (China)

2013-04-15

Compared with other fluorescent probes, ratiometric fluorescence responses are more attractive because the ratio between the two emission intensities can be used to measure the analyte concentration and provide a built-in correction for environmental effects. A highly selective and sensitive ratiometric fluorescent probe for Fe{sup 3+} was synthesized, which exhibits an enhanced fluorescence with a large red-shift in emission from 361 to 455 nm upon addition of Fe{sup 3+}. The red-shift of the emission peak can be ascribed to the reformed orbital, and the increase of emission intensity may be ascribed to the inhibition of the rotation of C–C bonds between each two aromatic rings. -- Graphical abstract: A highly selective and sensitive ratiometric fluorescent probe for Fe{sup 3+} was synthesized, which exhibits an enhanced fluorescence with a large red-shift in emission from 361 to 455 nm upon addition of Fe{sup 3+}. Highlights: ► A ratiometric fluorescent probe for Fe{sup 3+} was synthesized. ► The probe exhibits an enhanced fluorescence with a red-shift upon addition of Fe{sup 3+}. ► Inhibition of the rotation of C–C bonds was possible detection mechanism for Fe{sup 3+}.

Determination of the lactone and lactone plus carboxylate forms of 9-aminocamptothecin in human plasma by sensitive high-performance liquid chromatography with fluorescence detection

NARCIS (Netherlands)

W.J. Loos (Walter); A. Sparreboom (Alex); J. Verweij (Jaap); K. Nooter (Kees); G. Stoter (Gerrit); J.H.M. Schellens (Jan)

1997-01-01

textabstractTwo sensitive reversed-phase high-performance liquid chromatographic fluorescence methods, with simple sample handling at the site of the patient, are described for the determination of the lactone and lactone plus carboxylate forms of g-aminocamptothecin (9AC). For 9AC lactone, the

Fluorescence ELISA for sensitive detection of ochratoxin A based on glucose oxidase-mediated fluorescence quenching of CdTe QDs

Energy Technology Data Exchange (ETDEWEB)

Liang, Yi [State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047 (China); Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang 330047 (China); Huang, Xiaolin [State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047 (China); Yu, Ruijin [College of Science, Northwest A& F University, Yangling, Shaanxi 712100 (China); Zhou, Yaofeng [State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047 (China); Xiong, Yonghua, E-mail: yhxiongchen@163.com [State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047 (China); Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang 330047 (China)

2016-09-14

The present study described a novel fluorescence enzyme-linked immunosorbent assay (ELISA) used to detect ochratoxin A (OTA) by using the glucose oxidase (GOx)-mediated fluorescence quenching of mercaptopropionic acid-capped CdTe quantum dots (MPA-QDs), in which GOx was used as an alternative to horseradish peroxidase (HRP) for the oxidization of glucose into hydrogen peroxide (H{sub 2}O{sub 2}) and gluconic acid. The MPA-QDs were used as a fluorescent signal output, whose fluorescence variation was extremely sensitive to the presence of H{sub 2}O{sub 2} or hydrogen ions in the solution. Under the optimized conditions, the proposed fluorescence ELISA demonstrated a good linear detection of OTA in corn extract from 2.4 pg mL{sup −1} to 625 pg mL{sup −1} with a limit of detection of 2.2 pg mL{sup −1}, which was approximately 15-fold lower than that of conventional HRP-based ELISA. Our developed fluorescence immunoassay was also similar to HRP-based ELISA in terms of selectivity, accuracy, and reproducibility. In summary, this study was the first to use the GOx-mediated fluorescence quenching of QDs in immunoassay to detect OTA, offering a new possibility for the analysis of other mycotoxins and biomolecules. - Highlights: • A novel fluorescence ELISA was first developed for the detection of OTA by using GOx-mediated fluorescence quenching of QDs. • The pH- and H{sub 2}O{sub 2}-sensitive MPA-capped CdTe QDs were used as a fluorescent signal output to improve the detection sensitivity. • This novel method open up a different vision to detect other mycotoxins and biomolecules.

A highly selective and sensitive fluorescent chemosensor and its application for rapid on-site detection of Al3 +

Science.gov (United States)

Yue, Xiao-li; Wang, Zhao-qing; Li, Chao-rui; Yang, Zheng-yin

2018-03-01

In this paper, a simple naphthalene-based derivative (HL) has been designed and synthesized as a Al3 +-selective fluorescent chemosensor based on the PET mechanism. HL exhibited high selectivity and sensitivity towards Al3 + over other commonly coexisting metal ions in ethanol with a detection limit of 2.72 nM. The 1:1 binding stoichiometry of the complex (HL-Al3 +) was determined from the Job's plot based on fluorescence titrations and the ESI-MS spectrum data. Moreover, the binding site of HL with Al3 + was assured by the 1H NMR titration experiment. The binding constant (Ka) of the complex (HL-Al3 +) was calculated to be 5.06 × 104 M- 1 according to the Benesi-Hildebrand equation. In addition, the recognizing process of HL towards Al3 + was chemically reversible by adding Na2EDTA. Importantly, HL could directly and rapidly detect aluminum ion through the filter paper without resorting to additional instrumental analysis.

High-resolution and high sensitivity mesoscopic fluorescence tomography based on de-scanning EMCCD: System design and thick tissue imaging applications

Science.gov (United States)

Ozturk, Mehmet Saadeddin

Optical microscopy has been one of the essential tools for biological studies for decades, however, its application areas was limited to superficial investigation due to strong scattering in live tissues. Even though advanced techniques such as confocal or multiphoton methods have been recently developed to penetrate beyond a few hundreds of microns deep in tissues, they still cannot perform in the mesoscopic regime (millimeter scale) without using destructive sample preparation protocols such as clearing techniques. They provide rich cellular information; however, they cannot be readily employed to investigate the biological processes at larger scales. Herein, we will present our effort to establish a novel imaging approach that can quantify molecular expression in intact tissues, well beyond the current microscopy depth limits. Mesoscopic Fluorescence Molecular Tomography (MFMT) is an emerging imaging modality that offers unique potential for the non-invasive molecular assessment of thick in-vitro and in-vivo live tissues. This novel imaging modality is based on an optical inverse problem that allows for retrieval of the quantitative spatial distribution of fluorescent tagged bio-markers at millimeter depth. MFMT is well-suited for in-vivo subsurface tissue imaging and thick bio-printed specimens due to its high sensitivity and fast acquisition times, as well as relatively large fields of view. Herein, we will first demonstrate the potential of this technique using our first generation MFMT system applied to multiplexed reporter gene imaging (in-vitro) and determination of Photodynamic Therapy (PDT) agent bio-distribution in a mouse model (in-vivo). Second, we will present the design rationale, in silico benchmarking, and experimental validation of a second generation MFMT (2GMFMT) system. We will demonstrate the gain in resolution and sensitivity achieved due to the de-scanned dense detector configuration implemented. The potential of this novel platform will be

Sensitive determination of nucleic acids using organic nanoparticle fluorescence probes

Science.gov (United States)

Zhou, Yunyou; Bian, Guirong; Wang, Leyu; Dong, Ling; Wang, Lun; Kan, Jian

2005-06-01

This paper describes the preparation of organic nanoparticles by reprecipitation method under sonication and vigorous stirring. Transmission electron microscopy (TEM) was used to characterize the size and size distribution of the luminescent nanoparticles. Their average diameter was about 25 nm with a size variation of ±18%. The fluorescence decay lifetime of the nanoparticles also was determined on a self-equipped fluorospectrometer with laser light source. The lifetime (˜0.09 μs) of nanoparticles is about three times long as that of the monomer. The nanoparticles were in abundant of hydrophilic groups, which increased their miscibility in aqueous solution. These organic nanoparticles have high photochemical stability, excellent resistance to chemical degradation and photodegradation, and a good fluorescence quantum yield (25%). The fluorescence can be efficiently quenched by nucleic acids. Based on the fluorescence quenching of nanoparticles, a fluorescence quenching method was developed for determination of microamounts of nucleic acids by using the nanoparticles as a new fluorescent probe. Under optimal conditions, maximum fluorescence quenching is produced, with maximum excitation and emission wavelengths of 345 and 402 nm, respectively. Under optimal conditions, the calibration graphs are linear over the range 0.4-19.0 μg ml -1 for calf thymus DNA (ct-DNA) and 0.3-19.0 μg ml -1 for fish sperm DNA (fs-DNA). The corresponding detection limits are 0.25 μg ml -1 for ct-DNA and 0.17 μg ml -1 for fs-DNA. The relative standard deviation of six replicate measurements is 1.3-2.1%. The method is simple, rapid and sensitive with wide linear range. The recovery and relative standard deviation are very satisfactory.

A Conjugated Aptamer-Gold Nanoparticle Fluorescent Probe for Highly Sensitive Detection of rHuEPO-α

Directory of Open Access Journals (Sweden)

Zhaoyang Zhang

2011-11-01

Full Text Available We present here a novel conjugated aptamer-gold nanoparticle (Apt-AuNPs fluorescent probe and its application for specific detection of recombinant human erythropoietin-α (rHuEPO-α. In this nanobiosensor, 12 nm AuNPs function as both a nano-scaffold and a nano-quencher (fluorescent energy acceptor, on the surface of which the complementary sequences are linked (as cODN-AuNPs and pre-hybridized with carboxymethylfluorescein (FAM-labeled anti-rHuEPO-α aptamers. Upon target protein binding, the aptamers can be released from the AuNP surface and the fluorescence signal is restored. Key variables such as the length of linker, the hybridization site and length have been designed and optimized. Full performance evaluation including sensitivity, linear range and interference substances are also described. This nanobiosensor provides a promising approach for a simple and direct quantification of rHuEPO-α concentrations as low as 0.92 nM within a few hours.

High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators.

Science.gov (United States)

Bullen, A; Patel, S S; Saggau, P

1997-07-01

The design and implementation of a high-speed, random-access, laser-scanning fluorescence microscope configured to record fast physiological signals from small neuronal structures with high spatiotemporal resolution is presented. The laser-scanning capability of this nonimaging microscope is provided by two orthogonal acousto-optic deflectors under computer control. Each scanning point can be randomly accessed and has a positioning time of 3-5 microseconds. Sampling time is also computer-controlled and can be varied to maximize the signal-to-noise ratio. Acquisition rates up to 200k samples/s at 16-bit digitizing resolution are possible. The spatial resolution of this instrument is determined by the minimal spot size at the level of the preparation (i.e., 2-7 microns). Scanning points are selected interactively from a reference image collected with differential interference contrast optics and a video camera. Frame rates up to 5 kHz are easily attainable. Intrinsic variations in laser light intensity and scanning spot brightness are overcome by an on-line signal-processing scheme. Representative records obtained with this instrument by using voltage-sensitive dyes and calcium indicators demonstrate the ability to make fast, high-fidelity measurements of membrane potential and intracellular calcium at high spatial resolution (2 microns) without any temporal averaging.

Quantum dots (QDs) based fluorescence probe for the sensitive determination of kaempferol

Science.gov (United States)

Tan, Xuanping; Liu, Shaopu; Shen, Yizhong; He, Youqiu; Yang, Jidong

2014-12-01

In this work, using the quenching of fluorescence of thioglycollic acid (TGA)-capped CdTe quantum dots (QDs), a novel method for the determination of kaempferol (KAE) has been developed. Under optimum conditions, a linear calibration plot of the quenched fluorescence intensity at 552 nm against the concentration of KAE was observed in the range of 4-44 μg mL-1 with a detection limit (3σ/K) of 0.79 μg mL-1. In addition, the detailed reaction mechanism has also been proposed on the basis of electron transfer supported by ultraviolet-visible (UV-vis) absorption and fluorescence (FL) spectroscopy. The method has been applied for the determination of KAE in pharmaceutical preparations with satisfactory results. The proposed method manifested several advantages such as high sensitivity, short analysis time, low cost and ease of operation.

Determination of paraquat in water samples using a sensitive fluorescent probe titration method.

Science.gov (United States)

Yao, Feihu; Liu, Hailong; Wang, Guangquan; Du, Liming; Yin, Xiaofen; Fu, Yunlong

2013-06-01

Paraquat (PQ), a nonselective herbicide, is non-fluorescent in aqueous solutions. Thus, its determination through direct fluorescent methods is not feasible. The supramolecular inclusion interaction of PQ with cucurbit[7]uril was studied by a fluorescent probe titration method. Significant quenching of the fluorescence intensity of the cucurbit[7]uril-coptisine fluorescent probe was observed with the addition of PQ. A new fluorescent probe titration method with high selectivity and sensitivity at the ng/mL level was developed to determine PQ in aqueous solutions with good precision and accuracy based on the significant quenching of the supramolecular complex fluorescence intensity. The proposed method was successfully used in the determination of PQ in lake water, tap water, well water, and ditch water in an agricultural area, with recoveries of 96.73% to 105.77%. The fluorescence quenching values (deltaF) showed a good linear relationship with PQ concentrations from 1.0 x 10(-8) to 1.2 x 10(-5) mol/L with a detection limit of 3.35 x 10(-9) mol/L. In addition, the interaction models of the supramolecular complexes formed between the host and the guest were established using theoretical calculations. The interaction mechanism between the cucurbit[7]uril and PQ was confirmed by 1H NMR spectroscopy.

Highly Specific and Sensitive Fluorescent Nanoprobes for Image-Guided Resection of Sub-Millimeter Peritoneal Tumors.

Science.gov (United States)

Colby, Aaron H; Berry, Samantha M; Moran, Ann M; Pasion, Kristine Amber; Liu, Rong; Colson, Yolonda L; Ruiz-Opazo, Nelson; Grinstaff, Mark W; Herrera, Victoria L M

2017-02-28

A current challenge in the treatment of peritoneal carcinomatosis is the inability to detect, visualize, and resect small or microscopic tumors of pancreatic, ovarian, or mesothelial origin. In these diseases, the completeness of primary tumor resection is directly correlated with patient survival, and hence, identifying small sub-millimeter tumors (i.e., disseminated disease) is critical. Thus, new imaging techniques and probes are needed to improve cytoreductive surgery and patient outcomes. Highly fluorescent rhodamine-labeled expansile nanoparticles (HFR-eNPs) are described for use as a visual aid during cytoreductive surgery of pancreatic carcinomatosis. The covalent incorporation of rhodamine into ∼30 nm eNPs increases the fluorescent signal compared to free rhodamine, thereby affording a brighter and more effective probe than would be achieved by a single rhodamine molecule. Using the intraperitoneal route of administration, HFR-eNPs localize to regions of large (∼1 cm), sub-centimeter, and sub-millimeter intraperitoneal tumor in three different animal models, including pancreatic, mesothelioma, and ovarian carcinoma. Tumoral localization of the HFR-eNPs depends on both the material property (i.e., eNP polymer) as well as the surface chemistry (anionic surfactant vs PEGylated noncharged surfactant). In a rat model of pancreatic carcinomatosis, HFR-eNP identification of tumor is validated against gold-standard histopathological analysis to reveal that HFR-eNPs possess high specificity (99%) and sensitivity (92%) for tumors, in particular, sub-centimeter and microscopic sub-millimeter tumors, with an overall accuracy of 95%. Finally, as a proof-of-concept, HFR-eNPs are used to guide the resection of pancreatic tumors in a rat model of peritoneal carcinomatosis.

Highly fluorescent and morphology-controllable graphene quantum dots-chitosan hybrid xerogels for in vivo imaging and pH-sensitive drug carrier

Energy Technology Data Exchange (ETDEWEB)

Lv, Ouyang; Tao, Yongxin; Qin, Yong [Advanced Catalysis and Green Manufacturing Collaborative Innovation Center, School of Petrochemical Engineering, Changzhou University, Changzhou 213164 (China); Chen, Chuanxiang [School of Environmental and Chemical Engineering, Jiangsu University of Science and Technology, Zhenjiang 212003 (China); Pan, Yan; Deng, Linhong [Institute of Biomedical Engineering and Health Sciences, Changzhou University, Changzhou 213164 (China); Liu, Li [School of pharmaceutical Engineering & Life Science, Changzhou University, Changzhou 213164 (China); Kong, Yong, E-mail: yzkongyong@126.com [Advanced Catalysis and Green Manufacturing Collaborative Innovation Center, School of Petrochemical Engineering, Changzhou University, Changzhou 213164 (China)

2016-10-01

Highly fluorescent graphene quantum dots (GQDs)-chitosan (CS) hybrid xerogels (GQDs-CS) were facilely synthesized, and the morphology of GQDs-CS was controllable by varying the content of GQDs in the xerogel. The GQDs-CS exhibited a porous and three-dimensional (3D) network structure when the content of GQDs reached 43% (wt%) in the xerogel, which was beneficial for drug loading and sustained release. The as-prepared GQDs-CS could also be applied for in vivo imaging since it showed strong blue, green and red luminescence under excitation of varying wavelengths. Moreover, the pH-induced protonation/deprotonation of the –NH{sub 2} groups on CS chains can result in a pH-dependent drug delivery behavior of the GQDs-CS hybrid xerogel. - Graphical abstract: Highly fluorescent and morphology-controllable graphene quantum dots-chitosan hybrid xerogels for in vivo imaging and pH-sensitive drug carrier. Display Omitted - Highlights: • Highly fluorescent GQDs-CS hybrid xerogels were facilely synthesized. • The as-made xerogels exhibited various morphologies with different GQDs contents. • The GQDs-CS exhibited a porous and 3D network when the content of GQDs reached 43%. • The GQDs-CS could be applied for in vivo imaging since it showed strong luminescence. • The protonation/deprotonation of –NH{sub 2} on CS result in a pH-dependent drug delivery.

Highly fluorescent and morphology-controllable graphene quantum dots-chitosan hybrid xerogels for in vivo imaging and pH-sensitive drug carrier

International Nuclear Information System (INIS)

Lv, Ouyang; Tao, Yongxin; Qin, Yong; Chen, Chuanxiang; Pan, Yan; Deng, Linhong; Liu, Li; Kong, Yong

2016-01-01

Highly fluorescent graphene quantum dots (GQDs)-chitosan (CS) hybrid xerogels (GQDs-CS) were facilely synthesized, and the morphology of GQDs-CS was controllable by varying the content of GQDs in the xerogel. The GQDs-CS exhibited a porous and three-dimensional (3D) network structure when the content of GQDs reached 43% (wt%) in the xerogel, which was beneficial for drug loading and sustained release. The as-prepared GQDs-CS could also be applied for in vivo imaging since it showed strong blue, green and red luminescence under excitation of varying wavelengths. Moreover, the pH-induced protonation/deprotonation of the –NH_2 groups on CS chains can result in a pH-dependent drug delivery behavior of the GQDs-CS hybrid xerogel. - Graphical abstract: Highly fluorescent and morphology-controllable graphene quantum dots-chitosan hybrid xerogels for in vivo imaging and pH-sensitive drug carrier. Display Omitted - Highlights: • Highly fluorescent GQDs-CS hybrid xerogels were facilely synthesized. • The as-made xerogels exhibited various morphologies with different GQDs contents. • The GQDs-CS exhibited a porous and 3D network when the content of GQDs reached 43%. • The GQDs-CS could be applied for in vivo imaging since it showed strong luminescence. • The protonation/deprotonation of –NH_2 on CS result in a pH-dependent drug delivery.

High speed fluorescence imaging with compressed ultrafast photography

Science.gov (United States)

Thompson, J. V.; Mason, J. D.; Beier, H. T.; Bixler, J. N.

2017-02-01

Fluorescent lifetime imaging is an optical technique that facilitates imaging molecular interactions and cellular functions. Because the excited lifetime of a fluorophore is sensitive to its local microenvironment,1, 2 measurement of fluorescent lifetimes can be used to accurately detect regional changes in temperature, pH, and ion concentration. However, typical state of the art fluorescent lifetime methods are severely limited when it comes to acquisition time (on the order of seconds to minutes) and video rate imaging. Here we show that compressed ultrafast photography (CUP) can be used in conjunction with fluorescent lifetime imaging to overcome these acquisition rate limitations. Frame rates up to one hundred billion frames per second have been demonstrated with compressed ultrafast photography using a streak camera.3 These rates are achieved by encoding time in the spatial direction with a pseudo-random binary pattern. The time domain information is then reconstructed using a compressed sensing algorithm, resulting in a cube of data (x,y,t) for each readout image. Thus, application of compressed ultrafast photography will allow us to acquire an entire fluorescent lifetime image with a single laser pulse. Using a streak camera with a high-speed CMOS camera, acquisition rates of 100 frames per second can be achieved, which will significantly enhance our ability to quantitatively measure complex biological events with high spatial and temporal resolution. In particular, we will demonstrate the ability of this technique to do single-shot fluorescent lifetime imaging of cells and microspheres.

Rapid and sensitive detection of ketamine in blood using novel fluorescence genosensor.

Science.gov (United States)

Ding, Yanjun; Li, Xingmei; Guo, Yadong; Yan, Jie; Ling, Jiang; Li, Weichen; Lan, Lingmei; Chang, Yunfeng; Cai, Jifeng; Zha, Lagabaiyla

2017-12-01

In recent years, drug abuse has been considered as a most challenging social problem that aroused public attention. Ketamine has increased in unregulated use as a 'recreational drug' in teenagers. However, there is no suitable and maneuverable detection method for ketamine in situ at the moment. Fluorescence sensor technique, with predominant recognition and simple operation, is a good potential application in drug detection. Here, we first reported a highly sensitive and selective fluorescence genosensor for rapid detection of ketamine based on DNA-templated silver nanoclusters (DNA-AgNCs) probes, in which the DNA sequence could specially recognize ketamine with high affinity. Parameters affecting detection efficiency were investigated and optimized. Under optimum conditions, the as-prepared genosensor can allow for the determination of ketamine in the concentration range of 0.0001-20 μg/mL with two linear equations: one is y = 2.84x-7.139 (R 2 = 0.987) for 0.0001-0.1 μg/mL, and the other is y = 1.87x-0.091 (R 2 = 0.962) for 0.1-20 μg/mL, and the estimated detection limit of ketamine is 0.06 ng/mL. Moreover, the feasibility of this proposed method was also demonstrated by analyzing forensic blood samples. Compared with official gas chromatography/mass spectrometry (GC/MS), this fluorescence genosensor is simple, rapid, and accurate for quantitative determination of ketamine in blood for pharmaceutical and forensic analysis. Overall, it is the first report on a fluorescence genosensor for detecting ketamine directly in blood. This research may provide a new insight for the analyst to band fluorescence genosensor technology together with drug monitoring in the battle against drug abuse and forensic examination. Graphical abstract High selectively detection of ketamine using a novel fluorescence genosensor based on DNA-AgNCs probe.

A nanocluster-based fluorescent sensor for sensitive hemoglobin detection.

Science.gov (United States)

Yang, Dongqin; Meng, Huijie; Tu, Yifeng; Yan, Jilin

2017-08-01

In this report, a fluorescence sensor for sensitive detection of hemoglobin was developed. Gold nanoclusters were first synthesized with bovine serum albumin. It was found that both hydrogen peroxide and hemoglobin could weakly quench the fluorescence from the gold nanoclusters, but when these two were applied onto the nanolcusters simultaneously, a much improved quenching was resulted. This enhancing effect was proved to come from the catalytic generation of hydroxyl radical by hemoglobin. Under an optimized condition, the quenching linearly related to the concentration of hemoglobin in the range of 1-250nM, and a limit of detection as low as 0.36nM could be obtained. This provided a sensitive means for the quantification of Hb. The sensor was then successfully applied for blood analyses with simple sample pretreatment. Copyright © 2017 Elsevier B.V. All rights reserved.

Highly sensitive enzymatic determination of urea based on the pH-dependence of the fluorescence of graphene quantum dots

International Nuclear Information System (INIS)

Shao, Taili; Zhang, Ping; Zhuo, Shujuan; Zhu, Changqing; Tang, Lin

2015-01-01

We report on a nanoparticle-based fluorescent sensing scheme for urea. It is based on the finding that graphene quantum dots (GQDs) display pH-sensitive green fluorescence if photoexcited at 460 nm. Fluorescence is gradually quenched due to an increase in the local pH value as a result of the hydrolysis of urea as catalyzed by urease. The effect was used to quantify urea in the 0.1–100 mM concentration range, with a limit of detection of 0.01 mM. The method was successfully applied to the determination of urea in human serum samples. The method is simple, effective, and therefore holds promise as a platform for sensing urea in blood. (author)

A highly selective and sensitive photoswitchable fluorescent probe for Hg2+ based on bisthienylethene-rhodamine 6G dyad and for live cells imaging.

Science.gov (United States)

Xu, Li; Wang, Sheng; Lv, Yingnian; Son, Young-A; Cao, Derong

2014-07-15

A new photochromic diarylethene derivative bearing rhodamine 6G dimmer as a fluorescent molecular probe is designed and synthesized successfully. All the compounds are characterized by nuclear magnetic resonance and mass spectrometry. The bisthienylethene-rhodamine 6G dyad exhibit excellent phtochromism with reversibly color and fluorescence changes alternating irradiation with ultraviolet and visible light. Upon addition of Hg(2+), its color changes from colorless to red and its fluorescence is remarkably enhanced. Whereas other ions including K(+), Na(+), Ca(2+), Mg(2+), Fe(2+), Co(2+), Ni(2+), Cu(2+), Zn(2+), Mn(2+), Pb(2+), Ni(2+), Fe(3+), Al(3+), Cr(3+) and so on induce basically no spectral changes, which constitute a highly selective and sensitive photoswitchable fluorescent probe toward Hg(2+). Furthermore, by means of laser confocal scanning microscopy experiments, it is demonstrated that this probe can be applied for live cell imaging and monitoring Hg(2+) in living lung cancer cells with satisfying results, which shows its value of potential application in environmental and biological systems. Copyright © 2014 Elsevier B.V. All rights reserved.

Sensitive detection of strong acidic condition by a novel rhodamine-based fluorescent pH chemosensor.

Science.gov (United States)

Tan, Jia-Lian; Yang, Ting-Ting; Liu, Yu; Zhang, Xue; Cheng, Shu-Jin; Zuo, Hua; He, Huawei

2016-05-01

A novel rhodamine-based fluorescent pH probe responding to extremely low pH values has been synthesized and characterized. This probe showed an excellent photophysical response to pH on the basis that the colorless spirocyclic structure under basic conditions opened to a colored and highly fluorescent form under extreme acidity. The quantitative relationship between fluorescence intensity and pH value (1.75-2.62) was consistent with the equilibrium equation pH = pKa + log[(Imax - I)/(I - Imin)]. This sensitive pH probe was also characterized with good reversibility and no interaction with interfering metal ions, and was successfully applied to image Escherichia coli under strong acidity. Copyright © 2015 John Wiley & Sons, Ltd.

Terbium fluorescence as a sensitive, inexpensive probe for UV-induced damage in nucleic acids

International Nuclear Information System (INIS)

El-Yazbi, Amira F.; Loppnow, Glen R.

2013-01-01

Graphical abstract: -- Highlights: •Simple, inexpensive, mix-and-read assay for positive detection of DNA damage. •Recognition of undamaged DNA via hybridization to a hairpin probe. •Terbium(III) fluorescence reports the amount of damage by binding to ssDNA. •Tb/hairpin is a highly selective and sensitive fluorescent probe for DNA damage. -- Abstract: Much effort has been focused on developing methods for detecting damaged nucleic acids. However, almost all of the proposed methods consist of multi-step procedures, are limited, require expensive instruments, or suffer from a high level of interferences. In this paper, we present a novel simple, inexpensive, mix-and-read assay that is generally applicable to nucleic acid damage and uses the enhanced luminescence due to energy transfer from nucleic acids to terbium(III) (Tb 3+ ). Single-stranded oligonucleotides greatly enhance the Tb 3+ emission, but duplex DNA does not. With the use of a DNA hairpin probe complementary to the oligonucleotide of interest, the Tb 3+ /hairpin probe is applied to detect ultraviolet (UV)-induced DNA damage. The hairpin probe hybridizes only with the undamaged DNA. However, the damaged DNA remains single-stranded and enhances the intrinsic fluorescence of Tb 3+ , producing a detectable signal directly proportional to the amount of DNA damage. This allows the Tb 3+ /hairpin probe to be used for sensitive quantification of UV-induced DNA damage. The Tb 3+ /hairpin probe showed superior selectivity to DNA damage compared to conventional molecular beacons probes (MBs) and its sensitivity is more than 2.5 times higher than MBs with a limit of detection of 4.36 ± 1.2 nM. In addition, this probe is easier to synthesize and more than eight times cheaper than MBs, which makes its use recommended for high-throughput, quantitative analysis of DNA damage

Visual and sensitive fluorescent sensing for ultratrace mercury ions by perovskite quantum dots.

Science.gov (United States)

Lu, Li-Qiang; Tan, Tian; Tian, Xi-Ke; Li, Yong; Deng, Pan

2017-09-15

Mercury ions sensing is an important issue for human health and environmental safety. A novel fluorescence nanosensor was designed for rapid visual detection of ultratrace mercury ions (Hg 2+ ) by using CH 3 NH 3 PbBr 3 perovskite quantum dots (QDs) based on the surface ion-exchange mechanism. The synthesized CH 3 NH 3 PbBr 3 QDs can emitt intense green fluorescence with high quantum yield of 50.28%, and can be applied for Hg 2+ sensing with the detection limit of 0.124 nM (24.87 ppt) in the range of 0 nM-100 nM. Furthermore, the interfering metal ions have no any influence on the fluorescence intensity of QDs, showing the perovskite QDs possess the high selectivity and sensitivity for Hg 2+ detection. The sensing mechanism of perovskite QDs for Hg 2+ is has also been investigated by XPS, EDX studies, showing Pb 2+ on the surface of perovskite QDs has been partially replaced by Hg 2+ . Spot plate test shows that the perovskite QDs can also be used for visual detection of Hg 2+ . Our research indicated the perovskite QDs are promising candidates for the visual fluorescence detection of environmental micropollutants. Copyright © 2017 Elsevier B.V. All rights reserved.

Ratio-metric sensor to detect riboflavin via fluorescence resonance energy transfer with ultrahigh sensitivity

Science.gov (United States)

Wang, Jilong; Su, Siheng; Wei, Junhua; Bahgi, Roya; Hope-Weeks, Louisa; Qiu, Jingjing; Wang, Shiren

2015-08-01

In this paper, a novel fluorescence resonance energy transfer (FRET) ration-metric fluorescent probe based on heteroatom N, S doped carbon dots (N, S-CDs) was developed to determine riboflavin in aqueous solutions. The ratio of two emission intensities at different wavelengths is applied to determine the concentration of riboflavin (RF). This method is more effective in reducing the background interference and fluctuation of diverse conditions. Therefore, this probe obtains high sensitivity with a low limit of detection (LOD) of 1.9 nM (0.7 ng/ml) which is in the highest level of all riboflavin detection approaches and higher than single wavelength intensity detection (1.9 μM). In addition, this sensor has a high selectivity of detecting riboflavin in deionized water (pH=7) with other biochemical like amino acids. Moreover, riboflavin in aqueous solution is very sensitive to sunlight and can be degraded to lumiflavin, which is toxic. Because the N, S doped carbon dots cannot serve as an energy donor for N, S doped carbon dots and lumiflavin system, this system makes it easy to determine whether the riboflavin is degraded or not, which is first to be reported. This platform may provide possibilities to build a new and facile fluorescence resonance energy transfer based sensor to detect analytes and metamorphous analytes in aqueous solution.

Molecularly imprinted fluorescent probe based on FRET for selective and sensitive detection of doxorubicin

Energy Technology Data Exchange (ETDEWEB)

Xu, Zhifeng, E-mail: 897061147@qq.com [College of Chemistry and Materials Science, Hengyang Normal University, Key Laboratory of Functional Organometallic Materials of Hunan Province University, Hengyang 421008 (China); Deng, Peihong; Li, Junhua [College of Chemistry and Materials Science, Hengyang Normal University, Key Laboratory of Functional Organometallic Materials of Hunan Province University, Hengyang 421008 (China); Xu, Li [Department of Applied Chemistry, College of Materials and Energy, South China Agricultural University, Guangzhou 510642 (China); Tang, Siping [College of Chemistry and Materials Science, Hengyang Normal University, Key Laboratory of Functional Organometallic Materials of Hunan Province University, Hengyang 421008 (China)

2017-04-15

Highlights: • FRET-based molecularly imprinted probe for detection of doxorubicin was prepared. • The detection limit of the probe was 13.8 nM for doxorubicin. • The FRET-based probe had a higher selectivity for the template than ordinary MIMs. - Abstract: In this work, a new type of fluorescent probe for detection of doxorubicin has been constructed by the combined use of fluorescence resonance energy transfer (FRET) technology and molecular imprinting technique (MIT). Using doxorubicin as the template, the molecularly imprinted polymer thin layer was fabricated on the surfaces of carbon dot (CD) modified silica by sol-gel polymerization. The excitation energy of the fluorescent donor (CDs) could be transferred to the fluorescent acceptor (doxorubicin). The FRET based fluorescent probe demonstrated high sensitivity and selectivity for doxorubicin. The detection limit was 13.8 nM. The fluorescent probe was successfully applied for detecting doxorubicin in doxorubicin-spiked plasmas with a recovery of 96.8–103.8%, a relative standard deviation (RSD) of 1.3–2.8%. The strategy for construction of FRET-based molecularly imprinted materials developed in this work is very promising for analytical applications.

A label-free fluorescence biosensor for highly sensitive detection of lectin based on carboxymethyl chitosan-quantum dots and gold nanoparticles.

Science.gov (United States)

Liu, Ziping; Liu, Hua; Wang, Lei; Su, Xingguang

2016-08-17

In this work, we report a novel label-free fluorescence "turn off-on" biosensor for lectin detection. The highly sensitive and selective sensing system is based on the integration of carboxymethyl chitosan (CM-CHIT), CuInS2 quantum dots (QDs) and Au nanoparticles (NPs). Firstly, CuInS2 QDs featuring carboxyl groups were directly synthesized via a hydrothermal synthesis method. Then, the carboxyl groups on the CuInS2 QDs surface were interacted with the amino groups (NH2), carboxyl groups (COOH) and hydroxyl groups (OH) within CM-CHIT polymeric chains via electrostatic interactions and hydrogen bonding to form CM-CHIT-QDs assemblies. Introduction of Au NPs could quench the fluorescence of CM-CHIT-QDs through electron and energy transfer. In the presence of lectin, lectin could bind exclusively with CM-CHIT-QDs by means of specific multivalent carbohydrate-protein interaction. Thus, the electron and energy transfer process between CM-CHIT-QDs and Au NPs was inhibited, and as a result, the fluorescence of CM-CHIT-QDs was effectively "turned on". Under the optimum conditions, there was a good linear relationship between the fluorescence intensity ratio I/I0 (I and I0 were the fluorescence intensity of CM-CHIT-QDs-Au NPs in the presence and absence of lectin, respectively) and lectin concentration in the range of 0.2-192.5 nmol L(-1), And the detection limit could be down to 0.08 nmol L(-1). Furthermore, the proposed biosensor was employed for the determination of lectin in fetal bovine serum samples with satisfactory results. Copyright © 2016 Elsevier B.V. All rights reserved.

Simple and sensitive fluorescence assay of restriction endonuclease on graphene oxide

International Nuclear Information System (INIS)

Gang, Jong Back

2015-01-01

Restriction endonucleases hydrolyze internal phosphodiester bonds at specific sites in a DNA sequence. These enzymes are essential in a variety of fields, such as biotechnology and clinical diagnostics. It is of great importance and necessity for the scientific and biomedical use of enzymes to measure endonuclease activity. In this study, graphene oxide (GO) has been used as a platform to measure enzyme activity with high sensitivity. To increase the detection sensitivity of Hinf I, the endonuclease-digested reaction was treated with exonuclease III (Exo III) and a fluorescence assay was conducted to measure the emission. Results showed that Exo III treatment enhanced 2.7-fold signal-to-background ratio for the detection of Hinf I compared with that done without Exo III in the presence of GO

Interpenetrated Binary Supramolecular Nanofibers for Sensitive Fluorescence Detection of Six Classes of Explosives.

Science.gov (United States)

Xiong, Wei; Zhu, Qijian; Gong, Yanjun; Wang, Chen; Che, Yanke; Zhao, Jincai

2018-04-03

In this work, we develop a sequential self-assembly approach to fabricate interpenetrated binary supramolecular nanofibers consisting of carbazole oligomer 1-cobalt(II) (1-Co 2+ ) coordination nanofibers and oligomer 2 nanofibers for the sensitive detection of six classes of explosives. When exposed to peroxide explosives (e.g., H 2 O 2 ), Co 2+ in 1-Co 2+ coordination nanofibers can be reduced to Co + that can transfer an electron to the excited 2 nanofibers and thereby quench their fluorescence. On the other hand, when exposed to the other five classes of explosives, the excited 2 nanofibers can transfer an electron to explosives to quench their fluorescence. On the basis of the distinct fluorescence quenching mechanisms, six classes of explosives can be sensitively detected. Herein, we provide a new strategy to design broad-band fluorescence sensors for a rich identification of threats.

A novel detection platform for parallel monitoring of DNA hybridization with high sensitivity and specificity

DEFF Research Database (Denmark)

Yi, Sun; Perch-Nielsen, Ivan R.; Wang, Zhenyu

We developed a high-sensitive platform to monior multiple hybridization events in real time. By creating a microoptical array in a polymeric chip, the system combine the excellent discriminative power of supercritical angle fluorescence (SAF) microscopy with high-throughput capabilities of microa......We developed a high-sensitive platform to monior multiple hybridization events in real time. By creating a microoptical array in a polymeric chip, the system combine the excellent discriminative power of supercritical angle fluorescence (SAF) microscopy with high-throughput capabilities...

Soy flour-derived carbon dots: facile preparation, fluorescence enhancement, and sensitive Fe{sup 3+} detection

Energy Technology Data Exchange (ETDEWEB)

Fang, Liyang; Xu, Qian [China University of Petroleum, State Key Laboratory of Heavy Oil Processing (China); Zheng, Xing [Bei Jing Sinen En-Tech Co., Ltd (China); Zhang, Weina; Zheng, Jingtang, E-mail: jtzheng03@163.com; Wu, Mingbo, E-mail: wumb@upc.edu.cn; Wu, Wenting, E-mail: wuwt@upc.edu.cn [China University of Petroleum, State Key Laboratory of Heavy Oil Processing (China)

2016-08-15

Soy flour-derived carbon quantum dots (C-dots) were successfully synthesized via a facile one-step hydrothermal approach. The as-prepared C-dots exhibit an average diameter of 2.5 nm and the crystalline lattices are consistent with graphitic carbons. Meanwhile, they show strong photoluminescence (quantum yield is 7.85 %), good water solubility, and high photostability. Importantly, structural defects of the C-dots were designed to obtain controllable fluorescence, which was achieved by changing the contents of N defects and O defects of C-dots. Our results indicate that N defects can more effectively enhance the fluorescence emission than O defects. As the preparation temperature increases, the N defects are fine-tuned by substituting for partial O defects, reducing nonradiative recombination and enhancing fluorescence intensity, which is further confirmed by surface passivation. Due to its fine photostability, high sensitivity, and good selectivity for Fe{sup 3+}, the as-prepared C-dots were used as fluorescence probes for detection of ferric ion. The detection limitation comes to 0.021 µM.

Ultrahigh sensitivity endoscopic camera using a new CMOS image sensor: providing with clear images under low illumination in addition to fluorescent images.

Science.gov (United States)

Aoki, Hisae; Yamashita, Hiromasa; Mori, Toshiyuki; Fukuyo, Tsuneo; Chiba, Toshio

2014-11-01

We developed a new ultrahigh-sensitive CMOS camera using a specific sensor that has a wide range of spectral sensitivity characteristics. The objective of this study is to present our updated endoscopic technology that has successfully integrated two innovative functions; ultrasensitive imaging as well as advanced fluorescent viewing. Two different experiments were conducted. One was carried out to evaluate the function of the ultrahigh-sensitive camera. The other was to test the availability of the newly developed sensor and its performance as a fluorescence endoscope. In both studies, the distance from the endoscopic tip to the target was varied and those endoscopic images in each setting were taken for further comparison. In the first experiment, the 3-CCD camera failed to display the clear images under low illumination, and the target was hardly seen. In contrast, the CMOS camera was able to display the targets regardless of the camera-target distance under low illumination. Under high illumination, imaging quality given by both cameras was quite alike. In the second experiment as a fluorescence endoscope, the CMOS camera was capable of clearly showing the fluorescent-activated organs. The ultrahigh sensitivity CMOS HD endoscopic camera is expected to provide us with clear images under low illumination in addition to the fluorescent images under high illumination in the field of laparoscopic surgery.

A novel dansyl-based fluorescent probe for highly selective detection of ferric ions.

Science.gov (United States)

Yang, Min; Sun, Mingtai; Zhang, Zhongping; Wang, Suhua

2013-02-15

A novel dansyl-based fluorescent probe was synthesized and characterized. It exhibits high selectivity and sensitivity towards Fe(3+) ion. This fluorescent probe is photostable, water soluble and pH insensitive. The limit of detection is found to be 0.62 μM. These properties make it a good fluorescent probe for Fe(3+) ion detection in both chemical and biological systems. Spike recovery test confirms its practical application in tap water samples. Copyright © 2012 Elsevier B.V. All rights reserved.

Development of an Ultrasonication-Assisted Extraction Based HPLC With a Fluorescence Method for Sensitive Determination of Aflatoxins in Highly Acidic Hibiscus sabdariffa.

Science.gov (United States)

Liu, Xiaofei; Ying, Guangyao; Sun, Chaonan; Yang, Meihua; Zhang, Lei; Zhang, Shanshan; Xing, Xiaoyan; Li, Qian; Kong, Weijun

2018-01-01

The high acidity and complex components of Hibiscus sabdariffa have provided major challenges for sensitive determination of trace aflatoxins. In this study, sample pretreatment of H. sabdariffa was systematically developed for sensitive high performance liquid chromatography-fluorescence detection (HPLC-FLD) after ultrasonication-assisted extraction, immunoaffinity column (IAC) clean-up and on-line post-column photochemical derivatization (PCD). Aflatoxins B 1 , B 2 , G 1 , G 2 were extracted from samples by using methanol/water (70:30, v/v ) with the addition of NaCl. The solutions were diluted 1:8 with 0.1 M phosphate buffer (pH 8.0) to negate the issues of high acidity and matrix interferences. The established method was validated with satisfactory linearity ( R > 0.999), sensitivity (limits of detection (LODs) and limits of quantitation (LOQs) of 0.15-0.65 and 0.53-2.18 μg/kg, respectively), precision (RSD sabdariffa samples indicated that one sample incubated with Aspergillus flavus was positive with aflatoxin B 1 (AFB 1 ) at 3.11 μg/kg. The strategy developed in this study also has the potential to reliably extract and sensitively detect more mycotoxins in other complex acidic matrices, such as traditional Chinese medicines, foodstuffs, etc.

Fluorescent QDs-polystyrene composite nanospheres for highly efficient and rapid protein antigen detection

Energy Technology Data Exchange (ETDEWEB)

Zhou, Changhua; Mao, Mao [Henan University, Key Laboratory for Special Functional Materials of the Ministry of Education (China); Yuan, Hang [Tsinghua University, Life Science Division, Graduate School at Shenzhen (China); Shen, Huaibin [Henan University, Key Laboratory for Special Functional Materials of the Ministry of Education (China); Wu, Feng; Ma, Lan, E-mail: malan@sz.tsinghua.edu.cn [Tsinghua University, Life Science Division, Graduate School at Shenzhen (China); Li, Lin Song, E-mail: lsli@henu.edu.cn [Henan University, Key Laboratory for Special Functional Materials of the Ministry of Education (China)

2013-09-15

In this paper, high-quality carboxyl-functionalized fluorescent (red, green, and blue emitting) nanospheres (46-103 nm) consisting of hydrophobic quantum dots (QDs) and polystyrene were prepared by a miniemulsion polymerization approach. This miniemulsion polymerization approach induced a homogeneous distribution and high aqueous-phase transport efficiency of fluorescent QDs in composite nanospheres, which proved the success of our encoding QDs strategy. The obtained fluorescent nanospheres exhibited high stability in aqueous solution under a wide range of pH, different salt concentrations, PBS buffer, and thermal treatment at 80 Degree-Sign C. Based on the red emitting composite nanosphere, we performed fluorescent lateral flow immunoassay (LFIA) strips for high-sensitivity and rapid alpha-fetal protein detection. The detection limit reached 0.1 ng/mL, which was 200 times higher than commercial colloidal gold-labeled LFIA strips, and it reached similar detection level in enzyme-linked immunosorbent assay kit.

Second and third generation voltage-sensitive fluorescent proteins for monitoring membrane potential

Directory of Open Access Journals (Sweden)

Amelie Perron

2009-06-01

Full Text Available Over the last decade, optical neuroimaging methods have been enriched by engineered biosensors derived from fluorescent protein (FP reporters fused to protein detectors that convert physiological signals into changes of intrinsic FP fluorescence. These FP-based indicators are genetically encoded, and hence targetable to specific cell populations within networks of heterologous cell types. Among this class of biosensors, the development of optical probes for membrane potential is both highly desirable and challenging. A suitable FP voltage sensor would indeed be a valuable tool for monitoring the activity of thousands of individual neurons simultaneously in a non-invasive manner. Previous prototypic genetically-encoded FP voltage indicators achieved a proof of principle but also highlighted several difficulties such as poor cell surface targeting and slow kinetics. Recently, we developed a new series of FRET-based Voltage-Sensitive Fluorescent Proteins (VSFPs, referred to as VSFP2s, with efficient targeting to the plasma membrane and high responsiveness to membrane potential signaling in excitable cells. In addition to these FRET-based voltage sensors, we also generated a third series of probes consisting of single FPs with response kinetics suitable for the optical imaging of fast neuronal signals. These newly available genetically-encoded reporters for membrane potential will be instrumental for future experimental approaches directed toward the understanding of neuronal network dynamics and information processing in the brain. Here, we review the development and current status of these novel fluorescent probes.

[Sensitive Determination of Chondroitin Sulfate by Fluorescence Recovery of an Anionic Aluminum Phthalocyanine-Cationic Surfactant Ion-Association Complex Used as a Fluorescent Probe Emitting at Red Region].

Science.gov (United States)

Chen, Lin; Huang, Ping; Yang, Hui-qing; Deng, Ya-bin; Guo, Meng-lin; Li, Dong-hui

2015-08-01

Determination of chondroitin sulfate in the biomedical field has an important value. The conventional methods for the assay of chondroitin sulfate are still unsatisfactory in sensitivity, selectivity or simplicity. This work aimed at developing a novel method for sensitive and selective determination of chondroitin sulfate by fluorimetry. We found that some kinds of cationic surfactants have the ability to quench the fluorescence of tetrasulfonated aluminum phthalocyanine (AlS4Pc), a strongly fluorescent compound which emits at red region, with high efficiency. But, the fluorescence of the above-mentioned fluorescence quenching system recovered significantly when chondroitin sulfate (CS) exits. Tetradecyl dimethyl benzyl ammonium chloride(TDBAC) which was screened from all of the candidates of cationic surfactants was chosen as the quencher because it shows the most efficient quenching effect. It was found that the fluorescence of AlS4Pc was extremely quenched by TDBAC because of the formation of association complex between AlS4Pc and TDBAC. Fluorescence of the association complex recovered dramatically after the addition of chondroitin sulfate (CS) due to the ability of chondroitin sulfate to shift the association equilibrium of the association, leading to the release of AlS4Pc, thus resulting in an increase in the fluorescence of the reaction system. Based on this phenomenon, a novel method with simplicity, accuracy and sensitivity was developed for quantitative determination of CS. Factors including the reaction time, influencing factors and the effect of coexisting substances were investigated and discussed. Under optimum conditions the linear range of the calibration curve was 0.20~10.0 μg · mL(-1). The detection limit for CS was 0.070 μg · mL(-1). The method has been applied to the analysis of practical samples with satisfied results. This work expands the applications of AlS4Pc in biomedical area.

Smartphone-Based Fluorescent Diagnostic System for Highly Pathogenic H5N1 Viruses

Science.gov (United States)

Yeo, Seon-Ju; Choi, Kyunghan; Cuc, Bui Thi; Hong, Nguyen Ngoc; Bao, Duong Tuan; Ngoc, Nguyen Minh; Le, Mai Quynh; Hang, Nguyen Le Khanh; Thach, Nguyen Co; Mallik, Shyam Kumar; Kim, Hak Sung; Chong, Chom-Kyu; Choi, Hak Soo; Sung, Haan Woo; Yu, Kyoungsik; Park, Hyun

2016-01-01

Field diagnostic tools for avian influenza (AI) are indispensable for the prevention and controlled management of highly pathogenic AI-related diseases. More accurate, faster and networked on-site monitoring is demanded to detect such AI viruses with high sensitivity as well as to maintain up-to-date information about their geographical transmission. In this work, we assessed the clinical and field-level performance of a smartphone-based fluorescent diagnostic device with an efficient reflective light collection module using a coumarin-derived dendrimer-based fluorescent lateral flow immunoassay. By application of an optimized bioconjugate, a smartphone-based diagnostic device had a two-fold higher detectability as compared to that of the table-top fluorescence strip reader for three different AI subtypes (H5N3, H7N1, and H9N2). Additionally, in a clinical study of H5N1-confirmed patients, the smartphone-based diagnostic device showed a sensitivity of 96.55% (28/29) [95% confidence interval (CI): 82.24 to 99.91] and a specificity of 98.55% (68/69) (95% CI: 92.19 to 99.96). The measurement results from the distributed individual smartphones were wirelessly transmitted via short messaging service and collected by a centralized database system for further information processing and data mining. Smartphone-based diagnosis provided highly sensitive measurement results for H5N1 detection within 15 minutes. Because of its high sensitivity, portability and automatic reporting feature, the proposed device will enable agile identification of patients and efficient control of AI dissemination. PMID:26877781

Smartphone-Based Fluorescent Diagnostic System for Highly Pathogenic H5N1 Viruses.

Science.gov (United States)

Yeo, Seon-Ju; Choi, Kyunghan; Cuc, Bui Thi; Hong, Nguyen Ngoc; Bao, Duong Tuan; Ngoc, Nguyen Minh; Le, Mai Quynh; Hang, Nguyen Le Khanh; Thach, Nguyen Co; Mallik, Shyam Kumar; Kim, Hak Sung; Chong, Chom-Kyu; Choi, Hak Soo; Sung, Haan Woo; Yu, Kyoungsik; Park, Hyun

2016-01-01

Field diagnostic tools for avian influenza (AI) are indispensable for the prevention and controlled management of highly pathogenic AI-related diseases. More accurate, faster and networked on-site monitoring is demanded to detect such AI viruses with high sensitivity as well as to maintain up-to-date information about their geographical transmission. In this work, we assessed the clinical and field-level performance of a smartphone-based fluorescent diagnostic device with an efficient reflective light collection module using a coumarin-derived dendrimer-based fluorescent lateral flow immunoassay. By application of an optimized bioconjugate, a smartphone-based diagnostic device had a two-fold higher detectability as compared to that of the table-top fluorescence strip reader for three different AI subtypes (H5N3, H7N1, and H9N2). Additionally, in a clinical study of H5N1-confirmed patients, the smartphone-based diagnostic device showed a sensitivity of 96.55% (28/29) [95% confidence interval (CI): 82.24 to 99.91] and a specificity of 98.55% (68/69) (95% CI: 92.19 to 99.96). The measurement results from the distributed individual smartphones were wirelessly transmitted via short messaging service and collected by a centralized database system for further information processing and data mining. Smartphone-based diagnosis provided highly sensitive measurement results for H5N1 detection within 15 minutes. Because of its high sensitivity, portability and automatic reporting feature, the proposed device will enable agile identification of patients and efficient control of AI dissemination.

Highly selective and sensitive fluorescence detection of Zn(2+) and Cd(2+) ions by using an acridine sensor.

Science.gov (United States)

Visscher, A; Bachmann, S; Schnegelsberg, C; Teuteberg, T; Mata, R A; Stalke, D

2016-04-07

Fluorescence spectroscopy investigations of the new acridine derivative bis(N,N-dimethylaminemethylene)acridine (3) show remarkable selectivity and sensitivity towards Zn(2+) and Cd(2+) ions in methanol and for the latter even in water. Through the chelation of the metal ions the present PET effect is quenched, significantly enhancing the emission intensity of the fluorophore. In solution, the bonding situation is studied by fluorescence and NMR spectroscopy, as well as ESI-TOF mass-spectrometry measurements. The solid state environment is investigated by X-ray diffraction and computational calculations. Here, we can show the complexation of the zinc and cadmium ions by the methylene bridged amine receptors as well as by the nitrogen atom of the acridine system.

Toehold-mediated strand displacement reaction triggered isothermal DNA amplification for highly sensitive and selective fluorescent detection of single-base mutation.

Science.gov (United States)

Zhu, Jing; Ding, Yongshun; Liu, Xingti; Wang, Lei; Jiang, Wei

2014-09-15

Highly sensitive and selective detection strategy for single-base mutations is essential for risk assessment of malignancy and disease prognosis. In this work, a fluorescent detection method for single-base mutation was proposed based on high selectivity of toehold-mediated strand displacement reaction (TSDR) and powerful signal amplification capability of isothermal DNA amplification. A discrimination probe was specially designed with a stem-loop structure and an overhanging toehold domain. Hybridization between the toehold domain and the perfect matched target initiated the TSDR along with the unfolding of the discrimination probe. Subsequently, the target sequence acted as a primer to initiate the polymerization and nicking reactions, which released a great abundant of short sequences. Finally, the released strands were annealed with the reporter probe, launching another polymerization and nicking reaction to produce lots of G-quadruplex DNA, which could bind the N-methyl mesoporphyrin IX to yield an enhanced fluorescence response. However, when there was even a single base mismatch in the target DNA, the TSDR was suppressed and so subsequent isothermal DNA amplification and fluorescence response process could not occur. The proposed approach has been successfully implemented for the identification of the single-base mutant sequences in the human KRAS gene with a detection limit of 1.8 pM. Furthermore, a recovery of 90% was obtained when detecting the target sequence in spiked HeLa cells lysate, demonstrating the feasibility of this detection strategy for single-base mutations in biological samples. Copyright © 2014 Elsevier B.V. All rights reserved.

DNA-functionalized gold nanoparticle-based fluorescence polarization for the sensitive detection of silver ions.

Science.gov (United States)

Wang, Gongke; Wang, Shuangli; Yan, Changling; Bai, Guangyue; Liu, Yufang

2018-04-05

Despite their practical applications, Ag + ions are environmental pollutants and affect human health. So the effective detection methods of Ag + ions are imperative. Herein, we developed a simple, sensitive, selective, and cost-effective fluorescence polarization sensor for Ag + detection in aqueous solution using thiol-DNA-functionalized gold nanoparticles (AuNPs). In this sensing strategy, Ag + ions can specifically interact with a cytosine-cytosine (CC) mismatch in DNA duplexes and form stable metal-mediated cytosine-Ag + -cytosine (C-Ag + -C) base pairs. The formation of the C-Ag + -C complex results in evident changes in the molecular volume and fluorescence polarization signal. To achieve our aims, we prepared two complementary DNA strands containing C-base mismatches (probe A: 5'-SH-A 10 -TACCACTCCTCAC-3' and probe B: 5'-TCCTCACCAGTCCTA-FAM-3'). The stable hybridization between probe A and probe B occurs with the formation of the C-Ag + -C complex in the presence of Ag + ions, leading to obvious fluorescence quenching in comparison to the system without AuNP enhancement. The assay can be used to identify nanomolar levels of Ag + within 6 min at room temperature, and has extremely high specificity for Ag + , even in the presence of higher concentrations of interfering metal ions. Furthermore, the sensor was successfully applied to the detection of Ag + ions in environmental water samples and showed excellent selectivity and high sensitivity, implying its promising application in the future. Copyright © 2018 Elsevier B.V. All rights reserved.

Sensitivity and specificity of fluorescence microlymphography for detecting lymphedema of the lower extremity.

Science.gov (United States)

Keo, Hong H; Schilling, Marianne; Büchel, Roland; Gröchenig, Ernst; Engelberger, Rolf P; Willenberg, Torsten; Baumgartner, Iris; Gretener, Silvia B

2013-06-01

Fluorescence microlymphography (FML) is used to visualize the lymphatic capillaries. A maximum spread of the fluorescence dye of ≥ 12 mm has been suggested for the diagnosis of lymphedema. However, data on sensitivity and specificity are lacking. The aim of this study was to investigate the accuracy of FML for diagnosing lymphedema in patients with leg swelling. Patients with lower extremity swelling were clinically assessed and separated into lymphedema and non-lymphatic edema groups. FML was studied in all affected legs and the maximum spread of lymphatic capillaries was measured. Test accuracy and receiver operator characteristic (ROC) analysis was performed to assess possible threshold values that predict lymphedema. Between March 2008 and August 2011 a total of 171 patients (184 legs) with a median age of 43.5 (IQR 24, 54) years were assessed. Of those, 94 (51.1%) legs were diagnosed with lymphedema. The sensitivity, specificity, positive and negative likelihood ratio and positive and negative predictive value were 87%, 64%, 2.45, 0.20, 72% and 83% for the 12-mm cut-off level and 79%, 83%, 4.72, 0.26, 83% and 79% for the 14-mm cut-off level, respectively. The area under the ROC curve was 0.82 (95% CI: 0.76, 0.88). Sensitivity was higher in the secondary versus primary lymphedema (95.0% vs 74.3%, p = 0.045). No major adverse events were observed. In conclusion, FML is a simple and safe technique for detecting lymphedema in patients with leg swelling. A cut-off level of ≥ 14-mm maximum spread has a high sensitivity and high specificity of detecting lymphedema and should be chosen.

Tungsten disulfide nanosheet and exonuclease III co-assisted amplification strategy for highly sensitive fluorescence polarization detection of DNA glycosylase activity

International Nuclear Information System (INIS)

Zhao, Jingjin; Ma, Yefei; Kong, Rongmei; Zhang, Liangliang; Yang, Wen; Zhao, Shulin

2015-01-01

Herein, we introduced a tungsten disulfide (WS 2 ) nanosheet and exonuclease III (Exo III) co-assisted signal amplification strategy for highly sensitive fluorescent polarization (FP) assay of DNA glycosylase activity. Two DNA glycosylases, uracil-DNA glycosylase (UDG) and human 8-oxoG DNA glycosylase 1 (hOGG1), were tested. A hairpin-structured probe (HP) which contained damaged bases in the stem was used as the substrate. The removal of damaged bases from substrate by DNA glycosylase would lower the melting temperature of HP. The HP was then opened and hybridized with a FAM dye-labeled single strand DNA (DP), generating a duplex with a recessed 3′-terminal of DP. This design facilitated the Exo III-assisted amplification by repeating the hybridization and digestion of DP, liberating numerous FAM fluorophores which could not be adsorbed on WS 2 nanosheet. Thus, the final system exhibited a small FP signal. However, in the absence of DNA glycosylases, no hybridization between DP and HP was occurred, hampering the hydrolysis of DP by Exo III. The intact DP was then adsorbed on the surface of WS 2 nanosheet that greatly amplified the mass of the labeled-FAM fluorophore, resulting in a large FP value. With the co-assisted amplification strategy, the sensitivity was substantially improved. In addition, this method was applied to detect UDG activity in cell extracts. The study of the inhibition of UDG was also performed. Furthermore, this method is simple in design, easy in implementation, and selective, which holds potential applications in the DNA glycosylase related mechanism research and molecular diagnostics. - Highlights: • A fluorescence polarization strategy for DNA glycosylase activity detection was developed. • The present method was based on WS 2 nanosheet and exonuclease III co-assisted signal amplification. • A high sensitivity and desirable selectivity were achieved. • This method provides a promising universal platform for DNA glycosylase

Tungsten disulfide nanosheet and exonuclease III co-assisted amplification strategy for highly sensitive fluorescence polarization detection of DNA glycosylase activity

Energy Technology Data Exchange (ETDEWEB)

Zhao, Jingjin; Ma, Yefei [Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources of Education Ministry, Guangxi Normal University, Guilin, 541004 (China); Kong, Rongmei [The Key Laboratory of Life-Organic Analysis, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu, Shandong 273165 (China); Zhang, Liangliang, E-mail: liangzhang319@163.com [Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources of Education Ministry, Guangxi Normal University, Guilin, 541004 (China); Yang, Wen; Zhao, Shulin [Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources of Education Ministry, Guangxi Normal University, Guilin, 541004 (China)

2015-08-05

Herein, we introduced a tungsten disulfide (WS{sub 2}) nanosheet and exonuclease III (Exo III) co-assisted signal amplification strategy for highly sensitive fluorescent polarization (FP) assay of DNA glycosylase activity. Two DNA glycosylases, uracil-DNA glycosylase (UDG) and human 8-oxoG DNA glycosylase 1 (hOGG1), were tested. A hairpin-structured probe (HP) which contained damaged bases in the stem was used as the substrate. The removal of damaged bases from substrate by DNA glycosylase would lower the melting temperature of HP. The HP was then opened and hybridized with a FAM dye-labeled single strand DNA (DP), generating a duplex with a recessed 3′-terminal of DP. This design facilitated the Exo III-assisted amplification by repeating the hybridization and digestion of DP, liberating numerous FAM fluorophores which could not be adsorbed on WS{sub 2} nanosheet. Thus, the final system exhibited a small FP signal. However, in the absence of DNA glycosylases, no hybridization between DP and HP was occurred, hampering the hydrolysis of DP by Exo III. The intact DP was then adsorbed on the surface of WS{sub 2} nanosheet that greatly amplified the mass of the labeled-FAM fluorophore, resulting in a large FP value. With the co-assisted amplification strategy, the sensitivity was substantially improved. In addition, this method was applied to detect UDG activity in cell extracts. The study of the inhibition of UDG was also performed. Furthermore, this method is simple in design, easy in implementation, and selective, which holds potential applications in the DNA glycosylase related mechanism research and molecular diagnostics. - Highlights: • A fluorescence polarization strategy for DNA glycosylase activity detection was developed. • The present method was based on WS{sub 2} nanosheet and exonuclease III co-assisted signal amplification. • A high sensitivity and desirable selectivity were achieved. • This method provides a promising universal platform for DNA

Photosynthesis and chlorophyll fluorescence reaction to different shade stresses of weak light sensitive maize

International Nuclear Information System (INIS)

Wang, J.; Li, F.; Shi, Z.; Huang, H.; Jia, S.

2017-01-01

A split-plot experimental study was conducted to evaluate the effect of different shade stresses on photosynthesis and chlorophyll fluorescence of maize leaves.The experiment was designed on the south farm of Special Corn Institute, Shenyang Agricultural University, China.Data was collected from the day maize tasseled (Jul. 21) to the beginning of grouting (Aug.12 ) under 18%, 28%, 38%, 60%, and 75% shade stress to determine indexes such as photosynthesis and chlorophyll fluorescence after 15 days of shade treatment. Pairs of near-isogenic lines (NILs) of Shennong 98A (a barren stalk inbred line) and Shennong 98B (an un-barren stalk inbred line) were used as experimental materials to further reveal photosynthetic mechanisms of weak light sensitive maize when exposed to weak light conditions. Thus, a foundation was established for high density-resistant (shade resistant) corn breeding,while identifying weak light sensitive varieties. After shading treatment, chlorophyll a and total chlorophyll content of both varieties increased, chlorophyll b content first increased, followed by a decrease, while the net photosynthetic rate and stomatal conductance showed a gradually decreasing trend. The changing trends of photochemical quenching coefficient(qp) and effective quantum yield of PSII photochemistry (FPSII)were similar, FPSII and qP increased significantly as shading stress increased from 18% to 38%;however, FPSII and qP declined significantly under 60% and 75% shading stresses. The changing trend of NPQ was opposite to FPSII and qP. A comparison of both inbred lines showed that photosynthesis and chlorophyll fluorescence characteristics of Shennong 98B were superior to Shennong 98A. This study revealed the relationships between weak light sensitive lines and shade intensities by comparing differences in photosynthesis and chlorophyll fluorescence parameters. (author)

Automatic and integrated micro-enzyme assay (AIμEA) platform for highly sensitive thrombin analysis via an engineered fluorescence protein-functionalized monolithic capillary column.

Science.gov (United States)

Lin, Lihua; Liu, Shengquan; Nie, Zhou; Chen, Yingzhuang; Lei, Chunyang; Wang, Zhen; Yin, Chao; Hu, Huiping; Huang, Yan; Yao, Shouzhuo

2015-04-21

Nowadays, large-scale screening for enzyme discovery, engineering, and drug discovery processes require simple, fast, and sensitive enzyme activity assay platforms with high integration and potential for high-throughput detection. Herein, a novel automatic and integrated micro-enzyme assay (AIμEA) platform was proposed based on a unique microreaction system fabricated by a engineered green fluorescence protein (GFP)-functionalized monolithic capillary column, with thrombin as an example. The recombinant GFP probe was rationally engineered to possess a His-tag and a substrate sequence of thrombin, which enable it to be immobilized on the monolith via metal affinity binding, and to be released after thrombin digestion. Combined with capillary electrophoresis-laser-induced fluorescence (CE-LIF), all the procedures, including thrombin injection, online enzymatic digestion in the microreaction system, and label-free detection of the released GFP, were integrated in a single electrophoretic process. By taking advantage of the ultrahigh loading capacity of the AIμEA platform and the CE automatic programming setup, one microreaction column was sufficient for many times digestion without replacement. The novel microreaction system showed significantly enhanced catalytic efficiency, about 30 fold higher than that of the equivalent bulk reaction. Accordingly, the AIμEA platform was highly sensitive with a limit of detection down to 1 pM of thrombin. Moreover, the AIμEA platform was robust and reliable to detect thrombin in human serum samples and its inhibition by hirudin. Hence, this AIμEA platform exhibits great potential for high-throughput analysis in future biological application, disease diagnostics, and drug screening.

Expression of pH-sensitive green fluorescent protein in Arabidopsis thaliana

Science.gov (United States)

Moseyko, N.; Feldman, L. J.

2001-01-01

This is the first report on using green fluorescent protein (GFP) as a pH reporter in plants. Proton fluxes and pH regulation play important roles in plant cellular activity and therefore, it would be extremely helpful to have a plant gene reporter system for rapid, non-invasive visualization of intracellular pH changes. In order to develop such a system, we constructed three vectors for transient and stable transformation of plant cells with a pH-sensitive derivative of green fluorescent protein. Using these vectors, transgenic Arabidopsis thaliana and tobacco plants were produced. Here the application of pH-sensitive GFP technology in plants is described and, for the first time, the visualization of pH gradients between different developmental compartments in intact whole-root tissues of A. thaliana is reported. The utility of pH-sensitive GFP in revealing rapid, environmentally induced changes in cytoplasmic pH in roots is also demonstrated.

Analysis of Light Gathering Abilities of Dynamically Solidified Micro-lenses, and Their Implementation to Improve Sensitivity of Fluorescent PCR Micro-detectors.

Science.gov (United States)

Wu, Jian; Guo, Wei; Wang, Chunyan; Yu, Kuanxin; Chen, Tao; Li, Yinghui

2015-06-01

Fluorescent polymerase chain reaction (PCR) is becoming the preferred method of quantitative analysis due to its high specificity and sensitivity. We propose to use a new kind of micro-lens, dynamically solidified with optic glue, to improve the sensitivity of fluorescent PCR micro-detector. We developed light ray track equations for these lenses and used them to calculate relative light intensity distribution curve for stimulation lenses and illumination point probability distribution curve for detection lenses. We manufactured dynamically solidified micro-lenses using optic glue NOA61, and measured their light gathering ability. Lenses with radius/thickness (R/H) ratio of 4 reached light focusing ratio of 3.85 (stimulation lens) and photon collection efficiency of 0.86 (detection lens). We then used dynamically solidified lenses in PCR fluorescence micro-detector and analyzed their effect on the detector sensitivity. We showed that the use of dynamically solidified micro-lenses with R/H = 4 resulted in over 4.4-fold increased sensitivity of the detector.

Sensitive and Selective Ratiometric Fluorescence Probes for Detection of Intracellular Endogenous Monoamine Oxidase A.

Science.gov (United States)

Wu, Xiaofeng; Li, Lihong; Shi, Wen; Gong, Qiuyu; Li, Xiaohua; Ma, Huimin

2016-01-19

Monoamine oxidase A (MAO-A) is known to widely exist in most cell lines in the body, and its dysfunction (unusually high or low levels of MAO-A) is thought to be responsible for several psychiatric and neurological disorders. Thus, a sensitive and selective method for evaluating the relative MAO-A levels in different live cells is urgently needed to better understand the function of MAO-A, but to our knowledge such a method is still lacking. Herein, we rationally design two new ratiometric fluorescence probes (1 and 2) that can sensitively and selectively detect MAO-A. The probes are constructed by incorporating a recognition group of propylamine into the fluorescent skeleton of 1,8-naphthalimide, and the detection mechanism is based on amine oxidation and β-elimination to release the fluorophore (4-hydroxy-N-butyl-1,8-naphthalimide), which is verified by HPLC analysis. Reaction of the probes with MAO-A produces a remarkable fluorescence change from blue to green, and the ratio of fluorescence intensity at 550 and 454 nm is directly proportional to the concentration of MAO-A in the ranges of 0.5-1.5 and 0.5-2.5 μg/mL with detection limits of 1.1 and 10 ng/mL (k = 3) for probes 1 and 2, respectively. Surprisingly, these probes show strong fluorescence responses to MAO-A but almost none to MAO-B (one of two isoforms of MAO), indicating superior ability to distinguish MAO-A from MAO-B. The high specificity of the probes for MAO-A over MAO-B is further supported by different inhibitor experiments. Moreover, probe 1 displays higher sensitivity than probe 2 and is thus investigated to image the relative MAO-A levels in different live cells, such as HeLa and NIH-3T3 cells. It is found that the concentration of endogenous MAO-A in HeLa cells is approximately 1.8 times higher than that in NIH-3T3 cells, which is validated by the result from an ELISA kit. Additionally, the proposed probes may find more uses in the specific detection of MAO-A between the two isoforms of MAO

Silver Nanoparticle-Based Fluorescence-Quenching Lateral Flow Immunoassay for Sensitive Detection of Ochratoxin A in Grape Juice and Wine

Science.gov (United States)

Jiang, Hu; Li, Xiangmin; Xiong, Ying; Pei, Ke; Nie, Lijuan; Xiong, Yonghua

2017-01-01

A silver nanoparticle (AgNP)-based fluorescence-quenching lateral flow immunoassay with competitive format (cLFIA) was developed for sensitive detection of ochratoxin A (OTA) in grape juice and wine samples in the present study. The Ru(phen)32+-doped silica nanoparticles (RuNPs) were sprayed on the test and control line zones as background fluorescence signals. The AgNPs were designed as the fluorescence quenchers of RuNPs because they can block the exciting light transferring to the RuNP molecules. The proposed method exhibited high sensitivity for OTA detection, with a detection limit of 0.06 µg/L under optimized conditions. The method also exhibited a good linear range for OTA quantitative analysis from 0.08 µg/L to 5.0 µg/L. The reliability of the fluorescence-quenching cLFIA method was evaluated through analysis of the OTA-spiked red grape wine and juice samples. The average recoveries ranged from 88.0% to 110.0% in red grape wine and from 92.0% to 110.0% in grape juice. Meanwhile, less than a 10% coefficient variation indicated an acceptable precision of the cLFIA method. In summary, the new AgNP-based fluorescence-quenching cLFIA is a simple, rapid, sensitive, and accurate method for quantitative detection of OTA in grape juice and wine or other foodstuffs. PMID:28264472

Silver Nanoparticle-Based Fluorescence-Quenching Lateral Flow Immunoassay for Sensitive Detection of Ochratoxin A in Grape Juice and Wine

Directory of Open Access Journals (Sweden)

Hu Jiang

2017-02-01

Full Text Available A silver nanoparticle (AgNP-based fluorescence-quenching lateral flow immunoassay with competitive format (cLFIA was developed for sensitive detection of ochratoxin A (OTA in grape juice and wine samples in the present study. The Ru(phen 3 2 + -doped silica nanoparticles (RuNPs were sprayed on the test and control line zones as background fluorescence signals. The AgNPs were designed as the fluorescence quenchers of RuNPs because they can block the exciting light transferring to the RuNP molecules. The proposed method exhibited high sensitivity for OTA detection, with a detection limit of 0.06 µg/L under optimized conditions. The method also exhibited a good linear range for OTA quantitative analysis from 0.08 µg/L to 5.0 µg/L. The reliability of the fluorescence-quenching cLFIA method was evaluated through analysis of the OTA-spiked red grape wine and juice samples. The average recoveries ranged from 88.0% to 110.0% in red grape wine and from 92.0% to 110.0% in grape juice. Meanwhile, less than a 10% coefficient variation indicated an acceptable precision of the cLFIA method. In summary, the new AgNP-based fluorescence-quenching cLFIA is a simple, rapid, sensitive, and accurate method for quantitative detection of OTA in grape juice and wine or other foodstuffs.

Facile and sensitive determination of N-nitrosamines in food samples by high-performance liquid chromatography via combining fluorescent labeling with dispersive liquid-liquid microextraction.

Science.gov (United States)

Lu, Shuaimin; Wu, Di; Li, Guoliang; Lv, Zhengxian; Gong, Peiwei; Xia, Lian; Sun, Zhiwei; Chen, Guang; Chen, Xuefeng; You, Jinmao; Wu, Yongning

2017-11-01

The intake of N-nitrosamines (NAs) from foodstuffs is considered to be an important influence factor for several cancers. But the rapid and sensitive screening of NAs remains a challenge in the field of food safety. Inspired by that, a sensitive and rapid method was demonstrated for determination of five NAs (Nitrosopyrrolidine, Nitrosodimethylamine, Nitrosodiethylamine, Nitrosodipropylamine and Nitrosodibutylamine) using dispersive liquid-liquid microextraction (DLLME) followed by high-performance liquid chromatography with fluorescence detection (HPLC-FLD). The NAs were firstly denitrosated and labeled by 2-(11H-benzo[a]carbazol-11-yl) ethyl carbonochloridate (BCEC-Cl) and finally enriched by DLLME. Furthermore, the main DLLME conditions were optimized systematically. Under the optimal conditions, satisfactory limits of detection (LODs) were obtained with a range of 0.01-0.07ngg -1 , which were significantly lower than the reported methods. The developed method showed many merits including rapidity, simplicity, high sensitivity and excellent selectivity, which shows a broad prospect in food safety analysis. Copyright © 2017 Elsevier Ltd. All rights reserved.

DHA-fluorescent probe is sensitive to membrane order and reveals molecular adaptation of DHA in ordered lipid microdomains☆

Science.gov (United States)

Teague, Heather; Ross, Ron; Harris, Mitchel; Mitchell, Drake C.; Shaikh, Saame Raza

2012-01-01

Docosahexaenoic acid (DHA) disrupts the size and order of plasma membrane lipid microdomains in vitro and in vivo. However, it is unknown how the highly disordered structure of DHA mechanistically adapts to increase the order of tightly packed lipid microdomains. Therefore, we studied a novel DHA-Bodipy fluorescent probe to address this issue. We first determined if the DHA-Bodipy probe localized to the plasma membrane of primary B and immortal EL4 cells. Image analysis revealed that DHA-Bodipy localized into the plasma membrane of primary B cells more efficiently than EL4 cells. We then determined if the probe detected changes in plasma membrane order. Quantitative analysis of time-lapse movies established that DHA-Bodipy was sensitive to membrane molecular order. This allowed us to investigate how DHA-Bodipy physically adapted to ordered lipid microdomains. To accomplish this, we employed steady-state and time-resolved fluorescence anisotropy measurements in lipid vesicles of varying composition. Similar to cell culture studies, the probe was highly sensitive to membrane order in lipid vesicles. Moreover, these experiments revealed, relative to controls, that upon incorporation into highly ordered microdomains, DHA-Bodipy underwent an increase in its fluorescence lifetime and molecular order. In addition, the probe displayed a significant reduction in its rotational diffusion compared to controls. Altogether, DHA-Bodipy was highly sensitive to membrane order and revealed for the first time that DHA, despite its flexibility, could become ordered with less rotational motion inside ordered lipid microdomains. Mechanistically, this explains how DHA acyl chains can increase order upon formation of lipid microdomains in vivo. PMID:22841541

Highly sensitive and selective detection of Pb2+ using a turn-on fluorescent aptamer DNA silver nanoclusters sensor.

Science.gov (United States)

Zhang, Baozhu; Wei, Chunying

2018-05-15

A novel turn-on fluorescent biosensor has been constructed using C-PS2.M-DNA-templated silver nanoclusters (Ag NCs) with an average diameter of about 1 nm. The proposed approach presents a low-toxic, simple, sensitive, and selective detection for Pb 2+ . The fluorescence intensity of C-PS2.M-DNA-Ag NCs enhances significantly in the presence of Pb 2+ , which is attributed to the special interaction between Pb 2+ and its aptamer DNA PS2.M. Pb 2+ induces the aptamer to form G-quadruplex and makes two darkish DNA/Ag NCs located at the 3' and 5' terminus close, resulting in the fluorescence light-up. Moreover, Pb 2+ can be detected as low as 3.0 nM within a good linear range from 5 to 50 nM (R = 0.9862). Furthermore, the application for detection of Pb 2+ in real water samples further demonstrates the reliability of the sensor. Thus, this sensor system shows a potential application for monitoring Pb 2+ in environmental samples. Copyright © 2018 Elsevier B.V. All rights reserved.

A sensitive fluorescent sensor for quantification of alpha-fetoprotein based on immunosorbent assay and click chemistry.

Science.gov (United States)

Xie, Qunfang; Weng, Xiuhua; Lu, Lijun; Lin, Zhenyu; Xu, Xiongwei; Fu, Caili

2016-03-15

A novel fluoresencent immunosensor for determination of cancer biomarkers such as alpha-fetoprotein (AFP) was designed by utilizing both the high specificity of antigen-antibody sandwich structure and the high sensitivity of the click chemistry based fluorescence detection. Instead of an enzyme or fluorophore, the CuO nanoparticles are labeled on the detection antibody, which was not susceptible to the change of the external environments. The CuO nanoparticles which were modified on the sandwich structure can be dissolved to produce Cu(2+) ions with the help of HCl and then the Cu(2+) ions were reduced by sodium ascorbate to produce Cu(+) ions which triggered the Cu(+) catalyzed alkyne-azide cycloaddition (CuAAC) reaction between the weak fluorescent compound (3-azido-7-hydroxycoumarin) and propargyl alcohol to form a strong fluorescent compound. A good linear relationship was observed between the fluorescence increase factor of the system and the concentration of AFP in the range of 0.025-5.0 ng/mL with a detection limit of 12 pg/mL (S/N=3). The proposed fluorescent sensor had been applied to detect AFP in the human serum samples and gave satisfactory results. Copyright © 2015 Elsevier B.V. All rights reserved.

High yield fabrication of fluorescent nanodiamonds

International Nuclear Information System (INIS)

Boudou, Jean-Paul; Curmi, Patrick A; Jelezko, Fedor; Wrachtrup, Joerg; Balasubramanian, Gopalakrischnan; Reuter, Rolf; Aubert, Pascal; Sennour, Mohamed; Thorel, Alain; Gaffet, Eric

2009-01-01

A new fabrication method to produce homogeneously fluorescent nanodiamonds with high yields is described. The powder obtained by high energy ball milling of fluorescent high pressure, high temperature diamond microcrystals was converted in a pure concentrated aqueous colloidal dispersion of highly crystalline ultrasmall nanoparticles with a mean size less than or equal to 10 nm. The whole fabrication yield of colloidal quasi-spherical nanodiamonds was several orders of magnitude higher than those previously reported starting from microdiamonds. The results open up avenues for the industrial cost-effective production of fluorescent nanodiamonds with well-controlled properties.

High yield fabrication of fluorescent nanodiamonds

Energy Technology Data Exchange (ETDEWEB)

Boudou, Jean-Paul; Curmi, Patrick A [Structure and Activity of Normal and Pathological Biomolecules-INSERM/UEVE U829, Universite d' Evry-Val d' Essonne, Batiment Maupertuis, Rue du pere Andre Jarlan, F-91025 Evry (France); Jelezko, Fedor; Wrachtrup, Joerg; Balasubramanian, Gopalakrischnan; Reuter, Rolf [3.Physikalisches Institut, University of Stuttgart, Pfaffenwaldring 57, D-70550 Stuttgart (Germany); Aubert, Pascal [Nanometric Media Laboratory, Universite d' Evry-Val d' Essonne, Batiment Maupertuis, Rue du pere Andre Jarlan, F-91025 Evry (France); Sennour, Mohamed; Thorel, Alain [Centre des Materiaux, Mines Paris, ParisTech, BP 87, F-91000 Evry (France); Gaffet, Eric [Nanomaterials Research Group-UMR 5060, CNRS, UTBM, Site de Sevenans, F-90010 Belfort (France)], E-mail: jpb.cnrs@free.fr, E-mail: pcurmi@univ-evry.fr, E-mail: f.jelezko@physik.uni-stuttgart.de

2009-06-10

A new fabrication method to produce homogeneously fluorescent nanodiamonds with high yields is described. The powder obtained by high energy ball milling of fluorescent high pressure, high temperature diamond microcrystals was converted in a pure concentrated aqueous colloidal dispersion of highly crystalline ultrasmall nanoparticles with a mean size less than or equal to 10 nm. The whole fabrication yield of colloidal quasi-spherical nanodiamonds was several orders of magnitude higher than those previously reported starting from microdiamonds. The results open up avenues for the industrial cost-effective production of fluorescent nanodiamonds with well-controlled properties.

A Low-Cost, High-Performance System for Fluorescence Lateral Flow Assays

Directory of Open Access Journals (Sweden)

Linda G. Lee

2013-10-01

Full Text Available We demonstrate a fluorescence lateral flow system that has excellent sensitivity and wide dynamic range. The illumination system utilizes an LED, plastic lenses and plastic and colored glass filters for the excitation and emission light. Images are collected on an iPhone 4. Several fluorescent dyes with long Stokes shifts were evaluated for their signal and nonspecific binding in lateral flow. A wide range of values for the ratio of signal to nonspecific binding was found, from 50 for R-phycoerythrin (R-PE to 0.15 for Brilliant Violet 605. The long Stokes shift of R-PE allowed the use of inexpensive plastic filters rather than costly interference filters to block the LED light. Fluorescence detection with R-PE and absorbance detection with colloidal gold were directly compared in lateral flow using biotinylated bovine serum albumen (BSA as the analyte. Fluorescence provided linear data over a range of 0.4–4,000 ng/mL with a 1,000-fold signal change while colloidal gold provided non-linear data over a range of 16–4,000 ng/mL with a 10-fold signal change. A comparison using human chorionic gonadotropin (hCG as the analyte showed a similar advantage in the fluorescent system. We believe our inexpensive yet high-performance platform will be useful for providing quantitative and sensitive detection in a point-of-care setting.

Highly selective rhodamine-based fluorescence turn-on chemosensor for Al3+ ion

Science.gov (United States)

Manjunath, Rangasamy; Kannan, Palaninathan

2018-05-01

A new rhodamine-based colorimetric and fluorescent turn-on chemosensor (L) has been designed and synthesized for selective and sensitive detection of Al3+ ion. The sensing behavior toward metal ion was investigated by UV/Vis and fluorescence spectroscopy. Upon addition of Al3+ ion to solution of L provided a visual color change as well as significantly fluorescent enhancement, while other metal ions including Na+, Mg2+, K+, Mn2+, Fe3+, Ni2+, Cu2+, Zn2+, Pb2+, Cd2+ and Hg2+ ions fails to generate a distinct color and spectral changes, the distinct color change and rapid switch-on fluorescence also provide naked eye detection for Al3+ ion. The mechanism involved equilibrium between non-fluorescent spirocyclic form and highly fluorescent ring open form process was utilized and 1:2 stoichiometry for L-Al3+ complex formed with an association constant of 1.42 × 103 M-1. Moreover, chemosensor L was applied for living cell imaging and confirmed that can be used as a fluorescent probe for monitoring Al3+ ion in living cells.

Highly efficient fluorescence sensing with hollow core photonic crystal fibers

DEFF Research Database (Denmark)

Smolka, Stephan; Barth, Michael; Benson, Oliver

2008-01-01

We investigate hollow core photonic crystal fibers for ultra-sensitive fluorescence detection by selectively infiltrating the central hole with fluorophores. Dye concentrations down to 10(-9) M can be detected using only nanoliter sample volumes.......We investigate hollow core photonic crystal fibers for ultra-sensitive fluorescence detection by selectively infiltrating the central hole with fluorophores. Dye concentrations down to 10(-9) M can be detected using only nanoliter sample volumes....

Sensitive and background-free determination of thiols from wastewater samples by MOF-5 extraction coupled with high-performance liquid chromatography with fluorescence detection using a novel fluorescence probe of carbazole-9-ethyl-2-maleimide.

Science.gov (United States)

Lv, Zhengxian; Sun, Zhiwei; Song, Cuihua; Lu, Shuaimin; Chen, Guang; You, Jinmao

2016-12-01

A sensitive and background-free pre-column derivatization method for the determination of thiol compounds using metal-organic framework material (MOF-5) as dispersive solid-phase extraction (DSPE) adsorbent followed by high-performance liquid chromatography fluorescence detection (HPLC-FLD) has been developed. In this paper, a novel labeling reagent, carbazole-9-ethyl-2-maleimide(CAEM), was synthesized and reacted with thiols at 40°C for 10min in the presence of PBS buffer (0.02mol/L, pH 7.5). Interestingly, CAEM itself had no fluorescence, while its derivatives exhibited intense fluorescence with an excitation maximum at λ ex 274nm and an emission maximum at λ em 363nm, which greatly reduced the background interference and improved the sensitivity of the method. Furthermore, the MOF-5 was prepared and used as DSPE adsorbent for the selective adsorption of thiols from wastewater sample. Under the optimized experimental conditions, an excellent linearity for all analytes over their concentration ranges of 0.01-1.0μmol/L (R 2 >0.9986)were obtained with the limit of detection (LOD) ranging from 8 to 17.1pmol/L for nine tested thiols. The feasibility of this method for the determination of thiols in wastewater samples had been evaluated and satisfactory average recoveries (n=3) were achieved with the range of 86.6-98.5%. Copyright © 2016 Elsevier B.V. All rights reserved.

A nanoscale Zr-based fluorescent metal-organic framework for selective and sensitive detection of hydrogen sulfide

Science.gov (United States)

Li, Yanping; Zhang, Xin; Zhang, Ling; Jiang, Ke; Cui, Yuanjing; Yang, Yu; Qian, Guodong

2017-11-01

Hydrogen sulfide (H2S) has been commonly viewed as a gas signaling molecule in various physiological and pathological processes. However, the highly efficient H2S detection still remains challenging. Herein, we designed a new robust nano metal-organic framework (MOF) UiO-66-CH=CH2 as a fluorescent probe for rapid, sensitive and selective detection of biological H2S. UiO-66-CH=CH2 was prepared by heating ZrCl4 and 2-vinylterephthalic acid via a simple method. UiO-66-CH=CH2 displayed fluorescence quenching to H2S and kept excellent selectivity in the presence of biological relevant analytes especially the cysteine and glutathione. This MOF-based probe also exhibited fast response (10 s) and high sensitivity with a detection limit of 6.46 μM which was within the concentration range of biological H2S in living system. Moreover, this constructed MOF featured water-stability, nanoscale (20-30 nm) and low toxicity, which made it a promising candidate for biological H2S sensing.

Laser-induced fluorescence detection strategies for sodium atoms and compounds in high-pressure combustors

Science.gov (United States)

Weiland, Karen J. R.; Wise, Michael L.; Smith, Gregory P.

1993-01-01

A variety of laser-induced fluorescence schemes were examined experimentally in atmospheric pressure flames to determine their use for sodium atom and salt detection in high-pressure, optically thick environments. Collisional energy transfer plays a large role in fluorescence detection. Optimum sensitivity, at the parts in 10 exp 9 level for a single laser pulse, was obtained with the excitation of the 4p-3s transition at 330 nm and the detection of the 3d-3p fluorescence at 818 nm. Fluorescence loss processes, such as ionization and amplified spontaneous emission, were examined. A new laser-induced atomization/laser-induced fluorescence detection technique was demonstrated for NaOH and NaCl. A 248-nm excimer laser photodissociates the salt molecules present in the seeded flames prior to atom detection by laser-induced fluorescence.

Development of an Ultrasonication-Assisted Extraction Based HPLC With a Fluorescence Method for Sensitive Determination of Aflatoxins in Highly Acidic Hibiscus sabdariffa

Directory of Open Access Journals (Sweden)

Xiaofei Liu

2018-04-01

Full Text Available The high acidity and complex components of Hibiscus sabdariffa have provided major challenges for sensitive determination of trace aflatoxins. In this study, sample pretreatment of H. sabdariffa was systematically developed for sensitive high performance liquid chromatography-fluorescence detection (HPLC-FLD after ultrasonication-assisted extraction, immunoaffinity column (IAC clean-up and on-line post-column photochemical derivatization (PCD. Aflatoxins B1, B2, G1, G2 were extracted from samples by using methanol/water (70:30, v/v with the addition of NaCl. The solutions were diluted 1:8 with 0.1 M phosphate buffer (pH 8.0 to negate the issues of high acidity and matrix interferences. The established method was validated with satisfactory linearity (R > 0.999, sensitivity (limits of detection (LODs and limits of quantitation (LOQs of 0.15–0.65 and 0.53–2.18 μg/kg, respectively, precision (RSD <11%, stability (RSD of 0.2–3.6%, and accuracy (recovery rates of 86.0–102.3%, which all met the stipulated analytical requirements. Analysis of 28 H. sabdariffa samples indicated that one sample incubated with Aspergillus flavus was positive with aflatoxin B1 (AFB1 at 3.11 μg/kg. The strategy developed in this study also has the potential to reliably extract and sensitively detect more mycotoxins in other complex acidic matrices, such as traditional Chinese medicines, foodstuffs, etc.

“Turn-off” fluorescent data array sensor based on double quantum dots coupled with chemometrics for highly sensitive and selective detection of multicomponent pesticides

Energy Technology Data Exchange (ETDEWEB)

Fan, Yao; Liu, Li; Sun, Donglei; Lan, Hanyue [The Modernization Engineering Technology Research Center of Ethnic Minority Medicine of Hubei Province, College of Pharmacy, South-Central University for Nationalities, Wuhan 430074 (China); Fu, Haiyan, E-mail: fuhaiyan@mail.scuec.edu.cn [The Modernization Engineering Technology Research Center of Ethnic Minority Medicine of Hubei Province, College of Pharmacy, South-Central University for Nationalities, Wuhan 430074 (China); Yang, Tianming, E-mail: tmyang@mail.scuec.edu.cn [The Modernization Engineering Technology Research Center of Ethnic Minority Medicine of Hubei Province, College of Pharmacy, South-Central University for Nationalities, Wuhan 430074 (China); She, Yuanbin, E-mail: sheyb@zjut.edu.cn [State Key Laboratory Breeding Base of Green Chemistry-Synthesis Technology, College of Chemical Engineering, Zhejiang University of Technology, Hangzhou 310032 (China); Ni, Chuang [The Modernization Engineering Technology Research Center of Ethnic Minority Medicine of Hubei Province, College of Pharmacy, South-Central University for Nationalities, Wuhan 430074 (China)

2016-04-15

As a popular detection model, the fluorescence “turn-off” sensor based on quantum dots (QDs) has already been successfully employed in the detections of many materials, especially in the researches on the interactions between pesticides. However, the previous studies are mainly focused on simple single track or the comparison based on similar concentration of drugs. In this work, a new detection method based on the fluorescence “turn-off” model with water-soluble ZnCdSe and CdSe QDs simultaneously as the fluorescent probes is established to detect various pesticides. The fluorescence of the two QDs can be quenched by different pesticides with varying degrees, which leads to the differences in positions and intensities of two peaks. By combining with chemometrics methods, all the pesticides can be qualitative and quantitative respectively even in real samples with the limit of detection was 2 × 10{sup −8} mol L{sup −1} and a recognition rate of 100%. This work is, to the best of our knowledge, the first report on the detection of pesticides based on the fluorescence quenching phenomenon of double quantum dots combined with chemometrics methods. What's more, the excellent selectivity of the system has been verified in different mediums such as mixed ion disruption, waste water, tea and water extraction liquid drugs. - Highlights: • A new model based on double QDs is established for pesticide residues detection. • The fluorescent data array sensor is coupled with chmometrics methods. • The sensor can be highly sensitive and selective detection in actual samples.

“Turn-off” fluorescent data array sensor based on double quantum dots coupled with chemometrics for highly sensitive and selective detection of multicomponent pesticides

International Nuclear Information System (INIS)

Fan, Yao; Liu, Li; Sun, Donglei; Lan, Hanyue; Fu, Haiyan; Yang, Tianming; She, Yuanbin; Ni, Chuang

2016-01-01

As a popular detection model, the fluorescence “turn-off” sensor based on quantum dots (QDs) has already been successfully employed in the detections of many materials, especially in the researches on the interactions between pesticides. However, the previous studies are mainly focused on simple single track or the comparison based on similar concentration of drugs. In this work, a new detection method based on the fluorescence “turn-off” model with water-soluble ZnCdSe and CdSe QDs simultaneously as the fluorescent probes is established to detect various pesticides. The fluorescence of the two QDs can be quenched by different pesticides with varying degrees, which leads to the differences in positions and intensities of two peaks. By combining with chemometrics methods, all the pesticides can be qualitative and quantitative respectively even in real samples with the limit of detection was 2 × 10"−"8 mol L"−"1 and a recognition rate of 100%. This work is, to the best of our knowledge, the first report on the detection of pesticides based on the fluorescence quenching phenomenon of double quantum dots combined with chemometrics methods. What's more, the excellent selectivity of the system has been verified in different mediums such as mixed ion disruption, waste water, tea and water extraction liquid drugs. - Highlights: • A new model based on double QDs is established for pesticide residues detection. • The fluorescent data array sensor is coupled with chmometrics methods. • The sensor can be highly sensitive and selective detection in actual samples.

Fluorescence enhancement of CdTe/CdS quantum dots by coupling of glyphosate and its application for sensitive detection of copper ion

International Nuclear Information System (INIS)

Liu Zhengqing; Liu Shaopu; Yin Pengfei; He Youqiu

2012-01-01

Graphical abstract: Glyphosate (Glyp) had been used to modify the surface of CdTe/CdS QDs, resulting in the enhancement of fluorescence intensity. The Glyp-functionalized QDs fluorescent probe offers good sensitivity and selectivity for detecting Cu 2+ based on the fluorescence quenching. Highlights: ► Water soluble CdTe/CdS quantum dots capped with glyphosate were firstly synthesized. ► The fluorescence of the Glyp-functionalized QDs was quenched by copper ion. ► A new fluorescent sensor for copper ion was developed based on the prepared QDs. ► The sensor exhibited high sensitivity and good selectivity for copper ion. - Abstract: A novel fluorescent probe for Cu 2+ determination based on the fluorescence quenching of glyphosate (Glyp)-functionalized quantum dots (QDs) was firstly reported. Glyp had been used to modify the surface of QDs to form Glyp-functionalized QDs following the capping of thioglycolic acid on the core–shell CdTe/CdS QDs. Under the optimal conditions, the response was linearly proportional to the concentration of Cu 2+ between 2.4 × 10 −2 μg mL −1 and 28 μg mL −1 , with a detection limit of 1.3 × 10 −3 μg mL −1 (3δ). The Glyp-functionalized QDs fluorescent probe offers good sensitivity and selectivity for detecting Cu 2+ . The fluorescent probe was successfully used for the determination of Cu 2+ in environmental samples. The mechanism of reaction was also discussed.

In-vacuum scattered light reduction with black cupric oxide surfaces for sensitive fluorescence detection.

Science.gov (United States)

Norrgard, E B; Sitaraman, N; Barry, J F; McCarron, D J; Steinecker, M H; DeMille, D

2016-05-01

We demonstrate a simple and easy method for producing low-reflectivity surfaces that are ultra-high vacuum compatible, may be baked to high temperatures, and are easily applied even on complex surface geometries. Black cupric oxide (CuO) surfaces are chemically grown in minutes on any copper surface, allowing for low-cost, rapid prototyping, and production. The reflective properties are measured to be comparable to commercially available products for creating optically black surfaces. We describe a vacuum apparatus which uses multiple blackened copper surfaces for sensitive, low-background detection of molecules using laser-induced fluorescence.

TU-G-207-03: High Spatial Resolution and High Sensitivity X-Ray Fluorescence Imaging

International Nuclear Information System (INIS)

Xing, L.

2015-01-01

Last few years has witnessed the development of novel of X-ray imaging modalities, such as spectral CT, phase contrast CT, and X-ray acoustic/fluorescence/luminescence imaging. This symposium will present the recent advances of these emerging X-ray imaging modalities and update the attendees with knowledge in various related topics, including X-ray photon-counting detectors, X-ray physics underlying the emerging applications beyond the traditional X-ray imaging, image reconstruction for the novel modalities, characterization and evaluation of the systems, and their practical implications. In addition, the concept and practical aspects of X-ray activatable targeted nanoparticles for molecular X-ray imaging will be discussed in the context of X-ray fluorescence and luminescence CT. Learning Objectives: Present background knowledge of various emerging X-ray imaging techniques, such as spectral CT, phase contrast CT and X-ray fluorescence/luminescence CT. Discuss the practical need, technical aspects and current status of the emerging X-ray imaging modalities. Describe utility and future impact of the new generation of X-ray imaging applications

Toehold strand displacement-driven assembly of G-quadruplex DNA for enzyme-free and non-label sensitive fluorescent detection of thrombin.

Science.gov (United States)

Xu, Yunying; Zhou, Wenjiao; Zhou, Ming; Xiang, Yun; Yuan, Ruo; Chai, Yaqin

2015-02-15

Based on a new signal amplification strategy by the toehold strand displacement-driven cyclic assembly of G-quadruplex DNA, the development of an enzyme-free and non-label aptamer sensing approach for sensitive fluorescent detection of thrombin is described. The target thrombin associates with the corresponding aptamer of the partial dsDNA probes and liberates single stranded initiation sequences, which trigger the toehold strand displacement assembly of two G-quadruplex containing hairpin DNAs. This toehold strand displacement reaction leads to the cyclic reuse of the initiation sequences and the production of DNA assemblies with numerous G-quadruplex structures. The fluorescent dye, N-Methyl mesoporphyrin IX, binds to these G-quadruplex structures and generates significantly amplified fluorescent signals to achieve highly sensitive detection of thrombin down to 5 pM. Besides, this method shows high selectivity towards the target thrombin against other control proteins. The developed thrombin sensing method herein avoids the modification of the probes and the involvement of any enzyme or nanomaterial labels for signal amplification. With the successful demonstration for thrombin detection, our approach can be easily adopted to monitor other target molecules in a simple, low-cost, sensitive and selective way by choosing appropriate aptamer/ligand pairs. Copyright © 2014 Elsevier B.V. All rights reserved.

High sensitivity detection of selenium by laser excited atomic fluorescence spectrometry using electrothermal atomization

International Nuclear Information System (INIS)

Heitmann, U.; Hese, A.; Schoknecht, G.; Gries, W.

1995-01-01

The high sensitivity detection of the trace element selenium is reported. The analytical method applied is Laser Excited Atomic Fluorescence Spectrometry using Electrothermal Atomization within a graphite furnace atomizer. For the production of tunable laser radiation in the VUV spectral region a laser system was developed which consists of two dye lasers pumped by a Nd:YAG laser. The laser radiations are subsequently frequency doubled and sum frequency mixed by nonlinear optical KDP or BBO crystals, respectively. The system works with a repetition rate of 20 Hz and provides output energies of up to 100 μJ in the VUV at a pulse duration of 5 ns. The analytical investigations were focused on the detection of selenium in aqueous solutions and samples of human whole blood. From measurements on aqueous standards detection limits of 1.5 ng/l for selenium were obtained, with corresponding absolute detected masses of only 15 fg. The linear dynamic range spanned six orders of magnitude and good precision was achieved. In case of human whole blood samples the recovery was found to be within the range of 96% to 104%. The determination of the selenium content yielded medians of [119.5 ± 17.3] μg/l for 200 frozen blood samples taken in 1988 and [109.1 ± 15.6] μg/l for 103 fresh blood samples. (author)

Comparison of sensitivities and detection limits between direct excitation and secondary excitation modes in energy dispersive x-ray fluorescence analysis

International Nuclear Information System (INIS)

Artz, B.E.; Short, M.A.

1976-01-01

A comparison was made between the direct tube excitation mode and the secondary target excitation mode using a Kevex 0810 energy dispersive x-ray fluorescence system. Relative sensitivities and detection limits were determined with two system configurations. The first configuration used a standard, high power, x-ray fluorescence tube to directly excite the specimen. Several x-ray tubes, including chromium, molybdenum, and tungsten, both filtered and not filtered, were employed. The second configuration consisted of using the x-ray tube to excite a secondary target which in turn excited the specimen. Appropriate targets were compared to the direct excitation results. Relative sensitivities and detection limits were determined for K-series lines for elements from magnesium to barium contained in a low atomic number matrix and in a high atomic number matrix

Selective and sensitive fluorescence-shift probes based on two dansyl groups for mercury(ii) ion detection.

Science.gov (United States)

Ma, Li-Jun; Liu, Jialun; Deng, Lefang; Zhao, Meili; Deng, Zhifu; Li, Xutian; Tang, Jian; Yang, Liting

2014-11-01

Two probes ( and ) bearing two dansyl fluorophores were synthesized and applied to the detection of mercury(ii) ions in aqueous solution. These probes exhibited a selective response to Hg(2+) in a buffered solution, with high sensitivity and a unique fluorescence response signal which displayed a blue-shift effect in the fluorescence emission peak. The Hg(2+) recognition mechanisms of the probes were determined by NMR spectroscopy, ESI-MS and UV-vis spectroscopy. The results showed that probe and mercury(ii) ions formed an unusual 2:2 stoichiometric ratio complex, while probe and Hg(2+) formed a multidentate complex with a stoichiometric ratio of 2:1.

Highly fluorescent carbon dots as nanoprobes for sensitive and selective determination of 4-nitrophenol in surface waters

International Nuclear Information System (INIS)

Ahmed, Gaber Hashem Gaber; Laíño, Rosana Badía; Calzón, Josefa Angela García; García, Marta Elena Díaz

2015-01-01

We report on the synthesis of carbon dots (C-dots) by thermal carbonization of a mixture of ethyleneglycol bis-(2-aminoethyl ether)-N,N,N’,N’-tetraacetic acid (EGTA) and tris(hydroxymethyl)aminomethane (Tris). The resulting C-dots were characterized by X-ray diffraction, proton and carbon nuclear magnetic resonance, FTIR and fluorescence spectroscopy, and high-resolution TEM. The data reveal that the C-dots are mainly capped with hydroxy and carbonyl groups and are highly fluorescent with an emission peak that shifts from 427 to 438 nm if the excitation wavelength is increased from 310 to 360–370 nm. Fluorescence is quenched by 4-nitrophenol (4-NP), and this effect was exploited to design a simple and rapid protocol for the determination of 4-NP. The detection limit is 28 nM and the linear range extends from 0.1 to 50 μM. The method was successfully applied to the determination of 4-NP in spiked river and sea waters. (author)

A highly sensitive fluorescence quenching method for perphenazine detection based on its catalysis of K{sub 2}S{sub 2}O{sub 8} oxidizing rhodamine 6G

Energy Technology Data Exchange (ETDEWEB)

Zhang, Lihong; Huang, Qitong; Lin, Changqing [Department of Food and Biological Engineering, Zhangzhou Institute of Technology, Zhangzhou, 363000 (China); Lin, Xiaofeng [College of Chemistry and Environment, Minnan Normal University, Zhangzhou, 363000 (China); Huang, Yiqun [Zhangzhou Affiliated Hospital of Fujian Medical University, Zhangzhou 363000 (China); Liu, Jiaming, E-mail: mnsdljm@163.com [College of Chemistry and Environment, Minnan Normal University, Zhangzhou, 363000 (China); Ma, Xudong, E-mail: maxudong005@hotmail.com [Zhangzhou Affiliated Hospital of Fujian Medical University, Zhangzhou 363000 (China)

2014-12-15

In this paper, the fluorescence spectra of Rhod 6G (rhodamine 6G)–K{sub 2}S{sub 2}O{sub 8}–PPH (perphenazine) were studied. We found that Rhod 6G existed in the form of Rhod 6G{sup +} under the conditions of 60 °C, 10 min and pH 5.42, and Rhod 6G{sup +} can emit strong and stable fluorescence. Further study showed that when PPH and Rhod 6G{sup +} coexisted, the ester exchange reaction carried out between -OH of PPH and -COOC{sub 2}H{sub 5} of Rhod 6G{sup +} to produced Rhod 6G{sup +}–PPH compound. More interestingly, K{sub 2}S{sub 2}O{sub 8} could oxidize Rhod 6G{sup +} and quench its RTP signal, while PPH was oxidized to red compound PPH′ by K{sub 2}S{sub 2}O{sub 8}, and Rhod 6G{sup +}–PPH′ and PPH were produced in the ester exchange reaction between the -OH of PPH′ and the -COOC{sub 2}H{sub 5} of Rhod 6G{sup +}–PPH. In the above process, PPH catalyzed K{sub 2}S{sub 2}O{sub 8} oxidizing Rhod 6G, which caused the fluorescence signal of the system to quench sharply. Hence, a catalytic fluorescence quenching method for the determination of residual PPH has been developed based on the its catalyzing K{sub 2}S{sub 2}O{sub 8} oxidize rhodamine 6G. This sensitive, accurate, simple and selective fluorescence quenching method was used to determine residual PPH in biological samples with the results consisting with those obtained by high performance liquid chromatography (HPLC), showing good accuracy. The structures of Rhod 6G{sup +}, PPH and Rhod 6G{sup +}–PPH were characterized by infrared spectra. The reaction mechanism of the determination of PPH was also discussed. - Highlights: • Fluorescence for the determination of perphenazine (PPH) had been established. • This method had high sensitivity (limit of detection was 3.3×10{sup −14} g mL{sup −1}). • This method had been applied to determination of PPH in biological samples. • Structures of Rhod 6G{sup +}, PPH and Rhod 6G{sup +}–PPH were characterized by infrared spectra. • Mechanism

Highly selective and sensitive fluorescent chemosensor for femtomolar detection of silver ion in aqueous medium

Directory of Open Access Journals (Sweden)

Abraham Daniel Arulraj

2015-12-01

Full Text Available The chemical sensing for the trace level detection of silver ion in aqueous solution still remains a challenge using simple, rapid, and inexpensive method. We report that thionine can be used as a fluorescent probe for the detection of Ag+ ion. The successive addition of Ag+ ion to the solution containing thionine quenches (turns-off the fluorescence intensity of thionine. Association and quenching constants have been estimated by the Benesi–Hildebrand method and Stern–Volmer plot, respectively. From the plot, the nature of the fluorescence quenching was confirmed as static quenching. An important feature of our chemosensor is high selectivity towards the determination of silver ion in aqueous solution over the other competitive metal ions. The detection limit of the sensor achieved 5 fM for Ag+ ion, which is superior to all previously reported chemosensors. The NMR and FT-IR studies were also carried out to support the complex formation between thionine and Ag+ ion. The practicality of the proposed chemosensor for determination of Ag+ ion was carried in untreated water samples.

Fluorescence enhancement of CdTe/CdS quantum dots by coupling of glyphosate and its application for sensitive detection of copper ion

Energy Technology Data Exchange (ETDEWEB)

Liu Zhengqing; Liu Shaopu; Yin Pengfei [Key Laboratory on Luminescence and Real-Time Analysis, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China); He Youqiu, E-mail: heyq@swu.edu.cn [Key Laboratory on Luminescence and Real-Time Analysis, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China)

2012-10-01

Graphical abstract: Glyphosate (Glyp) had been used to modify the surface of CdTe/CdS QDs, resulting in the enhancement of fluorescence intensity. The Glyp-functionalized QDs fluorescent probe offers good sensitivity and selectivity for detecting Cu{sup 2+} based on the fluorescence quenching. Highlights: Black-Right-Pointing-Pointer Water soluble CdTe/CdS quantum dots capped with glyphosate were firstly synthesized. Black-Right-Pointing-Pointer The fluorescence of the Glyp-functionalized QDs was quenched by copper ion. Black-Right-Pointing-Pointer A new fluorescent sensor for copper ion was developed based on the prepared QDs. Black-Right-Pointing-Pointer The sensor exhibited high sensitivity and good selectivity for copper ion. - Abstract: A novel fluorescent probe for Cu{sup 2+} determination based on the fluorescence quenching of glyphosate (Glyp)-functionalized quantum dots (QDs) was firstly reported. Glyp had been used to modify the surface of QDs to form Glyp-functionalized QDs following the capping of thioglycolic acid on the core-shell CdTe/CdS QDs. Under the optimal conditions, the response was linearly proportional to the concentration of Cu{sup 2+} between 2.4 Multiplication-Sign 10{sup -2} {mu}g mL{sup -1} and 28 {mu}g mL{sup -1}, with a detection limit of 1.3 Multiplication-Sign 10{sup -3} {mu}g mL{sup -1} (3{delta}). The Glyp-functionalized QDs fluorescent probe offers good sensitivity and selectivity for detecting Cu{sup 2+}. The fluorescent probe was successfully used for the determination of Cu{sup 2+} in environmental samples. The mechanism of reaction was also discussed.

Kinetic analysis of DAF-FM activation by NO: toward calibration of a NO-sensitive fluorescent dye.

Science.gov (United States)

Namin, Shabnam M; Nofallah, Sara; Joshi, Mahesh S; Kavallieratos, Konstantinos; Tsoukias, Nikolaos M

2013-01-15

Nitric oxide (NO) research in biomedicine has been hampered by the absence of a method that will allow quantitative measurement of NO in biological tissues with high sensitivity and selectivity, and with adequate spatial and temporal resolution. 4-amino-5-methylamino-2',7'-difluorofluorescein (DAF-FM) is a NO sensitive fluorescence probe that has been used widely for qualitative assessment of cellular NO production. However, calibration of the fluorescent signal and quantification of NO concentration in cells and tissues using fluorescent probes, have provided significant challenge. In this study we utilize a combination of mathematical modeling and experimentation to elucidate the kinetics of NO/DAF-FM reaction in solution. Modeling and experiments suggest that the slope of fluorescent intensity (FI) can be related to NO concentration according to the equation: ddtFI=2αk(1)NO(2)O(2)DAF-FMkNO+DAF-FM where α is a proportionality coefficient that relates FI to unit concentration of activated DAF-FM, k(1) is the NO oxidation rate constant, and k was estimated to be 4.3±0.6. The FI slope exhibits saturation kinetics with DAF-FM concentration. Interestingly, the effective half-maximum constant (EC(50)) increases proportionally to NO concentration. This result is not in agreement with the proposition that N(2)O(3) is the NO oxidation byproduct that activates DAF-FM. Kinetic analysis suggests that the reactive intermediate should exhibit NO-dependent consumption and thus NO(2)() is a more likely candidate. The derived rate law can be used for the calibration of DAF-FM fluorescence and the quantification of NO concentration in biological tissues. Copyright © 2012 Elsevier Inc. All rights reserved.

Nitrogen-Doped Carbon Quantum Dots as Fluorescent Probes for Sensitive and Selective Detection of Nitrite

Directory of Open Access Journals (Sweden)

Zhibiao Feng

2017-11-01

Full Text Available Nitrites are the upstream precursors of the carcinogenic nitrosamines, which are widely found in the natural environment and many food products. It is important to develop a simple and sensitive sensor for detecting nitrites. In this work, a fluorescence probe based on nitrogen-doped carbon quantum dots (N-CQDs was developed for the sensitive and selective determination of nitrites. At pH 2, the fluorescence of N-CQDs can be selectively quenched by nitrite due to the fact N-nitroso compounds can be formed in the reaction of amide groups with nitrous acid, which results in fluorescence static quenching. Under optimal conditions, fluorescence intensity quenching upon addition of nitrite gives a satisfactory linear relationship covering the linear range of 0.2–20 μM, and the limit of detection (LOD is 40 nM. Moreover, this method has been successfully applied to the determination of nitrites in tap water, which indicates its great potential for monitoring of nitrites in environmental samples.

Determination of aristolochic acids by high-performance liquid chromatography with fluorescence detection.

Science.gov (United States)

Wang, Yinan; Chan, Wan

2014-06-25

Nephrotoxic and carcinogenic aristolochic acids (AAs) are naturally occurring nitrophenanthrene carboxylic acids in the herbal genus Aristolochia. The misuse of AA-containing herbs in preparing slimming drugs has caused hundred of cases of kidney disease in Belgium women in a slimming regime in the early 1990s. Accumulating evidence also suggested that prolong dietary intake of AA-contaminated food is one of the major causes to the Balkan endemic nephropathy that was first observed in the late 1950s. Therefore, analytical methods of high sensitivity are extremely important for safeguarding human exposure to AA-containing herbal medicines, herbal remedies, and food composites. In this paper, we describe the development of a new high-performance liquid chromatography coupled fluorescence detector (HPLC-FLD) method for the sensitive determination of AAs. The method makes use of a novel cysteine-induced denitration reaction that "turns on" the fluorescence of AAs for fluorometric detections. Our results showed that the combination of cysteine-induced denitration and HPLC-FLD analysis allows for sensitive quantification of AA-I and AA-II at detection limits of 27.1 and 25.4 ng/g, respectively. The method was validated and has been successfully applied in quantifying AAs in Chinese herbal medicines.

Two rhodamine lactam modulated lysosome-targetable fluorescence probes for sensitively and selectively monitoring subcellular organelle pH change

Energy Technology Data Exchange (ETDEWEB)

Li, Hongmei [Ministry of Education Key Laboratory of Synthetic and Natural Functional Molecule Chemistry, College of Chemistry & Materials Science, Northwest University, Xi' an 710069 (China); Wang, Cuiling [Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University, Xi' an 710069 (China); She, Mengyao; Zhu, Yuelu; Zhang, Jidong; Yang, Zheng [Ministry of Education Key Laboratory of Synthetic and Natural Functional Molecule Chemistry, College of Chemistry & Materials Science, Northwest University, Xi' an 710069 (China); Liu, Ping, E-mail: liuping@nwu.edu.cn [Ministry of Education Key Laboratory of Synthetic and Natural Functional Molecule Chemistry, College of Chemistry & Materials Science, Northwest University, Xi' an 710069 (China); Wang, Yaoyu [Ministry of Education Key Laboratory of Synthetic and Natural Functional Molecule Chemistry, College of Chemistry & Materials Science, Northwest University, Xi' an 710069 (China); Li, Jianli, E-mail: lijianli@nwu.edu.cn [Ministry of Education Key Laboratory of Synthetic and Natural Functional Molecule Chemistry, College of Chemistry & Materials Science, Northwest University, Xi' an 710069 (China)

2015-11-05

Be a powerful technique for convenient detection of pH change in living cells, especially at subcellular level, fluorescent probes has attracted more and more attention. In this work, we designed and synthesized three rhodamine lactam modulated fluorescent probes RS1, RS2 and RS3, which all respond sensitively toward weak acidity (pH range 4–6) via the photophysical property in buffer solution without interference from the other metal ions, and they also show ideal pKa values and excellent reversibility. Particularly, by changing the lone pair electrons distribution of lactam-N atom with different conjugations, RS2 and RS3 exhibit high quantum yield, negligible cytotoxicity and excellent permeability. They are suitable to stain selectively lysosomes of tumor cells and monitor its pH changes sensitively via optical molecular imaging. The above findings suggest that the probes we designed could act as ideal and easy method for investigating the pivotal role of H{sup +} in lysosomes and are potential pH detectors in disease diagnosis through direct intracellular imaging. - Highlights: • Two probes for sensitively and selectively monitoring weak acidic pH change. • The pKa of the probes was highly suitable for staining lysosomes in tumor cells. • The properties of those probes were changed by different conjugate system. • These probes have negligible cytotoxicity and good sensitivity in vivo.

Two rhodamine lactam modulated lysosome-targetable fluorescence probes for sensitively and selectively monitoring subcellular organelle pH change

International Nuclear Information System (INIS)

Li, Hongmei; Wang, Cuiling; She, Mengyao; Zhu, Yuelu; Zhang, Jidong; Yang, Zheng; Liu, Ping; Wang, Yaoyu; Li, Jianli

2015-01-01

Be a powerful technique for convenient detection of pH change in living cells, especially at subcellular level, fluorescent probes has attracted more and more attention. In this work, we designed and synthesized three rhodamine lactam modulated fluorescent probes RS1, RS2 and RS3, which all respond sensitively toward weak acidity (pH range 4–6) via the photophysical property in buffer solution without interference from the other metal ions, and they also show ideal pKa values and excellent reversibility. Particularly, by changing the lone pair electrons distribution of lactam-N atom with different conjugations, RS2 and RS3 exhibit high quantum yield, negligible cytotoxicity and excellent permeability. They are suitable to stain selectively lysosomes of tumor cells and monitor its pH changes sensitively via optical molecular imaging. The above findings suggest that the probes we designed could act as ideal and easy method for investigating the pivotal role of H + in lysosomes and are potential pH detectors in disease diagnosis through direct intracellular imaging. - Highlights: • Two probes for sensitively and selectively monitoring weak acidic pH change. • The pKa of the probes was highly suitable for staining lysosomes in tumor cells. • The properties of those probes were changed by different conjugate system. • These probes have negligible cytotoxicity and good sensitivity in vivo.

Oligonucleotide-stabilized fluorescent silver nanoclusters for the specific and sensitive detection of biotin.

Science.gov (United States)

Xiong, Xiaoli; Tang, Yan; Zhao, Jingjin; Zhao, Shulin

2016-02-21

A novel biotin fluorescent probe based on oligonucleotide-stabilized silver nanoclusters (DNA-AgNCs) was synthesized by employing a biotinylated cytosine-rich sequence as a synthesized template. The fluorescence properties of the DNA-AgNCs are related to the modified position of the DNA. When biotin is linked to the middle thymine base of the DNA sequence, the DNA-AgNCs emit the strongest fluorescence. Moreover, the stability of the DNA-AgNCs was affected by avidin through biotin-avidin binding, quenching the fluorescence of the DNA-AgNCs. In contrast, if free biotin is further introduced into this system, the quenching is apparently weakened by competition, leading to the restoration of fluorescence. This phenomenon can be utilized for the detection of biotin. Under the optimal conditions, the fluorescence recovery is linearly proportional to the concentration of biotin in the range of 10 nM-1.0 μM with a detection limit of 6.0 nM. This DNA-AgNCs probe with excellent fluorescent properties is sensitive and selective for the detection of biotin and has been applied for the determination of biotin in wheat flour.

Improving the Sensitivity and Functionality of Mobile Webcam-Based Fluorescence Detectors for Point-of-Care Diagnostics in Global Health

Science.gov (United States)

Rasooly, Reuven; Bruck, Hugh Alan; Balsam, Joshua; Prickril, Ben; Ossandon, Miguel; Rasooly, Avraham

2016-01-01

Resource-poor countries and regions require effective, low-cost diagnostic devices for accurate identification and diagnosis of health conditions. Optical detection technologies used for many types of biological and clinical analysis can play a significant role in addressing this need, but must be sufficiently affordable and portable for use in global health settings. Most current clinical optical imaging technologies are accurate and sensitive, but also expensive and difficult to adapt for use in these settings. These challenges can be mitigated by taking advantage of affordable consumer electronics mobile devices such as webcams, mobile phones, charge-coupled device (CCD) cameras, lasers, and LEDs. Low-cost, portable multi-wavelength fluorescence plate readers have been developed for many applications including detection of microbial toxins such as C. Botulinum A neurotoxin, Shiga toxin, and S. aureus enterotoxin B (SEB), and flow cytometry has been used to detect very low cell concentrations. However, the relatively low sensitivities of these devices limit their clinical utility. We have developed several approaches to improve their sensitivity presented here for webcam based fluorescence detectors, including (1) image stacking to improve signal-to-noise ratios; (2) lasers to enable fluorescence excitation for flow cytometry; and (3) streak imaging to capture the trajectory of a single cell, enabling imaging sensors with high noise levels to detect rare cell events. These approaches can also help to overcome some of the limitations of other low-cost optical detection technologies such as CCD or phone-based detectors (like high noise levels or low sensitivities), and provide for their use in low-cost medical diagnostics in resource-poor settings. PMID:27196933

Improving the Sensitivity and Functionality of Mobile Webcam-Based Fluorescence Detectors for Point-of-Care Diagnostics in Global Health

Directory of Open Access Journals (Sweden)

Reuven Rasooly

2016-05-01

Full Text Available Resource-poor countries and regions require effective, low-cost diagnostic devices for accurate identification and diagnosis of health conditions. Optical detection technologies used for many types of biological and clinical analysis can play a significant role in addressing this need, but must be sufficiently affordable and portable for use in global health settings. Most current clinical optical imaging technologies are accurate and sensitive, but also expensive and difficult to adapt for use in these settings. These challenges can be mitigated by taking advantage of affordable consumer electronics mobile devices such as webcams, mobile phones, charge-coupled device (CCD cameras, lasers, and LEDs. Low-cost, portable multi-wavelength fluorescence plate readers have been developed for many applications including detection of microbial toxins such as C. Botulinum A neurotoxin, Shiga toxin, and S. aureus enterotoxin B (SEB, and flow cytometry has been used to detect very low cell concentrations. However, the relatively low sensitivities of these devices limit their clinical utility. We have developed several approaches to improve their sensitivity presented here for webcam based fluorescence detectors, including (1 image stacking to improve signal-to-noise ratios; (2 lasers to enable fluorescence excitation for flow cytometry; and (3 streak imaging to capture the trajectory of a single cell, enabling imaging sensors with high noise levels to detect rare cell events. These approaches can also help to overcome some of the limitations of other low-cost optical detection technologies such as CCD or phone-based detectors (like high noise levels or low sensitivities, and provide for their use in low-cost medical diagnostics in resource-poor settings.

Calcium Sensitive Fluorescent Dyes Fluo-4 and Fura Red under Pressure: Behaviour of Fluorescence and Buffer Properties under Hydrostatic Pressures up to 200 MPa.

Science.gov (United States)

Schneidereit, D; Vass, H; Reischl, B; Allen, R J; Friedrich, O

2016-01-01

The fluorescent Ca2+ sensitive dyes Fura Red (ratiometric) and Fluo-4 (non-ratiometric) are widely utilized for the optical assessment of Ca2+ fluctuations in vitro as well as in situ. The fluorescent behavior of these dyes is strongly depends on temperature, pH, ionic strength and pressure. It is crucial to understand the response of these dyes to pressure when applying calcium imaging technologies in the field of high pressure bioscience. Therefore, we use an optically accessible pressure vessel to pressurize physiological Ca2+-buffered solutions at different fixed concentrations of free Ca2+ (1 nM to 25.6 μM) and a specified dye concentration (12 μM) to pressures of 200 MPa, and record dye fluorescence intensity. Our results show that Fluo-4 fluorescence intensity is reduced by 31% per 100 MPa, the intensity of Fura Red is reduced by 10% per 100 MPa. The mean reaction volume for the dissociation of calcium from the dye molecules [Formula: see text] is determined to -17.8 ml mol-1 for Fluo-4 and -21.3 ml mol-1 for Fura Red. Additionally, a model is presented that is used to correct for pressure-dependent changes in pH and binding affinity of Ca2+ to EGTA, as well as to determine the influence of these changes on dye fluorescence.

Calcium Sensitive Fluorescent Dyes Fluo-4 and Fura Red under Pressure: Behaviour of Fluorescence and Buffer Properties under Hydrostatic Pressures up to 200 MPa

Science.gov (United States)

Vass, H.; Reischl, B.; Allen, R. J.; Friedrich, O.

2016-01-01

The fluorescent Ca2+ sensitive dyes Fura Red (ratiometric) and Fluo-4 (non-ratiometric) are widely utilized for the optical assessment of Ca2+ fluctuations in vitro as well as in situ. The fluorescent behavior of these dyes is strongly depends on temperature, pH, ionic strength and pressure. It is crucial to understand the response of these dyes to pressure when applying calcium imaging technologies in the field of high pressure bioscience. Therefore, we use an optically accessible pressure vessel to pressurize physiological Ca2+-buffered solutions at different fixed concentrations of free Ca2+ (1 nM to 25.6 μM) and a specified dye concentration (12 μM) to pressures of 200 MPa, and record dye fluorescence intensity. Our results show that Fluo-4 fluorescence intensity is reduced by 31% per 100 MPa, the intensity of Fura Red is reduced by 10% per 100 MPa. The mean reaction volume for the dissociation of calcium from the dye molecules Δdv¯ is determined to -17.8 ml mol-1 for Fluo-4 and -21.3 ml mol-1 for Fura Red. Additionally, a model is presented that is used to correct for pressure-dependent changes in pH and binding affinity of Ca2+ to EGTA, as well as to determine the influence of these changes on dye fluorescence. PMID:27764134

Highly-sensitive aptasensor based on fluorescence resonance energy transfer between l-cysteine capped ZnS quantum dots and graphene oxide sheets for the determination of edifenphos fungicide.

Science.gov (United States)

Arvand, Majid; Mirroshandel, Aazam A

2017-10-15

With the advantages of excellent optical properties and biocompatibility, single-strand DNA-functionalized quantum dots have been widely applied in biosensing and bioimaging. A new aptasensor with easy operation, high sensitivity, and high selectivity was developed by immobilizing the aptamer on water soluble l-cysteine capped ZnS quantum dots (QDs). Graphene oxide (GO) sheets are mixed with the aptamer-QDs. Consequently, the aptamer-conjugated QDs bind to the GO sheets to form a GO/aptamer-QDs ensemble. This aptasensor enables the energy transfer based on a fluorescence resonance energy transfer (FRET) from the QDs to the GO sheets, quenching the fluorescence of QDs. The GO/aptamer-QDs ensemble assay acts as a "turn-on'' fluorescent sensor for edifenphos (EDI) detection. When GO was replaced by EDI, the fluorescence of QDs was restored and its intensity was proportional to the EDI concentration. This GO-based aptasensor under the optimum conditions exhibited excellent analytical performance for EDI determination, ranging from 5×10 -4 to 6×10 -3 mg L -1 with the detection limit of 1.3×10 -4 mgL -1 . Furthermore, the designed aptasensor exhibited excellent selectivity toward EDI compared to other pesticides and herbicides with similar structures such as diazinon, heptachlor, endrin, dieldrin, butachlor and chlordane. Good reproducibility and precision (RSD =3.9%, n =10) of the assay indicates the high potential of the aptasensor for quantitative trace analysis of EDI. Moreover, the results demonstrate the applicability of the aptasensor for monitoring EDI fungicide in spiked real samples. Copyright © 2017 Elsevier B.V. All rights reserved.

Multiplex and high-throughput DNA detection using surface plasmon mediated fluorescence

Science.gov (United States)

Mei, Zhong

The overall objective of this research project was to develop a user-friendly and sensitive biosensor for nucleic acid aptamers with multiplexing and high-throughput capability. The sensing was based on the fluorescence signals emitted by the fluorophores coupling with plamonic nanoparticle (gold nanorod) deposited on a patterned substrate. Gold nanorods (GNRs) were synthesized using a binary mixture of hexadecyltrimethylammonium bromide (CTAB) and sodium oleate (NaOL) in seed mediated growth method. Polytetrafluoroethylene (PTFE) printed glass slides were selectively coated with a gold thin-film to define hydrophilic areas for GNR deposition. Due to the wettablity contrast, GNR solution dropped on the slide was induced to assemble exclusively in the hydrophilic spots. By controlling temperature and humidity of the evaporation process, vertically-standing GNR arrays were achieved on the pattered slide. Fluorescence was conjugated to GNR surface via DNA double strand with tunable length. Theoretical simulation predicted a flat layer ( 30 nm thick) of uniform "hot spots" presented on the GNR tips, which could modify the nearby fluorescence. Experimentally, the vertical GNR arrays yielded metallic enhanced fluorescence (MEF) effect, which was dependent on the spectrum overlap and GNR-fluorophore distance. Specifically, the maximum enhancement of Quasar 670 and Alexa 750 was observed when it was coupled with GNR664 (plasmonic wavelength 664 nm) and GNR778 respectively at a distance of 16 nm, while the carboxyfluorescein (FAM) was at maximal intensity when attached to gold nanosphere520. This offers an opportunity for multiplexed DNA sensing. Based on this, we developed a novel GNR mediated fluorescence biosensor for DNA detection. Fluorescence labeled haipin-DNA probes were introduced to designated spots of GNR array with the matching LSPR wavelengths on the substrate. The fluorescence was quenched originally because of Forster resonance energy transfer (FRET) effect

Colorimetric and fluorescent chemosensor for highly selective and sensitive relay detection of Cu2 + and H2PO4- in aqueous media

Science.gov (United States)

Su, Jun-Xia; Wang, Xiao-Ting; Chang, Jing; Wu, Gui-Yuan; Wang, Hai-Ming; Yao, Hong; Lin, Qi; Zhang, You-Ming; Wei, Tai-Bao

2017-07-01

In this manuscript, a new colorimetric and fluorescent chemosensor (T) was designed and synthesized, it could successively detect Cu2 + and H2PO4- in DMSO/H2O (v/v = 9:1, pH = 7.2) buffer solution with high selectivity and sensitivity. When added Cu2 + ions into the solution of T, it showed a color changes from yellow to colorless, meanwhile, the green fluorescence of sensor T quenched. This recognition behavior was not affected in the presence of other cations, including Hg2 +, Ag+, Ca2 +, Co2 +, Ni2 +, Cd2 +, Pb2 +, Zn2 +, Cr3 +, and Mg2 + ions. More interestingly, the Cu2 + ions contain sensor T solution could recover the color and fluorescence upon the addition of H2PO4- anions in the same medium. And other surveyed anions (including F-, Cl-, Br-, I-, AcO-, HSO4-, ClO4-, CN- and SCN-) had nearly no influence on the recognition behavior. The detection limits of T to Cu2 + and T-Cu2 + to H2PO4- were evaluated to be 1.609 × 10- 8 M and 0.994 × 10- 7 M, respectively. In addition, the sensor T also could be served as a recyclable component and the logic gate output was also defined in sensing materials. The test strips based on sensor T were fabricated, which acted as a convenient and efficient Cu2 + and H2PO4- test kits.

Bead-based competitive fluorescence immunoassay for sensitive and rapid diagnosis of cyanotoxin risk in drinking water.

Science.gov (United States)

Yu, Hye-Weon; Jang, Am; Kim, Lan Hee; Kim, Sung-Jo; Kim, In S

2011-09-15

Due to the increased occurrence of cyanobacterial blooms and their toxins in drinking water sources, effective management based on a sensitive and rapid analytical method is in high demand for security of safe water sources and environmental human health. Here, a competitive fluorescence immunoassay of microcystin-LR (MCYST-LR) is developed in an attempt to improve the sensitivity, analysis time, and ease-of-manipulation of analysis. To serve this aim, a bead-based suspension assay was introduced based on two major sensing elements: an antibody-conjugated quantum dot (QD) detection probe and an antigen-immobilized magnetic bead (MB) competitor. The assay was composed of three steps: the competitive immunological reaction of QD detection probes against analytes and MB competitors, magnetic separation and washing, and the optical signal generation of QDs. The fluorescence intensity was found to be inversely proportional to the MCYST-LR concentration. Under optimized conditions, the proposed assay performed well for the identification and quantitative analysis of MCYST-LR (within 30 min in the range of 0.42-25 μg/L, with a limit of detection of 0.03 μg/L). It is thus expected that this enhanced assay can contribute both to the sensitive and rapid diagnosis of cyanotoxin risk in drinking water and effective management procedures.

Sensitive fluorescence on-off probes for the fast detection of a chemical warfare agent mimic.

Science.gov (United States)

Khan, Muhammad Shar Jhahan; Wang, Ya-Wen; Senge, Mathias O; Peng, Yu

2018-01-15

Two highly sensitive probes bearing a nucleophilic imine moiety have been utilized for the selective detection of chemical warfare agent (CWA) mimics. Diethyl chlorophosphate (DCP) was used as mimic CWAs. Both iminocoumarin-benzothiazole-based probes not only demonstrated a remarkable fluorescence ON-OFF response and good recognition, but also exhibited fast response times (10s) along with color changes upon addition of DCP. Limits of detection for the two sensors 1 and 2 were calculated as 0.065μM and 0.21μM, respectively, which are much lower than most other reported probes. These two probes not only show high sensitivity and selectivity in solution, but can also be applied for the recognition of DCP in the gas state, with significant color changes easily observed by the naked eye. Copyright © 2017 Elsevier B.V. All rights reserved.

Sensitized fluorescence in thallium induced in collisions with Hg(6/sup 3/P/sub 1/) atoms

Energy Technology Data Exchange (ETDEWEB)

Wade, M K; Czajkowski, M; Krause, L [Windsor Univ., Ontario (Canada). Dept. of Physics

1978-07-01

The transfer of excitation from excited mercury atoms to ground-state thallium atoms was investigated using techniques of sensitized fluorescence. A Hg-Tl vapor mixture contained in a quartz cell was irradiated with Hg 2537 A resonance radiation which caused the mercury atoms to become excited to the 6/sup 3/P/sub 1/ state. Subsequent collisions between the Hg(6/sup 3/P/sub 1/) and Tl(6/sup 2/Psub(1/2)) atoms resulted in the population of the 8/sup 2/Ssub(1/2), 6/sup 2/D, and 7/sup 2/Ssub(1/2) thallium states, whose decay gave rise to sensitized fluorescence of wavelengths 3231, 3520, 3776, and 5352 A. Intensity measurements on the sensitized fluorescence and on the Hg 2537 A resonance fluorescence, observed at right angles to the direction of excitation, yielded cross sections of 3.0, 0.3, and 0.05 A/sup 2/ for collisional excitation transfer from Hg(6/sup 3/P/sub 1/) to the 8/sup 2/Ssub(1/2), 6/sup 2/D, and 7/sup 2/Ssub(1/2) states in thallium, respectively. The results are fully consistent with previously determined cross sections for excitation transfer in other binary metallic vapor systems.

Sensitive determination of enoxacin in pharmaceutical formulations by its quench effect on the fluorescence of glutathione-capped CdTe quantum dots.

Science.gov (United States)

Yang, Qiong; Tan, Xuanping; Yang, Jidong

2016-02-01

A sensitive and simple method for the determination of enoxacin (ENX) was developed based on the fluorescence quenching effect of ENX for glutathione (GSH)-capped CdTe quantum dots (QDs). Under optimum conditions, a good linear relationship was obtained from 4.333 × 10(-9)  mol⋅L(-1) to 1.4 × 10(-5)  mol⋅L(-1) with a correlation coefficient (R) of 0.9987, and the detection limit (3σ/K) was 1.313 × 10(-9)  mol⋅L(-1). The corresponding mechanism has been proposed on the basis of electron transfer supported by ultraviolet-visible (UV) light absorption, fluorescence spectroscopy, and the measurement of fluorescence lifetime. The method has been applied to the determination of ENX in pharmaceutical formulations (enoxacin gluconate injections and commercial tablets) with satisfactory results. The proposed method manifested several advantages such as high sensitivity, short analysis time, low cost and ease of operation. Copyright © 2015 John Wiley & Sons, Ltd.

Luminescent Lanthanide Reporters for High-Sensitivity Novel Bioassays

Energy Technology Data Exchange (ETDEWEB)

Anstey, Mitchell R. [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Fruetel, Julia A. [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Foster, Michael E. [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Hayden, Carl C. [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Buckley, Heather L. [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Arnold, John [Sandia National Lab. (SNL-CA), Livermore, CA (United States)

2013-09-01

Biological imaging and assay technologies rely on fluorescent organic dyes as reporters for a number of interesting targets and processes. However, limitations of organic dyes such as small Stokes shifts, spectral overlap of emission signals with native biological fluorescence background, and photobleaching have all inhibited the development of highly sensitive assays. To overcome the limitations of organic dyes for bioassays, we propose to develop lanthanide-based luminescent dyes and demonstrate them for molecular reporting applications. This relatively new family of dyes was selected for their attractive spectral and chemical properties. Luminescence is imparted by the lanthanide atom and allows for relatively simple chemical structures that can be tailored to the application. The photophysical properties offer unique features such as narrow and non-overlapping emission bands, long luminescent lifetimes, and long wavelength emission, which enable significant sensitivity improvements over organic dyes through spectral and temporal gating of the luminescent signal.Growth in this field has been hindered due to the necessary advanced synthetic chemistry techniques and access to experts in biological assay development. Our strategy for the development of a new lanthanide-based fluorescent reporter system is based on chelation of the lanthanide metal center using absorbing chromophores. Our first strategy involves "Click" chemistry to develop 3-fold symmetric chelators and the other involves use of a new class of tetrapyrrole ligands called corroles. This two-pronged approach is geared towards the optimization of chromophores to enhance light output.

Graphene oxide-sensitized molecularly imprinted opto-polymers for charge-transfer fluorescent sensing of cyanoguanidine.

Science.gov (United States)

Liu, Huilin; Zhou, Kaiwen; Chen, Xiaomo; Wang, Jing; Wang, Shuo; Sun, Baoguo

2017-11-15

The hierarchical structuring of materials offers exciting opportunities to construct functional sensors. Multiple processes were combined to create complex materials for the selective detection of cyanoguanidine (CYA) using graphene oxide-sensitized molecularly imprinted opto-polymers (MIOP). Molecular imprinting was used to construct molecular-scale analyte-selective cavities, graphene oxide was introduced to provide a platform for the polymerization, and increase the stability and binding kinetic properties, and 3-methacryloxy propyl trimethoxy silane-modified quantum dots were combined with a functional monomer to increase the fluorescence quantum yield. Polymer cross-linking and fluorescence intensity were optimized for molecular recognition and opto-sensing detection. Selective and sensitive, fluorescence sensing of CYA was possible at concentrations as low as to 1.6μM. It could be applied to the rapid and cost-effective monitoring of CYA in infant formula. The approach is generic and applicable to many molecules and conventional opto-sensors, based on molecularly imprinted polymer formulations, individually or in multiplexed arrays. Copyright © 2017 Elsevier Ltd. All rights reserved.

Use of selenium to detect mercury in water and cells: an enhancement of the sensitivity and specificity of a seleno fluorescent probe.

Science.gov (United States)

Tang, Bo; Ding, Baiyu; Xu, Kehua; Tong, Lili

2009-01-01

Seleno fluorescent probe: An organoselenium fluorescent probe (FSe-1) for mercury was designed based on the irreversible deselenation mechanism. FSe-1 exhibits an ultrahigh selectivity and sensitivity for Hg(2+) detection only for reactive selenium atom sites, due the strong affinity between Se and Hg. Furthermore, the new probe has been successfully used for imaging mercury ions in RAW 264.7 cells (a mouse macrophage cell line; see figure).Inspired by the antitoxic function of selenium towards heavy-metal ions, we designed an organoselenium fluorescent probe (FSe-1) for mercury. The reaction of FSe-1 and Hg(2+) is an irreversible deselenation mechanism based on the selenophilic character of mercury. FSe-1 exhibits an ultrahigh selectivity and sensitivity for Hg(2+) detection only for reactive selenium atom sites due to the strong affinity between Se and Hg. The experimental results proved that FSe-1 was selective for Hg(2+) ions over other relevant metal ions and bioanalytes, and also showed an enhancement in sensitivity of up to 1.0 nM, which is lower than the current Environmental Protection Agency standard for drinking water. Furthermore, the new probe has been successfully applied to the imaging of mercury ions in RAW 264.7 cells (a mouse macrophage cell line) with high sensitivity and selectivity.

Smartphone-Based Fluorescent Diagnostic System for Highly Pathogenic H5N1 Viruses

OpenAIRE

Yeo, Seon-Ju; Choi, Kyunghan; Cuc, Bui Thi; Hong, Nguyen Ngoc; Bao, Duong Tuan; Ngoc, Nguyen Minh; Le, Mai Quynh; Hang, Nguyen Le Khanh; Thach, Nguyen Co; Mallik, Shyam Kumar; Kim, Hak Sung; Chong, Chom-Kyu; Choi, Hak Soo; Sung, Haan Woo; Yu, Kyoungsik

2016-01-01

Field diagnostic tools for avian influenza (AI) are indispensable for the prevention and controlled management of highly pathogenic AI-related diseases. More accurate, faster and networked on-site monitoring is demanded to detect such AI viruses with high sensitivity as well as to maintain up-to-date information about their geographical transmission. In this work, we assessed the clinical and field-level performance of a smartphone-based fluorescent diagnostic device with an efficient reflect...

Fluorescent Biosensors Based on Single-Molecule Counting.

Science.gov (United States)

Ma, Fei; Li, Ying; Tang, Bo; Zhang, Chun-Yang

2016-09-20

Biosensors for highly sensitive, selective, and rapid quantification of specific biomolecules make great contributions to biomedical research, especially molecular diagnostics. However, conventional methods for biomolecular assays often suffer from insufficient sensitivity and poor specificity. In some case (e.g., early disease diagnostics), the concentration of target biomolecules is too low to be detected by these routine approaches, and cumbersome procedures are needed to improve the detection sensitivity. Therefore, there is an urgent need for rapid and ultrasensitive analytical tools. In this respect, single-molecule fluorescence approaches may well satisfy the requirement and hold promising potential for the development of ultrasensitive biosensors. Encouragingly, owing to the advances in single-molecule microscopy and spectroscopy over past decades, the detection of single fluorescent molecule comes true, greatly boosting the development of highly sensitive biosensors. By in vitro/in vivo labeling of target biomolecules with proper fluorescent tags, the quantification of certain biomolecule at the single-molecule level is achieved. In comparison with conventional ensemble measurements, single-molecule detection-based analytical methods possess the advantages of ultrahigh sensitivity, good selectivity, rapid analysis time, and low sample consumption. Consequently, single-molecule detection may be potentially employed as an ideal analytical approach to quantify low-abundant biomolecules with rapidity and simplicity. In this Account, we will summarize our efforts for developing a series of ultrasensitive biosensors based on single-molecule counting. Single-molecule counting is a member of single-molecule detection technologies and may be used as a very simple and ultrasensitive method to quantify target molecules by simply counting the individual fluorescent bursts. In the fluorescent sensors, the signals of target biomolecules may be translated to the

Selective and sensitive fluorescent sensor for Pd2+ using coumarin 460 for real-time and biological applications.

Science.gov (United States)

Ashwin, Bosco Christin Maria Arputham; Sivaraman, Gandhi; Stalin, Thambusamy; Yuvakkumar, Rathinam; Muthu Mareeswaran, Paulpandian

2018-05-03

The efficient fluorescent property of coumarin 460 (C460) is utilized to sense the Pd 2+ selectively and sensitively. Fabrication of a sensor strip using commercial adhesive tape is achieved and the detection of Pd 2+ is attempted using a handy UV torch. The naked eye detection in solution state using UV chamber is also attempted. The calculated high binding constant values support the strong stable complex formation of Pd 2+ with C460. The detection limit up to 2.5 × 10 -7  M is achieved using fluorescence spectrometer, which is considerably low from the WHO's recommendation. The response of coumarin 460 with various cations also studied. The quenching is further studied by the lifetime measurements. The binding mechanism is clearly explained by the 1 H NMR titration. The sensing mechanism is established as ICT. C460 strip's Pd 2+ quenching detection is further confirmed by solid-state PL study. The in-vitro response of Pd 2+ in a living cell is also studied using fluorescent imaging studies by means of HeLa cell lines and this probe is very compatible with biological environments. It could be applicable to sense trace amounts of a Pd 2+ ion from various industries. Compared with previous reports, this one is very cheap, sensitive, selective and suitable for biological systems. Copyright © 2018 Elsevier B.V. All rights reserved.

Sensitive and selective detection of Hg2+ and Cu2+ ions by fluorescent Ag nanoclusters synthesized via a hydrothermal method

Science.gov (United States)

Liu, Jing; Ren, Xiangling; Meng, Xianwei; Fang, Zheng; Tang, Fangqiong

2013-09-01

An easily prepared fluorescent Ag nanoclusters (Ag NCs) probe for the sensitive and selective detection of Hg2+ and Cu2+ ions was developed here. The Ag NCs were synthesized by using polymethacrylic acid sodium salt as a template via a convenient hydrothermal process. The as-prepared fluorescent Ag NCs were monodispersed, uniform and less than 2 nm in diameter, and can be quenched in the presence of mercury (Hg2+) or copper (Cu2+) ions. Excellent linear relationships existed between the quenching degree of the Ag NCs and the concentrations of Hg2+ or Cu2+ ions in the range of 10 nM to 20 μM or 10 nM to 30 μM, respectively. By using ethylenediaminetetraacetate (EDTA) as the masking agent of Cu2+, Hg2+ was exclusively detected in coexistence with Cu2+ with high sensitivity (LOD = 10 nM), which also provided a reusable detection method for Cu2+. Furthermore, the different quenching phenomena caused by the two metals ions such as changes in visible colour, shifts of UV absorbance peaks and changes in size of Ag NCs make it easy to distinguish between them. Therefore the easily synthesized fluorescent Ag NCs may have great potential as Hg2+ and Cu2+ ions sensors.An easily prepared fluorescent Ag nanoclusters (Ag NCs) probe for the sensitive and selective detection of Hg2+ and Cu2+ ions was developed here. The Ag NCs were synthesized by using polymethacrylic acid sodium salt as a template via a convenient hydrothermal process. The as-prepared fluorescent Ag NCs were monodispersed, uniform and less than 2 nm in diameter, and can be quenched in the presence of mercury (Hg2+) or copper (Cu2+) ions. Excellent linear relationships existed between the quenching degree of the Ag NCs and the concentrations of Hg2+ or Cu2+ ions in the range of 10 nM to 20 μM or 10 nM to 30 μM, respectively. By using ethylenediaminetetraacetate (EDTA) as the masking agent of Cu2+, Hg2+ was exclusively detected in coexistence with Cu2+ with high sensitivity (LOD = 10 nM), which also provided a

A Comparison of the Capability of Sensitivity Level 3 and Sensitivity Level 4 Fluorescent Penetrants to Detect Fatigue Cracks in Aluminum

Science.gov (United States)

Parker, Bradford, H.

2009-01-01

Historically both sensitivity level 3 and sensitivity level 4 fluorescent penetrants have been used to perform NASA Standard Level inspections of aerospace hardware. In April 2008, NASA-STD-5009 established a requirement that only sensitivity level 4 penetrants were acceptable for inspections of NASA hardware. Having NASA contractors change existing processes or perform demonstration tests to certify sensitivity level 3 penetrants posed a potentially huge cost to the Agency. This study was conducted to directly compare the probability of detection sensitivity level 3 and level 4 penetrants using both Method A and Method D inspection processes. The study results strongly support the conclusion that sensitivity level 3 penetrants are acceptable for NASA Standard Level inspections

Sensitive determination of pipecolic acid in serum by high-performance liquid chromatography using 4-(5,6-dimethoxy-2-phthalimidinyl)-2-methoxyphenylsulfonyl chloride as a fluorescent labelling reagent

International Nuclear Information System (INIS)

Inoue, Hirofumi; Sakata, Yasuhiko; Fukunaga, Keiko; Nishio, Hiroaki; Tsuruta, Yasuto

2004-01-01

A simple and highly sensitive high-performance liquid chromatography (HPLC) for the determination of pipecolic acid (PA) in serum was developed. Pipecolic acid and nipecotic acid (internal standard (IS)) were derivatised with 4-(5,6-dimethoxy-2-phthalimidinyl)-2-methoxyphenylsulfonyl chloride (DMS-Cl) to produce fluorescent sulfonamides. The labelling reaction was carried out at 70 degree sign C for 15 min at pH 9.0. The fluorescent derivatives were separated on a reversed-phase column (45 degree sign C) with a stepwise elution using methanol/acetonitrile/10 mmol l -1 acetic acid (42:5:53) and methanol at a flow rate of 1.0 ml/min and detected at excitation and emission wavelengths of 316 and 403 nm, respectively. The labelling yield was 100.8%. The detection limit of pipecolic acid was 4 fmol at signal-to-noise ratio of 3. The within-day and day-to-day relative standard deviations were 3.3-8.1 and 1.4-6.4%, respectively. The concentration of pipecolic acid in normal human serum was 1.09±0.37 μmol l -1

Highly sensitive and selective detection of dopamine using one-pot synthesized highly photoluminescent silicon nanoparticles.

Science.gov (United States)

Zhang, Xiaodong; Chen, Xiaokai; Kai, Siqi; Wang, Hong-Yin; Yang, Jingjing; Wu, Fu-Gen; Chen, Zhan

2015-03-17

A simple and highly efficient method for dopamine (DA) detection using water-soluble silicon nanoparticles (SiNPs) was reported. The SiNPs with a high quantum yield of 23.6% were synthesized by using a one-pot microwave-assisted method. The fluorescence quenching capability of a variety of molecules on the synthesized SiNPs has been tested; only DA molecules were found to be able to quench the fluorescence of these SiNPs effectively. Therefore, such a quenching effect can be used to selectively detect DA. All other molecules tested have little interference with the dopamine detection, including ascorbic acid, which commonly exists in cells and can possibly affect the dopamine detection. The ratio of the fluorescence intensity difference between the quenched and unquenched cases versus the fluorescence intensity without quenching (ΔI/I) was observed to be linearly proportional to the DA analyte concentration in the range from 0.005 to 10.0 μM, with a detection limit of 0.3 nM (S/N = 3). To the best of our knowledge, this is the lowest limit for DA detection reported so far. The mechanism of fluorescence quenching is attributed to the energy transfer from the SiNPs to the oxidized dopamine molecules through Förster resonance energy transfer. The reported method of SiNP synthesis is very simple and cheap, making the above sensitive and selective DA detection approach using SiNPs practical for many applications.

Label-free tungsten disulfide quantum dots as a fluorescent sensing platform for highly efficient detection of copper (II) ions

International Nuclear Information System (INIS)

Zhao Xuan; He Da-Wei; Wang Yong-Sheng; Hu Yin; Fu Chen; Li Xue

2017-01-01

A fluorescent probe for the sensitive and selective determination of copper ion (Cu 2+ ) is presented. It is based on the use of tungsten disulfide quantum dots (WS 2 QDs) which is independent of the pH of solution and emits strong blue fluorescence. Copper ions could cause aggregation of the WS 2 QDs and lead to fluorescence quenching of WS 2 QDs. The change of fluorescence intensity is proportional to the concentration of Cu 2+ , and the limit of detection is 0.4 μM. The fluorescent probe is highly selective for Cu 2+ over some potentially interfering ions. These results indicate that WS 2 QDs, as a fluorescent sensing platform, can meet the selective requirements for biomedical and environmental application. (paper)

Enhanced fluorescence sensitivity by coupling yttrium-analyte complexes and three-way fast high-performance liquid chromatography data modeling

Energy Technology Data Exchange (ETDEWEB)

Alcaraz, Mirta R.; Culzoni, María J., E-mail: mculzoni@fbcb.unl.edu.ar; Goicoechea, Héctor C., E-mail: hgoico@fbcb.unl.edu.ar

2016-01-01

The present study reports a sensitive chromatographic method for the analysis of seven fluoroquinolones (FQs) in environmental water samples, by coupling yttrium-analyte complex and three-way chromatographic data modeling. This method based on the use of HPLC-FSFD does not require complex or tedious sample treatments or enrichment processes before the analysis, due to the significant fluorescence increments of the analytes reached by the presence of Y{sup 3+}. Enhancement achieved for the FQs signals obtained after Y{sup 3+} addition reaches 103- to 1743-fold. Prediction results corresponding to the application of MCR-ALS to the validation set showed relative error of prediction (REP%) values below 10% in all cases. A recovery study that includes the simultaneous determination of the seven FQs in three different environmental aqueous matrices was conducted. The recovery studies assert the efficiency and the accuracy of the proposed method. The LOD values calculated are in the order of part per trillion (below 0.5 ng mL{sup −1} for all the FQs, except for enoxacin). It is noteworthy to mention that the method herein proposed, which does not include pre-concentration steps, allows reaching LOD values in the same order of magnitude than those achieved by more sophisticated methods based on SPE and UHPLC-MS/MS. - Highlights: • Highly sensitive method for the analysis of seven fluoroquinolones. • Coupling of yttrium-analyte complex and three-way modeling. • Complex or tedious sample treatments or enrichment processes are nor required. • Accuracy on the quantitation of fluoroquinolones in real water river samples.

A Microneedle Functionalized with Polyethyleneimine and Nanotubes for Highly Sensitive, Label-Free Quantification of DNA.

Science.gov (United States)

Saadat-Moghaddam, Darius; Kim, Jong-Hoon

2017-08-16

The accurate measure of DNA concentration is necessary for many DNA-based biological applications. However, the current methods are limited in terms of sensitivity, reproducibility, human error, and contamination. Here, we present a microneedle functionalized with polyethyleneimine (PEI) and single-walled carbon nanotubes (SWCNTs) for the highly sensitive quantification of DNA. The microneedle was fabricated using ultraviolet (UV) lithography and anisotropic etching, and then functionalized with PEI and SWCNTs through a dip coating process. The electrical characteristics of the microneedle change with the accumulation of DNA on the surface. Current-voltage measurements in deionized water were conducted to study these changes in the electrical properties of the sensor. The sensitivity test found the signal to be discernable from the noise level down to 100 attomolar (aM), demonstrating higher sensitivity than currently available UV fluorescence and UV absorbance based methods. A microneedle without any surface modification only had a 100 femtomolar (fM) sensitivity. All measurement results were consistent with fluorescence microscopy.

Non-Covalent Fluorescent Labeling of Hairpin DNA Probe Coupled with Hybridization Chain Reaction for Sensitive DNA Detection.

Science.gov (United States)

Song, Luna; Zhang, Yonghua; Li, Junling; Gao, Qiang; Qi, Honglan; Zhang, Chengxiao

2016-04-01

An enzyme-free signal amplification-based assay for DNA detection was developed using fluorescent hairpin DNA probes coupled with hybridization chain reaction (HCR). The hairpin DNAs were designed to contain abasic sites in the stem moiety. Non-covalent labeling of the hairpin DNAs was achieved when a fluorescent ligand was bound to the abasic sites through hydrogen bonding with the orphan cytosine present on the complementary strand, accompanied by quench of ligand fluorescence. As a result, the resultant probes, the complex formed between the hairpin DNA and ligand, showed almost no fluorescence. Upon hybridization with target DNA, the probe underwent a dehybridization of the stem moiety containing an abasic site. The release of ligand from the abasic site to the solution resulted in an effective fluorescent enhancement, which can be used as a signal. Compared with a sensing system without HCR, a 20-fold increase in the sensitivity was achieved using the sensing system with HCR. The fluorescent intensity of the sensing system increased with the increase in target DNA concentration from 0.5 nM to 100 nM. A single mismatched target ss-DNA could be effectively discriminated from complementary target DNA. Genotyping of a G/C single-nucleotide polymorphism of polymerase chain reaction (PCR) products was successfully demonstrated with the sensing system. Therefore, integrating HCR strategy with non-covalent labeling of fluorescent hairpin DNA probes provides a sensitive and cost-effective DNA assay. © The Author(s) 2016.

Nanozyme-based bio-barcode assay for high sensitive and logic-controlled specific detection of multiple DNAs.

Science.gov (United States)

Lin, Xiaodong; Liu, Yaqing; Tao, Zhanhui; Gao, Jinting; Deng, Jiankang; Yin, Jinjin; Wang, Shuo

2017-08-15

Since HCV and HIV share a common transmission path, high sensitive detection of HIV and HCV gene is of significant importance to improve diagnosis accuracy and cure rate at early stage for HIV virus-infected patients. In our investigation, a novel nanozyme-based bio-barcode fluorescence amplified assay is successfully developed for simultaneous detection of HIV and HCV DNAs with excellent sensitivity in an enzyme-free and label-free condition. Here, bimetallic nanoparticles, PtAu NPs , present outstanding peroxidase-like activity and act as barcode to catalyze oxidation of nonfluorescent substrate of amplex red (AR) into fluorescent resorufin generating stable and sensitive "Turn On" fluorescent output signal, which is for the first time to be integrated with bio-barcode strategy for fluorescence detection DNA. Furthermore, the provided strategy presents excellent specificity and can distinguish single-base mismatched mutant from target DNA. What interesting is that cascaded INHIBIT-OR logic gate is integrated with biosensors for the first time to distinguish individual target DNA from each other under logic function control, which presents great application in development of rapid and intelligent detection. Copyright © 2017. Published by Elsevier B.V.

Sensitization of uranium fluorescence using 2,6-pyridinedicarboxylic acid: Application for the determination of uranium in the presence of lanthanides

International Nuclear Information System (INIS)

Maji, S.; Viswanathan, K.S.

2009-01-01

The 2,6-pyridinedicarboxylic acid (PDA) has been shown to efficiently sensitize and enhance the fluorescence of uranium in aqueous medium. Interestingly, this ligand stabilizes the UO 2 2+ species, which without the ligand is known to be in a negligible concentration, in aqueous medium at pH 6. The ligand sensitized enhancement of UO 2 2+ fluorescence by PDA, provides an analytical tool for the determination of uranium at trace levels, in aqueous medium. Furthermore, PDA is also known to enhance the fluorescence of lanthanides; consequently, the simultaneous determination of uranium and lanthanides, using PDA as a fluorescence sensitizing agent, becomes a possibility, which has been demonstrated in this work. We have shown that the use of PDA yields detection limits of 2.2x10 -7 M for UO 2 2+ , 1x10 -8 M for Tb 3+ and 5x10 -9 M for Eu 3+ in the simultaneous determination of these analytes.

Synchronous fluorescence based biosensor for albumin determination by cooperative binding of fluorescence probe in a supra-biomolecular host-protein assembly.

Science.gov (United States)

Patra, Digambara

2010-01-15

A synchronous fluorescence probe based biosensor for estimation of albumin with high sensitivity and selectivity was developed. Unlike conventional fluorescence emission or excitation spectral measurements, synchronous fluorescence measurement offered exclusively a new synchronous fluorescence peak in the shorter wavelength range upon binding of chrysene with protein making it an easy identification tool for albumin determination. The cooperative binding of a fluorescence probe, chrysene, in a supramolecular host-protein assembly during various albumin assessments was investigated. The presence of supramolecular host molecules such as beta-cyclodextrin, curucurbit[6]uril or curucurbit[7]uril have little influence on sensitivity or limit of detection during albumin determination but reduced dramatically interference from various coexisting metal ion quenchers/enhancers. Using the present method the limit of detection for BSA and gamma-Globulin was found to be 0.005 microM which is more sensitive than reported values. Copyright 2009 Elsevier B.V. All rights reserved.

Label-free fluorescence strategy for sensitive microRNA detection based on isothermal exponential amplification and graphene oxide.

Science.gov (United States)

Li, Wei; Hou, Ting; Wu, Min; Li, Feng

2016-01-01

MicroRNAs (miRNAs) play an important role in many biological processes, and have been regarded as potential targets and biomarkers in cancer diagnosis and therapy. Also, to meet the big challenge imposed by the characteristics of miRNAs, such as small size and vulnerability to enzymatic digestion, it is of great importance to develop accurate, sensitive and simple miRNA assays. Herein, we developed a label-free fluorescence strategy for sensitive miRNA detection by combining isothermal exponential amplification and the unique features of SYBR Green I (SG) and graphene oxide (GO), in which SG gives significantly enhanced fluorescence upon intercalation into double-stranded DNAs (dsDNAs), and GO selectively adsorbs miRNA, single-stranded DNA and SG, to protect miRNA from enzymatic digestion, and to quench the fluorescence of the adsorbed SG. In the presence of the target miRNA, the ingeniously designed hairpin probe (HP) is unfolded and the subsequent polymerization and strand displacement reaction takes place to initiate the target recycling process. The newly formed dsDNAs are then recognized and cleaved by the nicking enzyme, generating new DNA triggers with the same sequence as the target miRNA, which hybridize with intact HPs to initiate new extension reactions. As a result, the circular exponential amplification for target miRNA is achieved and large amount of dsDNAs are formed to generate significantly enhanced fluorescence upon the intercalation of SG. Thus sensitive and selective fluorescence miRNA detection is realized, and the detection limit of 3 fM is obtained. Besides, this method exhibits additional advantages of simplicity and low cost, since expensive and tedious labeling process is avoided. Therefore, the as-proposed label-free fluorescence strategy has great potential in the applications in miRNA-related clinical practices and biochemical researches. Copyright © 2015 Elsevier B.V. All rights reserved.

RNA-ID, a highly sensitive and robust method to identify cis-regulatory sequences using superfolder GFP and a fluorescence-based assay.

Science.gov (United States)

Dean, Kimberly M; Grayhack, Elizabeth J

2012-12-01

We have developed a robust and sensitive method, called RNA-ID, to screen for cis-regulatory sequences in RNA using fluorescence-activated cell sorting (FACS) of yeast cells bearing a reporter in which expression of both superfolder green fluorescent protein (GFP) and yeast codon-optimized mCherry red fluorescent protein (RFP) is driven by the bidirectional GAL1,10 promoter. This method recapitulates previously reported progressive inhibition of translation mediated by increasing numbers of CGA codon pairs, and restoration of expression by introduction of a tRNA with an anticodon that base pairs exactly with the CGA codon. This method also reproduces effects of paromomycin and context on stop codon read-through. Five key features of this method contribute to its effectiveness as a selection for regulatory sequences: The system exhibits greater than a 250-fold dynamic range, a quantitative and dose-dependent response to known inhibitory sequences, exquisite resolution that allows nearly complete physical separation of distinct populations, and a reproducible signal between different cells transformed with the identical reporter, all of which are coupled with simple methods involving ligation-independent cloning, to create large libraries. Moreover, we provide evidence that there are sequences within a 9-nt library that cause reduced GFP fluorescence, suggesting that there are novel cis-regulatory sequences to be found even in this short sequence space. This method is widely applicable to the study of both RNA-mediated and codon-mediated effects on expression.

Imaging Lysosomal pH Alteration in Stressed Cells with a Sensitive Ratiometric Fluorescence Sensor.

Science.gov (United States)

Xue, Zhongwei; Zhao, Hu; Liu, Jian; Han, Jiahuai; Han, Shoufa

2017-03-24

The organelle-specific pH is crucial for cell homeostasis. Aberrant pH of lysosomes has been manifested in myriad diseases. To probe lysosome responses to cell stress, we herein report the detection of lysosomal pH changes with a dual colored probe (CM-ROX), featuring a coumarin domain with "always-on" blue fluorescence and a rhodamine-lactam domain activatable to lysosomal acidity to give red fluorescence. With sensitive ratiometric signals upon subtle pH changes, CM-ROX enables discernment of lysosomal pH changes in cells undergoing autophagy, cell death, and viral infection.

Solid state photon upconversion utilizing thermally activated delayed fluorescence molecules as triplet sensitizer

Energy Technology Data Exchange (ETDEWEB)

Wu, Tony C.; Congreve, Daniel N.; Baldo, Marc A., E-mail: baldo@mit.edu [Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 (United States)

2015-07-20

The ability to upconvert light is useful for a range of applications, from biological imaging to solar cells. But modern technologies have struggled to upconvert incoherent incident light at low intensities. Here, we report solid state photon upconversion employing triplet-triplet exciton annihilation in an organic semiconductor, sensitized by a thermally activated-delayed fluorescence (TADF) dye. Compared to conventional phosphorescent sensitizers, the TADF dye maximizes the wavelength shift in upconversion due to its small singlet-triplet splitting. The efficiency of energy transfer from the TADF dye is 9.1%, and the conversion yield of sensitizer exciton pairs to singlet excitons in the annihilator is 1.1%. Our results demonstrate upconversion in solid state geometries and with non-heavy metal-based sensitizer materials.

A novel fluorescence immunoassay for the sensitive detection of Escherichia coli O157:H7 in milk based on catalase-mediated fluorescence quenching of CdTe quantum dots

International Nuclear Information System (INIS)

Chen, Rui; Huang, Xiaolin; Li, Juan; Shan, Shan; Lai, Weihua; Xiong, Yonghua

2016-01-01

Immunoassay is a powerful tool for rapid detection of food borne pathogens in food safety monitoring. However, conventional immunoassay always suffers from low sensitivity when it employs enzyme-catalyzing chromogenic substrates to generate colored molecules as signal outputs. In the present study, we report a novel fluorescence immunoassay for the sensitive detection of E. coli O157:H7 through combination of the ultrahigh bioactivity of catalase to hydrogen peroxide (H_2O_2) and H_2O_2-sensitive mercaptopropionic acid modified CdTe QDs (MPA-QDs) as a signal transduction. Various parameters, including the concentrations of anti-E. coli O157:H7 polyclonal antibody and biotinylated monoclonal antibody, the amounts of H_2O_2 and streptavidin labeled catalase (CAT), the hydrolysis temperature and time of CAT to H_2O_2, as well as the incubation time between H_2O_2 and MPA-QDs, were systematically investigated and optimized. With optimal conditions, the catalase-mediated fluorescence quenching immunoassay exhibits an excellent sensitivity for E. coli O157:H7 with a detection limit of 5 × 10"2 CFU/mL, which was approximately 140 times lower than that of horseradish peroxidase-based colorimetric immunoassay. The reliability of the proposed method was further evaluated using E. coli O157:H7 spiked milk samples. The average recoveries of E. coli O157:H7 concentrations from 1.18 × 10"3 CFU/mL to 1.18 × 10"6 CFU/mL were in the range of 65.88%–105.6%. In brief, the proposed immunoassay offers a great potential for rapid and sensitive detection of other pathogens in food quality control. - Highlights: • A novel fluorescence immunoassay was developed for the ultrasensitive detection of E. coli O157:H7. • This detection was achieved through the combination of the high bioactivity of CAT and H_2O_2-sensitive QDs. • The activity of CAT to H_2O_2 is 1000 folds higher than that of the HRP to tetramethylbenzidine. • The limit of detection of the proposed method could

Impurity monitoring by laser-induced fluorescence techniques

International Nuclear Information System (INIS)

Gelbwachs, J.A.

1984-01-01

Laser-induced fluorescence spectroscopy can provide a highly sensitive and selective means of detecting atomic and ionic impurities. Because the photodetector can be physically isolated from the laser-excited region, these techniques can be applied to monitoring in hostile environments. The basic concepts behind fluorescence detection are reviewed. Saturated optical excitation is shown to maximize impurity atom emission yield while mitigating effects of laser intensity fluctuations upon absolute density calibration. Monitoring in high- and low-pressure monitoring environments is compared. Methods to improve detection sensitivity by luminescence background suppression are presented

Selective and Sensitive Detection of Cyanide Based on the Displacement Strategy Using a Water-Soluble Fluorescent Probe

Science.gov (United States)

La, Ming; Hao, Yuanqiang; Wang, Zhaoyang; Han, Guo-Cheng; Qu, Lingbo

2016-01-01

A water-soluble fluorescent probe (C-GGH) was used for the highly sensitive and selective detection of cyanide (CN−) in aqueous media based on the displacement strategy. Due to the presence of the recognition unit GGH (Gly-Gly-His), the probe C-GGH can coordinate with Cu2+ and consequently display ON-OFF type fluorescence response. Furthermore, the in situ formed nonfluorescent C-GGH-Cu2+ complex can act as an effective OFF-ON type fluorescent probe for sensing CN− anion. Due to the strong binding affinity of CN− to Cu2+, CN− can extract Cu2+ from C-GGH-Cu2+ complex, leading to the release of C-GGH and the recovery of fluorescent emission of the system. The probe C-GGH-Cu2+ allowed detection of CN− in aqueous solution with a LOD (limit of detection) of 0.017 μmol/L which is much lower than the maximum contaminant level (1.9 μmol/L) for CN− in drinking water set by the WHO (World Health Organization). The probe also displayed excellent specificity for CN− towards other anions, including F−, Cl−, Br−, I−, SCN−, PO4 3−, N3 −, NO3 −, AcO−, SO4 2−, and CO3 2−. PMID:26881185

Selective and Sensitive Detection of Cyanide Based on the Displacement Strategy Using a Water-Soluble Fluorescent Probe

Directory of Open Access Journals (Sweden)

Ming La

2016-01-01

Full Text Available A water-soluble fluorescent probe (C-GGH was used for the highly sensitive and selective detection of cyanide (CN− in aqueous media based on the displacement strategy. Due to the presence of the recognition unit GGH (Gly-Gly-His, the probe C-GGH can coordinate with Cu2+ and consequently display ON-OFF type fluorescence response. Furthermore, the in situ formed nonfluorescent C-GGH-Cu2+ complex can act as an effective OFF-ON type fluorescent probe for sensing CN− anion. Due to the strong binding affinity of CN− to Cu2+, CN− can extract Cu2+ from C-GGH-Cu2+ complex, leading to the release of C-GGH and the recovery of fluorescent emission of the system. The probe C-GGH-Cu2+ allowed detection of CN− in aqueous solution with a LOD (limit of detection of 0.017 μmol/L which is much lower than the maximum contaminant level (1.9 μmol/L for CN− in drinking water set by the WHO (World Health Organization. The probe also displayed excellent specificity for CN− towards other anions, including F−, Cl−, Br−, I−, SCN−, PO43-, N3-, NO3-, AcO−, SO42-, and CO32-.

A new simple phthalimide-based fluorescent probe for highly selective cysteine and bioimaging for living cells

Science.gov (United States)

Shen, Youming; Zhang, Xiangyang; Zhang, Youyu; Zhang, Chunxiang; Jin, Junling; Li, Haitao

2017-10-01

A new turn-on phthalimide fluorescent probe has designed and synthesized for sensing cysteine (Cys) based on excited state intramolecular proton transfer (ESIPT) process. It is consisted of a 3-hydroxyphthalimide derivative moiety as the fluorophore and an acrylic ester group as a recognition receptor. The acrylic ester acts as an ESIPT blocking agent. Upon addition of cystein, intermolecular nucleophilic attack of cysteine on acrylic ester releases the fluorescent 3-hydroxyphthalimide derivative, thereby enabling the ESIPT process and leading to enhancement of fluorescence. The probe displays high sensitivity, excellent selectivity and with large Stokes shift toward cysteine. The linear interval range of the fluorescence titration ranged from 0 to 1.0 × 10- 5 M and detection limit is low (6 × 10- 8 M). In addition, the probe could be used for bio-imaging in living cells.

Highly sensitive FRET-based fluorescence immunoassay for aflatoxin B1 using cadmium telluride quantum dots

International Nuclear Information System (INIS)

Zekavati, Roya; Bayat, Mansour; Safi, Shahabeddin; Hashemi, Seyed Jamal; Rahmani-Cherati, Tavoos; Tabatabaei, Meisam; Mohsenifar, Afshin

2013-01-01

We report on a competitive immunoassay for the determination of aflatoxin B1 using fluorescence resonance energy transfer (FRET) from anti-aflatoxin B1 antibody (immobilized on the shell of CdTe quantum dots) to Rhodamine 123 (Rho 123-labeled aflatoxin B1 bound to albumin). The highly specific immuno reaction between the antibody against aflatoxin B1 on the QDs and the labeled-aflatoxin B1 brings the Rho 123 fluorophore (acting as the acceptor) and the QDs (acting as the donor) in close spatial proximity and causes FRET to occur upon photoexcitation of the QDs. In the absence of unlabeled aflatoxin B1, the antigen-antibody complex is stable, and strong emission resulting from the FRET from QDs to labeled aflatoxin B1 is observed. In the presence of aflatoxin B1, it will compete with the labeled aflatoxin B1-albumin complex for binding to the antibody-QDs conjugate so that FRET will be increasingly suppressed. The reduction in the fluorescence intensity of the acceptor correlates well with the concentration of aflatoxin B1. The feasibility of the method was established by the detection of aflatoxin B1 in spiked human serum. There is a linear relationship between the increased fluorescence intensity of Rho 123 with increasing concentration of aflatoxin B1 in spike human serum, over the range of 0.1–0.6 μmol·mL −1 . The limit of detection is 2 × 10 −11 M. This homogeneous competitive detection scheme is simple, rapid and efficient, and does not require excessive washing and separation steps. (author)

Riboflavin enhanced fluorescence of highly reduced graphene oxide

Science.gov (United States)

Iliut, Maria; Gabudean, Ana-Maria; Leordean, Cosmin; Simon, Timea; Teodorescu, Cristian-Mihail; Astilean, Simion

2013-10-01

The improvement of graphene derivates' fluorescence properties is a challenging topic and very few ways were reported up to now. In this Letter we propose an easy method to enhance the fluorescence of highly reduced graphene oxide (rGO) through non-covalent binding to a molecular fluorophore, namely the riboflavin (Rb). While the fluorescence of Rb is quenched, the Rb - decorated rGO exhibits strong blue fluorescence and significantly increased fluorescence lifetime, as compared to its pristine form. The data reported here represent a promising start towards tailoring the optical properties of rGOs, having utmost importance in optical applications.

Problems of fluorescent imaging and its solution using nanofluorophores. Part I: Advantages of fluorescent nanoparticles over conventional organic fluorophores

International Nuclear Information System (INIS)

Zhelev, Z.; Hadjidekov, G.; Zlateva, G.; Spasov, L.; Bakalova, R.

2011-01-01

The application of fluorescence in deep-tissue imaging is rapidly expanding in fast several years. The progress in fluorescent molecular probes and fluorescent imaging techniques gives an opportunity to detect single cells and even molecules in live organisms. The highly sensitive and high-speed fluorescent molecular sensors and detection devices allow the application of fluorescence in functional imaging. With development of novel bright fluorophores based on nano-technologies and fluorescence scanners with high spatial and temporal resolution, the fluorescent imaging has a potential to become an alternative of the other non-invasive imaging techniques as magnetic resonance imaging, positron-emission tomography, X-ray, computing tomography. This review outlines the current status and future trends of fluorescent nanoparticles - quantum dots (QDs), as a new generation of fluorophores in experimental and pre-clinical fluorescent imaging diagnostic. Part 1 focuses on the advantages of quantum dots over conventional organic fluorophores and defines the major requirements to the 'perfect' fluorophore for fluorescent deep-tissue imaging diagnostic. The analysis is based on the limitations of fluorescent imaging in vivo and overcome by using quantum dots

Sensitization of uranium fluorescence using 2,6-pyridinedicarboxylic acid: Application for the determination of uranium in the presence of lanthanides

Energy Technology Data Exchange (ETDEWEB)

Maji, S. [Materials Chemistry Division, Chemistry Group, Indira Gandhi Centre for Atomic Research, Kalpakkam 603 102 (India); Viswanathan, K.S., E-mail: vish@igcar.gov.i [Materials Chemistry Division, Chemistry Group, Indira Gandhi Centre for Atomic Research, Kalpakkam 603 102 (India)

2009-11-15

The 2,6-pyridinedicarboxylic acid (PDA) has been shown to efficiently sensitize and enhance the fluorescence of uranium in aqueous medium. Interestingly, this ligand stabilizes the UO{sub 2}{sup 2+} species, which without the ligand is known to be in a negligible concentration, in aqueous medium at pH 6. The ligand sensitized enhancement of UO{sub 2}{sup 2+} fluorescence by PDA, provides an analytical tool for the determination of uranium at trace levels, in aqueous medium. Furthermore, PDA is also known to enhance the fluorescence of lanthanides; consequently, the simultaneous determination of uranium and lanthanides, using PDA as a fluorescence sensitizing agent, becomes a possibility, which has been demonstrated in this work. We have shown that the use of PDA yields detection limits of 2.2x10{sup -7} M for UO{sub 2}{sup 2+}, 1x10{sup -8} M for Tb{sup 3+} and 5x10{sup -9} M for Eu{sup 3+} in the simultaneous determination of these analytes.

Highly selective detection of p-nitrophenol using fluorescence assay based on boron, nitrogen co-doped carbon dots.

Science.gov (United States)

Xiao, Na; Liu, Shi Gang; Mo, Shi; Li, Na; Ju, Yan Jun; Ling, Yu; Li, Nian Bing; Luo, Hong Qun

2018-07-01

p-Nitrophenol (p-NP) contaminants seriously endanger environmental and living beings health, hence to establish a sensitive and selective method is of great importance for the determination of p-NP. In this work, boron and nitrogen co-doped carbon dots (B,N-CDs) were synthesized by one-step hydrothermal method using 3-aminophenylboronic acid as the sole precursor. The product was characterized through high-resolution transmission electron microscopy, fluorescence spectroscopy, UV-visible absorption spectroscopy, X-ray photoelectron spectroscopy, and Fourier transform infrared spectroscopy. Without any functionalized modification, B,N-CDs can be directly applied as a 'turn-off' fluorescent probe for rapid, highly selective, and sensitive detection of p-NP. The fluorescent sensor based on the B,N-CDs exhibited a broad linear response to the concentration of p-NP in the range of 0.5 - 60 μM and 60 - 200 μM, respectively, and provided a detection limit of 0.2 μM. It was found that only the absorption spectrum of p-NP has a wide overlap with the fluorescence excitation and emission spectra of B,N-CDs compared to those of other representative analogues. The response mechanism was due to the inner filter effect and the formation of dynamic covalent B-O bonds between B,N-CDs and p-NP, which endowed the sensing platform with the rapid response and high selectivity to p-NP. Finally, the sensor showed the practicability of p-NP determination in environmental water samples. Copyright © 2018 Elsevier B.V. All rights reserved.

Sensitive detection of biothiols and histidine based on the recovered fluorescence of the carbon quantum dots–Hg(II) system

International Nuclear Information System (INIS)

Hou, Juan; Zhang, Fengshuang; Yan, Xu; Wang, Long; Yan, Jin; Ding, Hong; Ding, Lan

2015-01-01

Highlights: • Carbon quantum dots-based probe was used for detection of GSH, Cys or His. • The fluorescence of CQDs was quenched by Hg(II) and then recovered by GSH, Cys or His. • No further surface modification or purification of CQDs was required. • This sensor exhibits superior accuracy and sensitivity. • The proposed method was simple in design, fast in operation. - Abstract: In this paper, we presented a novel, rapid and highly sensitive sensor for glutathione (GSH), cysteine (Cys) and histidine (His) based on the recovered fluorescence of the carbon quantum dots (CQDs)–Hg(II) system. The CQDs were synthesized by microwave-assisted approach in one pot according to our previous report. The fluorescence of CQDs could be quenched in the presence of Hg(II) due to the coordination occurring between Hg(II) and functional groups on the surface of CQDs. Subsequently, the fluorescence of the CQDs–Hg(II) system was recovered gradually with the addition of GSH, Cys or His due to their stronger affinity with Hg(II). A good linear relationship was obtained from 0.10 to 20 μmol L −1 for GSH, from 0.20 to 45 μmol L −1 for Cys and from 0.50 to 60 μmol L −1 for His, respectively. This method has been successfully applied to the trace detection of GSH, Cys or His in human serum samples with satisfactory results. The proposed method was simple in design and fast in operation, which demonstrated great potential in bio-sensing fields

Sensitive detection of biothiols and histidine based on the recovered fluorescence of the carbon quantum dots–Hg(II) system

Energy Technology Data Exchange (ETDEWEB)

Hou, Juan; Zhang, Fengshuang; Yan, Xu; Wang, Long; Yan, Jin [College of Chemistry, Jilin University, 2699 Qianjin Street, Changchun 130012 (China); Ding, Hong [State Key Laboratory of Inorganic Synthesis and Preparative Chemistry, College of Chemistry, Jilin University, Changchun 130012 (China); Ding, Lan, E-mail: dinglan@jlu.edu.cn [College of Chemistry, Jilin University, 2699 Qianjin Street, Changchun 130012 (China)

2015-02-15

Highlights: • Carbon quantum dots-based probe was used for detection of GSH, Cys or His. • The fluorescence of CQDs was quenched by Hg(II) and then recovered by GSH, Cys or His. • No further surface modification or purification of CQDs was required. • This sensor exhibits superior accuracy and sensitivity. • The proposed method was simple in design, fast in operation. - Abstract: In this paper, we presented a novel, rapid and highly sensitive sensor for glutathione (GSH), cysteine (Cys) and histidine (His) based on the recovered fluorescence of the carbon quantum dots (CQDs)–Hg(II) system. The CQDs were synthesized by microwave-assisted approach in one pot according to our previous report. The fluorescence of CQDs could be quenched in the presence of Hg(II) due to the coordination occurring between Hg(II) and functional groups on the surface of CQDs. Subsequently, the fluorescence of the CQDs–Hg(II) system was recovered gradually with the addition of GSH, Cys or His due to their stronger affinity with Hg(II). A good linear relationship was obtained from 0.10 to 20 μmol L{sup −1} for GSH, from 0.20 to 45 μmol L{sup −1} for Cys and from 0.50 to 60 μmol L{sup −1} for His, respectively. This method has been successfully applied to the trace detection of GSH, Cys or His in human serum samples with satisfactory results. The proposed method was simple in design and fast in operation, which demonstrated great potential in bio-sensing fields.

Cytotoxicity Test Based on Human Cells Labeled with Fluorescent Proteins: Fluorimetry, Photography, and Scanning for High-Throughput Assay.

Science.gov (United States)

Kalinina, Marina A; Skvortsov, Dmitry A; Rubtsova, Maria P; Komarova, Ekaterina S; Dontsova, Olga A

2018-06-01

High- and medium-throughput assays are now routine methods for drug screening and toxicology investigations on mammalian cells. However, a simple and cost-effective analysis of cytotoxicity that can be carried out with commonly used laboratory equipment is still required. The developed cytotoxicity assays are based on human cell lines stably expressing eGFP, tdTomato, mCherry, or Katushka2S fluorescent proteins. Red fluorescent proteins exhibit a higher signal-to-noise ratio, due to less interference by medium autofluorescence, in comparison to green fluorescent protein. Measurements have been performed on a fluorescence scanner, a plate fluorimeter, and a camera photodocumentation system. For a 96-well plate assay, the sensitivity per well and the measurement duration were 250 cells and 15 min for the scanner, 500 cells and 2 min for the plate fluorimeter, and 1000 cells and less than 1 min for the camera detection. These sensitivities are similar to commonly used MTT (tetrazolium dye) assays. The used scanner and the camera had not been previously applied for cytotoxicity evaluation. An image processing scheme for the high-resolution scanner is proposed that significantly diminishes the number of control wells, even for a library containing fluorescent substances. The suggested cytotoxicity assay has been verified by measurements of the cytotoxicity of several well-known cytotoxic drugs and further applied to test a set of novel bacteriotoxic compounds in a medium-throughput format. The fluorescent signal of living cells is detected without disturbing them and adding any reagents, thus allowing to investigate time-dependent cytotoxicity effects on the same sample of cells. A fast, simple and cost-effective assay is suggested for cytotoxicity evaluation based on mammalian cells expressing fluorescent proteins and commonly used laboratory equipment.

The sensitivity and selectivity properties of a fluorescence sensor based on quinoline-Bodipy

Energy Technology Data Exchange (ETDEWEB)

Nuri Kursunlu, Ahmed, E-mail: ankursunlu@gmail.com; Guler, Ersin

2014-01-15

A novel florescence sensor (Q-BODIPY) based on quinoline-Bodipy (quinoline-boradiazaindacene) was prepared by ‘click chemistry’ in several stages. The sensing actions of Q-BODIPY were confirmed by UV–vis titration, emission and excitation spectroscopic studies in presence of Mn{sup 2+}, Co{sup 2+}, Ni{sup 2+}, Cu{sup 2+}, Zn{sup 2+}, Cd{sup 2+}, Sn{sup 2+}, Hg{sup 2+}, Pb{sup 2+}, La{sup 3+}, Ga{sup 3+}, Er{sup 3+} and Yb{sup 3+} ions in methanol:H{sub 2}O (1:1) medium. Whereas some metal ions can only cause quenching effect on the fluorescence intensity of Q-BODIPY, some of them show an increase in fluorescence intensity. The stoichiometry of host–guest complexes formed was determined by Job′s plot method. The binding constants were calculated by Stern–Volmer method. As a fluorescence sensor, Q-BODIPY shows the best selectivity performance against Zn{sup 2+} ions in according to all spectroscopic data. -- Highlights: • Q-BODIPY prepared by several techniques shows a fluorescent behavior toward p, d and f block metal ions. • Q-BODIPY has both a more sensitivity and more effective ability for the detection of Zn(II) ion. • The synthesis strategies to produce Bodipy′s with metal coordinating offer a new approach for the design of novel fluorescence sensors.

Fluorescence photooxidation with eosin: a method for high resolution immunolocalization and in situ hybridization detection for light and electron microscopy

Science.gov (United States)

1994-01-01

A simple method is described for high-resolution light and electron microscopic immunolocalization of proteins in cells and tissues by immunofluorescence and subsequent photooxidation of diaminobenzidine tetrahydrochloride into an insoluble osmiophilic polymer. By using eosin as the fluorescent marker, a substantial improvement in sensitivity is achieved in the photooxidation process over other conventional fluorescent compounds. The technique allows for precise correlative immunolocalization studies on the same sample using fluorescence, transmitted light and electron microscopy. Furthermore, because eosin is smaller in size than other conventional markers, this method results in improved penetration of labeling reagents compared to gold or enzyme based procedures. The improved penetration allows for three-dimensional immunolocalization using high voltage electron microscopy. Fluorescence photooxidation can also be used for high resolution light and electron microscopic localization of specific nucleic acid sequences by in situ hybridization utilizing biotinylated probes followed by an eosin-streptavidin conjugate. PMID:7519623

Sensitive and rapid detection of endogenous hydrogen sulfide distributing in different mouse viscera via a two-photon fluorescent probe

International Nuclear Information System (INIS)

Chen, Qian; Yang, Jinfeng; Li, Yinhui; Zheng, Jing; Yang, Ronghua

2015-01-01

Development of efficient methods for detection of endogenous H 2 S in living cells and tissues is of considerable significance for better understanding the biological and pathological functions of H 2 S. Two-photon (TP) fluorescent probes are favorable as powerful molecular tools for studying physiological process due to its non-invasiveness, high spatiotemporal resolution and deep-tissues imaging. Up to date, several TP probes for intracellular H 2 S imaging have been designed, but real-time imaging of endogenous H 2 S-related biological processes in tissues is hampered due to low sensitivity, long response time and interference from other biothiols. To address this issue, we herein report a novel two-photon fluorescent probe (TPP-H 2 S) for highly sensitive and fast monitoring and imaging H 2 S levels in living cells and tissues. In the presence of H 2 S, it exhibits obviously improved sensitivity (LOD: 0.12 μM) and fast response time (about 2 min) compared with the reported two-photon H 2 S probes. With two-photon excitation, TPP-H 2 S displays high signal-to-noise ratio and sensitivity even no interference in cell growth media. As further application, TPP-H 2 S is applied for fast imaging of H 2 S in living cells and different fresh tissues by two-photon confocal microscope. Most importantly we first measured the endogenous H 2 S level in different viscera by vivisection and found that the distribution of endogenous H 2 S mostly in brain, liver and lung. The excellent sensing properties of TPP-H 2 S make it a practically useful tool for further studying biological roles of H 2 S. - Highlights: • This two-photon probe exhibits an improved sensitivity and response time to H 2 S. • This probe shows excellent membrane permeability and fast visualization of H 2 S in living cells and tissues. • This probe is successfully applied to measure the endogenously produced H 2 S levels in different viscera of mouse.

NIR-induced highly sensitive detection of latent finger-marks by NaYF4:Yb,Er upconversion nanoparticles in a dry powder state

Science.gov (United States)

Wang, Meng; Li, Ming; Yang, Mingying; Zhang, Xiaomei; Yu, Aoyang; Zhu, Ye; Qiu, Penghe; Mao, Chuanbin

2016-01-01

The most commonly found fingermarks at crime scenes are latent and, thus, an efficient method for detecting latent fingermarks is very important. However, traditional developing techniques have drawbacks such as low detection sensitivity, high background interference, complicated operation, and high toxicity. To tackle this challenge, we employed fluorescent NaYF4:Yb,Er upconversion nanoparticles (UCNPs), which can fluoresce visible light when excited by 980 nm human-safe near-infrared light, to stain the latent fingermarks on various substrate surfaces. The UCNPs were successfully used as a novel fluorescent label for the detection of latent fingermarks with high sensitivity, low background, high efficiency, and low toxicity on various substrates including non-infiltrating materials (glass, marble, aluminum alloy sheets, stainless steel sheets, aluminum foils, and plastic cards), semi-infiltrating materials (floor leathers, ceramic tiles, wood floor, and painted wood), and infiltrating materials such as various types of papers. This work shows that UCNPs are a versatile fluorescent label for the facile detection of fingermarks on virtually any material, enabling their practical applications in forensic sciences. PMID:27818741

High resolution x-ray fluorescence spectroscopy - a new technique for site- and spin-selectivity

International Nuclear Information System (INIS)

Wang, Xin

1996-12-01

X-ray spectroscopy has long been used to elucidate electronic and structural information of molecules. One of the weaknesses of x-ray absorption is its sensitivity to all of the atoms of a particular element in a sample. Through out this thesis, a new technique for enhancing the site- and spin-selectivity of the x-ray absorption has been developed. By high resolution fluorescence detection, the chemical sensitivity of K emission spectra can be used to identify oxidation and spin states; it can also be used to facilitate site-selective X-ray Absorption Near Edge Structure (XANES) and site-selective Extended X-ray Absorption Fine Structure (EXAFS). The spin polarization in K fluorescence could be used to generate spin selective XANES or spin-polarized EXAFS, which provides a new measure of the spin density, or the nature of magnetic neighboring atoms. Finally, dramatic line-sharpening effects by the combination of absorption and emission processes allow observation of structure that is normally unobservable. All these unique characters can enormously simplify a complex x-ray spectrum. Applications of this novel technique have generated information from various transition-metal model compounds to metalloproteins. The absorption and emission spectra by high resolution fluorescence detection are interdependent. The ligand field multiplet model has been used for the analysis of Kα and Kβ emission spectra. First demonstration on different chemical states of Fe compounds has shown the applicability of site selectivity and spin polarization. Different interatomic distances of the same element in different chemical forms have been detected using site-selective EXAFS

Laser-Induced Fluorescence Detection in High-Throughput Screening of Heterogeneous Catalysts and Single Cells Analysis

Energy Technology Data Exchange (ETDEWEB)

Su, Hui [Iowa State Univ., Ames, IA (United States)

2001-01-01

Laser-induced fluorescence detection is one of the most sensitive detection techniques and it has found enormous applications in various areas. The purpose of this research was to develop detection approaches based on laser-induced fluorescence detection in two different areas, heterogeneous catalysts screening and single cell study. First, we introduced laser-induced imaging (LIFI) as a high-throughput screening technique for heterogeneous catalysts to explore the use of this high-throughput screening technique in discovery and study of various heterogeneous catalyst systems. This scheme is based on the fact that the creation or the destruction of chemical bonds alters the fluorescence properties of suitably designed molecules. By irradiating the region immediately above the catalytic surface with a laser, the fluorescence intensity of a selected product or reactant can be imaged by a charge-coupled device (CCD) camera to follow the catalytic activity as a function of time and space. By screening the catalytic activity of vanadium pentoxide catalysts in oxidation of naphthalene, we demonstrated LIFI has good detection performance and the spatial and temporal resolution needed for high-throughput screening of heterogeneous catalysts. The sample packing density can reach up to 250 x 250 subunits/cm₂ for 40-μm wells. This experimental set-up also can screen solid catalysts via near infrared thermography detection.

Laser-Induced Fluorescence Detection in High-Throughput Screening of Heterogeneous Catalysts and Single Cells Analysis

International Nuclear Information System (INIS)

Hui Su

2001-01-01

Laser-induced fluorescence detection is one of the most sensitive detection techniques and it has found enormous applications in various areas. The purpose of this research was to develop detection approaches based on laser-induced fluorescence detection in two different areas, heterogeneous catalysts screening and single cell study. First, we introduced laser-induced imaging (LIFI) as a high-throughput screening technique for heterogeneous catalysts to explore the use of this high-throughput screening technique in discovery and study of various heterogeneous catalyst systems. This scheme is based on the fact that the creation or the destruction of chemical bonds alters the fluorescence properties of suitably designed molecules. By irradiating the region immediately above the catalytic surface with a laser, the fluorescence intensity of a selected product or reactant can be imaged by a charge-coupled device (CCD) camera to follow the catalytic activity as a function of time and space. By screening the catalytic activity of vanadium pentoxide catalysts in oxidation of naphthalene, we demonstrated LIFI has good detection performance and the spatial and temporal resolution needed for high-throughput screening of heterogeneous catalysts. The sample packing density can reach up to 250 x 250 subunits/cm(sub 2) for 40-(micro)m wells. This experimental set-up also can screen solid catalysts via near infrared thermography detection

A novel fluorescence immunoassay for the sensitive detection of Escherichia coli O157:H7 in milk based on catalase-mediated fluorescence quenching of CdTe quantum dots

Energy Technology Data Exchange (ETDEWEB)

Chen, Rui [College of Life Science, Nanchang University, Nanchang, 330031 (China); State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047 (China); Huang, Xiaolin; Li, Juan; Shan, Shan; Lai, Weihua [State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047 (China); Xiong, Yonghua, E-mail: yhxiongchen@163.com [College of Life Science, Nanchang University, Nanchang, 330031 (China); State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047 (China)

2016-12-01

Immunoassay is a powerful tool for rapid detection of food borne pathogens in food safety monitoring. However, conventional immunoassay always suffers from low sensitivity when it employs enzyme-catalyzing chromogenic substrates to generate colored molecules as signal outputs. In the present study, we report a novel fluorescence immunoassay for the sensitive detection of E. coli O157:H7 through combination of the ultrahigh bioactivity of catalase to hydrogen peroxide (H{sub 2}O{sub 2}) and H{sub 2}O{sub 2}-sensitive mercaptopropionic acid modified CdTe QDs (MPA-QDs) as a signal transduction. Various parameters, including the concentrations of anti-E. coli O157:H7 polyclonal antibody and biotinylated monoclonal antibody, the amounts of H{sub 2}O{sub 2} and streptavidin labeled catalase (CAT), the hydrolysis temperature and time of CAT to H{sub 2}O{sub 2}, as well as the incubation time between H{sub 2}O{sub 2} and MPA-QDs, were systematically investigated and optimized. With optimal conditions, the catalase-mediated fluorescence quenching immunoassay exhibits an excellent sensitivity for E. coli O157:H7 with a detection limit of 5 × 10{sup 2} CFU/mL, which was approximately 140 times lower than that of horseradish peroxidase-based colorimetric immunoassay. The reliability of the proposed method was further evaluated using E. coli O157:H7 spiked milk samples. The average recoveries of E. coli O157:H7 concentrations from 1.18 × 10{sup 3} CFU/mL to 1.18 × 10{sup 6} CFU/mL were in the range of 65.88%–105.6%. In brief, the proposed immunoassay offers a great potential for rapid and sensitive detection of other pathogens in food quality control. - Highlights: • A novel fluorescence immunoassay was developed for the ultrasensitive detection of E. coli O157:H7. • This detection was achieved through the combination of the high bioactivity of CAT and H{sub 2}O{sub 2}-sensitive QDs. • The activity of CAT to H{sub 2}O{sub 2} is 1000 folds higher than that of the HRP

High-level fluorescence labeling of gram-positive pathogens.

Directory of Open Access Journals (Sweden)

Simone Aymanns

Full Text Available Fluorescence labeling of bacterial pathogens has a broad range of interesting applications including the observation of living bacteria within host cells. We constructed a novel vector based on the E. coli streptococcal shuttle plasmid pAT28 that can propagate in numerous bacterial species from different genera. The plasmid harbors a promoterless copy of the green fluorescent variant gene egfp under the control of the CAMP-factor gene (cfb promoter of Streptococcus agalactiae and was designated pBSU101. Upon transfer of the plasmid into streptococci, the bacteria show a distinct and easily detectable fluorescence using a standard fluorescence microscope and quantification by FACS-analysis demonstrated values that were 10-50 times increased over the respective controls. To assess the suitability of the construct for high efficiency fluorescence labeling in different gram-positive pathogens, numerous species were transformed. We successfully labeled Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus dysgalactiae subsp. equisimilis, Enterococcus faecalis, Enterococcus faecium, Streptococcus mutans, Streptococcus anginosus and Staphylococcus aureus strains utilizing the EGFP reporter plasmid pBSU101. In all of these species the presence of the cfb promoter construct resulted in high-level EGFP expression that could be further increased by growing the streptococcal and enterococcal cultures under high oxygen conditions through continuous aeration.

Use of zero order diffraction of a grating monochromator towards convenient and sensitive detection of fluorescent analytes in multi fluorophoric systems

Science.gov (United States)

Panigrahi, Suraj Kumar; Mishra, Ashok Kumar

2018-02-01

White light excitation fluorescence (WLEF) is known to possess analytical advantage in terms of enhanced sensitivity and facile capture of the entire fluorescence spectral signature of multi component fluorescence systems. Using the zero order diffraction of the grating monochromator on the excitation side of a commercial spectrofluorimeter, it has been shown that WLEF spectral measurements can be conveniently carried out. Taking analyte multi-fluorophoric systems like (i) drugs and vitamins spiked in urine sample, (ii) adulteration of extra virgin olive oil with olive pomace oil and (iii) mixture of fabric dyes, it was observed that there is a significant enhancement of measurement sensitivity. The total fluorescence spectral response could be conveniently analysed using PLS2 regression. This work brings out the ease of the use of a conventional fluorimeter for WLEF measurements.

Synthesis of photoluminescent o-phenylenediamine–m-phenylenediamine copolymer nanospheres: An effective fluorescent sensing platform for selective and sensitive detection of chromium(VI) ion

Energy Technology Data Exchange (ETDEWEB)

Song, Xun [Chemical Synthesis and Pollution Control, Key Laboratory of Sichuan Province, School of Chemistry and Chemical Industry, China West Normal University, Nanchong 637002 (China); Sun, Huaiyu [Applied Technique College of Southwest Peteoleum University, Nanchong 637002 (China); Yang, Siwei; Zhao, Shizhen [Chemical Synthesis and Pollution Control, Key Laboratory of Sichuan Province, School of Chemistry and Chemical Industry, China West Normal University, Nanchong 637002 (China); Liao, Fang, E-mail: liaozhang2003@163.com [Chemical Synthesis and Pollution Control, Key Laboratory of Sichuan Province, School of Chemistry and Chemical Industry, China West Normal University, Nanchong 637002 (China)

2016-01-15

In this paper, we demonstrated a fluorescent o-phenylenediamine–m-phenylenediamine copolymer sensing system, which was synthesized by a facile and one-step hydrothermal method. The copolymer was first used as fluorescent probe for the detection of Chromium(VI) ion (Cr{sup 6+}) and showed high selectivity and sensitivity. The detection limit was 1×10{sup −11} M. It showed excellent linear relationships in wide range of 7×10{sup −11}–6×10{sup −10} M. Moreover, the addition of ethylenediaminetetraacetate (EDTA) to the detection system could successfully combine with Cr{sup 6+} to form metal chelates, making the fluorescence recovery of o-phenylenediamine–m-phenylenediamine copolymer. What is important, the prepared process had no addition of initiating agent.

Folic acid functionalized silver nanoparticles with sensitivity and selectivity colorimetric and fluorescent detection for Hg2+ and efficient catalysis.

Science.gov (United States)

Su, Dongyue; Yang, Xin; Xia, Qingdong; Zhang, Qi; Chai, Fang; Wang, Chungang; Qu, Fengyu

2014-09-05

In this research, folic acid functionalized silver nanoparticles (FA-AgNPs) were selected as a colorimetric and a 'turn on' fluorescent sensor for detecting Hg(2+). After being added into Hg(2+), AgNPs can emit stable fluorescence at 440 nm when the excitation wavelength is selected at 275 nm. The absorbance and fluorescence of the FA-AgNPs could reflect the concentration of the Hg(2+) ions. Thus, we developed a simple, sensitive analytical method to detect Hg(2+) based on the colorimetric and fluorescence enhancement of FA-AgNPs. The sensor exhibits two linear response ranges between absorbance and fluorescence intensity with Hg(2+) concentration, respectively. Meanwhile, a detection limit of 1 nM is estimated based on the linear relationship between responses with a concentration of Hg(2+). The high specificity of Hg(2+) with FA-AgNPs interactions provided the excellent selectivity towards detecting Hg(2+) over other metal ions (Pb(2+), Mg(2+), Zn(2+), Ni(2+), Cu(2+), Co(2+), Ca(2+), Mn(2+), Fe(2+), Cd(2+), Ba(2+), Cr(6+) and Cr(3+)). This will provide a simple, effective and multifunctional colorimetric and fluorescent sensor for on-site and real-time Hg(2+) ion detection. The proposed method can be applied to the analysis of trace Hg(2+) in lake water. Additionally, the FA-AgNPs can be used as efficient catalyst for the reduction of 4-nitrophenol and potassium hexacyanoferrate (III).

A Fluorescent Probe for Sensitive Detection of Hydrazine and Its Application in Red Wine and Water.

Science.gov (United States)

Wang, Jialin; Wang, Hao; Yang, Shaoxiang; Tian, Hongyu; Liu, Yongguo; Hao, Yanfeng; Zhang, Jie; Sun, Baoguo

2018-01-01

A fluorescent probe, 7-(diethylamino)-2-oxo-2H-chromene-4-carbaldehyde (probe 1), was designed and synthesized for the sensitive detection of hydrazine. The addition of N 2 H 4 caused the fluorescence intensity of probe 1 to decrease. The probe's fluorescence was turn-off after adding N 2 H 4 , which could be observed under UV light at 365 nm. Moreover, once treated with different concentrations N 2 H 4 solutions, the solution color change could be distinguished, which indicates that probe 1 could be used as a visual sensor for hydrazine. Moreover, probe 1 can be used as a signal tool to determine hydrazine levels in solutions, such as red wine and water.

A highly sensitive fluorescence resonance energy transfer aptasensor for staphylococcal enterotoxin B detection based on exonuclease-catalyzed target recycling strategy

International Nuclear Information System (INIS)

Wu, Shijia; Duan, Nuo; Ma, Xiaoyuan; Xia, Yu; Wang, Hongxin; Wang, Zhouping

2013-01-01

Graphical abstract: -- Highlights: •An ultrasensitive FRET aptasensor was developed for staphylococcal enterotoxin B determination. •SEB was recognized by SEB aptamer with high affinity and specificity. •The Mn 2+ doped NaYF 4 :Yb/Er UCNPs used as donor to quencher dye (BHQ 3 ) in new FRET. •The fluorescence intensity was prominently amplified using an exonuclease-catalyzed target recycling strategy. -- Abstract: An ultrasensitive fluorescence resonance energy transfer (FRET) bioassay was developed to detect staphylococcal enterotoxin B (SEB), a low molecular exotoxin, using an aptamer-affinity method coupled with upconversion nanoparticles (UCNPs)-sensing, and the fluorescence intensity was prominently enhanced using an exonuclease-catalyzed target recycling strategy. To construct this aptasensor, both fluorescence donor probes (complementary DNA 1 –UCNPs) and fluorescence quencher probes (complementary DNA 2 –Black Hole Quencher 3 (BHQ 3 )) were hybridized to an SEB aptamer, and double-strand oligonucleotides were fabricated, which quenched the fluorescence of the UCNPs via FRET. The formation of an aptamer–SEB complex in the presence of the SEB analyte resulted in not only the dissociation of aptamer from the double-strand DNA but also both the disruption of the FRET system and the restoration of the UCNPs fluorescence. In addition, the SEB was liberated from the aptamer–SEB complex using exonuclease I, an exonuclease specific to single-stranded DNA, for analyte recycling by selectively digesting a particular DNA (SEB aptamer). Based on this exonuclease-catalyzed target recycling strategy, an amplified fluorescence intensity could be produced using different SEB concentrations. Using optimized experimental conditions produced an ultrasensitive aptasensor for the detection of SEB, with a wide linear range of 0.001–1 ng mL −1 and a lower detection limit (LOD) of 0.3 pg mL −1 SEB (at 3σ). The fabricated aptasensor was used to measure SEB in a

Stabilizing Protein Effects on the Pressure Sensitivity of Fluorescent Gold Nanoclusters

Science.gov (United States)

2016-01-13

affected by the environment of the stabilizing protein, allowing these hybrid systems to act as sensors in many applications.2,9,14–19 This has led...Biosens Bioelectron. 2012;32:297–299. 8. Joseph D, Geckeler KE. Synthesis of highly fluorescent gold nanoclusters using egg white proteins. Colloids Surf...Chang HW, Chien YC, Hsiao JK, Cheng JT, Chou PT. Insulin -directed synthesis of fluorescent gold nanoclusters: preservation of insulin bioactivity and

A two-step lyssavirus real-time polymerase chain reaction using degenerate primers with superior sensitivity to the fluorescent antigen test.

Science.gov (United States)

Suin, Vanessa; Nazé, Florence; Francart, Aurélie; Lamoral, Sophie; De Craeye, Stéphane; Kalai, Michael; Van Gucht, Steven

2014-01-01

A generic two-step lyssavirus real-time reverse transcriptase polymerase chain reaction (qRT-PCR), based on a nested PCR strategy, was validated for the detection of different lyssavirus species. Primers with 17 to 30% of degenerate bases were used in both consecutive steps. The assay could accurately detect RABV, LBV, MOKV, DUVV, EBLV-1, EBLV-2, and ABLV. In silico sequence alignment showed a functional match with the remaining lyssavirus species. The diagnostic specificity was 100% and the sensitivity proved to be superior to that of the fluorescent antigen test. The limit of detection was ≤ 1 50% tissue culture infectious dose. The related vesicular stomatitis virus was not recognized, confirming the selectivity for lyssaviruses. The assay was applied to follow the evolution of rabies virus infection in the brain of mice from 0 to 10 days after intranasal inoculation. The obtained RNA curve corresponded well with the curves obtained by a one-step monospecific RABV-qRT-PCR, the fluorescent antigen test, and virus titration. Despite the presence of degenerate bases, the assay proved to be highly sensitive, specific, and reproducible.

Evaluation of a fluorescence polarographic immunoassay with increased sensitivity for measurement of low concentrations of tobramycin in serum

NARCIS (Netherlands)

Touw, D J; de Graaf, A I; de Goede, P

The limits of quantitation of the assay of tobramycin in serum by the fluorescence polarization immunoassay system marketed by Abbott Laboratories (TDxFLx system) are 0.1 and 10.0 mg/L. For some pharmacokinetic studies, however, a more sensitive analysis is needed. The sensitivity of the TDxFLx

Laser-Induced Fluorescence Detection in High-Throughput Screening of Heterogeneous Catalysts and Single Cells Analysis

Energy Technology Data Exchange (ETDEWEB)

Su, Hui [Iowa State Univ., Ames, IA (United States)

2001-01-01

Laser-induced fluorescence detection is one of the most sensitive detection techniques and it has found enormous applications in various areas. The purpose of this research was to develop detection approaches based on laser-induced fluorescence detection in two different areas, heterogeneous catalysts screening and single cell study. First, the author introduced laser-induced imaging (LIFI) as a high-throughput screening technique for heterogeneous catalysts to explore the use of this high-throughput screening technique in discovery and study of various heterogeneous catalyst systems. This scheme is based on the fact that the creation or the destruction of chemical bonds alters the fluorescence properties of suitably designed molecules. By irradiating the region immediately above the catalytic surface with a laser, the fluorescence intensity of a selected product or reactant can be imaged by a charge-coupled device (CCD) camera to follow the catalytic activity as a function of time and space. By screening the catalytic activity of vanadium pentoxide catalysts in oxidation of naphthalene, they demonstrated LIFI has good detection performance and the spatial and temporal resolution needed for high-throughput screening of heterogeneous catalysts. The sample packing density can reach up to 250 x 250 subunits/cm² for 40-μm wells. This experimental set-up also can screen solid catalysts via near infrared thermography detection. In the second part of this dissertation, the author used laser-induced native fluorescence coupled with capillary electrophoresis (LINF-CE) and microscope imaging to study the single cell degranulation. On the basis of good temporal correlation with events observed through an optical microscope, they have identified individual peaks in the fluorescence electropherograms as serotonin released from the granular core on contact with the surrounding fluid.

Quantum Dot-Fullerene Based Molecular Beacon Nanosensors for Rapid, Highly Sensitive Nucleic Acid Detection.

Science.gov (United States)

Liu, Ye; Kannegulla, Akash; Wu, Bo; Cheng, Li-Jing

2018-05-15

Spherical fullerene (C 60 ) can quench the fluorescence of a quantum dot (QD) through energy transfer and charge transfer processes, with the quenching efficiency regulated by the number of proximate C 60 on each QD. With the quenching property and its small size compared with other nanoparticle-based quenchers, it is advantageous to group a QD reporter and multiple C 60 -labeled oligonucleotide probes to construct a molecular beacon (MB) probe for sensitive, robust nucleic acid detection. We demonstrated a rapid, high-sensitivity DNA detection method using the nanosensors composed of QD-C 60 based MBs carried by magnetic nanoparticles (MNPs). The assay was accelerated by first dispersing the nanosensors in analytes for highly efficient DNA capture resulting from short-distance 3-dimensional diffusion of targets to the sensor surface and then concentrating the nanosensors to a substrate by magnetic force to amplify the fluorescence signal for target quantification. The enhanced mass transport enabled a rapid detection (< 10 min) with a small sample volume (1-10 µl). The high signal-to-noise ratio produced by the QD-C 60 pairs and magnetic concentration yielded a detection limit of 100 fM (~106 target DNA copies for a 10 µl analyte). The rapid, sensitive, label-free detection method will benefit the applications in point-of-care molecular diagnostic technologies.

Highly fluorescent and superparamagnetic nanosystem for biomedical applications

Science.gov (United States)

Cabrera, Mariana P.; E Cabral Filho, Paulo; Silva, Camila M. C. M.; Oliveira, Rita M.; Geraldes, Carlos F. G. C.; Castro, M. Margarida C. A.; Costa, Benilde F. O.; Henriques, Marta S. C.; Paixão, José A.; Carvalho, Luiz B., Jr.; Santos, Beate S.; Hallwass, Fernando; Fontes, Adriana; Pereira, Giovannia A. L.

2017-07-01

This work reports on highly fluorescent and superparamagnetic bimodal nanoparticles (BNPs) obtained by a simple and efficient method as probes for fluorescence analysis and/or contrast agents for MRI. These promising BNPs with small dimensions (ca. 17 nm) consist of superparamagnetic iron oxide nanoparticles (SPIONs) covalently bound with CdTe quantum dots (ca. 3 nm). The chemical structure of the magnetic part of BNPs is predominantly magnetite, with minor goethite and maghemite contributions, as shown by Mössbauer spectroscopy, which is compatible with the x-ray diffraction data. Their size evaluation by different techniques showed that the SPION derivatization process, in order to produce the BNPs, does not lead to a large size increase. The BNPs saturation magnetization, when corrected for the organic content of the sample, is ca. 68 emu g-1, which is only slightly reduced relative to the bare nanoparticles. This indicates that the SPION surface functionalization does not change considerably the magnetic properties. The BNP aqueous suspensions presented stability, high fluorescence, high relaxivity ratio (r 2/r 1 equal to 25) and labeled efficiently HeLa cells as can be seen by fluorescence analysis. These BNP properties point to their applications as fluorescent probes as well as negative T 2-weighted MRI contrast agents. Moreover, their potential magnetic response could also be used for fast bioseparation applications.

Study on fluorescence-sensitization of 3 variations of scintillators in high speed autoradiography

International Nuclear Information System (INIS)

Wang Zhenli; Liu Guimin

1993-01-01

The sensitizing effects of POPOP, PBD and PPO were compared in 3 H-TdR incorporation experiment, 3 H-TdR low and high concentration twice labelling experiment and a cell migration tracer experiment. The results indicate that 7.5% PPO in xylene with an exposure time of 48 h is most satisfactory. The efficiency was increased for 15-20 times

l-Tryptophan-capped carbon quantum dots for the sensitive and selective fluorescence detection of mercury ion in aqueous solution

Energy Technology Data Exchange (ETDEWEB)

Wan, Xuejuan; Li, Shifeng; Zhuang, Lulu; Tang, Jiaoning, E-mail: tjn@szu.edu.cn [Shenzhen University, Shenzhen Key Laboratory of Special Functional Materials, College of Materials Science and Engineering (China)

2016-07-15

l-Tryptophan-capped carbon quantum dots (l-CQDs) were facilely synthesized through “green” methodology, and the obtained material was utilized as a sensitive and selective fluorescence sensor for mercury ion (Hg{sup 2+}) in pure aqueous solutions. Carboxyl-functionalized CQDs were first green synthesized by a one-step hydrothermal route, and l-tryptophan was then attached to CQDs via direct surface condensation reaction in aqueous solution at room temperature. The as-synthesized l-CQDs had an average size of ca. 5 nm with a good dispersity in water, and exhibited a favorable selectivity for Hg{sup 2+} ions over a range of other common metal cations in aqueous solution (10 mM PBS buffer, pH 6.0). Upon the addition of Hg{sup 2+}, a complete fluorescence quenching (ON–OFF switching) of l-CQDs was evident from the fluorescence titration experiment, and the fluorescence detection limit of Hg{sup 2+} was calculated to be 11 nM, which indicated that the obtained environmentally friendly l-CQDs had sensitive detection capacity for Hg{sup 2+} in aqueous solution.

Highly selective and sensitive fluorescent chemosensor for femtomolar detection of silver ion in aqueous medium

OpenAIRE

Arulraj, Abraham Daniel; Devasenathipathy, Rajkumar; Chen, Shen-Ming; Vasantha, Vairathevar Sivasamy; Wang, Sea-Fue

2015-01-01

The chemical sensing for the trace level detection of silver ion in aqueous solution still remains a challenge using simple, rapid, and inexpensive method. We report that thionine can be used as a fluorescent probe for the detection of Ag+ ion. The successive addition of Ag+ ion to the solution containing thionine quenches (turns-off) the fluorescence intensity of thionine. Association and quenching constants have been estimated by the Benesi–Hildebrand method and Stern–Volmer plot, respectiv...

Rotationally cooled laser induced fluorescence determination of polycyclic aromatic hydrocarbons

International Nuclear Information System (INIS)

Warren, J.A.; Hayes, J.M.; Small, G.J.

1982-01-01

In recent years the development of new highly selective and sensitive methods for the characterization and determination of polycyclic aromatic hydrocarbons (PAHs) and their derivatives in complex mixtures has received considerable attention. High selectivity is associated here with the ability to distinguish between substitutional isomers of PAHs. Attainment of this selectivity with capillary column-gas chromatography-mass spectrometry for complex mixtures is very difficult and time-consuming. Alternative approaches are, therefore, required. Given that the majority of PAHs fluoresce with reasonable quantum yields and that high sensitivities are afforded by fluorescence detection, the possibility of developing high-resolution fluorescence based techniques is attractive. This is all the more so if the technique's selectivity does not rely on physical separation, e.g., chromatography. In this paper discussion is limited to such techniques

A simple and sensitive method for L-cysteine detection based on the fluorescence intensity increment of quantum dots

International Nuclear Information System (INIS)

Huang Shan; Xiao Qi; Li Ran; Guan Hongliang; Liu Jing; Liu Xiaorong; He Zhike; Liu Yi

2009-01-01

In this contribution, a simple and sensitive method for L-cysteine detection was established based on the increment of the fluorescence intensity of mercaptoacetic acid-capped CdSe/ZnS quantum dots (QDs) in aqueous solution. Meanwhile, the fluorescence characteristics and the optimal conditions were investigated in detail. Under the optimized conditions, the linear range of QDs fluorescence intensity versus the concentration of L-cysteine was 10-800 nmol L -1 , with a correlation coefficient (R) of 0.9969 and a limit of detection (3σ black) of 3.8 nmol L -1 . The relative standard deviation (R.S.D.) for 0.5 μmol L -1 L-cysteine was 1.1% (n = 5). There was no interference to coexisting foreign substances including common ions, carbohydrates, nucleotide acids and other 19 amino acids. The proposed method possessed the advantages of simplicity, rapidity and sensitivity. Synthetic amino acid samples, medicine sample together with human urine samples were analyzed by the methodology and the results were satisfying.

Sensitive and selective detection of Cu(II) ion: A new effective 1,8-naphthalimide-based fluorescence 'turn off' sensor.

Science.gov (United States)

Huang, Guozhen; Li, Chuang; Han, Xintong; Aderinto, Stephen Opeyemi; Shen, Kesheng; Mao, Shanshan; Wu, Huilu

2018-06-01

The present study reports the development of a new 1,8-naphthalimide-based fluorescent sensor V for monitoring Cu(II) ions. The sensor exhibited pH independence over a wide pH range 2.52-9.58, and indicated its possible use for monitoring Cu(II) ions in a competitive pH medium. The sensor also showed high selectivity and sensitivity towards the Cu(II) ions over other competitive metal ions in DMSO-HEPES buffer (v/v, 1:1; pH 7.4) with a fluorescence 'turn off' mode of 79.79% observed. A Job plot indicated the formation of a 1:1 binding mode of the sensor with Cu(II) ions. The association constant and detection limit were 1.14 × 10 6  M -1 and 4.67 × 10 -8 M, respectively. The fluorescence spectrum of the sensor was quenched due to the powerful paramagnetic nature of the Cu(II) ions. Potential application of this sensor was also demonstrated when determining Cu(II) ion levels in two different water samples. Copyright © 2018 John Wiley & Sons, Ltd.

Cysteine detection using a high-fluorescence sensor based on a nitrogen-doped graphene quantum dot–mercury(II) system

Energy Technology Data Exchange (ETDEWEB)

Liu, Zhenzhen; Gong, Yan; Fan, Zhefeng, E-mail: zhefengfan@126.com

2016-07-15

A novel and highly sensitive fluorescence sensor, which was based on the recovered fluorescence of a nitrogen-doped graphene quantum dot–Hg(II) system, was developed for cysteine detection. An easy, green, one-pot synthesis of nitrogen-doped graphene quantum dots was established by using citric acid and urea as carbon and nitrogen sources, respectively. The fluorescence of nitrogen-doped graphene quantum dots was significantly quenched by Hg(II) because of the efficient electron transfer between nitrogen-doped graphene quantum dots and Hg(II). Subsequently, fluorescence was recovered gradually upon cysteine addition to form a stable complex with Hg(II). The fluorescence sensor showed a response to cysteine within a wide concentration range of 0.05–30 μmol L{sup −1}, with a detection limit of 1.3 nmol L{sup −1}. The sensor was successfully applied to detect cysteine in honey and beer samples, with a recovery range of 98–105%.

Cysteine detection using a high-fluorescence sensor based on a nitrogen-doped graphene quantum dot–mercury(II) system

International Nuclear Information System (INIS)

Liu, Zhenzhen; Gong, Yan; Fan, Zhefeng

2016-01-01

A novel and highly sensitive fluorescence sensor, which was based on the recovered fluorescence of a nitrogen-doped graphene quantum dot–Hg(II) system, was developed for cysteine detection. An easy, green, one-pot synthesis of nitrogen-doped graphene quantum dots was established by using citric acid and urea as carbon and nitrogen sources, respectively. The fluorescence of nitrogen-doped graphene quantum dots was significantly quenched by Hg(II) because of the efficient electron transfer between nitrogen-doped graphene quantum dots and Hg(II). Subsequently, fluorescence was recovered gradually upon cysteine addition to form a stable complex with Hg(II). The fluorescence sensor showed a response to cysteine within a wide concentration range of 0.05–30 μmol L −1 , with a detection limit of 1.3 nmol L −1 . The sensor was successfully applied to detect cysteine in honey and beer samples, with a recovery range of 98–105%.

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

Science.gov (United States)

Lindvold, Lars R.; Hermann, Gregers G.

2016-12-01

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 this light source also is useful for exciting autofluorescence in healthy bladder mucosa. This autofluorescence then provides a contrast to the sensitized fluorescence (PDD) of tumours in the bladder.

Ultra-sensitive DNA assay based on single-molecule detection coupled with fluorescent quantum dot-labeling and its application to determination of messenger RNA

International Nuclear Information System (INIS)

Li Li; Li Xincang; Li Lu; Wang Jinxing; Jin Wenrui

2011-01-01

An ultra-sensitive single-molecule detection (SMD) method for quantification of DNA using total internal reflection fluorescence microscopy (TIRFM) coupled with fluorescent quantum dot (QD)-labeling was developed. In this method, the target DNA (tDNA) was captured by the capture DNA immobilized on the silanized coverslip blocked with ethanolamine and bovine serum albumin. Then, the QD-labeled probe DNA was hybridized to the tDNA. Ten fluorescent images of the QD-labeled sandwich DNA hybrids on the coverslip were taken by a high-sensitive CCD. The tDNA was quantified by counting the bright spots on the images using a calibration curve. The LOD of the method was 1 x 10 -14 mol L -1 . Several key factors, including image acquirement, fluorescence probe, substrate preparation, noise elimination from solutions and glass coverslips, and nonspecific adsorption and binding of solution-phase detection probes were discussed in detail. The method could be applied to quantify messenger RNA (mRNA) in cells. In order to determine mRNA, double-stranded RNA-DNA hybrids consisting of mRNA and corresponding cDNA were synthesized from the cellular mRNA template using reverse transcription in the presence of reverse transcriptase. After removing the mRNA in the double-stranded hybrids using ribonuclease, cDNA was quantified using the SMD-based TIRFM. Osteopontin mRNA in decidual stromal cells was chosen as the model analyte.

Ion beam induced fluorescence imaging in biological systems

International Nuclear Information System (INIS)

Bettiol, Andrew A.; Mi, Zhaohong; Vanga, Sudheer Kumar; Chen, Ce-belle; Tao, Ye; Watt, Frank

2015-01-01

Imaging fluorescence generated by MeV ions in biological systems such as cells and tissue sections requires a high resolution beam (<100 nm), a sensitive detection system and a fluorescent probe that has a high quantum efficiency and low bleaching rate. For cutting edge applications in bioimaging, the fluorescence imaging technique needs to break the optical diffraction limit allowing for sub-cellular structure to be visualized, leading to a better understanding of cellular function. In a nuclear microprobe this resolution requirement can be readily achieved utilizing low beam current techniques such as Scanning Transmission Ion Microscopy (STIM). In recent times, we have been able to extend this capability to fluorescence imaging through the development of a new high efficiency fluorescence detection system, and through the use of new novel fluorescent probes that are resistant to ion beam damage (bleaching). In this paper we demonstrate ion beam induced fluorescence imaging in several biological samples, highlighting the advantages and challenges associated with using this technique

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.

Highly sensitive rapid fluorescence detection of protein residues on surgical instruments

International Nuclear Information System (INIS)

Kovalev, Valeri I; Bartona, James S; Richardson, Patricia R; Jones, Anita C

2006-01-01

There is a risk of contamination of surgical instruments by infectious protein residues, in particular, prions which are the agents for Creutzfeldt-Jakob Disease in humans. They are exceptionally resistant to conventional sterilization, therefore it is important to detect their presence as contaminants so that alternative cleaning procedures can be applied. We describe the development of an optimized detection system for fluorescently labelled protein, suitable for in-hospital use. We show that under optimum conditions the technique can detect ∼10 attomole/cm 2 with a scan speed of ∼3-10 cm 2 /s of the test instrument's surface. A theoretical analysis and experimental measurements will be discussed

Highly Sensitive DNA Sensor Based on Upconversion Nanoparticles and Graphene Oxide.

Science.gov (United States)

Alonso-Cristobal, P; Vilela, P; El-Sagheer, A; Lopez-Cabarcos, E; Brown, T; Muskens, O L; Rubio-Retama, J; Kanaras, A G

2015-06-17

In this work we demonstrate a DNA biosensor based on fluorescence resonance energy transfer (FRET) between NaYF4:Yb,Er nanoparticles and graphene oxide (GO). Monodisperse NaYF4:Yb,Er nanoparticles with a mean diameter of 29.1 ± 2.2 nm were synthesized and coated with a SiO2 shell of 11 nm, which allowed the attachment of single strands of DNA. When these DNA-functionalized NaYF4:Yb,Er@SiO2 nanoparticles were in the proximity of the GO surface, the π-π stacking interaction between the nucleobases of the DNA and the sp(2) carbons of the GO induced a FRET fluorescence quenching due to the overlap of the fluorescence emission of the NaYF4:Yb,Er@SiO2 and the absorption spectrum of GO. By contrast, in the presence of the complementary DNA strands, the hybridization leads to double-stranded DNA that does not interact with the GO surface, and thus the NaYF4:Yb,Er@SiO2 nanoparticles remain unquenched and fluorescent. The high sensitivity and specificity of this sensor introduces a new method for the detection of DNA with a detection limit of 5 pM.

Fluorescence Behavior and Dural Infiltration of Meningioma Analyzed by 5-Aminolevulinic Acid-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

2017-12-01

To compare fluorescence intensity of tumor specimens, as measured by a fluorescence-guided surgery microscope and a spectrometer, to evaluate tumor infiltration of dura mater around meningiomas with help of these 2 different 5-aminolevulinic acid (5-ALA)-based fluorescence tools, and to correlate fluorescence intensity with histopathologic data. In a clinical series, meningiomas were resected by 5-ALA fluorescence-guided surgery. Fluorescence intensity was semiquantitatively rated by the surgeon at predefined points. Biopsies were harvested and fluorescence intensity measured by a spectrometer and histopathologically analyzed. Sampling was realized at the level of the dura in a centrifugal direction. A total of 104 biopsies (n = 13 tumors) were analyzed. 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 one half of the cases. The complementary use of both fluorescence tools may improve resection quality. Copyright © 2017 Elsevier Inc. All rights reserved.

High-sensitive portable ASE-2 X-ray analyzer of sulfur in mineral oil

International Nuclear Information System (INIS)

Anchugov, I.S.; Goganov, A.D.; Plotnikov, R.I.

2007-01-01

The high-sensitivity ASE-2 analyzer of sulfur on the basis of existing ASE-I device is designed. ASE-2 analyzer realizes a standard method of energy dispersion X-ray fluorescent determinations of a sulfur mass fraction in mineral oil and allows to carry out the quantitative determination of sulfur in hydrocarbonic raw material and fuel in a 0.002-5 mass.% range [ru

Increasing of sensitivity of fluorescent immunoassay analysis of alpha-fetoprotein by means of plasmonical silver nanoparticles

International Nuclear Information System (INIS)

Vashchenko, S.V.; Min'ko, A.A.; Romanenko, A.A.; Gaponenko, S.V.; Kulakovich, O.S.

2014-01-01

A test system is proposed based on metal enhanced fluorescence to analyze low concentrations of alpha-fetoprotein (AFP), a tumor marker. Antigen-antibody reaction was performed on polystyrene plates coated with silver nanoparticles to increase sensitivity of fluorescent immunoassay and signal-to-noise ratio as compared to silver-free system. As compared to widely used ELISA technique and other immunoassay techniques the proposed approach is characterized by smaller probe volume, fast analysis and simplicity. The proposed test system uses layer-by-layer assembly approach, LED excitation and nanowatt photodetection set-up. The proposed test system offers AFP detection at concentrations used in clinical practice. Fluorescence enhancement for labeled AFP antibodies on a silver substrate was found to depend on antibodies concentration and was up to 6 times. (authors)

Fluorescent probe based on heteroatom containing styrylcyanine: pH-sensitive properties and bioimaging in vivo

International Nuclear Information System (INIS)

Yang, Xiaodong; Gao, Ya; Huang, Zhibing; Chen, Xiaohui; Ke, Zhiyong; Zhao, Peiliang; Yan, Yichen; Liu, Ruiyuan; Qu, Jinqing

2015-01-01

A novel fluorescent probe based on heteroatom containing styrylcyanine is synthesized. The fluorescence of probe is bright green in basic and neutral media but dark orange in strong acidic environments, which could be reversibly switched. Such behavior enables it to work as a fluorescent pH sensor in the solution state and a chemosensor for detecting acidic and basic volatile organic compounds. Analyses by NMR spectroscopy confirm that the protonation or deprotonation of pyridinyl moiety is responsible for the sensing process. In addition, the fluorescent microscopic images of probe in live cells and zebrafish are achieved successfully, suggesting that the probe has good cell membrane permeability and low cytotoxicity. - Graphical abstract: A novel styrylcyanine-based fluorescent pH probe was designed and synthesized, the fluorescence of which is bright green in basic and neutral media but dark orange in strong acidic environments. Such behavior enables it to work as a fluorescent pH sensor in solution states, and a chemosensor for detecting volatile organic compounds with high acidity and basicity in solid state. In addition, it can be used for fluorescent imaging in living cell and living organism. - Highlights: • Bright green fluorescence was observed in basic and neutral media. • Dark orange fluorescence was found in strong acidic environments. • Volatile organic compounds with high acidity and basicity could be detected. • Bioimaging in living cell and living organism was achieved successfully

Highly sensitive rapid fluorescence detection of protein residues on surgical instruments

Energy Technology Data Exchange (ETDEWEB)

Kovalev, Valeri I [School of Engineering and Physical Sciences, Heriot-Watt University, Edinburgh EH14 4AS (United Kingdom); Bartona, James S [School of Engineering and Physical Sciences, Heriot-Watt University, Edinburgh EH14 4AS (United Kingdom); Richardson, Patricia R [School of Chemistry, University of Edinburgh, Edinburgh, EH9 3JJ (United Kingdom); Jones, Anita C [School of Chemistry, University of Edinburgh, Edinburgh, EH9 3JJ (United Kingdom)

2006-07-15

There is a risk of contamination of surgical instruments by infectious protein residues, in particular, prions which are the agents for Creutzfeldt-Jakob Disease in humans. They are exceptionally resistant to conventional sterilization, therefore it is important to detect their presence as contaminants so that alternative cleaning procedures can be applied. We describe the development of an optimized detection system for fluorescently labelled protein, suitable for in-hospital use. We show that under optimum conditions the technique can detect {approx}10 attomole/cm{sup 2} with a scan speed of {approx}3-10 cm{sup 2}/s of the test instrument's surface. A theoretical analysis and experimental measurements will be discussed.

Evaluation of diatomea algae Thalassiosira weissflogii sensitivity to chloride mercury and methylmercury by chlorophyll fluorescence analysis

Science.gov (United States)

Graevskaya, E. E.; Antal, T. K.; Matorin, D. N.; Voronova, E. N.; Pogosyan, S. I.; Rubin, A. B.

2003-05-01

Measurement of chlorophyll fluorescence has been shown to be a rapid, non-invasive, and reliable method to assess photosynthetic performance in a changing environment. In our study, the pulseamplitude-modulation (PAM) - fluorometric method was used to evaluate the sensitivity to chloride mercury and methylmercury chloride of diatomea microalgae Thalassiosira weissflogii. We found that 10^{-6} and 10^{-7} M MeHg led to a slow decrease in the PS II activity following for prolonged lag phase, whereas the algae was not sensitive to the same concentrations of HgCl2. However observed PS II inactivation by methylmercury was not complete and about 10 percents ofthe cells kept the high level of PS II activity as it was shown by microfluorometric analysis. These cells could determine adaptation of algae to methylmercury effect. Both toxicants decreased the rate of PS II reparation, as well as increased a heat pathway of excitation dissipation in PS II antennae complex.

A rhodamine B-based fluorescent sensor toward highly selective mercury (II) ions detection.

Science.gov (United States)

Jiao, Yang; Zhang, Lei; Zhou, Peng

2016-04-01

This work presented the design, syntheses and photophysical properties of a rhodamine B-based fluorescence probe, which exhibited a sensitive and selective recognition towards mercury (II). The chemosensor RA (Rhodamine- amide- derivative) contained a 5-aminoisophthalic acid diethyl ester and a rhodamine group, and the property of spirolactone of this chemosensor RA was detected by X-ray crystal structure analyses. Chemosensor RA afforded turn-on fluorescence enhancement and displayed high brightness for Hg(2+), which leaded to the opening of the spirolactone ring and consequently caused the appearance of strong absorption at visible range, moreover, the obvious and characteristic color changed from colorless to pink was observed. We envisioned that the chemosensor RA exhibited a considerable specificity with two mercury (II) ions which was attributed to the open of spirolactone over other interference metal ions. Copyright © 2015 Elsevier B.V. All rights reserved.

Sets of RNA repeated tags and hybridization-sensitive fluorescent probes for distinct images of RNA in a living cell.

Directory of Open Access Journals (Sweden)

Takeshi Kubota

Full Text Available BACKGROUND: Imaging the behavior of RNA in a living cell is a powerful means for understanding RNA functions and acquiring spatiotemporal information in a single cell. For more distinct RNA imaging in a living cell, a more effective chemical method to fluorescently label RNA is now required. In addition, development of the technology labeling with different colors for different RNA would make it easier to analyze plural RNA strands expressing in a cell. METHODOLOGY/PRINCIPAL FINDINGS: Tag technology for RNA imaging in a living cell has been developed based on the unique chemical functions of exciton-controlled hybridization-sensitive oligonucleotide (ECHO probes. Repetitions of selected 18-nucleotide RNA tags were incorporated into the mRNA 3'-UTR. Pairs with complementary ECHO probes exhibited hybridization-sensitive fluorescence emission for the mRNA expressed in a living cell. The mRNA in a nucleus was detected clearly as fluorescent puncta, and the images of the expression of two mRNAs were obtained independently and simultaneously with two orthogonal tag-probe pairs. CONCLUSIONS/SIGNIFICANCE: A compact and repeated label has been developed for RNA imaging in a living cell, based on the photochemistry of ECHO probes. The pairs of an 18-nt RNA tag and the complementary ECHO probes are highly thermostable, sequence-specifically emissive, and orthogonal to each other. The nucleotide length necessary for one tag sequence is much shorter compared with conventional tag technologies, resulting in easy preparation of the tag sequences with a larger number of repeats for more distinct RNA imaging.

Sensitive detection and separation of fluorescent derivatives using capillary electrophoresis with laser-induced fluorescence detection with 532nm Nd:YAG laser

International Nuclear Information System (INIS)

Vrabel, Patrik; Taborsky, Petr; Ryvolova, Marketa; Havel, Josef; Preisler, Jan

2006-01-01

Capillary electrophoresis with laser-induced fluorescence detection (CELIF) is a powerful tool for separation and sensitive determination of fluorescent species. Biologically active compounds, such as amino acids, peptides and proteins may exhibit native fluorescence, which is however often low and/or an expensive laser is required for excitation in UV. Therefore, labelling of the analytes with a fluorescent dye is usually necessary. In this work, a home-built CELIF instrument with diode pumped frequency-doubled continuous wave Nd:YAG excitation laser with feedback power regulation (532nm) was constructed. The suitability of this type of laser for LIF detection in a separation method was found excellent. A limit of detection (LOD) (S/N=3) of 2x10 -13 mol/l was achieved with rhodamine B, which is comparable to those obtained using similar instruments with Ar + laser [Y.F. Cheng, N.J. Dovichi, Science 242 (1988) 562, E.S. Yeung et al., J. Chromatogr. 608 (1992) 73]. LOD of a protein derivatized according to modified procedures [M.J. Little et al., Anal. Chim. Acta 339 (1997) 279, A. Chersi et al., Biochim. Biophys. Acta 1336 (1997) 83] was determined. Detection of the derivatives was found to be limited by insufficient reaction recovery at low analyte concentration, chemical noise, separation efficiency and quality of the derivatizing reagent rather than by the detector performance. As a consequence, a huge gap between the detection ability of CELIF instruments and LOD determined in real samples is revealed

Subunits of highly Fluorescent Protein R-Phycoerythrin as Probes for Cell Imaging and Single-Molecule Detection

Energy Technology Data Exchange (ETDEWEB)

Isailovic, Dragan [Iowa State Univ., Ames, IA (United States)

2005-01-01

The purposes of our research were: (1) To characterize subunits of highly fluorescent protein R-Phycoerythrin (R-PE) and check their suitability for single-molecule detection (SMD) and cell imaging, (2) To extend the use of R-PE subunits through design of similar proteins that will be used as probes for microscopy and spectral imaging in a single cell, and (3) To demonstrate a high-throughput spectral imaging method that will rival spectral flow cytometry in the analysis of individual cells. We first demonstrated that R-PE subunits have spectroscopic and structural characteristics that make them suitable for SMD. Subunits were isolated from R-PE by high-performance liquid chromatography (HPLC) and detected as single molecules by total internal reflection fluorescence microscopy (TIRFM). In addition, R-PE subunits and their enzymatic digests were characterized by several separation and detection methods including HPLC, capillary electrophoresis, sodium dodecyl sulfate-polyacrilamide gel electrophoresis (SDS-PAGE) and HPLC-electrospray ionization mass spectrometry (ESI-MS). Favorable absorption and fluorescence of the R-PE subunits and digest peptides originate from phycoerythrobilin (PEB) and phycourobilin (PUB) chromophores that are covalently attached to cysteine residues. High absorption coefficients and strong fluorescence (even under denaturing conditions), broad excitation and emission fluorescence spectra in the visible region of electromagnetic spectrum, and relatively low molecular weights make these molecules suitable for use as fluorescence labels of biomolecules and cells. We further designed fluorescent proteins both in vitro and in vivo (in Escherichia coli) based on the highly specific attachment of PEB chromophore to genetically expressed apo-subunits of R-PE. In one example, apo-alpha and apo-beta R-PE subunits were cloned from red algae Polisiphonia boldii (P. boldii), and expressed in E. coli. Although expressed apo-subunits formed inclusion

A sensitive fluorescent probe for the polar solvation dynamics at protein-surfactant interfaces.

Science.gov (United States)

Singh, Priya; Choudhury, Susobhan; Singha, Subhankar; Jun, Yongwoong; Chakraborty, Sandipan; Sengupta, Jhimli; Das, Ranjan; Ahn, Kyo-Han; Pal, Samir Kumar

2017-05-17

Relaxation dynamics at the surface of biologically important macromolecules is important taking into account their functionality in molecular recognition. Over the years it has been shown that the solvation dynamics of a fluorescent probe at biomolecular surfaces and interfaces account for the relaxation dynamics of polar residues and associated water molecules. However, the sensitivity of the dynamics depends largely on the localization and exposure of the probe. For noncovalent fluorescent probes, localization at the region of interest in addition to surface exposure is an added challenge compared to the covalently attached probes at the biological interfaces. Here we have used a synthesized donor-acceptor type dipolar fluorophore, 6-acetyl-(2-((4-hydroxycyclohexyl)(methyl)amino)naphthalene) (ACYMAN), for the investigation of the solvation dynamics of a model protein-surfactant interface. A significant structural rearrangement of a model histone protein (H1) upon interaction with anionic surfactant sodium dodecyl sulphate (SDS) as revealed from the circular dichroism (CD) studies is nicely corroborated in the solvation dynamics of the probe at the interface. The polarization gated fluorescence anisotropy of the probe compared to that at the SDS micellar surface clearly reveals the localization of the probe at the protein-surfactant interface. We have also compared the sensitivity of ACYMAN with other solvation probes including coumarin 500 (C500) and 4-(dicyanomethylene)-2-methyl-6-(p-dimethylamino-styryl)-4H-pyran (DCM). In comparison to ACYMAN, both C500 and DCM fail to probe the interfacial solvation dynamics of a model protein-surfactant interface. While C500 is found to be delocalized from the protein-surfactant interface, DCM becomes destabilized upon the formation of the interface (protein-surfactant complex). The timescales obtained from this novel probe have also been compared with other femtosecond resolved studies and molecular dynamics simulations.

A sensitive fluorescence quenching method for the detection of tartrazine with acriflavine in soft drinks.

Science.gov (United States)

Yang, Huan; Ran, Guihua; Yan, Jingjing; Zhang, Hui; Hu, Xiaoli

2018-03-01

In this work, a simple, rapid, sensitive, selective spectrofluorimetric method was applied to detect tartrazine. The fluorescence of acriflavine could be efficiently quenched by tartrazine. The method manifested real time response as well as presented satisfied linear relationship to tartrazine. The linear response range of tartrazine (R 2 = 0.9995) was from 0.056 to 5 μmol L -1 . The detection limit (3σ/k) was 0.017 μmol L -1 , indicating that this method could be applied to detect traces of tartrazine. The accuracy and precision of the method was further assured by recovery studies via a standard addition method, with percentage recoveries in the range of 96.0% to 103.0%. Moreover, a quenching mechanism was investigated systematically by the linear plots at varying temperatures based on the Stern-Volmer equation, fluorescence lifetime and UV-visible absorption spectra, all of which proved to be static quenching. This sensitive, selective assay possessed a great application prospect for the food industry owing to its simplicity and rapidity for the detection of tartrazine. Copyright © 2017 John Wiley & Sons, Ltd.

A Two-Step Lyssavirus Real-Time Polymerase Chain Reaction Using Degenerate Primers with Superior Sensitivity to the Fluorescent Antigen Test

Directory of Open Access Journals (Sweden)

Vanessa Suin

2014-01-01

Full Text Available A generic two-step lyssavirus real-time reverse transcriptase polymerase chain reaction (qRT-PCR, based on a nested PCR strategy, was validated for the detection of different lyssavirus species. Primers with 17 to 30% of degenerate bases were used in both consecutive steps. The assay could accurately detect RABV, LBV, MOKV, DUVV, EBLV-1, EBLV-2, and ABLV. In silico sequence alignment showed a functional match with the remaining lyssavirus species. The diagnostic specificity was 100% and the sensitivity proved to be superior to that of the fluorescent antigen test. The limit of detection was ≤1 50% tissue culture infectious dose. The related vesicular stomatitis virus was not recognized, confirming the selectivity for lyssaviruses. The assay was applied to follow the evolution of rabies virus infection in the brain of mice from 0 to 10 days after intranasal inoculation. The obtained RNA curve corresponded well with the curves obtained by a one-step monospecific RABV-qRT-PCR, the fluorescent antigen test, and virus titration. Despite the presence of degenerate bases, the assay proved to be highly sensitive, specific, and reproducible.

A simple and sensitive fluorescent sensor for methyl parathion based on L-tyrosine methyl ester functionalized carbon dots.

Science.gov (United States)

Hou, Juying; Dong, Jing; Zhu, Haishuang; Teng, Xue; Ai, Shiyun; Mang, Minglin

2015-06-15

In this paper, a simple and sensitive fluorescent sensor for methyl parathion is developed based on L-tyrosine methyl ester functionalized carbon dots (Tyr-CDs) and tyrosinase system. The carbon dots are obtained by simple hydrothermal reaction using citric acid as carbon resource and L-tyrosine methyl ester as modification reagent. The carbon dots are characterized by transmission electron microscope, high resolution transmission electron microscopy, X-ray diffraction spectrum, Fourier transform infrared spectroscopy, and X-ray photoelectron spectroscopy. The carbon dots show strong and stable photoluminescence with a quantum yield of 3.8%. Tyrosinase can catalyze the oxidation of tyrosine methyl ester on the surface of carbon dots to corresponding quinone products, which can quench the fluorescence of carbon dots. When organophosphorus pesticides (OPs) are introduced in system, they can decrease the enzyme activity, thus decrease the fluorescence quenching rate. Methyl parathion, as a model of OPs, was detected. Experimental results show that the enzyme inhibition rate is proportional to the logarithm of the methyl parathion concentration in the range 1.0×10(-10)-1.0×10(-4) M with the detection limit (S/N=3) of 4.8×10(-11) M. This determination method shows a low detection limit, wide linear range, good selectivity and high reproducibility. This sensing system has been successfully used for the analysis of cabbage, milk and fruit juice samples. Copyright © 2014 Elsevier B.V. All rights reserved.

Ultra-sensitive DNA assay based on single-molecule detection coupled with fluorescent quantum dot-labeling and its application to determination of messenger RNA

Energy Technology Data Exchange (ETDEWEB)

Li Li [School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100 (China); Li Xincang [School of Life Sciences, Shandong University, Jinan 250100 (China); Li Lu [School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100 (China); Wang Jinxing [School of Life Sciences, Shandong University, Jinan 250100 (China); Jin Wenrui, E-mail: jwr@sdu.edu.cn [School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100 (China)

2011-01-24

An ultra-sensitive single-molecule detection (SMD) method for quantification of DNA using total internal reflection fluorescence microscopy (TIRFM) coupled with fluorescent quantum dot (QD)-labeling was developed. In this method, the target DNA (tDNA) was captured by the capture DNA immobilized on the silanized coverslip blocked with ethanolamine and bovine serum albumin. Then, the QD-labeled probe DNA was hybridized to the tDNA. Ten fluorescent images of the QD-labeled sandwich DNA hybrids on the coverslip were taken by a high-sensitive CCD. The tDNA was quantified by counting the bright spots on the images using a calibration curve. The LOD of the method was 1 x 10{sup -14} mol L{sup -1}. Several key factors, including image acquirement, fluorescence probe, substrate preparation, noise elimination from solutions and glass coverslips, and nonspecific adsorption and binding of solution-phase detection probes were discussed in detail. The method could be applied to quantify messenger RNA (mRNA) in cells. In order to determine mRNA, double-stranded RNA-DNA hybrids consisting of mRNA and corresponding cDNA were synthesized from the cellular mRNA template using reverse transcription in the presence of reverse transcriptase. After removing the mRNA in the double-stranded hybrids using ribonuclease, cDNA was quantified using the SMD-based TIRFM. Osteopontin mRNA in decidual stromal cells was chosen as the model analyte.

Simple and sensitive detection method for diprophylline using glutathione-capped CdTe quantum dots as fluorescence probes

Energy Technology Data Exchange (ETDEWEB)

Ying, Suyan; Cui, Shumin [College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004 (China); Wang, Weiping, E-mail: wangwp@zjnu.edu.cn [College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004 (China); Feng, Jiuju [College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004 (China); Chen, Jianrong [College of Geography and Environmental Science, Zhejiang Normal University, Jinhua 321004 (China)

2014-01-15

A simple and sensitive method for detecting diprophylline (DPP) was developed based on the fluorescence quenching of glutathione-capped CdTe quantum dots (GSH–CdTe QDs) by using diprophylline in a KH{sub 2}PO{sub 4}–Na{sub 2}HPO{sub 4} medium. Parameters affecting the quenching efficiency, including types and pH of buffer solutions as well as temperature, reaction time, adding sequence, and interfering substances, were investigated and optimized. In optimum conditions, the calibration plot of the quenched fluorescence intensity F{sub 0}/F with a DPP concentration range of 1.67×10{sup –6} mol L{sup −1} to 1.33×10{sup –5} mol L{sup −1} was linear. The detection limit (with signal to noise ratio of 3) for DPP was 2.24×10{sup –7} mol L{sup −1}. The proposed method was successfully applied for detecting DPP in human serum. The recovery of the method was in the range of 87.41% to 117.94%. Finally, the possible quenching mechanism of GSH–CdTe QDs and DPP was also discussed. -- Highlights: • Fluorescence of GSH/CdTe QDs was quenched by diprophylline in phosphate medium. • A simple and sensitive detection method for diprophylline based on fluorescence quenching was developed. • Quenching mechanism of GSH-capped CdTe QDs with diprophylline was discussed.

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.

Ratiometric fluorescent nanoparticles for sensing temperature

Energy Technology Data Exchange (ETDEWEB)

Peng, Hong-Shang, E-mail: hillphs@yahoo.com.cn; Huang, Shi-Hua [Beijing Jiaotong University, Key Laboratory of Luminescence and Optical Information, Ministry of Education, Institute of Optoelectronic Technology (China); Wolfbeis, Otto S. [University of Regensburg, Institute of Analytical Chemistry, Chemo- and Biosensors (Germany)

2010-10-15

A ratiometric type of fluorescent nanoparticle was prepared via an encapsulation-reprecipitation method. By introducing an alkoxysilanized dye as a reference, the nanoparticles (NPs) give both a green and a red fluorescence under one single-wavelength excitation. The resulted ratiometric fluorescence is found to be highly temperature-dependent in the physiological range (25-45 {sup o}C), with an intensity temperature sensitivity of -4.0%/{sup o}C. Given the small size (20-30 nm in diameter) and biocompatible nature (silica out layer), such kind of NPs were very promising as temperature nanosensors for cellular sensing and imaging.

A reduced graphene oxide-based fluorescence resonance energy transfer sensor for highly sensitive detection of matrix metalloproteinase 2.

Science.gov (United States)

Xi, Gaina; Wang, Xiaoping; Chen, Tongsheng

2016-01-01

A novel fluorescence nanoprobe (reduced nano-graphene oxide [nrGO]/fluorescein isothiocyanate-labeled peptide [Pep-FITC]) for ultrasensitive detection of matrix metalloproteinase 2 (MMP2) has been developed by engineering the Pep-FITC comprising the specific MMP2 substrate domain (PLGVR) onto the surface of nrGO particles through non-covalent linkage. The nrGO was obtained by water bathing nano-graphene oxide under 90°C for 4 hours. After mixing the nrGO and Pep-FITC for 30 seconds, the fluorescence from Pep-FITC was almost completely quenched due to the fluorescence resonance energy transfer between fluorescein isothiocyanate (FITC) and nrGO. Upon cleavage of the amide bond between Leu and Gly in the Pep-FITC by protease-MMP2, the FITC bound to nrGO was separated from nrGO surface, disrupting the fluorescence resonance energy transfer process and resulting in fluorescence recovery of FITC. Under optimal conditions, the fluorescence recovery of nrGO/Pep-FITC was found to be directly proportional to the concentration of MMP2 within 0.02-0.1 nM. The detection limit of the nrGO/Pep-FITC was determined to be 3 pM, which is approximately tenfold lower than that of the unreduced carboxylated nano-graphene oxide/Pep-FITC probe.

Validation of a high-performance liquid chromatography/fluorescence detection method for the simultaneous quantification of fifteen polycyclic aromatic hydrocarbons

DEFF Research Database (Denmark)

Hansen, Åse Marie; Olsen, I L; Holst, E

1991-01-01

A high-performance liquid chromatography/fluorescence method using multiple wavelength shift for simultaneous quantification of different PAH compounds was developed. The new method was superior to the methods of DONG and GREENBERG [J. Liquid Chromatogr. 11, 1887-1905 (1988)] and WISE et al...... that no systematic errors, and only small unsystematic errors, could be demonstrated. Furthermore, the method had a good reproducibility and a high sensitivity....

Highly sensitive and multiplexed platforms for allergy diagnostics

Science.gov (United States)

Monroe, Margo R.

Allergy is a disorder of the immune system caused by an immune response to otherwise harmless environmental allergens. Currently 20% of the US population is allergic and 90% of pediatric patients and 60% of adult patients with asthma have allergies. These percentages have increased by 18.5% in the past decade, with predicted similar trends for the future. Here we design sensitive, multiplexed platforms to detect allergen-specific IgE using the Interferometric Reflectance Imaging Sensor (IRIS) for various clinical settings. A microarray platform for allergy diagnosis allows for testing of specific IgE sensitivity to a multitude of allergens, while requiring only small volumes of patient blood sample. However, conventional fluorescent microarray technology is limited by i) the variation of probe immobilization, which hinders the ability to make quantitative, assertive, and statistically relevant conclusions necessary in immunodiagnostics and ii) the use of fluorophore labels, which is not suitable for some clinical applications due to the tendency of fluorophores to stick to blood particulates and require daily calibration methods. This calibrated fluorescence enhancement (CaFE) method integrates the low magnification modality of IRIS with enhanced fluorescence sensing in order to directly correlate immobilized probe (major allergens) density to allergen-specific IgE in patient serum. However, this platform only operates in processed serum samples, which is not ideal for point of care testing. Thus, a high magnification modality of IRIS was adapted as an alternative allergy diagnostic platform to automatically discriminate and size single nanoparticles bound to specific IgE in unprocessed, characterized human blood and serum samples. These features make IRIS an ideal candidate for clinical and diagnostic applications, such a POC testing. The high magnification (nanoparticle counting) modality in conjunction with low magnification of IRIS in a combined instrument

Cleavable DNA-protein hybrid molecular beacon: A novel efficient signal translator for sensitive fluorescence anisotropy bioassay.

Science.gov (United States)

Hu, Pan; Yang, Bin

2016-01-15

Due to its unique features such as high sensitivity, homogeneous format, and independence on fluorescent intensity, fluorescence anisotropy (FA) assay has become a hotspot of study in oligonucleotide-based bioassays. However, until now most FA probes require carefully customized structure designs, and thus are neither generalizable for different sensing systems nor effective to obtain sufficient signal response. To address this issue, a cleavable DNA-protein hybrid molecular beacon was successfully engineered for signal amplified FA bioassay, via combining the unique stable structure of molecular beacon and the large molecular mass of streptavidin. Compared with single DNA strand probe or conventional molecular beacon, the DNA-protein hybrid molecular beacon exhibited a much higher FA value, which was potential to obtain high signal-background ratio in sensing process. As proof-of-principle, this novel DNA-protein hybrid molecular beacon was further applied for FA bioassay using DNAzyme-Pb(2+) as a model sensing system. This FA assay approach could selectively detect as low as 0.5nM Pb(2+) in buffer solution, and also be successful for real samples analysis with good recovery values. Compatible with most of oligonucleotide probes' designs and enzyme-based signal amplification strategies, the molecular beacon can serve as a novel signal translator to expand the application prospect of FA technology in various bioassays. Copyright © 2015 Elsevier B.V. All rights reserved.

Sensitive and selective turn off-on fluorescence detection of heparin based on the energy transfer platform using the BSA-stabilized Au nanoclusters/amino-functionalized graphene oxide hybrids.

Science.gov (United States)

Lan, Jing; Zou, Hong Yan; Wang, Qiang; Zeng, Ping; Li, Yuan Fang; Huang, Cheng Zhi

2016-12-01

An ultra-sensitive and selective turn off-on fluorescence detection of heparin based on the energy transfer in the BSA-stabilized gold nanoclusters/amino-functionalized graphene oxide (BSA-AuNCs/NH 2 -GO) hybrids was successfully realized. The BSA-AuNCs containing amounts of carboxyl groups could be absorbed on the surface of NH 2 -GO through the electrostatic interaction, which resulted in the fluorescence quenching of BSA-AuNCs with high efficiency. However, heparin, possessing high density of negative charge, could compete with BSA-AuNCs to bind NH 2 -GO and block the energy transfer from BSA-AuNCs to NH 2 -GO. The fluorescence recovery of BSA-AuNCs was closely related to the amount of heparin and there was a good linear relationship between fluorescence recovery of BSA-AuNCs and heparin over the range of 100ng/mL to 30μg/mL with a detection limit of 40ng/mL. What's more, the fluorescence assay was successfully applied for heparin sensing in human serums and intracellular imaging. Copyright © 2016 Elsevier B.V. All rights reserved.

Label free selective detection of estriol using graphene oxide-based fluorescence sensor

Science.gov (United States)

Kushwaha, H. S.; Sao, Reshma; Vaish, Rahul

2014-07-01

Water-soluble and fluorescent Graphene oxide (GO) is biocompatible, easy, and economical to synthesize. Interestingly, GO is also capable of quenching fluorescence. On the basis of its fluorescence and quenching abilities, GO has been reported to serve as an energy acceptor in a fluorescence resonance energy transfer (FRET) sensor. GO-based FRET biosensors have been widely reported for sensing of proteins, nucleic acid, ATP (Adenosine triphosphate), etc. GO complexes with fluorescent dyes and enzymes have been used to sense metal ions. Graphene derivatives have been used for sensing endocrine-disrupting chemicals like bisphenols and chlorophenols with high sensitivity and good reproducibility. On this basis, a novel GO based fluorescent sensor has been successfully designed to detect estriol with remarkable selectivity and sensitivity. Estriol is one of the three estrogens in women and is considered to be medically important. Estriol content of maternal urine or plasma acts as an important screening marker for estimating foetal growth and development. In addition, estriol is also used as diagnostic marker for diseases like breast cancer, osteoporosis, neurodegenerative and cardiovascular diseases, insulin resistance, lupus erythematosus, endometriosis, etc. In this present study, we report for the first time a rapid, sensitive with detection limit of 1.3 nM, selective and highly biocompatible method for label free detection of estriol under physiological conditions using fluorescence assay.

Strategies of molecular imprinting-based fluorescence sensors for chemical and biological analysis.

Science.gov (United States)

Yang, Qian; Li, Jinhua; Wang, Xiaoyan; Peng, Hailong; Xiong, Hua; Chen, Lingxin

2018-07-30

One pressing concern today is to construct sensors that can withstand various disturbances for highly selective and sensitive detecting trace analytes in complicated samples. Molecularly imprinted polymers (MIPs) with tailor-made binding sites are preferred to be recognition elements in sensors for effective targets detection, and fluorescence measurement assists in highly sensitive detection and user-friendly control. Accordingly, molecular imprinting-based fluorescence sensors (MI-FL sensors) have attracted great research interest in many fields such as chemical and biological analysis. Herein, we comprehensively review the recent advances in MI-FL sensors construction and applications, giving insights on sensing principles and signal transduction mechanisms, focusing on general construction strategies for intrinsically fluorescent or nonfluorescent analytes and improvement strategies in sensing performance, particularly in sensitivity. Construction strategies are well overviewed, mainly including the traditional indirect methods of competitive binding against pre-bound fluorescent indicators, employment of fluorescent functional monomers and embedding of fluorescence substances, and novel rational designs of hierarchical architecture (core-shell/hollow and mesoporous structures), post-imprinting modification, and ratiometric fluorescence detection. Furthermore, MI-FL sensor based microdevices are discussed, involving micromotors, test strips and microfluidics, which are more portable for rapid point-of-care detection and in-field diagnosing. Finally, the current challenges and future perspectives of MI-FL sensors are proposed. Copyright © 2018 Elsevier B.V. All rights reserved.

New highly fluorescent pH indicator for ratiometric RGB imaging of pCO2

International Nuclear Information System (INIS)

Schutting, Susanne; Klimant, Ingo; Borisov, Sergey M; De Beer, Dirk

2014-01-01

A new diketo-pyrrolo-pyrrole (DPP) indicator dye for optical sensing of carbon dioxide is prepared via a simple one step synthesis from commercially available low cost ‘Pigment Orange 73’. The pigment is modified via alkylation of one of the lactam nitrogens with a tert-butylbenzyl group. The indicator dye is highly soluble in organic solvents and in polymers and shows pH-dependent absorption (λ max 501 and 572 nm for the protonated and deprotonated forms, respectively) and emission spectra (λ max 524 and 605 nm for the protonated and deprotonated forms, respectively). Both the protonated and the deprotonated forms show high fluorescence quantum yields (Φ prot 0.86; Φ deprot 0.66). Hence, colorimetric read-out and ratiometric fluorescence intensity measurements are possible. The emission of the two forms of the indicator excellently matches the response of the green and the red channels of an RGB camera. This enables imaging of carbon dioxide distribution with a simple and low cost optical set-up. The sensor based on the new DPP dye shows very high sensitivity and is particularly promising for monitoring atmospheric levels of carbon dioxide. (paper)

Multiple temperature effects on up-conversion fluorescences of Er3+-Y b3+-Mo6+ codoped TiO2 and high thermal sensitivity

Directory of Open Access Journals (Sweden)

B. S. Cao

2015-08-01

Full Text Available We report multiple temperature effects on green and red up-conversion emissions in Er3+-Y b3+-Mo6+ codoped TiO2 phosphors. With increasing temperature, the decrease of the red emission from 4F9/2→4I15/2, the increase of green emission from 2H11/2→4I15/2 and another unchanged green emission from 4S3/2→4I15/2 were simultaneously observed, which are explained by steady-state rate equations analysis. Due to different evolution with temperature of the two green emissions, higher thermal sensitivity of optical thermal sensor was obtained based on the transitions with the largest fluorescence intensity ratio. Two parameters, maximum theoretical sensitivity (Smax and optimum operating temperature (Tmax are given to describe thermal sensing properties of the produced sensors. The intensity ratio and energy difference ΔE of a pair of energy levels are two main factors for the sensitivity and accuracy of sensors, which should be referred to design sensors with optimized sensing properties.

Recording membrane potential changes through photoacoustic voltage sensitive dye

DEFF Research Database (Denmark)

Zhang, Haichong K.; Kang, Jeeun; Yan, Ping

2017-01-01

Monitoring of the membrane potential is possible using voltage sensitive dyes (VSD), where fluorescence intensity changes in response to neuronal electrical activity. However, fluorescence imaging is limited by depth of penetration and high scattering losses, which leads to low sensitivity in vivo...... systems for external detection. In contrast, photoacoustic (PA) imaging, an emerging modality, is capable of deep tissue, noninvasive imaging by combining near infrared light excitation and ultrasound detection. In this work, we develop the theoretical concept whereby the voltage-dependent quenching...... the experimental PA intensity change depends on fluorescence and absorbance properties of the dye. These results not only demonstrate the voltage sensing capability of the dye, but also indicate the necessity of considering both fluorescence and absorbance spectral sensitivities in order to optimize...

Highly sensitive uric acid biosensor based on individual zinc oxide micro/nanowires

International Nuclear Information System (INIS)

Zhao, Yanguang; Yan, Xiaoqin; Kang, Zhuo; Lin, Pei; Fang, Xiaofei; Lei, Yang; Ma, Siwei; Zhang, Yue

2013-01-01

We describe the use of individual zinc oxide (ZnO) micro/nanowires in an electrochemical biosensor for uric acid. The wires were synthesized by chemical vapor deposition and possess uniform morphology and high crystallinity as revealed by scanning electron microscopy, X-ray diffraction, and photoluminescence studies. The enzyme uricase was then immobilized on the surface of the ZnO micro/nanowires by physical adsorption, and this was proven by Raman spectroscopy and fluorescence microscopy. The resulting uric acid biosensor undergoes fast electron transfer between the active site of the enzyme and the surface of the electrode. It displays high sensitivity (89.74 μA cm −2 mM −1 ) and a wide linear analytical range (between 0.1 mM and 0.59 mM concentrations of uric acid). This study also demonstrates the potential of the use of individual ZnO micro/nanowires for the construction of highly sensitive nano-sized biosensors. (author)

High-contrast fluorescence imaging based on the polarization dependence of the fluorescence enhancement using an optical interference mirror slide.

Science.gov (United States)

Yasuda, Mitsuru; Akimoto, Takuo

2015-01-01

High-contrast fluorescence imaging using an optical interference mirror (OIM) slide that enhances the fluorescence from a fluorophore located on top of the OIM surface is reported. To enhance the fluorescence and reduce the background light of the OIM, transverse-electric-polarized excitation light was used as incident light, and the transverse-magnetic-polarized fluorescence signal was detected. As a result, an approximate 100-fold improvement in the signal-to-noise ratio was achieved through a 13-fold enhancement of the fluorescence signal and an 8-fold reduction of the background light.

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

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.

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

Directory of Open Access Journals (Sweden)

Yoshichika Yoshioka

2008-10-01

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

A high-throughput fluorescence resonance energy transfer (FRET)-based endothelial cell apoptosis assay and its application for screening vascular disrupting agents

International Nuclear Information System (INIS)

Zhu, Xiaoming; Fu, Afu; Luo, Kathy Qian

2012-01-01

Highlights: ► An endothelial cell apoptosis assay using FRET-based biosensor was developed. ► The fluorescence of the cells changed from green to blue during apoptosis. ► This method was developed into a high-throughput assay in 96-well plates. ► This assay was applied to screen vascular disrupting agents. -- Abstract: In this study, we developed a high-throughput endothelial cell apoptosis assay using a fluorescence resonance energy transfer (FRET)-based biosensor. After exposure to apoptotic inducer UV-irradiation or anticancer drugs such as paclitaxel, the fluorescence of the cells changed from green to blue. We developed this method into a high-throughput assay in 96-well plates by measuring the emission ratio of yellow fluorescent protein (YFP) to cyan fluorescent protein (CFP) to monitor the activation of a key protease, caspase-3, during apoptosis. The Z′ factor for this assay was above 0.5 which indicates that this assay is suitable for a high-throughput analysis. Finally, we applied this functional high-throughput assay for screening vascular disrupting agents (VDA) which could induce endothelial cell apoptosis from our in-house compounds library and dioscin was identified as a hit. As this assay allows real time and sensitive detection of cell apoptosis, it will be a useful tool for monitoring endothelial cell apoptosis in living cell situation and for identifying new VDA candidates via a high-throughput screening.

Microcystin Detection Characteristics of Fluorescence Immunochromatography and High Performance Liquid Chromatography

International Nuclear Information System (INIS)

Pyo, Dong Jin; Park, Geun Young; Choi, Jong Chon; Oh, Chang Suk

2005-01-01

Different detection characteristics of fluorescence immunochromatography method and high performance liquid chromatography (HPLC) method for the analysis of cyanobacterial toxins were studied. In particular, low and high limits of detection, detection time and reproducibility and detectable microcystin species were compared when fluorescence immunochromatography method and high performance liquid chromatography method were applied for the detection of microcystin (MC), a cyclic peptide toxin of the freshwater cyanobacterium Microcystis aeruginosa. A Fluorescence immunochromatography assay system has the unique advantages of short detection time and low detection limit, and high performance liquid chromatography detection method has the strong advantage of individual quantifications of several species of microcystins

Fluorescence Molecular Tomography: Principles and Potential for Pharmaceutical Research

Directory of Open Access Journals (Sweden)

Florian Stuker

2011-04-01

Full Text Available Fluorescence microscopic imaging is widely used in biomedical research to study molecular and cellular processes in cell culture or tissue samples. This is motivated by the high inherent sensitivity of fluorescence techniques, the spatial resolution that compares favorably with cellular dimensions, the stability of the fluorescent labels used and the sophisticated labeling strategies that have been developed for selectively labeling target molecules. More recently, two and three-dimensional optical imaging methods have also been applied to monitor biological processes in intact biological organisms such as animals or even humans. These whole body optical imaging approaches have to cope with the fact that biological tissue is a highly scattering and absorbing medium. As a consequence, light propagation in tissue is well described by a diffusion approximation and accurate reconstruction of spatial information is demanding. While in vivo optical imaging is a highly sensitive method, the signal is strongly surface weighted, i.e., the signal detected from the same light source will become weaker the deeper it is embedded in tissue, and strongly depends on the optical properties of the surrounding tissue. Derivation of quantitative information, therefore, requires tomographic techniques such as fluorescence molecular tomography (FMT, which maps the three-dimensional distribution of a fluorescent probe or protein concentration. The combination of FMT with a structural imaging method such as X-ray computed tomography (CT or Magnetic Resonance Imaging (MRI will allow mapping molecular information on a high definition anatomical reference and enable the use of prior information on tissue’s optical properties to enhance both resolution and sensitivity. Today many of the fluorescent assays originally developed for studies in cellular systems have been successfully translated for experimental studies in animals. The opportunity of monitoring molecular

Fluorescent imaging of high-grade bladder cancer using a specific antagonist for chemokine receptor CXCR4.

Science.gov (United States)

Nishizawa, Koji; Nishiyama, Hiroyuki; Oishi, Shinya; Tanahara, Noriko; Kotani, Hirokazu; Mikami, Yoshiki; Toda, Yoshinobu; Evans, Barry J; Peiper, Stephen C; Saito, Ryoichi; Watanabe, Jun; Fujii, Nobutaka; Ogawa, Osamu

2010-09-01

We previously reported that the expression of CXC chemokine receptor-4 (CXCR4) was upregulated in invasive bladder cancers and that the small peptide T140 was a highly sensitive antagonist for CXCR4. In this study, we identified that CXCR4 expression was induced in high-grade superficial bladder tumors, including carcinoma in situ and invasive bladder tumors. To visualize the bladder cancer cells using urinary sediments from the patients and chemically induced mouse bladder cancer model, a novel fluorescent CXCR4 antagonist TY14003 was developed, that is a T140 derivative. TY14003 could label bladder cancer cell lines expressing CXCR4, whereas negative-control fluorescent peptides did not label them. When labeling urinary sediments from patients with invasive bladder cancer, positive-stained cells were identified in all patients with bladder cancer and positive urine cytology but not in controls. Although white blood cells in urine were also labeled with TY14003, they could be easily discriminated from urothelial cells by their shape and size. Finally, intravesical instillation of TY14003 into mouse bladder, using N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN)-induced bladder cancer model, demonstrated that fluorescent signals were detected in the focal areas of bladder of all mice examined at 12 weeks of BBN drinking by confocal microscopy and fluorescent endoscopy. On the contrary, all the normal bladders were found to be negative for TY14003 staining. In conclusion, these results indicate that TY14003 is a promising diagnostic tool to visualize small or flat high-grade superficial bladder cancer.

A novel portable fluorescence detection system for microfluidic card

International Nuclear Information System (INIS)

Shen, B; Xie, Y; Irawan, R

2008-01-01

Fluorescence based sensors are widely used in the field of biochemistry and medicine due to their high sensitivity and accuracy. But the cost and time required for each sample to be tested is high. If the diagnostic tools could be miniaturized, made simple to use and much less expensive, and readily available at the point of need such as emergency diagnosis, millions of people would be benefited from it. In this paper, we design a prototype of portable fluorescence detection system based on Fluorescence Filter Block and DAQ card which can emulate signal collection and processing functionalities. After the introduction of system structure and functional modules, we use a resolution approximation method to investigate the system performance. The evaluation shows that our prototype system has the sensitivity of 0.01 mMol/L (333.306 μg/mL) which meets most of the medical requirements.

Construction of a ColD cda Promoter-Based SOS-Green Fluorescent Protein Whole-Cell Biosensor with Higher Sensitivity toward Genotoxic Compounds than Constructs Based on recA, umuDC, or sulA Promoters

DEFF Research Database (Denmark)

Norman, Anders; Hansen, Lars Hestbjerg; Sørensen, Søren Johannes

2005-01-01

Four different green fluorescent protein (GFP)-based whole-cell biosensors were created based on the DNA damage inducible SOS response of Escherichia coli in order to evaluate the sensitivity of individual SOS promoters toward genotoxic substances. Treatment with the known carcinogen N-methyl-N'-......Four different green fluorescent protein (GFP)-based whole-cell biosensors were created based on the DNA damage inducible SOS response of Escherichia coli in order to evaluate the sensitivity of individual SOS promoters toward genotoxic substances. Treatment with the known carcinogen N......-cell biosensor which is not only able to detect minute levels of genotoxins but, due to its use of the green fluorescent protein, also a reporter system which should be applicable in high-throughput screening assays as well as a wide variety of in situ detection studies....

Fluorescence-based assay as a new screening tool for toxic chemicals

Science.gov (United States)

Moczko, Ewa; Mirkes, Evgeny M.; Cáceres, César; Gorban, Alexander N.; Piletsky, Sergey

2016-09-01

Our study involves development of fluorescent cell-based diagnostic assay as a new approach in high-throughput screening method. This highly sensitive optical assay operates similarly to e-noses and e-tongues which combine semi-specific sensors and multivariate data analysis for monitoring biochemical processes. The optical assay consists of a mixture of environmental-sensitive fluorescent dyes and human skin cells that generate fluorescence spectra patterns distinctive for particular physico-chemical and physiological conditions. Using chemometric techniques the optical signal is processed providing qualitative information about analytical characteristics of the samples. This integrated approach has been successfully applied (with sensitivity of 93% and specificity of 97%) in assessing whether particular chemical agents are irritating or not for human skin. It has several advantages compared with traditional biochemical or biological assays and can impact the new way of high-throughput screening and understanding cell activity. It also can provide reliable and reproducible method for assessing a risk of exposing people to different harmful substances, identification active compounds in toxicity screening and safety assessment of drugs, cosmetic or their specific ingredients.

Permethylated-β-Cyclodextrin Capped CdTe Quantum Dot and its Sensitive Fluorescence Analysis of Malachite Green.

Science.gov (United States)

Cao, Yujuan; Wei, Jiongling; Wu, Wei; Wang, Song; Hu, Xiaogang; Yu, Ying

2015-09-01

In the present work, the CdTe quantum dots were covalently conjugated with permethylated-β-cyclodextrin (OMe-β-CD) using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride as cross-linking reagent. The obtained functional quantum dots (OMe-β-CD/QDs) showed highly luminescent, water solubility and photostability as well as good inclusion ability to malachite green. A sensitive fluorescence method was developed for the analysis of malachite green in different samples. The good linearity was 2.0 × 10(-7)-1.0 × 10(-5) mol/L and the limit of detect was 1.7 × 10(-8) mol/L. The recoveries for three environmental water samples were 92.0-108.2 % with relative standard deviation (RSD) of 0.24-1.87 %, while the recovery for the fish sample was 94.3 % with RSD of 1.04 %. The results showed that the present method was sensitive and convenient to determine malachite green in complex samples. Graphical Abstract The analytical mechanism of OMe-β-CD/QDs and its linear response to MG.

Rapid, sensitive, and selective fluorescent DNA detection using iron-based metal-organic framework nanorods: Synergies of the metal center and organic linker.

Science.gov (United States)

Tian, Jingqi; Liu, Qian; Shi, Jinle; Hu, Jianming; Asiri, Abdullah M; Sun, Xuping; He, Yuquan

2015-09-15

Considerable recent attention has been paid to homogeneous fluorescent DNA detection with the use of nanostructures as a universal "quencher", but it still remains a great challenge to develop such nanosensor with the benefits of low cost, high speed, sensitivity, and selectivity. In this work, we report the use of iron-based metal-organic framework nanorods as a high-efficient sensing platform for fluorescent DNA detection. It only takes about 4 min to complete the whole "mix-and-detect" process with a low detection limit of 10 pM and a strong discrimination of single point mutation. Control experiments reveal the remarkable sensing behavior is a consequence of the synergies of the metal center and organic linker. This work elucidates how composition control of nanostructures can significantly impact their sensing properties, enabling new opportunities for the rational design of functional materials for analytical applications. Copyright © 2015 Elsevier B.V. All rights reserved.

Fluorescent and high intensity discharge lamp use in chambers and greenhouses

Science.gov (United States)

Langhans, Robert W.

1994-01-01

Fluorescent and High Intensity Discharge lamps have opened up great opportunities for researchers to study plant growth under controlled environment conditions and for commercial growers to increase plant production during low/light periods. Specific technical qualities of fluorescent and HID lamps have been critically reviewed. I will direct my remarks to fluorescent and high intensity discharge (HID) lamps in growth chambers, growth rooms, and greenhouses. I will discuss the advantages and disadvantages of using each lamp in growth chambers, growth rooms and greenhouses.

A novel fluorimetric sensing platform for highly sensitive detection of organophosphorus pesticides by using egg white-encapsulated gold nanoclusters.

Science.gov (United States)

Yan, Xu; Li, Hongxia; Hu, Tianyu; Su, Xingguang

2017-05-15

Assays for organophosphorus pesticides (OPs) with high sensitivity as well as on-site screening have been urgently required to protect ecosystem and prevent disease. Herein, a novel fluorimetric sensing platform was constructed for quantitative detection of OPs via tyrosinase (TYR) enzyme-controlled quenching of gold nanoclusters (AuNCs). One-step green synthetic approach was developed for the synthesis of AuNCs by using chicken egg white (CEW) as template and stabilizer. Initially, TYR can catalyze the oxidation of dopamine to dopaminechrome, which can efficiently quench the fluorescence intensity of AuNCs at 630nm based on dynamic quenching process. However, with the presence of OPs, the activity of TYR was inhibited, resulting in the fluorescence recovery of AuNCs. This proposed fluorescence platform was demonstrated to enable rapid detection for OPs (paraoxon as model) and to provide excellent sensitivity with a detection limit of 0.1ngmL -1 . Significantly, the fluorescence probe was used to prepare paper-based test strips for visual detection of OPs, which validated the excellent potential for real-time and on-site application. Copyright © 2016 Elsevier B.V. All rights reserved.

Fluorescent foci quantitation for high-throughput analysis

Directory of Open Access Journals (Sweden)

Elena Ledesma-Fernández

2015-06-01

Full Text Available A number of cellular proteins localize to discrete foci within cells, for example DNA repair proteins, microtubule organizing centers, P bodies or kinetochores. It is often possible to measure the fluorescence emission from tagged proteins within these foci as a surrogate for the concentration of that specific protein. We wished to develop tools that would allow quantitation of fluorescence foci intensities in high-throughput studies. As proof of principle we have examined the kinetochore, a large multi-subunit complex that is critical for the accurate segregation of chromosomes during cell division. Kinetochore perturbations lead to aneuploidy, which is a hallmark of cancer cells. Hence, understanding kinetochore homeostasis and regulation are important for a global understanding of cell division and genome integrity. The 16 budding yeast kinetochores colocalize within the nucleus to form a single focus. Here we have created a set of freely-available tools to allow high-throughput quantitation of kinetochore foci fluorescence. We use this ‘FociQuant’ tool to compare methods of kinetochore quantitation and we show proof of principle that FociQuant can be used to identify changes in kinetochore protein levels in a mutant that affects kinetochore function. This analysis can be applied to any protein that forms discrete foci in cells.

Ultrasensitive fluorescence immunoassay for detection of ochratoxin A using catalase-mediated fluorescence quenching of CdTe QDs

Science.gov (United States)

Huang, Xiaolin; Zhan, Shengnan; Xu, Hengyi; Meng, Xianwei; Xiong, Yonghua; Chen, Xiaoyuan

2016-04-01

Herein, for the first time we report an improved competitive fluorescent enzyme linked immunosorbent assay (ELISA) for the ultrasensitive detection of ochratoxin A (OTA) by using hydrogen peroxide (H2O2)-induced fluorescence quenching of mercaptopropionic acid-modified CdTe quantum dots (QDs). In this immunoassay, catalase (CAT) was labeled with OTA as a competitive antigen to connect the fluorescence signals of the QDs with the concentration of the target. Through the combinatorial use of H2O2-induced fluorescence quenching of CdTe QDs as a fluorescence signal output and the ultrahigh catalytic activity of CAT to H2O2, our proposed method could be used to perform a dynamic linear detection of OTA ranging from 0.05 pg mL-1 to 10 pg mL-1. The half maximal inhibitory concentration was 0.53 pg mL-1 and the limit of detection was 0.05 pg mL-1. These values were approximately 283- and 300-folds lower than those of horseradish peroxidase (HRP)-based conventional ELISA, respectively. The reported method is accurate, highly reproducible, and specific against other mycotoxins in agricultural products as well. In summary, the developed fluorescence immunoassay based on H2O2-induced fluorescence quenching of CdTe QDs can be used for the rapid and highly sensitive detection of mycotoxins or haptens in food safety monitoring.Herein, for the first time we report an improved competitive fluorescent enzyme linked immunosorbent assay (ELISA) for the ultrasensitive detection of ochratoxin A (OTA) by using hydrogen peroxide (H2O2)-induced fluorescence quenching of mercaptopropionic acid-modified CdTe quantum dots (QDs). In this immunoassay, catalase (CAT) was labeled with OTA as a competitive antigen to connect the fluorescence signals of the QDs with the concentration of the target. Through the combinatorial use of H2O2-induced fluorescence quenching of CdTe QDs as a fluorescence signal output and the ultrahigh catalytic activity of CAT to H2O2, our proposed method could be used to

A Label-Free and Sensitive Fluorescent Qualitative Assay for Bisphenol A Based on Rolling Circle Amplification/Exonuclease III-Combined Cascade Amplification

Directory of Open Access Journals (Sweden)

Xia Li

2016-10-01

Full Text Available Bisphenol A (BPA detection in drinking water and food packaging materials has attracted much attention since the discovery that BPA can interfere with normal physiological processes and cause adverse health effects. Here, we constructed a label-free aptamer fluorescent assay for selective and sensitive detection of BPA based on the rolling circle amplification (RCA/Exonuclease III (Exo III-combined cascade amplification strategy. First, the duplex DNA probe (RP with anti-BPA aptamer and trigger sequence was designed for BPA recognition and signal amplification. Next, under the action of BPA, the trigger probe was liberated from RP to initiate RCA reaction as primary amplification. Subsequently, the RCA products were used to trigger Exo III assisted secondary amplification with the help of hairpin probes, producing plenty of “G-quadruplex” in lantern-like structures. Finally, the continuously enriched “G-quadruplex lanterns” were lightened by zinc(II-protoporphyrin IX (ZnPPIX generating enhanced fluorescence signals. By integrating the primary RCA and secondary Exo III mediated cascade amplification strategy, this method displayed an excellent sensitivity with the detection limits of 5.4 × 10−17 M. In addition, the anti-BPA aptamer exhibits high recognition ability with BPA, guaranteeing the specificity of detection. The reporter signal probe (G-quadruplex with ZnPPIX provides a label-free fluorescence signals readout without complicated labeling procedures, making the method simple in design and cost-effective in operation. Moreover, environmental samples analysis was also performed, suggesting that our strategy was reliable and had a great potential application in environmental monitoring.

A Label-Free and Sensitive Fluorescent Qualitative Assay for Bisphenol A Based on Rolling Circle Amplification/Exonuclease III-Combined Cascade Amplification.

Science.gov (United States)

Li, Xia; Song, Juan; Xue, Qing-Wang; You, Fu-Heng; Lu, Xia; Kong, Yan-Cong; Ma, Shu-Yi; Jiang, Wei; Li, Chen-Zhong

2016-10-21

Bisphenol A (BPA) detection in drinking water and food packaging materials has attracted much attention since the discovery that BPA can interfere with normal physiological processes and cause adverse health effects. Here, we constructed a label-free aptamer fluorescent assay for selective and sensitive detection of BPA based on the rolling circle amplification (RCA)/Exonuclease III (Exo III)-combined cascade amplification strategy. First, the duplex DNA probe (RP) with anti-BPA aptamer and trigger sequence was designed for BPA recognition and signal amplification. Next, under the action of BPA, the trigger probe was liberated from RP to initiate RCA reaction as primary amplification. Subsequently, the RCA products were used to trigger Exo III assisted secondary amplification with the help of hairpin probes, producing plenty of "G-quadruplex" in lantern-like structures. Finally, the continuously enriched "G-quadruplex lanterns" were lightened by zinc(II)-protoporphyrin IX (ZnPPIX) generating enhanced fluorescence signals. By integrating the primary RCA and secondary Exo III mediated cascade amplification strategy, this method displayed an excellent sensitivity with the detection limits of 5.4 × 10 -17 M. In addition, the anti-BPA aptamer exhibits high recognition ability with BPA, guaranteeing the specificity of detection. The reporter signal probe (G-quadruplex with ZnPPIX) provides a label-free fluorescence signals readout without complicated labeling procedures, making the method simple in design and cost-effective in operation. Moreover, environmental samples analysis was also performed, suggesting that our strategy was reliable and had a great potential application in environmental monitoring.

Selective detection of heavy metal ions by calixarene-based fluorescent molecular sensors

Science.gov (United States)

Zhang, Haitao; Faye, Djibril; Zhang, Han; Lefevre, Jean-Pierre; Delaire, J. A.; Leray, Isabelle

2012-06-01

The synthesis, spectroscopic characterization and complexing properties of calixarene-based fluorescent sensors are reported. The calixarene bearing four dansyl fluorophores (Calix-DANS4) exhibits a very high affinity for the detection of lead. A fluorimetric micro-device based on the use of a Y-shape microchannel was developed and allows lead detection with a 5 ppb detection limit. For mercury detection, a fluorescent molecular sensor containing a calixarene anchored with four 8-quinolinoloxy groups (Calix-Q) has been synthesized. The absorption and fluorescence spectra of this sensor are sensitive to the presence of metal cations. An efficient fluorescence quenching is observed upon mercury complexation because of a photoinduced electron transfer from the fluorophore to the bound mercury. Calix-Q shows a high selectivity towards Hg2+ over interfering cations (Na+, K+, Ca2+, Cu2+, Zn2+, Cd2+ and Pb2+) and a 70 ppb sensitivity.

A novel fluorescence biosensor for sensitivity detection of tyrosinase and acid phosphatase based on nitrogen-doped graphene quantum dots.

Science.gov (United States)

Qu, Zhengyi; Na, Weidan; Liu, Xiaotong; Liu, Hua; Su, Xingguang

2018-01-02

In this paper, we developed a sensitive fluorescence biosensor for tyrosinase (TYR) and acid phosphatase (ACP) activity detection based on nitrogen-doped graphene quantum dots (N-GQDs). Tyrosine could be catalyzed by TYR to generate dopaquinone, which could efficiently quench the fluorescence of N-GQDs, and the degree of fluorescence quenching of N-GQDs was proportional to the concentration of TYR. In the presence of ACP, l-Ascorbic acid-2-phosphate (AAP) was hydrolyzed to generate ascorbic acid (AA), and dopaquinone was reduced to l-dopa, resulting in the fluorescence recovery of the quenched fluorescence by dopaquinone. Thus, a novel fluorescence biosensor for the detection of TYR and ACP activity based on N-GQDs was constructed. Under the optimized experimental conditions, the fluorescence intensity was linearly correlated with the concentration of TYR and ACP in the range of 0.43-3.85 U mL -1 and 0.04-0.7 mU mL -1 with a detection limit of 0.15 U mL -1 and 0.014 mU mL -1 , respectively. The feasibility of the proposed biosensor in real samples assay was also studied and satisfactory results were obtained. Copyright © 2017 Elsevier B.V. All rights reserved.

Europium-decorated graphene quantum dots as a fluorescent probe for label-free, rapid and sensitive detection of Cu{sup 2+} and L-cysteine

Energy Technology Data Exchange (ETDEWEB)

Lin, Liping [College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002 (China); Song, Xinhong; Chen, Yiying; Rong, Mingcong; Wang, Yiru [Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005 (China); Zhao, Li; Zhao, Tingting [Xiamen Huaxia College, Xiamen, 361024 (China); Chen, Xi, E-mail: xichen@xmu.edu.cn [Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005 (China); State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen, 361005 (China)

2015-09-03

In this work, europium-decorated graphene quantum dots (Eu-GQDs) were prepared by treating three-dimensional Eu-decorated graphene (3D Eu-graphene) via a strong acid treatment. Various characterizations revealed that Eu atoms were successfully complexed with the oxygen functional groups on the surface of graphene quantum dots (GQDs) with the atomic ratio of 2.54%. Compared with Eu free GQDs, the introduction of Eu atoms enhanced the electron density and improved the surface chemical activities of Eu-GQDs. Therefore, the obtained Eu-GQDs were used as a novel “off-on” fluorescent probe for the label-free determination of Cu{sup 2+} and L-cysteine (L-Cys) with high sensitivity and selectivity. The fluorescence intensity of Eu-GQDs was quenched in the presence of Cu{sup 2+} owing to the coordination reaction between Cu{sup 2+} and carboxyl groups on the surface of the Eu-GQDs. The fluorescence intensity of Eu-GQDs recovered with the subsequent addition of L-Cys because of the strong affinity of Cu{sup 2+} to L-Cys via the Cu–S bond. The experimental results showed that the fluorescence variation of the proposed approach had a good linear relationship in the range of 0.1–10 μM for Cu{sup 2+} and 0.5–50 μM for L-Cys with corresponding detection limits of 0.056 μM for Cu{sup 2+} and 0.31 μM for L-Cys. The current approach also displayed a special response to Cu{sup 2+} and L-Cys over the other co-existing metal ions and amino acids, and the results obtained from buffer-diluted serum samples suggested its applicability in biological samples. - Highlights: • The europium-decorated graphene quantum dots (Eu-GQDs) have been successfully prepared. • Various characterizations results proved that Eu atoms were successfully introduced into graphene quantum dots. • The introduced Eu atoms changed the electron density and surface chemical activities of Eu-GQDs. • Eu-GQDs were used as an “off-on” fluorescent probe for Cu{sup 2+} and L-cysteine detection

Multicolor fluorescent biosensor for multiplexed detection of DNA.

Science.gov (United States)

Hu, Rong; Liu, Tao; Zhang, Xiao-Bing; Huan, Shuang-Yan; Wu, Cuichen; Fu, Ting; Tan, Weihong

2014-05-20

Development of efficient methods for highly sensitive and rapid screening of specific oligonucleotide sequences is essential to the early diagnosis of serious diseases. In this work, an aggregated cationic perylene diimide (PDI) derivative was found to efficiently quench the fluorescence emission of a variety of anionic oligonucleotide-labeled fluorophores that emit at wavelengths from the visible to NIR region. This broad-spectrum quencher was then adopted to develop a multicolor biosensor via a label-free approach for multiplexed fluorescent detection of DNA. The aggregated perylene derivative exhibits a very high quenching efficiency on all ssDNA-labeled dyes associated with biosensor detection, having efficiency values of 98.3 ± 0.9%, 97 ± 1.1%, and 98.2 ± 0.6% for FAM, TAMRA, and Cy5, respectively. An exonuclease-assisted autocatalytic target recycling amplification was also integrated into the sensing system. High quenching efficiency combined with autocatalytic target recycling amplification afforded the biosensor with high sensitivity toward target DNA, resulting in a detection limit of 20 pM, which is about 50-fold lower than that of traditional unamplified homogeneous fluorescent assay methods. The quencher did not interfere with the catalytic activity of nuclease, and the biosensor could be manipulated in either preaddition or postaddition manner with similar sensitivity. Moreover, the proposed sensing system allows for simultaneous and multicolor analysis of several oligonucleotides in homogeneous solution, demonstrating its potential application in the rapid screening of multiple biotargets.

Oxygen sensitive polymeric nanocapsules for optical dissolved oxygen sensors

Science.gov (United States)

Sun, Zhijuan; Cai, Chenxin; Guo, Fei; Ye, Changhuai; Luo, Yingwu; Ye, Shuming; Luo, Jianchao; Zhu, Fan; Jiang, Chunyue

2018-04-01

Immobilization of the oxygen-sensitive probes (OSPs) in the host matrix greatly impacts the performance and long-term usage of the optical dissolved oxygen (DO) sensors. In this work, fluorescent dyes, as the OSPs, were encapsulated with a crosslinked fluorinated polymer shell by interfacial confined reversible addition fragmentation chain transfer miniemulsion polymerization to fabricate oxygen sensitive polymeric nanocapsules (NCs). The location of fluorescent dyes and the fluorescent properties of the NCs were fully characterized by fourier transform infrared spectrometer, x-ray photoelectron spectrometer and fluorescent spectrum. Dye-encapsulated capacity can be precisely tuned from 0 to 1.3 wt% without self-quenching of the fluorescent dye. The crosslinked fluorinated polymer shell is not only extremely high gas permeability, but also prevents the fluorescent dyes from leakage in aqueous as well as in various organic solvents, such as ethanol, acetone and tetrahydrofuran (THF). An optical DO sensor based on the oxygen sensitive NCs was fabricated, showing high sensitivity, short response time, full reversibility, and long-term operational stability of online monitoring DO. The sensitivity of the optical DO sensor is 7.02 (the ratio of the response value in fully deoxygenated and saturated oxygenated water) in the range 0.96-14.16 mg l-1 and the response time is about 14.3 s. The sensor’s work curve was fit well using the modified Stern-Volmer equation by two-site model, and its response values are hardly affected by pH ranging from 2 to 12 and keep constant during continuous measurement for 3 months. It is believed that the oxygen sensitive polymeric NCs-based optical DO sensor could be particularly useful in long-term online DO monitoring in both aqueous and organic solvent systems.

System for Gamma an X rays fluorescence spectrometric

International Nuclear Information System (INIS)

Alonso Abad, D.; Arista Romeu, E.; Bolanos Perez, L. and others

1997-01-01

A system for spectrometry of gamma or fluorescence X rays is presented. It sis composed by a Si(Li) semiconductors detector, a charge sensitive preamplifier, a high voltage power supply, a spectrometric amplifier and a monolithic 1024 channels multichannel analyzers or an IBM compatible 4096 channels add - on- card multichannel analyzer. The system can be configured as a 1024 or 4096 channels gamma or fluorescent X rays spectrometer

High-throughput screening with micro-x-ray fluorescence

International Nuclear Information System (INIS)

Havrilla, George J.; Miller, Thomasin C.

2005-01-01

Micro-x-ray fluorescence (MXRF) is a useful characterization tool for high-throughput screening of combinatorial libraries. Due to the increasing threat of use of chemical warfare (CW) agents both in military actions and against civilians by terrorist extremists, there is a strong push to improve existing methods and develop means for the detection of a broad spectrum of CW agents in a minimal amount of time to increase national security. This paper describes a combinatorial high-throughput screening technique for CW receptor discovery to aid in sensor development. MXRF can screen materials for elemental composition at the mesoscale level (tens to hundreds of micrometers). The key aspect of this work is the use of commercial MXRF instrumentation coupled with the inherent heteroatom elements within the target molecules of the combinatorial reaction to provide rapid and specific identification of lead species. The method is demonstrated by screening an 11-mer oligopeptide library for selective binding of the degradation products of the nerve agent VX. The identified oligopeptides can be used as selective molecular receptors for sensor development. The MXRF screening method is nondestructive, requires minimal sample preparation or special tags for analysis, and the screening time depends on the desired sensitivity

Fusions between green fluorescent protein and beta-glucuronidase as sensitive and vital bifunctional reporters in plants.

Science.gov (United States)

Quaedvlieg, N E; Schlaman, H R; Admiraal, P C; Wijting, S E; Stougaard, J; Spaink, H P

1998-11-01

By fusing the genes encoding green fluorescent protein (GFP) and beta-glucuronidase (GUS) we have created a set of bifunctional reporter constructs which are optimized for use in transient and stable expression studies in plants. This approach makes it possible to combine the advantage of GUS, its high sensitivity in histochemical staining, with the advantages of GFP as a vital marker. The fusion proteins were functional in transient expression studies in tobacco using either DNA bombardment or potato virus X as a vector, and in stably transformed Arabidopsis thaliana and Lotus japonicus plants. The results show that high level of expression does not interfere with efficient stable transformation in A. thaliana and L. japonicus. Using confocal laser scanning microscopy we show that the fusion constructs are very suitable for promoter expression studies in all organs of living plants, including root nodules. The use of these reporter constructs in the model legume L. japonicus offers exciting new possibilities for the study of the root nodulation process.

Fluorescence detection system for microfluidic droplets

Science.gov (United States)

Chen, Binyu; Han, Xiaoming; Su, Zhen; Liu, Quanjun

2018-05-01

In microfluidic detection technology, because of the universality of optical methods in laboratory, optical detection is an attractive solution for microfluidic chip laboratory equipment. In addition, the equipment with high stability and low cost can be realized by integrating appropriate optical detection technology on the chip. This paper reports a detection system for microfluidic droplets. Photomultiplier tubes (PMT) is used as a detection device to improve the sensitivity of detection. This system improves the signal to noise ratio by software filtering and spatial filter. The fluorescence intensity is proportional to the concentration of the fluorescence and intensity of the laser. The fluorescence micro droplets of different concentrations can be distinguished by this system.

Fluorescence-based high-throughput functional profiling of ligand-gated ion channels at the level of single cells.

Directory of Open Access Journals (Sweden)

Sahil Talwar

Full Text Available Ion channels are involved in many physiological processes and are attractive targets for therapeutic intervention. Their functional properties vary according to their subunit composition, which in turn varies in a developmental and tissue-specific manner and as a consequence of pathophysiological events. Understanding this diversity requires functional analysis of ion channel properties in large numbers of individual cells. Functional characterisation of ligand-gated channels involves quantitating agonist and drug dose-response relationships using electrophysiological or fluorescence-based techniques. Electrophysiology is limited by low throughput and high-throughput fluorescence-based functional evaluation generally does not enable the characterization of the functional properties of each individual cell. Here we describe a fluorescence-based assay that characterizes functional channel properties at single cell resolution in high throughput mode. It is based on progressive receptor activation and iterative fluorescence imaging and delivers >100 dose-responses in a single well of a 384-well plate, using α1-3 homomeric and αβ heteromeric glycine receptor (GlyR chloride channels as a model system. We applied this assay with transiently transfected HEK293 cells co-expressing halide-sensitive yellow fluorescent protein and different GlyR subunit combinations. Glycine EC50 values of different GlyR isoforms were highly correlated with published electrophysiological data and confirm previously reported pharmacological profiles for the GlyR inhibitors, picrotoxin, strychnine and lindane. We show that inter and intra well variability is low and that clustering of functional phenotypes permits identification of drugs with subunit-specific pharmacological profiles. As this method dramatically improves the efficiency with which ion channel populations can be characterized in the context of cellular heterogeneity, it should facilitate systems

Rapid quantification of live/dead lactic acid bacteria in probiotic products using high-sensitivity flow cytometry

Science.gov (United States)

He, Shengbin; Hong, Xinyi; Huang, Tianxun; Zhang, Wenqiang; Zhou, Yingxing; Wu, Lina; Yan, Xiaomei

2017-06-01

A laboratory-built high-sensitivity flow cytometer (HSFCM) was employed for the rapid and accurate detection of lactic acid bacteria (LAB) and their viability in probiotic products. LAB were stained with both the cell membrane-permeable SYTO 9 green-fluorescent nucleic acid stain and the red-fluorescent nucleic acid stain, propidium iodide, which penetrates only bacteria with compromised membranes. The side scatter and dual-color fluorescence signals of single bacteria were detected simultaneously by the HSFCM. Ultra-high temperature processing milk and skim milk spiked with Lactobacillus casei were used as the model systems for the optimization of sample pretreatment and staining. The viable LAB counts measured by the HSFCM were in good agreement with those of the plate count method, and the measured ratios between the live and dead LAB matched well with the theoretical ratios. The established method was successfully applied to the rapid quantification of live/dead LAB in yogurts and fermented milk beverages of different brands. Moreover, the concentration and viability status of LAB in ambient yogurt, a relatively new yet popular milk product in China, are also reported.

Hemoglobin detection using carbon dots as a fluorescence probe.

Science.gov (United States)

Barati, Ali; Shamsipur, Mojtaba; Abdollahi, Hamid

2015-09-15

Herein, we have described the application of high fluorescent carbon dots (CDs) without any surface modification as a simple and fast responding fluorescence probe for sensitive and selective determination of hemoglobin (Hb) in the presence of H2O2. Although Hb itself was able to quench the fluorescence of CDs, based on the inner filter effect (IFE) of the protein that affects both excitation and emission spectra of CDs, the presence of H2O2 resulted in further improvement of the sensitivity of Hb detection. The assay is based on the reaction of Hb with H2O2 that generates reactive oxygen species including hydroxyl (OH•) and superoxide (O2(•-)) radicals under heme degradation and/or iron release from Hb and the subsequent reaction of hydroxyl radicals, as strong oxidizing agents, with CDs resulting in high fluorescence quenching. The proposed probe was used for determination of Hb in concentration range of 1-100 nM with a detection limit of 0.4 nM. The method was successfully applied to the determination of Hb in human blood samples. Copyright © 2015 Elsevier B.V. All rights reserved.

Development of an underwater high sensitivity Cherenkov detector: Sea Urchin

International Nuclear Information System (INIS)

Camerini, U.; McGibney, D.; Roberts, A.

1982-01-01

The need for a high gain, high sensitivity Cherenkov light sensor to be used in a deep underwater muon and neutrino detector (DUMAND) array has led to the design of the Sea Urchin detector. In this design a spherical photocathode PMTis optically coupled through a glass hemisphere to a large number of glass spines, each of which is filled with a wavelength-shifting (WLS) solution of a high quantum efficiency phosphor. The Cherenkov radiation is absorbed in the spine, isotropically re-radiated at a longer wavelength, and a fraction of the fluorescent light is internally reflected in the spine, and guided to the photomultiplier concentrically located in the glass hemisphere. Experiments measuring the optical characteristics of the spines and computer programs simulating light transformation and detection cross sections are described. Overall optical gains in the range 5-10 are achieved. The WLS solution is inexpensive, and may have other applications. (orig.)

Bio-inspired photonic-crystal microchip for fluorescent ultratrace detection.

Science.gov (United States)

Hou, Jue; Zhang, Huacheng; Yang, Qiang; Li, Mingzhu; Song, Yanlin; Jiang, Lei

2014-06-02

Ultratrace detection attracts great interest because it is still a challenge to the early diagnosis and drug testing. Enriching the targets from highly diluted solutions to the sensitive area is a promising method. Inspired by the fog-collecting structure on Stenocara beetle's back, a photonic-crystal (PC) microchip with hydrophilic-hydrophobic micropattern was fabricated by inkjet printing. This device was used to realize high-sensitive ultratrace detection of fluorescence analytes and fluorophore-based assays. Coupled with the fluorescence enhancement effect of a PC, detection down to 10(-16) mol L(-1) was achieved. This design can be combined with biophotonic devices for the detection of drugs, diseases, and pollutions of the ecosystem. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A facile, sensitive, and highly specific trinitrophenol assay based on target-induced synergetic effects of acid induction and electron transfer towards DNA-templated copper nanoclusters.

Science.gov (United States)

Li, Haiyin; Chang, Jiafu; Hou, Ting; Ge, Lei; Li, Feng

2016-11-01

Reliable, selective and sensitive approaches for trinitrophenol (TNP) detection are highly desirable with respect to national security and environmental protection. Herein, a simple and novel fluorescent strategy for highly sensitive and specific TNP assay has been successfully developed, which is based on the quenching of the fluorescent poly(thymine)-templated copper nanoclusters (DNA-CuNCs), through the synergetic effects of acid induction and electron transfer. Upon the addition of TNP, donor-acceptor complexes between the electron-deficient nitro-groups in TNP and the electron-donating DNA templates are formed, resulting in the close proximity between TNP and CuNCs. Moreover, the acidity of TNP contributes to the pH decrease of the system. These factors combine to dramatically quench the fluorescence of DNA-CuNCs, providing a "signal-off" strategy for TNP sensing. The as-proposed strategy demonstrates high sensitivity for TNP assay, and a detection limit of 0.03μM is obtained, which is lower than those reported by using organic fluorescent materials. More significantly, this approach shows outstanding selectivity over a number of TNP analogues, such as 2,4,6-trinitrotoluene (TNT), 2,4-dinitrotoluene (DNT), 2,4-dinitrophenol (DNP), 3-nitrophenol (NP), nitrobenzene (NB), phenol (BP), and toluene (BT). Compared with previous studies, this method does not need complex DNA sequence design, fluorescent dye labeling, or sophisticated organic reactions, rendering the strategy with additional advantages of simplicity and cost-effectiveness. In addition, the as-proposed strategy has been adopted for the detection of TNP in natural water samples, indicating its great potential to be applied in the fields of public safety and environmental monitoring. Copyright © 2016 Elsevier B.V. All rights reserved.

Experimental design and quality assurance: in situ fluorescence instrumentation

Science.gov (United States)

Conmy, Robyn N.; Del Castillo, Carlos E.; Downing, Bryan D.; Chen, Robert F.

2014-01-01

Both instrument design and capabilities of fluorescence spectroscopy have greatly advanced over the last several decades. Advancements include solid-state excitation sources, integration of fiber optic technology, highly sensitive multichannel detectors, rapid-scan monochromators, sensitive spectral correction techniques, and improve data manipulation software (Christian et al., 1981, Lochmuller and Saavedra, 1986; Cabniss and Shuman, 1987; Lakowicz, 2006; Hudson et al., 2007). The cumulative effect of these improvements have pushed the limits and expanded the application of fluorescence techniques to numerous scientific research fields. One of the more powerful advancements is the ability to obtain in situ fluorescence measurements of natural waters (Moore, 1994). The development of submersible fluorescence instruments has been made possible by component miniaturization and power reduction including advances in light sources technologies (light-emitting diodes, xenon lamps, ultraviolet [UV] lasers) and the compatible integration of new optical instruments with various sampling platforms (Twardowski et at., 2005 and references therein). The development of robust field sensors skirt the need for cumbersome and or time-consuming filtration techniques, the potential artifacts associated with sample storage, and coarse sampling designs by increasing spatiotemporal resolution (Chen, 1999; Robinson and Glenn, 1999). The ability to obtain rapid, high-quality, highly sensitive measurements over steep gradients has revolutionized investigations of dissolved organic matter (DOM) optical properties, thereby enabling researchers to address novel biogeochemical questions regarding colored or chromophoric DOM (CDOM). This chapter is dedicated to the origin, design, calibration, and use of in situ field fluorometers. It will serve as a review of considerations to be accounted for during the operation of fluorescence field sensors and call attention to areas of concern when making

A novel pH-sensitive polymeric fluorescent probe: Synthesis, characterization and optical properties

Energy Technology Data Exchange (ETDEWEB)

Chen Dongyun; Xu Qingfeng; Xia Xuewei; Ge Jianfeng [Key Laboratory of Organic Synthesis of Jiangsu Province, College of Chemistry, Chemical Engineering and Material Science, Soochow University, Suzhou, Jiangsu, 215123 (China); Lu Jianmei, E-mail: lujm@suda.edu.cn [Key Laboratory of Organic Synthesis of Jiangsu Province, College of Chemistry, Chemical Engineering and Material Science, Soochow University, Suzhou, Jiangsu, 215123 (China); Li Najun, E-mail: linajun@sina.com [Key Laboratory of Organic Synthesis of Jiangsu Province, College of Chemistry, Chemical Engineering and Material Science, Soochow University, Suzhou, Jiangsu, 215123 (China)

2010-04-15

A novel initiator containing proflavine derivative moiety, 3,6-dibromo-isobutyramide acridine (DIA), was synthesized and initiated the atom transfer radical polymerization (ATRP) of methyl methacrylate (MMA). A water-soluble monomer, N,N-dimethylacrylamide (DMAA) was also initiated by DIA for comparison. The obtained fluorescent polymers, PMMA-DIA and PDMAA-DIA, were characterized by {sup 1}H NMR, GPC and TGA. The emission spectra of the fluorescent polymers exhibit obvious changes in color and fluorescence intensity along with pH varied in range of 3.0-9.0. In addition, the obtained polymers present good film-forming capacity and the films also have a high quantum yield and pH response. Both oil-soluble PMMA-DIA and water-soluble PDMAA-DIA have steady optical and chemical properties by containing proflavine moiety in the main chain.

Green synthesis of fluorescence carbon nanoparticles from yum and application in sensitive and selective detection of ATP.

Science.gov (United States)

Zhan, Zixuan; Cai, Jiao; Wang, Qi; Su, Yingying; Zhang, Lichun; Lv, Yi

2016-05-01

Fluorescent carbon nanoparticles (CPs), a fascinating class of recently discovered nanocarbons, have been widely known as some of the most promising sensing probes in biological or chemical analysis. In this study, we demonstrate a green synthetic methodology for generating water-soluble CPs with a quantum yield of approximately 24% via a simple heating process using yum mucilage as a carbon source. The prepared carbon nanoparticles with an ~10 nm size possessed excellent fluorescence properties, and the fluorescence of the CPs was strongly quenched by Fe(3+), and recovered by adenosine triphosphate (ATP), thus, an 'off' and 'on' system can be easily established. This 'CPs-Fe(3+)-ATP' strategy was sensitive and selective at detecting ATP with the linear range of 0.5 µmol L(-1) to 50 µmol L(-1) and with a detection limit of 0.48 µmol L(-1). Copyright © 2015 John Wiley & Sons, Ltd.

Design of remote laser-induced fluorescence system's acquisition circuit

Science.gov (United States)

Wang, Guoqing; Lou, Yue; Wang, Ran; Yan, Debao; Li, Xin; Zhao, Xin; Chen, Dong; Zhao, Qi

2017-10-01

Laser-induced fluorescence system(LIfS) has been found its significant application in identifying one kind of substance from another by its properties even it's thimbleful, and becomes useful in plenty of fields. Many superior works have reported LIfS' theoretical analysis , designs and uses. However, the usual LIPS is always constructed in labs to detect matter quite closely, for the system using low-power laser as excitation source and charge coupled device (CCD) as detector. Promoting the detectivity of LIfS is of much concern to spread its application. Here, we take a high-energy narrow-pulse laser instead of commonly used continuous wave laser to operate sample, thus we can get strong fluorescent. Besides, photomultiplier (PMT) with high sensitivity is adopted in our system to detect extremely weak fluorescence after a long flight time from the sample to the detector. Another advantage in our system, as the fluorescence collected into spectroscopy, multiple wavelengths of light can be converted to the corresponding electrical signals with the linear array multichannel PMT. Therefore, at the cost of high-powered incentive and high-sensitive detector, a remote LIFS is get. In order to run this system, it is of importance to turn light signal to digital signal which can be processed by computer. The pulse width of fluorescence is deeply associated with excitation laser, at the nanosecond(ns) level, which has a high demand for acquisition circuit. We design an acquisition circuit including, I/V conversion circuit, amplifying circuit and peak-holding circuit. The simulation of circuit shows that peak-holding circuit can be one effective approach to reducing difficulty of acquisition circuit.

Dual time-resolved temperature-jump fluorescence and infrared spectroscopy for the study of fast protein dynamics.

Science.gov (United States)

Davis, Caitlin M; Reddish, Michael J; Dyer, R Brian

2017-05-05

Time-resolved temperature-jump (T-jump) coupled with fluorescence and infrared (IR) spectroscopy is a powerful technique for monitoring protein dynamics. Although IR spectroscopy of the polypeptide amide I mode is more technically challenging, it offers complementary information because it directly probes changes in the protein backbone, whereas, fluorescence spectroscopy is sensitive to the environment of specific side chains. With the advent of widely tunable quantum cascade lasers (QCL) it is possible to efficiently probe multiple IR frequencies with high sensitivity and reproducibility. Here we describe a dual time-resolved T-jump fluorescence and IR spectrometer and its application to study protein folding dynamics. A Q-switched Ho:YAG laser provides the T-jump source for both time-resolved IR and fluorescence spectroscopy, which are probed by a QCL and Ti:Sapphire laser, respectively. The Ho:YAG laser simultaneously pumps the time-resolved IR and fluorescence spectrometers. The instrument has high sensitivity, with an IR absorbance detection limit of jump induced difference spectrum from 50ns to 0.5ms. This study demonstrates the power of the dual time-resolved T-jump fluorescence and IR spectroscopy to resolve complex folding mechanisms by complementary IR absorbance and fluorescence measurements of protein dynamics. Copyright © 2017 Elsevier B.V. All rights reserved.

Quantitative high dynamic range beam profiling for fluorescence microscopy

International Nuclear Information System (INIS)

Mitchell, T. J.; Saunter, C. D.; O’Nions, W.; Girkin, J. M.; Love, G. D.

2014-01-01

Modern developmental biology relies on optically sectioning fluorescence microscope techniques to produce non-destructive in vivo images of developing specimens at high resolution in three dimensions. As optimal performance of these techniques is reliant on the three-dimensional (3D) intensity profile of the illumination employed, the ability to directly record and analyze these profiles is of great use to the fluorescence microscopist or instrument builder. Though excitation beam profiles can be measured indirectly using a sample of fluorescent beads and recording the emission along the microscope detection path, we demonstrate an alternative approach where a miniature camera sensor is used directly within the illumination beam. Measurements taken using our approach are solely concerned with the illumination optics as the detection optics are not involved. We present a miniature beam profiling device and high dynamic range flux reconstruction algorithm that together are capable of accurately reproducing quantitative 3D flux maps over a large focal volume. Performance of this beam profiling system is verified within an optical test bench and demonstrated for fluorescence microscopy by profiling the low NA illumination beam of a single plane illumination microscope. The generality and success of this approach showcases a widely flexible beam amplitude diagnostic tool for use within the life sciences

CMOS Time-Resolved, Contact, and Multispectral Fluorescence Imaging for DNA Molecular Diagnostics

Directory of Open Access Journals (Sweden)

Nan Guo

2014-10-01

Full Text Available Instrumental limitations such as bulkiness and high cost prevent the fluorescence technique from becoming ubiquitous for point-of-care deoxyribonucleic acid (DNA detection and other in-field molecular diagnostics applications. The complimentary metal-oxide-semiconductor (CMOS technology, as benefited from process scaling, provides several advanced capabilities such as high integration density, high-resolution signal processing, and low power consumption, enabling sensitive, integrated, and low-cost fluorescence analytical platforms. In this paper, CMOS time-resolved, contact, and multispectral imaging are reviewed. Recently reported CMOS fluorescence analysis microsystem prototypes are surveyed to highlight the present state of the art.

An image fiber based fluorescent probe with associated signal processing scheme for biomedical diagnostics

International Nuclear Information System (INIS)

Vaishakh, M; Murukeshan, V M; Seah, L K

2008-01-01

A dual-modality image fiber based fluorescent probe that can be used for depth sensitive imaging and suppression of fluorescent emissions with nanosecond lifetime difference is proposed and illustrated in this paper. The system can give high optical sectioning and employs an algorithm for obtaining phase sensitive images. The system can find main application in in vivo biomedical diagnostics for detecting biochemical changes for distinguishing malignant tissue from healthy tissue

Listening to membrane potential: photoacoustic voltage-sensitive dye recording

Science.gov (United States)

Zhang, Haichong K.; Yan, Ping; Kang, Jeeun; Abou, Diane S.; Le, Hanh N. D.; Jha, Abhinav K.; Thorek, Daniel L. J.; Kang, Jin U.; Rahmim, Arman; Wong, Dean F.; Boctor, Emad M.; Loew, Leslie M.

2017-04-01

Voltage-sensitive dyes (VSDs) are designed to monitor membrane potential by detecting fluorescence changes in response to neuronal or muscle electrical activity. However, fluorescence imaging is limited by depth of penetration and high scattering losses, which leads to low sensitivity in vivo systems for external detection. By contrast, photoacoustic (PA) imaging, an emerging modality, is capable of deep tissue, noninvasive imaging by combining near-infrared light excitation and ultrasound detection. Here, we show that voltage-dependent quenching of dye fluorescence leads to a reciprocal enhancement of PA intensity. We synthesized a near-infrared photoacoustic VSD (PA-VSD), whose PA intensity change is sensitive to membrane potential. In the polarized state, this cyanine-based probe enhances PA intensity while decreasing fluorescence output in a lipid vesicle membrane model. A theoretical model accounts for how the experimental PA intensity change depends on fluorescence and absorbance properties of the dye. These results not only demonstrate PA voltage sensing but also emphasize the interplay of both fluorescence and absorbance properties in the design of optimized PA probes. Together, our results demonstrate PA sensing as a potential new modality for recording and external imaging of electrophysiological and neurochemical events in the brain.

Phase-sensitive flow cytometer

Energy Technology Data Exchange (ETDEWEB)

Steinkamp, J.A.

1992-12-31

This report describes phase-sensitive flow cytometer (FCM) which provides additional FCM capability to use the fluorescence lifetime of one or more fluorochromes bound to single cells to provide additional information regarding the cells. The resulting fluorescence emission can be resolved into individual fluorescence signals if two fluorochromes are present or can be converted directly to a decay lifetime from a single fluorochrome. The excitation light for the fluorochromes is modulated to produce an amplitude modulated fluorescence pulse as the fluorochrome is excited in the FCM. The modulation signal also forms a reference signal that is phase-shifted a selected amount for subsequent mixing with the output modulated fluorescence intensity signal in phase-sensitive detection circuitry. The output from the phase-sensitive circuitry is then an individual resolved fluorochrome signal or a single fluorochrome decay lifetime, depending on the applied phase shifts.

Highly sensitive and quantitative FRET-FLIM imaging in single dendritic spines using improved non-radiative YFP.

Science.gov (United States)

Murakoshi, Hideji; Lee, Seok-Jin; Yasuda, Ryohei

2008-08-01

Two-photon fluorescence lifetime imaging microscopy (TPFLIM) enables the quantitative measurements of fluorescence resonance energy transfer (FRET) in small subcellular compartments in light scattering tissue. We evaluated and optimized the FRET pair of mEGFP (monomeric EGFP with the A206K mutation) and REACh (non-radiative YFP variants) for TPFLIM. We characterized several mutants of REACh in terms of their "darkness," and their ability to act as a FRET acceptor for mEGFP in HeLa cells and hippocampal neurons. Since the commonly used monomeric mutation A206K increases the brightness of REACh, we introduced a different monomeric mutation (F223R) which does not affect the brightness. Also, we found that the folding efficiency of original REACh, as measured by the fluorescence lifetime of a mEGFP-REACh tandem dimer, was low and variable from cell to cell. Introducing two folding mutations (F46L, Q69M) into REACh increased the folding efficiency by approximately 50%, and reduced the variability of FRET signal. Pairing mEGFP with the new REACh (super-REACh, or sREACh) improved the signal-to-noise ratio compared to the mEGFP-mRFP or mEGFP-original REACh pair by approximately 50%. Using this new pair, we demonstrated that the fraction of actin monomers in filamentous and globular forms in single dendritic spines can be quantitatively measured with high sensitivity. Thus, the mEGFP-sREACh pair is suited for quantitative FRET measurement by TPFLIM, and enables us to measure protein-protein interactions in individual dendritic spines in brain slices with high sensitivity.

Target-induced structure switching of hairpin aptamers for label-free and sensitive fluorescent detection of ATP via exonuclease-catalyzed target recycling amplification.

Science.gov (United States)

Xu, Yunying; Xu, Jin; Xiang, Yun; Yuan, Ruo; Chai, Yaqin

2014-01-15

In this work, we described the development of a new label-free, simple and sensitive fluorescent ATP sensing platform based on exonuclease III (Exo III)-catalyzed target recycling (ECTR) amplification and SYBR Green I indicator. The hairpin aptamer probes underwent conformational structure switching and re-configuration in the presence of ATP, which led to catalytic cleavage of the re-configured aptamers by Exo III to release ATP and to initiate the ECTR process. Such ECTR process resulted in the digestion of a significant number of the hairpin aptamer probes, leading to much less intercalation of SYBR Green I to the hairpin stems and drastic suppression of the fluorescence emission for sensitive ATP detection down to the low nanomolar level. Due to the highly specific affinity bindings between aptamers and ATP, the developed method exhibited excellent selectivity toward ATP against other analogous molecules. Besides, our ATP sensing approach used un-modified aptamer probes and could be performed in a "mix-and-detect" fashion in homogenous solutions. All these distinct advantages of the developed method thus made it hold great potential for the development of simple and robust sensing strategies for the detection of other small molecules. © 2013 Elsevier B.V. All rights reserved.

Quantification of tumor fluorescence during intraoperative optical cancer imaging.

Science.gov (United States)

Judy, Ryan P; Keating, Jane J; DeJesus, Elizabeth M; Jiang, Jack X; Okusanya, Olugbenga T; Nie, Shuming; Holt, David E; Arlauckas, Sean P; Low, Phillip S; Delikatny, E James; Singhal, Sunil

2015-11-13

Intraoperative optical cancer imaging is an emerging technology in which surgeons employ fluorophores to visualize tumors, identify tumor-positive margins and lymph nodes containing metastases. This study compares instrumentation to measure tumor fluorescence. Three imaging systems (Spectropen, Glomax, Flocam) measured and quantified fluorescent signal-to-background ratios (SBR) in vitro, murine xenografts, tissue phantoms and clinically. Evaluation criteria included the detection of small changes in fluorescence, sensitivity of signal detection at increasing depths and practicality of use. In vitro, spectroscopy was superior in detecting incremental differences in fluorescence than luminescence and digital imaging (Ln[SBR] = 6.8 ± 0.6, 2.4 ± 0.3, 2.6 ± 0.1, p = 0.0001). In fluorescent tumor cells, digital imaging measured higher SBRs than luminescence (6.1 ± 0.2 vs. 4.3 ± 0.4, p = 0.001). Spectroscopy was more sensitive than luminometry and digital imaging in identifying murine tumor fluorescence (SBR = 41.7 ± 11.5, 5.1 ± 1.8, 4.1 ± 0.9, p = 0.0001), and more sensitive than digital imaging at detecting fluorescence at increasing depths (SBR = 7.0 ± 3.4 vs. 2.4 ± 0.5, p = 0.03). Lastly, digital imaging was the most practical and least time-consuming. All methods detected incremental differences in fluorescence. Spectroscopy was the most sensitive for small changes in fluorescence. Digital imaging was the most practical considering its wide field of view, background noise filtering capability, and sensitivity to increasing depth.

Glycine Insertion Makes Yellow Fluorescent Protein Sensitive to Hydrostatic Pressure

Science.gov (United States)

Watanabe, Tomonobu M.; Imada, Katsumi; Yoshizawa, Keiko; Nishiyama, Masayoshi; Kato, Chiaki; Abe, Fumiyoshi; Morikawa, Takamitsu J.; Kinoshita, Miki; Fujita, Hideaki; Yanagida, Toshio

2013-01-01

Fluorescent protein-based indicators for intracellular environment conditions such as pH and ion concentrations are commonly used to study the status and dynamics of living cells. Despite being an important factor in many biological processes, the development of an indicator for the physicochemical state of water, such as pressure, viscosity and temperature, however, has been neglected. We here found a novel mutation that dramatically enhances the pressure dependency of the yellow fluorescent protein (YFP) by inserting several glycines into it. The crystal structure of the mutant showed that the tyrosine near the chromophore flipped toward the outside of the β-can structure, resulting in the entry of a few water molecules near the chromophore. In response to changes in hydrostatic pressure, a spectrum shift and an intensity change of the fluorescence were observed. By measuring the fluorescence of the YFP mutant, we succeeded in measuring the intracellular pressure change in living cell. This study shows a new strategy of design to engineer fluorescent protein indicators to sense hydrostatic pressure. PMID:24014139

Sensitive detection of mercury and copper ions by fluorescent DNA/Ag nanoclusters in guanine-rich DNA hybridization.

Science.gov (United States)

Peng, Jun; Ling, Jian; Zhang, Xiu-Qing; Bai, Hui-Ping; Zheng, Liyan; Cao, Qiu-E; Ding, Zhong-Tao

2015-02-25

In this work, we designed a new fluorescent oligonucleotides-stabilized silver nanoclusters (DNA/AgNCs) probe for sensitive detection of mercury and copper ions. This probe contains two tailored DNA sequence. One is a signal probe contains a cytosine-rich sequence template for AgNCs synthesis and link sequence at both ends. The other is a guanine-rich sequence for signal enhancement and link sequence complementary to the link sequence of the signal probe. After hybridization, the fluorescence of hybridized double-strand DNA/AgNCs is 200-fold enhanced based on the fluorescence enhancement effect of DNA/AgNCs in proximity of guanine-rich DNA sequence. The double-strand DNA/AgNCs probe is brighter and stable than that of single-strand DNA/AgNCs, and more importantly, can be used as novel fluorescent probes for detecting mercury and copper ions. Mercury and copper ions in the range of 6.0-160.0 and 6-240 nM, can be linearly detected with the detection limits of 2.1 and 3.4 nM, respectively. Our results indicated that the analytical parameters of the method for mercury and copper ions detection are much better than which using a single-strand DNA/AgNCs. Copyright © 2014 Elsevier B.V. All rights reserved.

Highly selective and reversible chemosensor for Pd(2+) detected by fluorescence, colorimetry, and test paper.

Science.gov (United States)

Wang, Mian; Liu, Xiaomei; Lu, Huizhe; Wang, Hongmei; Qin, Zhaohai

2015-01-21

A "turn-on" fluorescent and colorimetric chemosensor (RBS) for Pd(2+) has been designed and synthesized through introduction of sulfur as a ligand atom to Rhodamine B. RBS exhibits high selectivity (freedom from the interference of Hg(2+ )in particular) and sensitivity toward Pd(2+) with a detection limit as low as 2.4 nM. RBS is also a reversible sensor, and it can be made into test paper to detect Pd(2+) in pure water. Compared to the chemosensors that introduced phosphorus to Rhodamine to detect Pd(2+), RBS can be synthesized more simply and economically.

Highly stable lipid-encapsulation of fluorescent nanodiamonds for bioimaging applications.

Science.gov (United States)

Sotoma, Shingo; Hsieh, Feng-Jen; Chen, Yen-Wei; Tsai, Pei-Chang; Chang, Huan-Cheng

2018-01-23

Highly stable lipid-encapsulated fluorescent nanodiamonds (FNDs) are produced by photo-crosslinking of diacetylene-containing lipids physically attached to the FND surface. Not only is this coating method simple and fast, but also it gives the FND-lipid hybrids favorable properties for bioapplications. The hybrids are useful as fluorescent biolabels as well as fiducial markers for correlative light and electron microscopy.

Simulating fluorescence light-canopy interaction in support of laser-induced fluorescence measurements

International Nuclear Information System (INIS)

Rosema, A.; Verhoef, W.; Schroote, J.; Snel, J.F.H.

1991-01-01

In the Netherlands an operational field instrument for the measurement of laser induced fluorescence of vegetation (LEAF) is developed. In addition, plant physiological and remote sensing research is done to support this new remote sensing instrument. This paper presents a general introduction on the subject of laser-induced fluorescence, including the relation between chlorophyll fluorescence and photosynthesis, spectral characteristics, and previous research. Also the LEAF system is briefly described. Subsequently, the development of a leaf fluorescence model (KMF) and a canopy fluorescence model (FLSAIL) are reported. With these simulation models a sensitivity study is carried out. Fluorescence of 685 nm appears to be most suitable to obtain information on photosynthesis and stress, but is also influenced by canopy structure. Separation of these two effects is studied

Volume labeling with Alexa Fluor dyes and surface functionalization of highly sensitive fluorescent silica (SiO2) nanoparticles

Science.gov (United States)

Wang, Wei; Nallathamby, Prakash D.; Foster, Carmen M.; Morrell-Falvey, Jennifer L.; Mortensen, Ninell P.; Doktycz, Mitchel J.; Gu, Baohua; Retterer, Scott T.

2013-10-01

A new synthesis approach is described that allows the direct incorporation of fluorescent labels into the volume or body of SiO2 nanoparticles. In this process, fluorescent Alexa Fluor dyes with different emission wavelengths were covalently incorporated into the SiO2 nanoparticles during their formation by the hydrolysis of tetraethoxysilane. The dye molecules were homogeneously distributed throughout the SiO2 nanoparticles. The quantum yields of the Alexa Fluor volume-labeled SiO2 nanoparticles were much higher than nanoparticles labeled using conventional organic dyes. The size of the resulting nanoparticles was controlled using microemulsion reaction media with sizes in the range of 20-100 nm and a polydispersity of cultured macrophages. Differences in particle agglomeration and cell association were clearly associated with differences in observed nanoparticle toxicity. The capacity to maintain particle fluorescence while making significant changes to surface chemistry makes these particles extremely versatile and useful for studies of particle agglomeration, uptake, and transport in environmental and biological systems.A new synthesis approach is described that allows the direct incorporation of fluorescent labels into the volume or body of SiO2 nanoparticles. In this process, fluorescent Alexa Fluor dyes with different emission wavelengths were covalently incorporated into the SiO2 nanoparticles during their formation by the hydrolysis of tetraethoxysilane. The dye molecules were homogeneously distributed throughout the SiO2 nanoparticles. The quantum yields of the Alexa Fluor volume-labeled SiO2 nanoparticles were much higher than nanoparticles labeled using conventional organic dyes. The size of the resulting nanoparticles was controlled using microemulsion reaction media with sizes in the range of 20-100 nm and a polydispersity of cultured macrophages. Differences in particle agglomeration and cell association were clearly associated with differences in

Microwave-assisted synthesis of highly luminescent N- and S-co-doped carbon dots as a ratiometric fluorescent probe for levofloxacin.

Science.gov (United States)

Li, Huiyu; Xu, Yuan; Ding, Jie; Zhao, Li; Zhou, Tianyu; Ding, Hong; Chen, Yanhua; Ding, Lan

2018-01-10

Uniform N- and S-co-doped carbon dots (NSCDs) with fluorescence quantum yields of up to 64% were synthesized via a one-step microwave-assisted method. Ammonium citrate and L-cysteine act as precursors, and synthesis is completed in 2.5 min using a 750 W microwave oven to give a 62% yield. The NSCDs show bright blue fluorescence (with excitation/emission peaks at 353/426 nm) and have narrow size distribution. On exposure to levofloxacin (LEV), the emission maximum shifts to 499 nm. This effect was used to design ratiometric (2-wavelength) assays for LEV. The fluorometric method (based on measurement of the fluorescence intensity ratio at 499 and 426 nm) has a detection limit of 5.1 μg·L -1 (3σ/k) and a linear range that extends from 0.01 to 70 mg·L -1 . The method was applied to the determination of LEV in three kinds of spiked water samples and has recoveries in the range from 98.6 to 106.8%. The fluorescent probe described here is highly selective and sensitive. Graphical Abstract Highly luminescent N- and S-co-doped carbon dots were synthesized using AC (ammonium citrate) and Cys (L-cysteine) by microwave-assisted method, and were applied to the visual and ratiometric fluorescence determination of LEV (levofloxacin).

Fluorescence-labeled methylation-sensitive amplified fragment length polymorphism (FL-MS-AFLP) analysis for quantitative determination of DNA methylation and demethylation status.

Science.gov (United States)

Kageyama, Shinji; Shinmura, Kazuya; Yamamoto, Hiroko; Goto, Masanori; Suzuki, Koichi; Tanioka, Fumihiko; Tsuneyoshi, Toshihiro; Sugimura, Haruhiko

2008-04-01

The PCR-based DNA fingerprinting method called the methylation-sensitive amplified fragment length polymorphism (MS-AFLP) analysis is used for genome-wide scanning of methylation status. In this study, we developed a method of fluorescence-labeled MS-AFLP (FL-MS-AFLP) analysis by applying a fluorescence-labeled primer and fluorescence-detecting electrophoresis apparatus to the existing method of MS-AFLP analysis. The FL-MS-AFLP analysis enables quantitative evaluation of more than 350 random CpG loci per run. It was shown to allow evaluation of the differences in methylation level of blood DNA of gastric cancer patients and evaluation of hypermethylation and hypomethylation in DNA from gastric cancer tissue in comparison with adjacent non-cancerous tissue.

Fluorescence detection of a protein-bound 2Fe2S cluster.

Science.gov (United States)

Hoff, Kevin G; Goodlitt, Rochelle; Li, Rui; Smolke, Christina D; Silberg, Jonathan J

2009-03-02

A fluorescent biosensor is described for 2Fe2S clusters that is composed of green fluorescent protein (GFP) fused to glutaredoxin 2 (Grx2), as illustrated here. 2Fe2S detection is based on the reduction of GFP fluorescence upon the 2Fe2S-induced dimerization of GFP-Grx2. This assay is sufficiently sensitive to detect submicromolar changes in 2Fe2S levels, thus making it suitable for high-throughput measurements of metallocluster degradation and synthesis reactions.

A new fluorescent pH probe for extremely acidic conditions

Energy Technology Data Exchange (ETDEWEB)

Xu, Yu [School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100 (China); Taishan College, Shandong University, Jinan 250100 (China); Jiang, Zheng [School of Life Science, Shandong University, Jinan 250100 (China); Taishan College, Shandong University, Jinan 250100 (China); Xiao, Yu [School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100 (China); Taishan College, Shandong University, Jinan 250100 (China); Bi, Fu-Zhen [School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100 (China); Miao, Jun-Ying, E-mail: miaojy@sdu.edu.cn [School of Life Science, Shandong University, Jinan 250100 (China); Zhao, Bao-Xiang, E-mail: bxzhao@sdu.edu.cn [School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100 (China)

2014-04-01

A new coumarin-based fluorescent probe can detect highly acidic conditions in both solution and bacteria with high selectivity and sensitivity. Highlights: • A new fluorescence probe for very low pH was synthesized and characterized. • The probe can monitor pH in solution and bacteria. • The two-step protonation of N atoms of the probe leads to fluorescence quenching. Abstract: A novel turn-off fluorescent probe based on coumarin and imidazole moiety for extremely acidic conditions was designed and developed. The probe with pK_a = 2.1 is able to respond to very low pH value (below 3.5) with high sensitivity relying on fluorescence quenching at 460 nm in fluorescence spectra or the ratios of absorbance maximum at 380 nm to that at 450 nm in UV–vis spectra. It can quantitatively detect pH value based on equilibrium equation, pH = pK_a -log[(I_x - I_b)/(I_a - I_x)]. It had very short response time that was less than 1 min, good reversibility and nearly no interference from common metal ions. Moreover, using ¹H NMR analysis and theoretical calculation of molecular orbital, we verified that a two-step protonation process of two N atoms of the probe leaded to photoinduced electron transfer (PET), which was actually the mechanism of the fluorescence quenching phenomenon under strongly acidic conditions. Furthermore, the probe was also applied to imaging strong acidity in bacteria, E.coli and had good effect. This work illustrates that the new probe could be a practical and ideal pH indicator for strongly acidic conditions with good biological significance.

Quantum dots versus organic fluorophores in fluorescent deep-tissue imaging--merits and demerits.

Science.gov (United States)

Bakalova, Rumiana; Zhelev, Zhivko; Gadjeva, Veselina

2008-12-01

The use of fluorescence in deep-tissue imaging is rapidly expanding in last several years. The progress in fluorescent molecular probes and fluorescent imaging techniques gives an opportunity to detect single cells and even molecular targets in live organisms. The highly sensitive and high-speed fluorescent molecular sensors and detection devices allow the application of fluorescence in functional imaging. With the development of novel bright fluorophores based on nanotechnologies and 3D fluorescence scanners with high spatial and temporal resolution, the fluorescent imaging has a potential to become an alternative of the other non-invasive imaging techniques as magnetic resonance imaging, positron-emission tomography, X-ray, computing tomography. The fluorescent imaging has also a potential to give a real map of human anatomy and physiology. The current review outlines the advantages of fluorescent nanoparticles over conventional organic dyes in deep-tissue imaging in vivo and defines the major requirements to the "perfect fluorophore". The analysis proceeds from the basic principles of fluorescence and major characteristics of fluorophores, light-tissue interactions, and major limitations of fluorescent deep-tissue imaging. The article is addressed to a broad readership - from specialists in this field to university students.

Herbicide impact on Hormosira banksii gametes measured by fluorescence and germination bioassays

International Nuclear Information System (INIS)

Seery, Cliff R.; Gunthorpe, Leanne; Ralph, Peter J.

2006-01-01

The innovative bioassay described here involves chlorophyll a fluorescence measurements of gametes from the macroalgae, Hormosira banksii, where gametes (eggs) were exposed to Diuron, Irgarol and Bromacil. Response was assessed as percent inhibition from control of effective quantum yield (ΔF/Fm') of photosystem II, herein referred to as % PSII Inhibition. This was measured with the dual-channelled pulse amplitude modulated (PAM) fluorometer, ToxY-PAM. The fluorescence bioassay was run simultaneously with an established H. banksii germination bioassay to compare sensitivity, precision, and time-to-result. The fluorescence bioassay gave highly sensitive results evidenced by EC 5 s (% PSII Inhibition) for Diuron, Irgarol and Bromacil being three, four and three orders of magnitude (respectively) lower than EC 5 s generated from the germination bioassays. Precision of the fluorescence bioassay was demonstrated with low coefficient of variations (<30%) for all three toxicants. With regard to time, the fluorescence bioassay gave results within 6 h, as opposed to more than 50 h for the germination bioassay. - Chlorophyll a fluorescence measurements form the basis of a macroalgal bioassay with many advantages over germination-based methods

A simple and sensitive fluorescent probe for specific detection of ...

Indian Academy of Sciences (India)

Yan-Fei Kang

A fluorescent probe, with simplicity of structure and convenience of synthesis, is capable of detecting ... Yan-Fei Kang et al. .... Pastore A, Federici G, Bertini E and Ptemonte F 2003 ... Urano Y 2010 New Strategies for Fluorescent Probe.

Controllable synthesis and characterization of highly fluorescent silver nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Li Junlin [Nanjing Normal University, School of Chemistry and Materials Science (China); An Xueqing, E-mail: anxueqin@ecust.edu.cn [East China University of Science and Technology, School of Chemistry and Molecular Engineering (China); Zhu Yinyan [Nanjing Normal University, School of Chemistry and Materials Science (China)

2012-12-15

Highly fluorescent silver nanoparticles (AgFNPs) have been prepared by microemulsion method and the sizes of AgFNPs were controlled by altering the molar ratio ({omega}) of water-to-surfactant in the water-in-oil microemulsion. The results were shown that the AgFNPs sizes increased with incremental molar ratio ({omega}) of water-to-surfactant. The AgFNPs have been characterized by transmission electron microscopy, dynamic light scattering, fluorescence and absorption spectroscopy, and fluorescence lifetime study. Study of the spectral characteristics was shown that the absorbance of AgFNPs increased significantly with the {omega}, and linear relationship between absorbance and the size of AgFNPs was observed. The increase of AgFNPs size caused a red shift of maximum absorption wavelength in the UV-Vis spectra, and the relationship between maximum absorption wavelength and AgFNPs size appeared linear dependence. The maximum fluorescence emission wavelength did not shift with the change of particles size, but the emission intensity increases with the {omega}. The results were shown that the other factors to affect the fluorescence properties of AgFNPs were the surface properties and microstructure, except the AgFNPs size. These surface properties depend upon the stabilizing agent, reactant concentration, and solvents and so on.

A selective and sensitive optical sensor for dissolved ammonia detection via agglomeration of fluorescent Ag nanoclusters and temperature gradient headspace single drop microextraction.

Science.gov (United States)

Dong, Jiang Xue; Gao, Zhong Feng; Zhang, Ying; Li, Bang Lin; Li, Nian Bing; Luo, Hong Qun

2017-05-15

In this paper, a simple sensor platform is presented for highly selective and sensitive detection of dissolved ammonia in aqueous solutions without pretreatment based on temperature gradient headspace single drop microextraction (HS-SDME) technique, and fluorescence and UV-vis spectrophotometry are utilized with the Ag nanoclusters (Ag NCs) functioned by citrate and glutathione as the probe. The sensing mechanism is based on the volatility of ammonia gas and the active response of Ag NCs to pH change caused by the introduction of ammonia. High pH can make the Ag NCs agglomerate and lead to the obvious decrease of fluorescence intensity and absorbance of Ag NCs solution. Moreover, the presented method exhibits a remarkably high selectivity toward dissolved ammonia over most of inorganic ions and amino acid, and shows a good linear range of 10-350μM (0.14-4.9mgNL -1 ) with a low detection limit of 336nM (4.70μgNL -1 ) at a signal-to-noise ratio of 3. In addition, the practical applications of the sensor have been successfully demonstrated by detecting dissolved ammonia in real samples. Copyright © 2016 Elsevier B.V. All rights reserved.

Fluorescence spectra of Rhodamine 6G for high fluence excitation laser radiation

CERN Document Server

Hung, J; Olaizola, A M

2003-01-01

Fluorescence spectral changes of Rhodamine 6G in ethanol and glycerol solutions and deposited as a film on a silica surface have been studied using a wide range of pumping field fluence at 532 nm at room temperature. Blue shift of the fluorescence spectra and fluorescence quenching of the dye molecule in solution are observed at high excitation fluence values. Such effects are not reported for the film sample. The effects are interpreted as the result of population redistribution in the solute-solvent molecular system induced by the high fluence field and the fluence dependence of the radiationless decay mechanism.

Development of fluorescent methods for DNA methyltransferase assay

Science.gov (United States)

Li, Yueying; Zou, Xiaoran; Ma, Fei; Tang, Bo; Zhang, Chun-yang

2017-03-01

DNA methylation modified by DNA methyltransferase (MTase) plays an important role in regulating gene transcription, cell growth and proliferation. The aberrant DNA MTase activity may lead to a variety of human diseases including cancers. Therefore, accurate and sensitive detection of DNA MTase activity is crucial to biomedical research, clinical diagnostics and therapy. However, conventional DNA MTase assays often suffer from labor-intensive operations and time-consuming procedures. Alternatively, fluorescent methods have significant advantages of simplicity and high sensitivity, and have been widely applied for DNA MTase assay. In this review, we summarize the recent advances in the development of fluorescent methods for DNA MTase assay. These emerging methods include amplification-free and the amplification-assisted assays. Moreover, we discuss the challenges and future directions of this area.

Portable X-ray fluorescence analyzer of high sensitivity using X-ray tube excitation

International Nuclear Information System (INIS)

Vatai, E.; Ando, L.

1982-01-01

A review of the three main methods of X-ray fluorescence analysis and their problems is given. The attainable accuracy and effectiveness of each method are discussed. The main properties of portable X-ray analyzers required by the industry are described. The results and experiences of R and D activities in ATOMKI (Debrecen, Hungary) for developing portable X-ray analyzers are presented. The only way for increasing the accuracy and decreasing the measuring time is the application of X-ray tube excitation instead of radioactive sources. The new ATOMKI equipment presently under construction and patenting uses X-ray tube excitation; it will increase the accuracy of concentration determination by one order of magnitude. (D.Gy.)

A new spectrometer for grazing incidence X-ray fluorescence for the characterization of Arsenic implants and Hf based high-k layers

International Nuclear Information System (INIS)

Ingerle, D.; Meirer, F.; Zoeger, N.; Pepponi, G.; Giubertoni, D.; Steinhauser, G.; Wobrauschek, P.; Streli, C.

2010-01-01

Grazing Incidence X-ray Fluorescence Analysis (GIXRF) is a powerful technique for depth-profiling and characterization of thin layers in depths up to a few hundred nanometers. By measurement of fluorescence signals at various incidence angles Grazing Incidence X-ray Fluorescence Analysis provides information on depth distribution and total dose of the elements in the layers. The technique is very sensitive even in depths of a few nanometers. As Grazing Incidence X-ray Fluorescence Analysis does not provide unambigous depth profile information and needs a realistic input depth profile for fitting, in the context of the EC funded European Integrated Activity of Excellence and Networking for Nano and Micro-Electronics Analysis (ANNA) Grazing Incidence X-ray Fluorescence Analysis is used as a complementary technique to Secondary Ion Mass Spectrometry (SIMS) for the characterization of Ultra Shallow Junctions (USJ). A measuring chamber was designed, constructed and tested to meet the requirements of Grazing Incidence X-ray Fluorescence Analysis. A measurement protocol was developed and tested. Some results for As implants as well as Hf based high k layers on Silicon are shown. For the determination of the bulk As content of the wafers, Instrumental Neutron Activation Analysis has also been applied for comparison.

Fluorescent chemosensor based on urea/thiourea moiety for sensing of Hg(II) ions in an aqueous medium with high sensitivity and selectivity: A comparative account on effect of molecular architecture on chemosensing

Science.gov (United States)

Mishra, Jayanti; Kaur, Harpreet; Ganguli, Ashok K.; Kaur, Navneet

2018-06-01

Mercury is a well-known heavy metal ion which is extremely poisonous to health but is still employed in the form of mercury salts and organomercury compounds in various industrial, anthropological and agricultural activities. Henceforth, its sensing in aqueous medium is an area of great interest in order to avoid its hazardous effect. In the present manuscript, urea/thiourea linkage bearing four organic ligands (1a, 1b, 2a and 2b) are synthesized by a three-step synthetic approach. The organic ligands were then employed to develop organic nanoparticles by re-precipitation method which was further probed for their selective recognition behavior in an aqueous medium using fluorescence spectroscopy. The fluorescence emission profile of the ONPs is used as a tool for the tracking of sensing behavior. The ONPs of 1b has shown selective recognition towards Hg(II) in aqueous medium evidenced by enhancement of fluorescence emission intensity after complexation of 1b ONP with Hg(II), among several alkali, alkaline earth and transition metal ions with a detection limit of the order of 0.84 μM. The ability of the proposed sensor to sense Hg(II) ions with high selectivity and sensitivity could be accounted to photo-induced electron transfer (PET) "OFF" mechanism at λem = 390 nm. This study reveals the application of the proposed thiourea-based sensor for the selective recognition of the Hg(II) ions in an aqueous medium.

Fluorescence molecular tomography in the presence of background fluorescence

International Nuclear Information System (INIS)

Soubret, Antoine; Ntziachristos, Vasilis

2006-01-01

Fluorescence molecular tomography is an emerging imaging technique that resolves the bio-distribution of engineered fluorescent probes developed for in vivo reporting of specific cellular and sub-cellular targets. The method can detect fluorochromes in picomole amounts or less, imaged through entire animals, but the detection sensitivity and imaging performance drop in the presence of background, non-specific fluorescence. In this study, we carried out a theoretical and an experimental investigation on the effect of background fluorescence on the measured signal and on the tomographic reconstruction. We further examined the performance of three subtraction methods based on physical models of photon propagation, using experimental data on phantoms and small animals. We show that the data pre-processing with subtraction schemes can improve image quality and quantification when non-specific background florescence is present

A fluorescence detection of D-penicillamine based on Cu(2+)-induced fluorescence quenching system of protein-stabilized gold nanoclusters.

Science.gov (United States)

Wang, Peng; Li, Bang Lin; Li, Nian Bing; Luo, Hong Qun

2015-01-25

In this contribution, a luminescent gold nanoclusters which were synthesized by bovine serum albumin as novel fluorescent probes were successfully utilized for the determination of D-penicillamine for the first time. Cupric ion was employed to quench the strong fluorescence of the gold nanoclusters, whereas the addition of D-penicillamine caused obvious restoration of fluorescence intensity of the Cu(2+)-gold nanoclusters system. Under optimum conditions, the increment in fluorescence intensity of Cu(2+)-gold nanoclusters system caused by D-penicillamine was linearly proportional to the concentration of D-penicillamine in the range of 2.0×10(-5)-2.39×10(-4) M. The detection limit for D-penicillamine was 5.4×10(-6) M. With the off-on fluorescence signal at 650 nm approaching the near-infrared region, the present sensor for D-penicillamine detection had high sensitivity and low spectral interference. Furthermore, the novel gold nanoclusters-based fluorescent sensor has been applied to the determination of D-penicillamine in real biological samples with satisfactory results. Copyright © 2014 Elsevier B.V. All rights reserved.

Effect of vesicle size on the prodan fluorescence in diheptadecanoylphosphatidylcholine bilayer membrane under atmospheric and high pressures.

Science.gov (United States)

Goto, Masaki; Sawaguchi, Hiroshi; Tamai, Nobutake; Matsuki, Hitoshi; Kaneshina, Shoji

2010-08-17

The bilayer phase behavior of diheptadecanoylphosphatidylcholine (C17PC) with different vesicle sizes (large multilamellar vesicle (LMV) and giant multilamellar vesicle (GMV)) was investigated by fluorescence spectroscopy using a polarity-sensitive fluorescent probe Prodan under atmospheric and high pressures. The difference in phase transitions and thermodynamic quantities of the transition was hardly observed between LMV and GMV used here. On the contrary, the Prodan fluorescence in the bilayer membranes changed depending on the size of vesicles as well as on the phase states. From the second derivative of fluorescence spectra, the three-dimensional image plots in which we can see the location of Prodan in the bilayer membrane as blue valleys were constructed for LMV and GMV under atmospheric pressure. The following characteristic behavior was found: (1) the Prodan molecules in GMV can be distributed to not only adjacent glycerol backbone region, but also near bulk-water region in the lamellar gel or ripple gel phase; (2) the blue valleys of GMV became deeper than those of LMV because of the greater surface density of the Prodan molecules per unit area of GMV than LMV; (3) the liquid crystalline phase of the bilayer excludes the Prodan molecules to a more hydrophilic region at the membrane surface with an increase in vesicle size; (4) the accurate information as to the phase transitions is gradually lost with increasing vesicle size. Under the high-pressure condition, the difference in Prodan fluorescence between LMV and GMV was essentially the same as the difference under atmospheric pressure except for the existence of the pressure-induced interdigitated gel phase. Further, we found that Prodan fluorescence spectra in the interdigitated gel phase were especially affected by the size of vesicles. This study revealed that the Prodan molecules can move around the headgroup region by responding not only to the phase state but also to the vesicle size, and they

Label-free fluorescence strategy for sensitive detection of adenosine triphosphate using a loop DNA probe with low background noise.

Science.gov (United States)

Lin, Chunshui; Cai, Zhixiong; Wang, Yiru; Zhu, Zhi; Yang, Chaoyong James; Chen, Xi

2014-07-15

A simple, rapid, label-free, and ultrasensitive fluorescence strategy for adenosine triphosphate (ATP) detection was developed using a loop DNA probe with low background noise. In this strategy, a loop DNA probe, which is the substrate for both ligation and digestion enzyme reaction, was designed. SYBR green I (SG I), a double-stranded specific dye, was applied for the readout fluorescence signal. Exonuclease I (Exo I) and exonuclease III (Exo III), sequence-independent nucleases, were selected to digest the loop DNA probe in order to minimize the background fluorescence signal. As a result, in the absence of ATP, the loop DNA was completely digested by Exo I and Exo III, leading to low background fluorescence owing to the weak electrostatic interaction between SG I and mononucleotides. On the other hand, ATP induced the ligation of the nicking site, and the sealed loop DNA resisted the digestion of Exo I and ExoIII, resulting in a remarkable increase of fluorescence response. Upon background noise reduction, the sensitivity of the ATP determination was improved significantly, and the detection limitation was found to be 1.2 pM, which is much lower than that in almost all the previously reported methods. This strategy has promise for wide application in the determination of ATP.

Hybrid system for in vivo real-time planar fluorescence and volumetric optoacoustic imaging

Science.gov (United States)

Chen, Zhenyue; Deán-Ben, Xosé Luís.; Gottschalk, Sven; Razansky, Daniel

2018-02-01

Fluorescence imaging is widely employed in all fields of cell and molecular biology due to its high sensitivity, high contrast and ease of implementation. However, the low spatial resolution and lack of depth information, especially in strongly-scattering samples, restrict its applicability for deep-tissue imaging applications. On the other hand, optoacoustic imaging is known to deliver a unique set of capabilities such as high spatial and temporal resolution in three dimensions, deep penetration and spectrally-enriched imaging contrast. Since fluorescent substances can generate contrast in both modalities, simultaneous fluorescence and optoacoustic readings can provide new capabilities for functional and molecular imaging of living organisms. Optoacoustic images can further serve as valuable anatomical references based on endogenous hemoglobin contrast. Herein, we propose a hybrid system for in vivo real-time planar fluorescence and volumetric optoacoustic tomography, both operating in reflection mode, which synergistically combines the advantages of stand-alone systems. Validation of the spatial resolution and sensitivity of the system were first carried out in tissue mimicking phantoms while in vivo imaging was further demonstrated by tracking perfusion of an optical contrast agent in a mouse brain in the hybrid imaging mode. Experimental results show that the proposed system effectively exploits the contrast mechanisms of both imaging modalities, making it especially useful for accurate monitoring of fluorescence-based signal dynamics in highly scattering samples.

Near-IR Two-Photon Fluorescent Sensor for K(+) Imaging in Live Cells.

Science.gov (United States)

Sui, Binglin; Yue, Xiling; Kim, Bosung; Belfield, Kevin D

2015-08-19

A new two-photon excited fluorescent K(+) sensor is reported. The sensor comprises three moieties, a highly selective K(+) chelator as the K(+) recognition unit, a boron-dipyrromethene (BODIPY) derivative modified with phenylethynyl groups as the fluorophore, and two polyethylene glycol chains to afford water solubility. The sensor displays very high selectivity (>52-fold) in detecting K(+) over other physiological metal cations. Upon binding K(+), the sensor switches from nonfluorescent to highly fluorescent, emitting red to near-IR (NIR) fluorescence. The sensor exhibited a good two-photon absorption cross section, 500 GM at 940 nm. Moreover, it is not sensitive to pH in the physiological pH range. Time-dependent cell imaging studies via both one- and two-photon fluorescence microscopy demonstrate that the sensor is suitable for dynamic K(+) sensing in living cells.

Ultra-sensitive detection of kanamycin for food safety using a reduced graphene oxide-based fluorescent aptasensor

Science.gov (United States)

Ha, Na-Reum; Jung, In-Pil; La, Im-Joung; Jung, Ho-Sup; Yoon, Moon-Young

2017-01-01

Overuse of antibiotics has caused serious problems, such as appearance of super bacteria, whose accumulation in the human body through the food chain is a concern. Kanamycin is a common antibiotic used to treat diverse infections; however, residual kanamycin can cause many side effects in humans. Thus, development of an ultra-sensitive, precise, and simple detection system for residual kanamycin in food products is urgently needed for food safety. In this study, we identified kanamycin-binding aptamers via a new screening method, and truncated variants were analyzed for optimization of the minimal sequence required for target binding. We found various aptamers with high binding affinity from 34.7 to 669 nanomolar Kdapp values with good specificity against kanamycin. Furthermore, we developed a reduced graphene oxide (RGO)-based fluorescent aptasensor for kanamycin detection. In this system, kanamycin was detected at a concentration as low as 1 pM (582.6 fg/mL). In addition, this method could detect kanamycin accurately in kanamycin-spiked blood serum and milk samples. Consequently, this simple, rapid, and sensitive kanamycin detection system with newly structural and functional analysis aptamer exhibits outstanding detection compared to previous methods and provides a new possibility for point of care testing and food safety.

Culture-free, highly sensitive, quantitative detection of bacteria from minimally processed samples using fluorescence imaging by smartphone.

Science.gov (United States)

Shrivastava, Sajal; Lee, Won-Il; Lee, Nae-Eung

2018-06-30

A critical unmet need in the diagnosis of bacterial infections, which remain a major cause of human morbidity and mortality, is the detection of scarce bacterial pathogens in a variety of samples in a rapid and quantitative manner. Herein, we demonstrate smartphone-based detection of Staphylococcus aureus in a culture-free, rapid, quantitative manner from minimally processed liquid samples using aptamer-functionalized fluorescent magnetic nanoparticles. The tagged S. aureus cells were magnetically captured in a detection cassette, and then fluorescence was imaged using a smartphone camera with a light-emitting diode as the excitation source. Our results showed quantitative detection capability with a minimum detectable concentration as low as 10 cfu/ml by counting individual bacteria cells, efficiently capturing S. aureus cells directly from a peanut milk sample within 10 min. When the selectivity of detection was investigated using samples spiked with other pathogenic bacteria, no significant non-specific detection occurred. Furthermore, strains of S. aureus from various origins showed comparable results, ensuring that the approach can be widely adopted. Therefore, the quantitative fluorescence imaging platform on a smartphone could allow on-site detection of bacteria, providing great potential assistance during major infectious disease outbreaks in remote and resource-limited settings. Copyright © 2018 Elsevier B.V. All rights reserved.

A novel fluorescence immunoassay for the sensitive detection of Escherichia coli O157:H7 in milk based on catalase-mediated fluorescence quenching of CdTe quantum dots.

Science.gov (United States)

Chen, Rui; Huang, Xiaolin; Li, Juan; Shan, Shan; Lai, Weihua; Xiong, Yonghua

2016-12-01

Immunoassay is a powerful tool for rapid detection of food borne pathogens in food safety monitoring. However, conventional immunoassay always suffers from low sensitivity when it employs enzyme-catalyzing chromogenic substrates to generate colored molecules as signal outputs. In the present study, we report a novel fluorescence immunoassay for the sensitive detection of E. coli O157:H7 through combination of the ultrahigh bioactivity of catalase to hydrogen peroxide (H 2 O 2 ) and H 2 O 2 -sensitive mercaptopropionic acid modified CdTe QDs (MPA-QDs) as a signal transduction. Various parameters, including the concentrations of anti-E. coli O157:H7 polyclonal antibody and biotinylated monoclonal antibody, the amounts of H 2 O 2 and streptavidin labeled catalase (CAT), the hydrolysis temperature and time of CAT to H 2 O 2 , as well as the incubation time between H 2 O 2 and MPA-QDs, were systematically investigated and optimized. With optimal conditions, the catalase-mediated fluorescence quenching immunoassay exhibits an excellent sensitivity for E. coli O157:H7 with a detection limit of 5 × 10 2  CFU/mL, which was approximately 140 times lower than that of horseradish peroxidase-based colorimetric immunoassay. The reliability of the proposed method was further evaluated using E. coli O157:H7 spiked milk samples. The average recoveries of E. coli O157:H7 concentrations from 1.18 × 10 3  CFU/mL to 1.18 × 10 6  CFU/mL were in the range of 65.88%-105.6%. In brief, the proposed immunoassay offers a great potential for rapid and sensitive detection of other pathogens in food quality control. Copyright © 2016 Elsevier B.V. All rights reserved.

Salicylimine-Based Colorimetric and Fluorescent Chemosensor for Selective Detection of Cyanide in Aqueous Buffer

Energy Technology Data Exchange (ETDEWEB)

Noh, Jin Young; Hwang, In Hong; Kim, Hyun; Song, Eun Joo; Kim, Kyung Beom; Kim, Cheal [Seoul National Univ., Seoul (Korea, Republic of)

2013-07-15

A simple colorimetric and fluorescent anion sensor 1 based on salicylimine showed a high selectivity and sensitivity for detection of cyanide in aqueous solution. The receptor 1 showed high selectivity toward CN{sup -} ions in a 1:1 stoichiometric manner, which induces a fast color change from colorless to orange and a dramatic enhancement in fluorescence intensity selectively for cyanide anions over other anions. Such selectivity resulted from the nucleophilic addition of CN{sup -} to the carbon atom of an electron-deficient imine group. The sensitivity of the fluorescence-based assay (0.06 μM) is below the 1.9 μM suggested by the World Health Organization (WHO) as the maximum allowable cyanide concentration in drinking water, capable of being a practical system for the monitoring of CN. concentrations in aqueous samples.

Recording membrane potential changes through photoacoustic voltage sensitive dye

Science.gov (United States)

Zhang, Haichong K.; Kang, Jeeun; Yan, Ping; Abou, Diane S.; Le, Hanh N. D.; Thorek, Daniel L. J.; Kang, Jin U.; Gjedde, Albert; Rahmim, Arman; Wong, Dean F.; Loew, Leslie M.; Boctor, Emad M.

2017-03-01

Monitoring of the membrane potential is possible using voltage sensitive dyes (VSD), where fluorescence intensity changes in response to neuronal electrical activity. However, fluorescence imaging is limited by depth of penetration and high scattering losses, which leads to low sensitivity in vivo systems for external detection. In contrast, photoacoustic (PA) imaging, an emerging modality, is capable of deep tissue, noninvasive imaging by combining near infrared light excitation and ultrasound detection. In this work, we develop the theoretical concept whereby the voltage-dependent quenching of dye fluorescence leads to a reciprocal enhancement of PA intensity. Based on this concept, we synthesized a novel near infrared photoacoustic VSD (PA-VSD) whose PA intensity change is sensitive to membrane potential. In the polarized state, this cyanine-based probe enhances PA intensity while decreasing fluorescence output in a lipid vesicle membrane model. With a 3-9 μM VSD concentration, we measured a PA signal increase in the range of 5.3 % to 18.1 %, and observed a corresponding signal reduction in fluorescence emission of 30.0 % to 48.7 %. A theoretical model successfully accounts for how the experimental PA intensity change depends on fluorescence and absorbance properties of the dye. These results not only demonstrate the voltage sensing capability of the dye, but also indicate the necessity of considering both fluorescence and absorbance spectral sensitivities in order to optimize the characteristics of improved photoacoustic probes. Together, our results demonstrate photoacoustic sensing as a potential new modality for sub-second recording and external imaging of electrophysiological and neurochemical events in the brain.

Determination of absolute Ba densities during dimming operation of fluorescent lamps by laser-induced fluorescence measurements

International Nuclear Information System (INIS)

Hadrath, S; Beck, M; Garner, R C; Lieder, G; Ehlbeck, J

2007-01-01

Investigations of fluorescent lamps (FL) are often focused on the electrodes, since the lifetime of the lamps is typically limited by the electrode lifetime and durability. During steady state operation, the work function lowering emitter material, in particular, barium, is lost. Greater barium losses occur under dimming conditions, in which reduced discharge currents lead to increased cathode falls, the result of the otherwise diminished heating of the electrode by the bombarding plasma ions. In this work the barium density near the electrodes of (FL), operating in high frequency dimming mode is investigated using the high-sensitivity method of laser-induced fluorescence. From these measurements we infer barium loss for a range of discharge currents and auxiliary coil heating currents. We show that the Ba loss can very easily be reduced by moderate auxiliary coil heating

High-performance fluorescence-encoded magnetic microbeads as microfluidic protein chip supports for AFP detection

Energy Technology Data Exchange (ETDEWEB)

Gong, Xiaoqun [School of Life Sciences, Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology, Collaborative Innovation Center of Chemical Science and Engineering, Tianjin University, Tianjin 300072 (China); Yan, Huan; Yang, Jiumin [Department of Laboratory Medicine, Tianjin Medical University General Hospital, Tianjin, 300052 (China); Wu, Yudong; Zhang, Jian; Yao, Yingyi [School of Life Sciences, Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology, Collaborative Innovation Center of Chemical Science and Engineering, Tianjin University, Tianjin 300072 (China); Liu, Ping [Bioscience (Tianjin) Diagnostic Technology CO., LTD, Tianjin, 300300 (China); Wang, Huiquan [Department of Biomedical Engineering, School of Electronics and Information Engineering, Tianjin Polytechnic University, Tianjin, 300387 (China); Hu, Zhidong, E-mail: huzhidong27@163.com [Department of Laboratory Medicine, Tianjin Medical University General Hospital, Tianjin, 300052 (China); Chang, Jin, E-mail: jinchang@tju.edu.cn [School of Life Sciences, Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology, Collaborative Innovation Center of Chemical Science and Engineering, Tianjin University, Tianjin 300072 (China)

2016-10-05

Fluorescence-encoded magnetic microbeads (FEMMs), with the fluorescence encoding ability of quantum dots (QDs) and magnetic enrichment and separation functions of Fe{sub 3}O{sub 4} nanoparticles, have been widely used for multiple biomolecular detection as microfluidic protein chip supports. However, the preparation of FEMMs with long-term fluorescent encoding and immunodetection stability is still a challenge. In this work, we designed a novel high-temperature chemical swelling strategy. The QDs and Fe{sub 3}O{sub 4} nanoparticles were effectively packaged into microbeads via the thermal motion of the polymer chains and the hydrophobic interaction between the nanoparticles and microbeads. The FEMMs obtained a highly uniform fluorescent property and long-term encoding and immunodetection stability and could be quickly magnetically separated and enriched. Then, the QD-encoded magnetic microbeads were applied to alpha fetoprotein (AFP) detection via sandwich immunoreaction. The properties of the encoded microspheres were characterized using a self-designed detecting apparatus, and the target molecular concentration in the sample was also quantified. The results suggested that the high-performance FEMMs have great potential in the field of biomolecular detection. - Graphical abstract: We designed a novel strategy to prepare a kind of high-performance fluorescence-encoded magnetic microbeads as microfluidic protein chip support with long-time fluorescent encoding and immunodetection stability for AFP detection. - Highlights: • A novel strategy combined the high temperature with chemical swelling technology is designed. • Based on hydrophobic interaction and polymer thermal motion, QDs and Fe{sub 3}O{sub 4} were effectively packaged into microbeads. • The fluorescence-encoded magnetic microbeads show long-term fluorescent encoding and immunodetection stability.

A novel graphene-based label-free fluorescence 'turn-on' nanosensor for selective and sensitive detection of phosphorylated species in biological samples and living cells.

Science.gov (United States)

Ke, Yaotang; Garg, Bhaskar; Ling, Yong-Chien

2016-02-28

A novel label-free fluorescence 'turn-on' nanosensor has been developed for highly selective and sensitive detection of phosphorylated species (Ps) in biological samples and living cells. The design strategy relies on the use of Ti(4+)-immobilized polydopamine (PDA) coated reduced graphene oxide (rGO@PDA-Ti(4+)) that serves as an attractive platform to bind riboflavin 5'-monophosphate molecules (FMNs) through ion-pair interactions between phosphate groups and Ti(4+). The as-prepared rGO@PDA-Ti(4+)-FMNs (nanosensor), fluoresce only weakly due to the ineffective Förster resonance energy transfer between the FMNs and rGO@PDA-Ti(4+). The experimental findings revealed that the microwave-assisted interaction of the nanosensor with α-, β-casein, ovalbumin, human serum, non-fat milk, egg white, and living cells (all containing Ps) releases FMNs (due to the high formation constant between phosphate groups and Ti(4+)), leading to an excellent fluorescence 'turn-on' response. The fluorescence spectroscopy, confocal microscopy, and MALDI-TOF MS spectrometry were used to detect Ps both qualitatively and quantitatively. Under the optimized conditions, the nanosensor showed a detection limit of ca. 118.5, 28.9, and 54.8 nM for the tryptic digests of α-, β-casein and ovalbumin, respectively. Furthermore, the standard addition method was used as a bench-mark proof for phosphopeptide quantification in egg white samples. We postulate that the present quantitative assay for Ps holds tremendous potential and may pave the way to disease diagnostics in the near future.

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.

High-resolution Imaging of pH in Alkaline Sediments and Water Based on a New Rapid Response Fluorescent Planar Optode

Science.gov (United States)

Han, Chao; Yao, Lei; Xu, Di; Xie, Xianchuan; Zhang, Chaosheng

2016-05-01

A new dual-lumophore optical sensor combined with a robust RGB referencing method was developed for two-dimensional (2D) pH imaging in alkaline sediments and water. The pH sensor film consisted of a proton-permeable polymer (PVC) in which two dyes with different pH sensitivities and emission colors: (1) chloro phenyl imino propenyl aniline (CPIPA) and (2) the coumarin dye Macrolex® fluorescence yellow 10 GN (MFY-10 GN) were entrapped. Calibration experiments revealed the typical sigmoid function and temperature dependencies. This sensor featured high sensitivity and fast response over the alkaline working ranges from pH 7.5 to pH 10.5. Cross-sensitivity towards ionic strength (IS) was found to be negligible for freshwater when IS applications.

An ultra-sensitive monoclonal antibody-based fluorescent microsphere immunochromatographic test strip assay for detecting aflatoxin M1 in milk

Science.gov (United States)

A rapid lateral flow fluorescent microspheres immunochromatography test strip (FMs-ICTS) has been developed for the detection of aflatoxin M1 (AFM1) residues in milk. For this purpose, an ultra-sensitive anti-AFM1 monoclonal antibody (MAb) 1D3 was prepared and identified. The IC50 value of the MA...

Sensitive and selective detection of adenine using fluorescent ZnS nanoparticles

International Nuclear Information System (INIS)

Meerabai Devi, L; Negi, Devendra P S

2011-01-01

We have used fluorescent ZnS nanoparticles as a probe for the determination of adenine. A typical 2 x 10 -7 M concentration of adenine quenches 39.3% of the ZnS fluorescence. The decrease in ZnS fluorescence as a function of adenine concentration was found to be linear in the concentration range 5 x 10 -9 -2 x 10 -7 M. The limit of detection (LOD) of adenine by this method is 3 nM. Among the DNA bases, only adenine quenched the fluorescence of ZnS nanoparticles in the submicromolar concentration range, thus adding selectivity to the method. The amino group of adenine was important in determining the quenching efficiency. Steady-state fluorescence experiments suggest that one molecule of adenine is sufficient to quench the emission arising from a cluster of ZnS consisting of about 20 molecules. Time-resolved fluorescence measurements indicate that the adenine molecules block the sites on the surface of ZnS responsible for emission with the longest lifetime component. This method may be applied for the determination of adenine in biological samples since the measurements have been carried out at pH 7.

A novel and sensitive method for determining vitamin B3 and B7 by pre-column derivatization and high-performance liquid chromatography method with fluorescence detection.

Directory of Open Access Journals (Sweden)

Baolei Fan

Full Text Available A new labeling reagent for vitamin analysis, 2-amino-10-ethyl acridine ketone (AEAO, has been synthesized and successfully applied to the analysis of vitamin B3 and vitamin B7 in different tea samples. The reaction of AEAO with vitamins could proceed easily and quickly in the presence of 1-ethyl-3-(3-dimethylaminopropyl-carbodiimide hydrochloride (EDC as condensing reagent within 45 min. The derivatives exhibited excellent fluorescence property with excitation and emission wavelengths of 290 nm and 430 nm, respectively. Response surface methodology (RSM was applied to the optimization of pre-column derivatization. Solid phase extraction with HLB cartridges was used for the extraction and purification of water-soluble vitamins in tea samples. The LODs for vitamin B3 and vitamin B7 were 2.56 and 2.22 ng mL-1, respectively. The proposed method was successfully applied to the analysis of vitamin B3 and vitamin B7 in different tea samples. The study provided a highly sensitive method for accurate analysis of trace vitamins from natural products.

Toehold-mediated strand displacement reaction-dependent fluorescent strategy for sensitive detection of uracil-DNA glycosylase activity.

Science.gov (United States)

Wu, Yushu; Wang, Lei; Jiang, Wei

2017-03-15

Sensitive detection of uracil-DNA glycosylase (UDG) activity is beneficial for evaluating the repairing process of DNA lesions. Here, toehold-mediated strand displacement reaction (TSDR)-dependent fluorescent strategy was constructed for sensitive detection of UDG activity. A single-stranded DNA (ssDNA) probe with two uracil bases and a trigger sequence were designed. A hairpin probe with toehold domain was designed, and a reporter probe was also designed. Under the action of UDG, two uracil bases were removed from ssDNA probe, generating apurinic/apyrimidinic (AP) sites. Then, the AP sites could inhibit the TSDR between ssDNA probe and hairpin probe, leaving the trigger sequence in ssDNA probe still free. Subsequently, the trigger sequence was annealed with the reporter probe, initiating the polymerization and nicking amplification reaction. As a result, numerous G-quadruplex (G4) structures were formed, which could bind with N-methyl-mesoporphyrin IX (NMM) to generate enhanced fluorescent signal. In the absence of UDG, the ssDNA probe could hybridize with the toehold domain of the hairpin probe to initiate TSDR, blocking the trigger sequence, and then the subsequent amplification reaction would not occur. The proposed strategy was successfully implemented for detecting UDG activity with a detection limit of 2.7×10 -5 U/mL. Moreover, the strategy could distinguish UDG well from other interference enzymes. Furthermore, the strategy was also applied for detecting UDG activity in HeLa cells lysate with low effect of cellular components. These results indicated that the proposed strategy offered a promising tool for sensitive quantification of UDG activity in UDG-related function study and disease prognosis. Copyright © 2016 Elsevier B.V. All rights reserved.

vuv fluorescence from selective high-order multiphoton excitation of N2

International Nuclear Information System (INIS)

Coffee, Ryan N.; Gibson, George N.

2004-01-01

Recent fluorescence studies suggest that ultrashort pulse laser excitation may be highly selective. Selective high-intensity laser excitation holds important consequences for the physics of multiphoton processes. To establish the extent of this selectivity, we performed a detailed comparative study of the vacuum ultraviolet fluorescence resulting from the interaction of N 2 and Ar with high-intensity infrared ultrashort laser pulses. Both N 2 and Ar reveal two classes of transitions, inner-valence ns ' l ' . From their pressure dependence, we associate each transition with either plasma or direct laser excitation. Furthermore, we qualitatively confirm such associations with the time dependence of the fluorescence signal. Remarkably, only N 2 presents evidence of direct laser excitation. This direct excitation produces ionic nitrogen fragments with inner-valence (2s) holes, two unidentified transitions, and one molecular transition, the N 2 + :X 2 Σ g + 2 Σ u + . We discuss these results in the light of a recently proposed model for multiphoton excitation

Self-interference fluorescence microscopy with three-phase detection for depth-resolved confocal epi-fluorescence imaging.

Science.gov (United States)

Braaf, Boy; de Boer, Johannes F

2017-03-20

Three-dimensional confocal fluorescence imaging of in vivo tissues is challenging due to sample motion and limited imaging speeds. In this paper a novel method is therefore presented for scanning confocal epi-fluorescence microscopy with instantaneous depth-sensing based on self-interference fluorescence microscopy (SIFM). A tabletop epi-fluorescence SIFM setup was constructed with an annular phase plate in the emission path to create a spectral self-interference signal that is phase-dependent on the axial position of a fluorescent sample. A Mach-Zehnder interferometer based on a 3 × 3 fiber-coupler was developed for a sensitive phase analysis of the SIFM signal with three photon-counter detectors instead of a spectrometer. The Mach-Zehnder interferometer created three intensity signals that alternately oscillated as a function of the SIFM spectral phase and therefore encoded directly for the axial sample position. Controlled axial translation of fluorescent microsphere layers showed a linear dependence of the SIFM spectral phase with sample depth over axial image ranges of 500 µm and 80 µm (3.9 × Rayleigh range) for 4 × and 10 × microscope objectives respectively. In addition, SIFM was in good agreement with optical coherence tomography depth measurements on a sample with indocyanine green dye filled capillaries placed at multiple depths. High-resolution SIFM imaging applications are demonstrated for fluorescence angiography on a dye-filled capillary blood vessel phantom and for autofluorescence imaging on an ex vivo fly eye.

Very High Spectral Resolution Imaging Spectroscopy: the Fluorescence Explorer (FLEX) Mission

Science.gov (United States)

Moreno, Jose F.; Goulas, Yves; Huth, Andreas; Middleton, Elizabeth; Miglietta, Franco; Mohammed, Gina; Nedbal, Ladislav; Rascher, Uwe; Verhoef, Wouter; Drusch, Matthias

2016-01-01

The Fluorescence Explorer (FLEX) mission has been recently selected as the 8th Earth Explorer by the European Space Agency (ESA). It will be the first mission specifically designed to measure from space vegetation fluorescence emission, by making use of very high spectral resolution imaging spectroscopy techniques. Vegetation fluorescence is the best proxy to actual vegetation photosynthesis which can be measurable from space, allowing an improved quantification of vegetation carbon assimilation and vegetation stress conditions, thus having key relevance for global mapping of ecosystems dynamics and aspects related with agricultural production and food security. The FLEX mission carries the FLORIS spectrometer, with a spectral resolution in the range of 0.3 nm, and is designed to fly in tandem with Copernicus Sentinel-3, in order to provide all the necessary spectral / angular information to disentangle emitted fluorescence from reflected radiance, and to allow proper interpretation of the observed fluorescence spatial and temporal dynamics.

Fabrication of high performance microlenses for an integrated capillary channel electrochromatograph with fluorescence detection

International Nuclear Information System (INIS)

Wendt, J. R.; Warren, M. E.; Sweatt, W. C.; Bailey, C. G.; Matzke, C. M.; Arnold, D. W.; Allerman, A. A.; Carter, T. R.; Asbill, R. E.; Samora, S.

1999-01-01

We describe the microfabrication of an extremely compact optical system as a key element in an integrated capillary channel electrochromatograph with fluorescence detection. The optical system consists of a vertical cavity surface-emitting laser (VCSEL), two high performance microlenses, and a commercial photodetector. The microlenses are multilevel diffractive optics patterned by electron beam lithography and etched by reactive ion etching in fused silica. The design uses substrate-mode propagation within the fused silica substrate. Two generations of optical subsystems are described. The first generation design has a 6 mm optical length and is integrated directly onto the capillary channel-containing substrate. The second generation design separates the optical system onto its own substrate module and the optical path length is further compressed to 3.5 mm. The first generation design has been tested using direct fluorescence detection with a 750 nm VCSEL pumping a 10 -4 M solution of CY-7 dye. The observed signal-to-noise ratio of better than 100:1 demonstrates that the background signal from scattered pump light is low despite the compact size of the optical system and is adequate for system sensitivity requirements. (c) 1999 American Vacuum Society

Fluorescent nanodiamond for biomedicine

International Nuclear Information System (INIS)

Milos Nesladek

2014-01-01

NV centers in diamond have gained strong interest as a novel tool for quantum information processing, quantum computing and quantum photonics. These applications are based on fluorescent and spin properties of NV-centres. However, in some conditions NV- can lose an electron and turn to NV0. The occupation of NV0 and NV- charge states depend on the position of their ground states with respect to the Fermi level and the mechanism of the charge transfer. Interestingly, that the charge switch has important implications on applications of fluorescent nanodiamond (fND) to nano-biology and nano-medicine. fND can be used for bio-marking and bio-tracking but also for the monitoring of targeted delivery to the cells. In this presentation we review the current state-of-the art for using fND particles for fluorescent bio imaging in cells and discuss the charge transfer and its luminescence stability by using ultra high sensitive spectroscopy methods to study the NV0 and NV- state occupation. (author)

Herbicide impact on Hormosira banksii gametes measured by fluorescence and germination bioassays

Energy Technology Data Exchange (ETDEWEB)

Seery, Cliff R. [Institute for Water and Environmental Resource Management, Department of Environmental Sciences, University of Technology, Sydney, Westbourne Street, Gore Hill, 2065 NSW (Australia); Gunthorpe, Leanne [Primary Industries Research Victoria (PIRVic), VIC (Australia); Ralph, Peter J. [Institute for Water and Environmental Resource Management, Department of Environmental Sciences, University of Technology, Sydney, Westbourne Street, Gore Hill, 2065 NSW (Australia)]. E-mail: peter.ralph@uts.edu.au

2006-03-15

The innovative bioassay described here involves chlorophyll a fluorescence measurements of gametes from the macroalgae, Hormosira banksii, where gametes (eggs) were exposed to Diuron, Irgarol and Bromacil. Response was assessed as percent inhibition from control of effective quantum yield ({delta}F/Fm') of photosystem II, herein referred to as % PSII Inhibition. This was measured with the dual-channelled pulse amplitude modulated (PAM) fluorometer, ToxY-PAM. The fluorescence bioassay was run simultaneously with an established H. banksii germination bioassay to compare sensitivity, precision, and time-to-result. The fluorescence bioassay gave highly sensitive results evidenced by EC{sub 5}s (% PSII Inhibition) for Diuron, Irgarol and Bromacil being three, four and three orders of magnitude (respectively) lower than EC{sub 5}s generated from the germination bioassays. Precision of the fluorescence bioassay was demonstrated with low coefficient of variations (<30%) for all three toxicants. With regard to time, the fluorescence bioassay gave results within 6 h, as opposed to more than 50 h for the germination bioassay. - Chlorophyll a fluorescence measurements form the basis of a macroalgal bioassay with many advantages over germination-based methods.

Toward robust high resolution fluorescence tomography: a hybrid row-action edge preserving regularization

Science.gov (United States)

Behrooz, Ali; Zhou, Hao-Min; Eftekhar, Ali A.; Adibi, Ali

2011-02-01

Depth-resolved localization and quantification of fluorescence distribution in tissue, called Fluorescence Molecular Tomography (FMT), is highly ill-conditioned as depth information should be extracted from limited number of surface measurements. Inverse solvers resort to regularization algorithms that penalize Euclidean norm of the solution to overcome ill-posedness. While these regularization algorithms offer good accuracy, their smoothing effects result in continuous distributions which lack high-frequency edge-type features of the actual fluorescence distribution and hence limit the resolution offered by FMT. We propose an algorithm that penalizes the total variation (TV) norm of the solution to preserve sharp transitions and high-frequency components in the reconstructed fluorescence map while overcoming ill-posedness. The hybrid algorithm is composed of two levels: 1) An Algebraic Reconstruction Technique (ART), performed on FMT data for fast recovery of a smooth solution that serves as an initial guess for the iterative TV regularization, 2) A time marching TV regularization algorithm, inspired by the Rudin-Osher-Fatemi TV image restoration, performed on the initial guess to further enhance the resolution and accuracy of the reconstruction. The performance of the proposed method in resolving fluorescent tubes inserted in a liquid tissue phantom imaged by a non-contact CW trans-illumination FMT system is studied and compared to conventional regularization schemes. It is observed that the proposed method performs better in resolving fluorescence inclusions at higher depths.

A trident dithienylethene-perylenemonoimide dyad with super fluorescence switching speed and ratio

Science.gov (United States)

Li, Chong; Yan, Hui; Zhao, Ling-Xi; Zhang, Guo-Feng; Hu, Zhe; Huang, Zhen-Li; Zhu, Ming-Qiang

2014-12-01

Photoswitchable fluorescent diarylethenes are promising in molecular optical memory and photonic devices. However, the performance of current diarylethenes is far from satisfactory because of the scarcity of high-speed switching capability and large fluorescence on-off ratio. Here we report a trident perylenemonoimide dyad modified by triple dithienylethenes whose photochromic fluorescence quenching ratio at the photostationary state exceeds 10,000 and the fluorescence quenching efficiency is close to 100% within seconds of ultraviolet irradiation. The highly sensitive fluorescence on/off switching of the trident dyad enables recyclable fluorescence patterning and all-optical transistors. The prototype optical device based on the trident dyad enables the optical switching of incident light and conversion from incident light wavelength to transmitted light wavelength, which is all-optically controlled, reversible and wavelength-convertible. In addition, the trident dyad-staining block copolymer vesicles are observed via optical nanoimaging with a sub-100 nm resolution, portending a potential prospect of the dithienylethene dyad in super-resolution imaging.

Feasibility of Using Fluorescence Spectrophotometry to Develop a Sensitive Dye Immersion Method for Container Closure Integrity Testing of Prefilled Syringes.

Science.gov (United States)

Lu, Xujin; Lloyd, David K; Klohr, Steven E

2016-01-01

A feasibility study was conducted for a sensitive and robust dye immersion method for the measurement of container closure integrity of unopened prefilled syringes using fluorescence spectrophotometry as the detection method. A Varian Cary Eclipse spectrofluorometer was used with a custom-made sample holder to position the intact syringe in the sample compartment for fluorescence measurements. Methylene blue solution was initially evaluated as the fluorophore in a syringe with excitation at 607 nm and emission at 682 nm, which generated a limit of detection of 0.05 μg/mL. Further studies were conducted using rhodamine 123, a dye with stronger fluorescence. Using 480 nm excitation and 525 nm emission, the dye in the syringe could be easily detected at levels as low as 0.001 μg/mL. The relative standard deviation for 10 measurements of a sample of 0.005 μg/mL (with repositioning of the syringe after each measurement) was less than 1.1%. A number of operational parameters were optimized, including the photomultiplier tube voltage, excitation, and emission slit widths. The specificity of the testing was challenged by using marketed drug products and a protein sample, which showed no interference to the rhodamine detection. Results obtained from this study demonstrated that using rhodamine 123 for container closure integrity testing with in-situ (in-syringe) fluorescence measurements significantly enhanced the sensitivity and robustness of the testing and effectively overcame limitations of the traditional methylene blue method with visual or UV-visible absorption detection. Ensuring container closure integrity of injectable pharmaceutical products is necessary to maintain quality throughout the shelf life of a sterile drug product. Container closure integrity testing has routinely been used to evaluate closure integrity during product development and production line qualification of prefilled syringes, vials, and devices. However, container closure integrity testing

High-sensitivity detection of cardiac troponin I with UV LED excitation for use in point-of-care immunoassay.

Science.gov (United States)

Rodenko, Olga; Eriksson, Susann; Tidemand-Lichtenberg, Peter; Troldborg, Carl Peder; Fodgaard, Henrik; van Os, Sylvana; Pedersen, Christian

2017-08-01

High-sensitivity cardiac troponin assay development enables determination of biological variation in healthy populations, more accurate interpretation of clinical results and points towards earlier diagnosis and rule-out of acute myocardial infarction. In this paper, we report on preliminary tests of an immunoassay analyzer employing an optimized LED excitation to measure on a standard troponin I and a novel research high-sensitivity troponin I assay. The limit of detection is improved by factor of 5 for standard troponin I and by factor of 3 for a research high-sensitivity troponin I assay, compared to the flash lamp excitation. The obtained limit of detection was 0.22 ng/L measured on plasma with the research high-sensitivity troponin I assay and 1.9 ng/L measured on tris-saline-azide buffer containing bovine serum albumin with the standard troponin I assay. We discuss the optimization of time-resolved detection of lanthanide fluorescence based on the time constants of the system and analyze the background and noise sources in a heterogeneous fluoroimmunoassay. We determine the limiting factors and their impact on the measurement performance. The suggested model can be generally applied to fluoroimmunoassays employing the dry-cup concept.

Synthesis and spectral properties of novel chlorinated pH fluorescent probes

International Nuclear Information System (INIS)

Wu Xianglong; Jin Xilang; Wang Yunxia; Mei Qibing; Li Jianli; Shi Zhen

2011-01-01

Eight chlorinated fluoresceins have been synthesized by the reaction of chlorinated resorcinols with 4, 5, 6, 7-tetrachlorophthalic anhydride or 3, 6-dichloro-4-carboxyphthalic anhydride in the presence of methanesulfonic acid. The spectral properties of the chlorinated fluoresceins were studied. It was found that they have absorption and emission maxima at long wavelengths and high fluorescence quantum yields. Emission spectra of chlorinated fluoresceins shifted towards long wavelength with increase in chlorine. pH-dependent properties of chlorinated fluoresceins were studied in detail. These compounds showed a strongly pH-sensitive range of 3.0-7.0. These chlorinated fluoresceins will be used as pH probes for pH measurement of the cell because of the high quantum yield and strong pH-sensitivity. - Research highlights: → Eight chlorinated fluoresceins have been synthesized in the presence of methanesulfonic acid. → Emission spectra of these compounds shifted towards long wavelength with increase in chlorine. → Eight chlorinated fluoresceins showed a strongly pH-sensitive range of 3.0-7.0. → They have emission maxima at long wavelengths and high fluorescence quantum yields.

X-ray fluorescence in Member States (Italy): Full field X-ray fluorescence imaging with high-energy and high-spatial resolution

Energy Technology Data Exchange (ETDEWEB)

Romano, F. P.; Masini, N.; Pappalardo, L., E-mail: romanop@lns.infn.it [IBAM, CNR, Via Biblioteca 4, 95124 Catania (Italy); Cosentino, L.; Gammino, S.; Mascali, D.; Rizzo, F. [INFN-LNS, Via S. Sofia 62, 95123 Catania (Italy)

2014-02-15

A full field X-ray camera for the X-Ray Fluorescence imaging of materials with high-energy and high-spatial resolution was designed and developed. The system was realized by coupling a pinhole collimator with a positionsensitive CCD detector. X-Ray fluorescence is induced on the samples by irradiation with an external X-ray tube. The characteristic X-ray spectra of the investigated materials are obtained by using a multi-frames acquisition in single-photon counting. The energy resolution measured at the Fe-Kα line was 157 eV. The spatial resolution of the system was determined by the analysis of a sharp-edge at different magnification values; it was estimated to be 90 μm at a magnification value of 3.2x and 190 μm at 0.8x. The present set-up of the system is suited to analyze samples with dimensions up to 5x4 cm{sup 2}. Typical measurement time is in the range between 1h to 4 h. (author)

Portable evanescent wave fiber biosensor for highly sensitive detection of Shigella

Science.gov (United States)

Xiao, Rui; Rong, Zhen; Long, Feng; Liu, Qiqi

2014-11-01

A portable evanescent wave fiber biosensor was developed to achieve the rapid and highly sensitive detection of Shigella. In this study, a DNA probe was covalently immobilized onto fiber-optic biosensors that can hybridize with a fluorescently labeled complementary DNA. The sensitivity of detection for synthesized oligonucleotides can reach 10-10 M. The surface of the sensor can be regenerated with 0.5% sodium dodecyl sulfate solution (pH 1.9) for over 30 times without significant deterioration of performance. The total analysis time for a single sample, including the time for measurement and surface regeneration, was less than 6 min. We employed real-time polymerase chain reaction (PCR) and compared the results of both methods to investigate the actual Shigella DNA detection capability of the fiber-optic biosensor. The fiber-optic biosensor could detect as low as 102 colony-forming unit/mL Shigella. This finding was comparable with that by real-time PCR, which suggests that this method is a potential alternative to existing detection methods.

A high-throughput direct fluorescence resonance energy transfer-based assay for analyzing apoptotic proteases using flow cytometry and fluorescence lifetime measurements.

Science.gov (United States)

Suzuki, Miho; Sakata, Ichiro; Sakai, Takafumi; Tomioka, Hiroaki; Nishigaki, Koichi; Tramier, Marc; Coppey-Moisan, Maïté

2015-12-15

Cytometry is a versatile and powerful method applicable to different fields, particularly pharmacology and biomedical studies. Based on the data obtained, cytometric studies are classified into high-throughput (HTP) or high-content screening (HCS) groups. However, assays combining the advantages of both are required to facilitate research. In this study, we developed a high-throughput system to profile cellular populations in terms of time- or dose-dependent responses to apoptotic stimulations because apoptotic inducers are potent anticancer drugs. We previously established assay systems involving protease to monitor live cells for apoptosis using tunable fluorescence resonance energy transfer (FRET)-based bioprobes. These assays can be used for microscopic analyses or fluorescence-activated cell sorting. In this study, we developed FRET-based bioprobes to detect the activity of the apoptotic markers caspase-3 and caspase-9 via changes in bioprobe fluorescence lifetimes using a flow cytometer for direct estimation of FRET efficiencies. Different patterns of changes in the fluorescence lifetimes of these markers during apoptosis were observed, indicating a relationship between discrete steps in the apoptosis process. The findings demonstrate the feasibility of evaluating collective cellular dynamics during apoptosis. Copyright © 2015 Elsevier Inc. All rights reserved.

Bioconjugated fluorescent silica nanoparticles for the rapid detection of Entamoeba histolytica.

Science.gov (United States)

Hemadi, Ahmad; Ekrami, Alireza; Oormazdi, Hormozd; Meamar, Ahmad Reza; Akhlaghi, Lame; Samarbaf-Zadeh, Ali Reza; Razmjou, Elham

2015-05-01

Rapid detection of Entamoeba histolytica based on fluorescent silica nanoparticle (FSNP) indirect immunofluorescence microscopy was evaluated. Silica nanoparticles were synthesized using Stöber's method, with their surface activated to covalently bind to, and immobilize, protein A. For biolabeling, FSNP was added to conjugated E. histolytica trophozoites with monoclonal anti-E. histolytica IgG1 for microscopic observation of fluorescence. Fluorescent silica nanoparticle sensitivity was determined with axenically cultured E. histolytica serially diluted to seven concentrations. Specificity was evaluated using other intestinal protozoa. Fluorescent silica nanoparticles detected E. histolytica at the lowest tested concentration with no cross-reaction with Entamoeba dispar, Entamoeba moshkovskii, Blastocystis sp., or Giardia lamblia. Visualization of E. histolytica trophozoites with anti-E. histolytica antibody labeled with fluorescein isothiocyanate (FITC) was compared with that using anti-E. histolytica antibody bioconjugated FSNP. Although FITC and FSNP produced similar results, the amount of specific antibody required for FITC to induce fluorescence of similar intensity was fivefold that for FSNP. Fluorescent silica nanoparticles delivered a rapid, simple, cost-effective, and highly sensitive and specific method of detecting E. histolytica. Further study is needed before introducing FSNP for laboratory diagnosis of amoebiasis. Copyright © 2015 Elsevier B.V. All rights reserved.

Highly selective detection of glutathione using a NIP/Cu2+ complex fluorescent probe

International Nuclear Information System (INIS)

Liang Wenrui; Zhao Zhi; Zhang Yang; Wang Qiusheng; Zhao Xin; Ouyang Jie

2012-01-01

A novel fluorescent compound, 4-(trimethyl ammonium chloride)acetamide-2-(1H-naphtho[2,3-d]imidazol-2-yl)phenol (TMACA-NIP), was synthesized and used as a fluorescent probe for detecting glutathione reduced (GSH). The new NIP-based probe exhibited high fluorescence in water, which was quenched during the presence of copper (II) due to the complexation between TMACA-NIP and Cu 2+ . But after adding GSH into the TMACA-NIP and Cu 2+ system, the fluorescence of TMACA-NIP was recovered because the binding force between GSH and Cu 2+ is stronger than that between TMACA-NIP and Cu 2+ , which destroys the equilibrium between NIP and copper (II) ions and releases the fluorescence probe of TMACA-NIP. This three-component competing system of NIP/Cu 2+ /GSH can be used to detect GSH simply and rapidly. - Highlights: ► A novel fluorescence probe was developed to detect GSH that operates in aqueous solution. ► TMACA-NIP was synthesized and employed as “read-out” units of NIP/Cu 2+ /GSH. ► NIP-based probe shows high selectivity over other sulfhydryl compounds.

Concentrations of Nicotinamide in Plasma by RP-HPLC With Fluorescence Detection

Directory of Open Access Journals (Sweden)

Pan Zhipeng

2016-01-01

Full Text Available The purpose of this study is to establish a new method for detecting nicotinamide concentration in plasma. In the experiment, the high performance liquid chromatography (HPLC method was used, with a fluorescence detector. The nicotinamide in the plasma was first converted to N1- methylnicotinamide, then reacted with acetophenone under certain conditions to produce fluorescent derivatives for testing. The method is a kind of highly sensitive detection, of which the lower limit is 10 ng/mL, the recovery rate is between 92.75% and 105.13%, and the relative standard deviation (RSD is between 3.76% and 4.43%. The results showed that this measurement method is accurate, sensitive and rapid. It meets the requirements of the experiment, and applies to the detection of nicotinamide concentration in plasma.

Sensitive Fluorescent Sensor for Recognition of HIV-1 dsDNA by Using Glucose Oxidase and Triplex DNA

Directory of Open Access Journals (Sweden)

Yubin Li

2018-01-01

Full Text Available A sensitive fluorescent sensor for sequence-specific recognition of double-stranded DNA (dsDNA was developed on the surface of silver-coated glass slide (SCGS. Oligonucleotide-1 (Oligo-1 was designed to assemble on the surface of SCGS and act as capture DNA, and oligonucleotide-2 (Oligo-2 was designed as signal DNA. Upon addition of target HIV-1 dsDNA (Oligo-3•Oligo-4, signal DNA could bind on the surface of silver-coated glass because of the formation of C•GoC in parallel triplex DNA structure. Biotin-labeled glucose oxidase (biotin-GOx could bind to signal DNA through the specific interaction of biotin-streptavidin, thereby GOx was attached to the surface of SCGS, which was dependent on the concentration of target HIV-1 dsDNA. GOx could catalyze the oxidation of glucose and yield H2O2, and the HPPA can be oxidized into a fluorescent product in the presence of HRP. Therefore, the concentration of target HIV-1 dsDNA could be estimated with fluorescence intensity. Under the optimum conditions, the fluorescence intensity was proportional to the concentration of target HIV-1 dsDNA over the range of 10 pM to 1000 pM, the detection limit was 3 pM. Moreover, the sensor had good sequence selectivity and practicability and might be applied for the diagnosis of HIV disease in the future.

New fluorescent pH sensors based on covalently linkable PET rhodamines

Science.gov (United States)

Aigner, Daniel; Borisov, Sergey M.; Orriach Fernández, Francisco J.; Fernández Sánchez, Jorge F.; Saf, Robert; Klimant, Ingo

2012-01-01

A new class of rhodamines for the application as indicator dyes in fluorescent pH sensors is presented. Their pH-sensitivity derives from photoinduced electron transfer between non-protonated amino groups and the excited chromophore which results in effective fluorescence quenching at increasing pH. The new indicator class carries a pentafluorophenyl group at the 9-position of the xanthene core where other rhodamines bear 2-carboxyphenyl substituents instead. The pentafluorophenyl group is used for covalent coupling to sensor matrices by “click” reaction with mercapto groups. Photophysical properties are similar to “classical” rhodamines carrying 2′-carboxy groups. pH sensors have been prepared with two different matrix materials, silica gel and poly(2-hydroxyethylmethacrylate). Both sensors show high luminescence brightness (absolute fluorescence quantum yield ΦF≈0.6) and high pH-sensitivity at pH 5–7 which makes them suitable for monitoring biotechnological samples. To underline practical applicability, a dually lifetime referenced sensor containing Cr(III)-doped Al2O3 as reference material is presented. PMID:22967541

Highly sensitive fiber-optic oxygen sensor based on palladium tetrakis (4-carboxyphenyl)porphyrin doped in ormosil

International Nuclear Information System (INIS)

Chu, Cheng-Shane; Chuang, Chih-Yung

2014-01-01

A simple, low-cost technique for fabrication of highly sensitive fiber-optic oxygen sensor is described. An organically modified silicate (ORMOSIL) as a matrix for the fabrication of oxygen sensing film was produced. The technique is based on coating the end of a plastic optical fiber with ormosil composite xerogel film sequestered with luminophore palladium (II) meso-tetra(4-carboxyphenyl)porphyrin (PdTCPP) prepared by a sol–gel process. The composite xerogel studied is tetraethylorthosilane (TEOS)/n-octyltriethoxysilane (Octyl-triEOS). Result shows that, expect for PdTCPP-doped TEOS/Octyl-triEOS composite xerogel show the high sensitivity and linear Stern–Volmer relationship which indicate the homogenous environment of the luminophore. The sensitivity of the optical oxygen sensor is quantified in terms of the ratio I N2 /I O2 , where I N2 and I O2 represent the detected fluorescence intensities in pure nitrogen and pure oxygen environments, respectively. The experimental result reveals that the PdTCPP-doped TEOS/Octyl-triEOS oxygen sensor has sensitivity of 153. - Highlights: • A simple, low-cost technique for fabrication of highly sensitive fiber-optic oxygen sensor is described. • ORMOSIL was produced to serve as a matrix for the fabrication of oxygen sensing film. • The fiber-optic oxygen sensor has sensitivity of I N2 /I 100O2 =153. • The stable and reproducible signals were obtained with the fiber-optic oxygen sensor

Novel fluorescent probe for highly sensitive bioassay using sequential enzyme-linked immunosorbent assay-capillary isoelectric focusing (ELISA-cIEF).

Science.gov (United States)

Henares, Terence G; Uenoyama, Yuta; Nogawa, Yuto; Ikegami, Ken; Citterio, Daniel; Suzuki, Koji; Funano, Shun-ichi; Sueyoshi, Kenji; Endo, Tatsuro; Hisamoto, Hideaki

2013-06-07

This paper presents a novel rhodamine diphosphate molecule that allows highly sensitive detection of proteins by employing sequential enzyme-linked immunosorbent assay and capillary isoelectric focusing (ELISA-cIEF). Seven-fold improvement in the immunoassay sensitivity and a 1-2 order of magnitude lower detection limit has been demonstrated by taking advantage of the combination of the enzyme-based signal amplification of ELISA and the concentration of enzyme reaction products by cIEF.

Quantum dot bio-conjugate: as a western blot probe for highly sensitive detection of cellular proteins

Energy Technology Data Exchange (ETDEWEB)

Kale, Sonia [Agharkar Research Institute (India); Kale, Anup [University of Alabama, Center for Materials for Information Technology (United States); Gholap, Haribhau; Rana, Abhimanyu [National Chemical Laboratory, Physical and Materials Chemistry Division (India); Desai, Rama [National Centre for Cell Science (India); Banpurkar, Arun [University of Pune, Department of Physics (India); Ogale, Satishchandra, E-mail: sb.ogale@ncl.res.in [National Chemical Laboratory, Physical and Materials Chemistry Division (India); Shastry, Padma, E-mail: padma@nccs.res.in [National Centre for Cell Science (India)

2012-03-15

In the present study, we report a quantum dot (QD)-tailored western blot analysis for a sensitive, rapid and flexible detection of the nuclear and cytoplasmic proteins. Highly luminescent CdTe and (CdTe)ZnS QDs are synthesized by aqueous method. High resolution transmission electron microscopy, Raman spectroscopy, fourier transform infrared spectroscopy, fluorescence spectroscopy and X-ray diffraction are used to characterize the properties of the quantum dots. The QDs are functionalized with antibodies of prostate apoptosis response-4 (Par-4), poly(ADP-ribose) polymerases and {beta} actin to specifically bind with the proteins localized in the nucleus and cytoplasm of the cells, respectively. The QD-conjugated antibodies are used to overcome the limitations of conventional western blot technique. The sensitivity and rapidity of protein detection in QD-based approach is very high, with detection limits up to 10 pg of protein. In addition, these labels provide the capability of enhanced identification and localization of marker proteins in intact cells by confocal laser scanning microscopy.

Imaging lysosomal highly reactive oxygen species and lighting up cancer cells and tumors enabled by a Si-rhodamine-based near-infrared fluorescent probe.

Science.gov (United States)

Zhang, Hongxing; Liu, Jing; Liu, Chenlu; Yu, Pengcheng; Sun, Minjia; Yan, Xiaohan; Guo, Jian-Ping; Guo, Wei

2017-07-01

Lysosomes have recently been regarded as the attractive pharmacological targets for selectively killing of cancer cells via lysosomal cell death (LCD) pathway that is closely associated with reactive oxygen species (ROS). However, the details on the ROS-induced LCD of cancer cells are still poorly understood, partially due to the absence of a lysosome-targetable, robust, and biocompatible imaging tool for ROS. In this work, we brought forward a Si-rhodamine-based fluorescent probe, named PSiR, which could selectively and sensitively image the pathologically more relavent highly reactive oxygen species (hROS: HClO, HO, and ONOO - ) in lysosomes of cancer cells. Compared with many of the existing hROS fluorescent probes, its superiorities are mainly embodied in the high stability against autoxidation and photoxidation, near-infrared exitation and emission, fast fluorescence off-on response, and specific lysosomal localization. Its practicality has been demonstrated by the real-time imaging of hROS generation in lysosomes of human non-small-cell lung cancer cells stimulated by anticancer drug β-lapachone. Moreover, the probe was sensitive enough for basal hROS in cancer cells, allowing its further imaging applications to discriminate not only cancer cells from normal cells, but also tumors from healthy tissues. Overall, our results strongly indicated that PSiR is a very promising imaging tool for the studies of ROS-related LCD of cancer cells, screening of new anticancer drugs, and early diagnosis of cancers. Copyright © 2017 Elsevier Ltd. All rights reserved.

A hybrid molecularly imprinted polymer coated quantum dot nanocomposite optosensor for highly sensitive and selective determination of salbutamol in animal feeds and meat samples.

Science.gov (United States)

Raksawong, Phannika; Chullasat, Kochaporn; Nurerk, Piyaluk; Kanatharana, Proespichaya; Davis, Frank; Bunkoed, Opas

2017-08-01

A hybrid molecularly imprinted polymer (MIP)-coated quantum dot (QD) nanocomposite was synthesized and applied as a fluorescence probe for the highly sensitive and selective determination of salbutamol. The hybrid MIP-coated QD nanocomposite was synthesized via a copolymerization process in the presence of thioglycolic acid capped CdTe QDs with salbutamol as a template, 3-aminopropyltriethoxysilane as the functional monomer, and tetraethyl orthosilicate as a cross-linker. The optimum molar ratio of template, monomer, and cross-linker was 1:6:20. The fluorescence intensity of the hybrid MIP-coated QDs was efficiently quenched after salbutamol rebound to the recognition sites, as a result of charge transfer from QDs to salbutamol. The synthesized hybrid MIP-coated QD nanocomposite showed high sensitivity and good selectivity toward salbutamol. Under the optimal recognition conditions, the fluorescence intensity was quenched linearly with increasing concentration of salbutamol in the range from 0.10 to 25.0 μg L -1 , with a detection limit of 0.034 μg L -1 . The hybrid optosensor developed was successfully applied in the determination of salbutamol in animal feeds and meat samples. Satisfactory recoveries were obtained in the range from 85% to 98%, with a standard deviation of less than 8%. Furthermore, the accuracy of the hybrid MIP-coated QD nanocomposite was investigated by comparison with a conventional high-performance liquid chromatography method, with the results obtained with two methods agreeing well with each other. The advantages of this sensing method are simplicity, rapidity, cost-effectiveness, high sensitivity, and good selectivity. Graphical Abstract The synthesis of hybrid MIP-coated QDs nanocomposite.

Size-selective QD@MOF core-shell nanocomposites for the highly sensitive monitoring of oxidase activities.

Science.gov (United States)

Wang, Ke; Li, Nan; Zhang, Jing; Zhang, Zhiqi; Dang, Fuquan

2017-01-15

In this work, we proposed a novel and facile method to monitor oxidase activities based on size-selective fluorescent quantum dot (QD)@metal-organic framework (MOF) core-shell nanocomposites (CSNCPs). The CSNCPs were synthesized from ZIF-8 and CdTe QDs in aqueous solution in 40min at room temperature with stirring. The prepared CdTe@ZIF-8 CSNCPs , which have excellent water dispersibility and stability, displays distinct fluorescence responses to hole scavengers of different molecular sizes (e.g., H 2 O 2 , substrate, and oxidase) due to the aperture limitation of the ZIF-8 shell. H 2 O 2 can efficiently quench the fluorescence of CdTe@ZIF-8 CSNCPs over a linearity range of 1-100nM with a detection limit of 0.29nM, whereas large molecules such as substrate and oxidase have very little effect on its fluorescence. Therefore, the highly sensitive detection of oxidase activities was achieved by monitoring the fluorescence quenching of CdTe@ZIF-8 CSNCPs by H 2 O 2 produced in the presence of substrate and oxidase, which is proportional to the oxidase activities. The linearity ranges of the uricase and glucose oxidase activity are 0.1-50U/L and 1-100U/L, respectively, and their detection limits are 0.024U/L and 0.26U/L, respectively. Therefore, the current QD@MOF CSNCPs based sensing system is a promising, widely applicable means of monitoring oxidase activities in biochemical research. Copyright © 2016 Elsevier B.V. All rights reserved.

Make Caffeine Visible: a Fluorescent Caffeine “Traffic Light” Detector

Science.gov (United States)

Xu, Wang; Kim, Tae-Hyeong; Zhai, Duanting; Er, Jun Cheng; Zhang, Liyun; Kale, Anup Atul; Agrawalla, Bikram Keshari; Cho, Yoon-Kyoung; Chang, Young-Tae

2013-07-01

Caffeine has attracted abundant attention due to its extensive existence in beverages and medicines. However, to detect it sensitively and conveniently remains a challenge, especially in resource-limited regions. Here we report a novel aqueous phase fluorescent caffeine sensor named Caffeine Orange which exhibits 250-fold fluorescence enhancement upon caffeine activation and high selectivity. Nuclear magnetic resonance spectroscopy and Fourier transform infrared spectroscopy indicate that π-stacking and hydrogen-bonding contribute to their interactions while dynamic light scattering and transmission electron microscopy experiments demonstrate the change of Caffeine Orange ambient environment induces its fluorescence emission. To utilize this probe in real life, we developed a non-toxic caffeine detection kit and tested it for caffeine quantification in various beverages. Naked-eye sensing of various caffeine concentrations was possible based on color changes upon irradiation with a laser pointer. Lastly, we performed the whole system on a microfluidic device to make caffeine detection quick, sensitive and automated.

DNA derived fluorescent bio-dots for sensitive detection of mercury and silver ions in aqueous solution

Energy Technology Data Exchange (ETDEWEB)

Song, Ting [Laboratory of Environmental Science and Technology, Xinjiang Technical Institute of Physics & Chemistry, Key Laboratory of Functional Materials and Devices for Special Environments, Chinese Academy of Sciences, Urumqi 830011 (China); University of Chinese Academy of Sciences, Beijing 100049 (China); Zhu, Xuefeng, E-mail: zhuxf@ms.xjb.ac.cn [Laboratory of Environmental Science and Technology, Xinjiang Technical Institute of Physics & Chemistry, Key Laboratory of Functional Materials and Devices for Special Environments, Chinese Academy of Sciences, Urumqi 830011 (China); Zhou, Shenghai [Laboratory of Environmental Science and Technology, Xinjiang Technical Institute of Physics & Chemistry, Key Laboratory of Functional Materials and Devices for Special Environments, Chinese Academy of Sciences, Urumqi 830011 (China); Yang, Guang [Electronic Materials Research Laboratory, Key Laboratory of the Ministry of Education and International Center for Dielectric Research, Xi’an Jiaotong University, Xi’an 710049 (China); Gan, Wei [Laboratory of Environmental Science and Technology, Xinjiang Technical Institute of Physics & Chemistry, Key Laboratory of Functional Materials and Devices for Special Environments, Chinese Academy of Sciences, Urumqi 830011 (China); Yuan, Qunhui, E-mail: yuanqh@ms.xjb.ac.cn [Laboratory of Environmental Science and Technology, Xinjiang Technical Institute of Physics & Chemistry, Key Laboratory of Functional Materials and Devices for Special Environments, Chinese Academy of Sciences, Urumqi 830011 (China)

2015-08-30

Highlights: • First application of a DNA derived fluorescent bio-dot for metal sensing. • Bio-dot was conveniently obtained via a mild thermal hydro-thermal synthesis. • Bio-dot was directly used for fluorescent sensing without further modification. • Bio-dot showed good fluorescent sensing property for Hg(II) and Ag(I). • Formation of T–Hg–T and C–Ag–C structures played key roles in sensing. - Abstract: Inspired by the high affinity between heavy metal ions and bio-molecules as well as the low toxicity of carbon-based quantum dots, we demonstrated the first application of a DNA derived carbonaceous quantum dots, namely bio-dots, in metal ion sensing. The present DNA-derived bio-dots contain graphitic carbon layers with 0.242 nm lattice fringes, exhibit excellent fluorescence property and can be obtained via a facile hydrothermal preparation procedure. Hg(II) and Ag(I) are prone to be captured by the bio-dots due to the existence of residual thymine (T) and cytosine (C) groups, resulting in a quenched fluorescence while other heavy metal ions would cause negligible changes on the fluorescent signals of the bio-dots. The bio-dots could be used as highly selective toxic-free biosensors, with two detecting linear ranges of 0–0.5 μM and 0.5–6 μM for Hg(II) and one linear range of 0–10 μM for Ag(I). The detection limits (at a signal-to-noise ratio of 3) were estimated to be 48 nM for Hg(II) and 0.31 μM for Ag(I), respectively. The detection of Hg(II) and Ag(I) could also be realized in the real water sample analyses, with satisfying recoveries ranging from 87% to 100%.

DNA derived fluorescent bio-dots for sensitive detection of mercury and silver ions in aqueous solution

International Nuclear Information System (INIS)

Song, Ting; Zhu, Xuefeng; Zhou, Shenghai; Yang, Guang; Gan, Wei; Yuan, Qunhui

2015-01-01

Highlights: • First application of a DNA derived fluorescent bio-dot for metal sensing. • Bio-dot was conveniently obtained via a mild thermal hydro-thermal synthesis. • Bio-dot was directly used for fluorescent sensing without further modification. • Bio-dot showed good fluorescent sensing property for Hg(II) and Ag(I). • Formation of T–Hg–T and C–Ag–C structures played key roles in sensing. - Abstract: Inspired by the high affinity between heavy metal ions and bio-molecules as well as the low toxicity of carbon-based quantum dots, we demonstrated the first application of a DNA derived carbonaceous quantum dots, namely bio-dots, in metal ion sensing. The present DNA-derived bio-dots contain graphitic carbon layers with 0.242 nm lattice fringes, exhibit excellent fluorescence property and can be obtained via a facile hydrothermal preparation procedure. Hg(II) and Ag(I) are prone to be captured by the bio-dots due to the existence of residual thymine (T) and cytosine (C) groups, resulting in a quenched fluorescence while other heavy metal ions would cause negligible changes on the fluorescent signals of the bio-dots. The bio-dots could be used as highly selective toxic-free biosensors, with two detecting linear ranges of 0–0.5 μM and 0.5–6 μM for Hg(II) and one linear range of 0–10 μM for Ag(I). The detection limits (at a signal-to-noise ratio of 3) were estimated to be 48 nM for Hg(II) and 0.31 μM for Ag(I), respectively. The detection of Hg(II) and Ag(I) could also be realized in the real water sample analyses, with satisfying recoveries ranging from 87% to 100%

Diffusion affected magnetic field effect in exciplex fluorescence

International Nuclear Information System (INIS)

Burshtein, Anatoly I.; Ivanov, Anatoly I.

2014-01-01

The fluorescence of the exciplex, 1 [D +δ A −δ ], formed at contact of photoexcited acceptor 1 A * with an electron donor 1 D, is known to be very sensitive to an external magnetic field, reducing the spin conversion efficiency in the resulting geminate radical ion pair, 1,3 [D + …A − ]. The relative increase of the exciplex fluorescence in the highest magnetic field compared to the lowest one, known as the magnetic field effect, crucially depends on the viscosity of the solvent. This phenomenon first studied experimentally is at first reproduced here theoretically. The magnetic field effect is shown to vanish in both limits of high and low solvent diffusivity reaching a maximum in between. It is also very sensitive to the solvent dielectric constant and to the exciplex and radical-ion pair conversion rates

Diffusion affected magnetic field effect in exciplex fluorescence

Science.gov (United States)

Burshtein, Anatoly I.; Ivanov, Anatoly I.

2014-07-01

The fluorescence of the exciplex, 1[D+δA-δ], formed at contact of photoexcited acceptor 1A* with an electron donor 1D, is known to be very sensitive to an external magnetic field, reducing the spin conversion efficiency in the resulting geminate radical ion pair, 1, 3[D+…A-]. The relative increase of the exciplex fluorescence in the highest magnetic field compared to the lowest one, known as the magnetic field effect, crucially depends on the viscosity of the solvent. This phenomenon first studied experimentally is at first reproduced here theoretically. The magnetic field effect is shown to vanish in both limits of high and low solvent diffusivity reaching a maximum in between. It is also very sensitive to the solvent dielectric constant and to the exciplex and radical-ion pair conversion rates.

Fluorescent and Colorimetric Electrospun Nanofibers for Heavy-Metal Sensing

Directory of Open Access Journals (Sweden)

Idelma A. A. Terra

2017-12-01

Full Text Available The accumulation of heavy metals in the human body and/or in the environment can be highly deleterious for mankind, and currently, considerable efforts have been made to develop reliable and sensitive techniques for their detection. Among the detection methods, chemical sensors appear as a promising technology, with emphasis on systems employing optically active nanofibers. Such nanofibers can be obtained by the electrospinning technique, and further functionalized with optically active chromophores such as dyes, conjugated polymers, carbon-based nanomaterials and nanoparticles, in order to produce fluorescent and colorimetric nanofibers. In this review we survey recent investigations reporting the use of optically active electrospun nanofibers in sensors aiming at the specific detection of heavy metals using colorimetry and fluorescence methods. The examples given in this review article provide sufficient evidence of the potential of optically electrospun nanofibers as a valid approach to fabricate highly selective and sensitive optical sensors for fast and low-cost detection of heavy metals.

Determination of acrolein in serum by high-performance liquid chromatography with fluorescence detection after pre-column fluorogenic derivatization using 1,2-diamino-4,5-dimethoxybenzene.

Science.gov (United States)

Imazato, Takahiro; Kanematsu, Mariko; Kishikawa, Naoya; Ohyama, Kaname; Hino, Takako; Ueki, Yukitaka; Maehata, Eisuke; Kuroda, Naotaka

2015-09-01

Acrolein is a major unsaturated aldehyde that is generated during the lipid peroxidation process. The measurement of acrolein in biological samples should be useful to estimate the degree of lipid peroxidation and to evaluate the effect of hazardous properties of acrolein on human health. In this study, a highly sensitive and selective high-performance liquid chromatography with fluorescence detection method was developed for the determination of acrolein in human serum. The proposed method involves the pre-column fluorogenic derivatization of acrolein with 1,2-diamino-4,5-dimethoxybenzene (DDB) as a reagent. The fluorescent derivative of acrolein could be detected clearly without any interfering reagent blank peaks because DDB does not have intrinsic fluorescence itself, and the detection limit was 10 nM (signal-to-noise ratio = 3). The proposed method could selectively detect acrolein in human serum with a simple protein precipitation treatment. Copyright © 2015 John Wiley & Sons, Ltd.

Synthesis of Novel Fluorescent Sensors Based on Naphthalimide Fluorophores for the Highly Selective Hg2+-Sensing

Directory of Open Access Journals (Sweden)

Yordkhuan Tachapermpon

2015-01-01

Full Text Available With an aim to develop the new sensors for optical detection of Hg2+ ions, two novel fluorometric sensors were designed and successfully prepared using 2-(3-(2-aminoethylsulfanylpropylsulfanylethanamine and one or two N-methylnaphthalimide moieties (1 and 2. Sensor 1 was obtained via N-alkylation, N-imidation and a one-pot nucleophilic aromatic substitution, and N-formylation of the amine, while sensor 2 was prepared via N-alkylation, N-imidation, and nucleophilic aromatic substitution. The characterization, including 1H NMR, 13C NMR, and mass spectrometry, was then performed for 1 and 2. The Hg2+-binding behaviors of the sensors were investigated in terms of sensitivity and selectivity by fluorescence spectroscopy. Sensor 1 especially provided the reversible and highly Hg2+-selective ON-OFF fluorescence behavior by discriminating various interfering ions such as Pb2+, Co2+, Cd2+, Mn2+, Fe2+, K+, Na+, and in particular Cu2+ and Ag+ with a detection limit of 22 ppb toward Hg2+ ions.

Fluorescent probes and nanoparticles for intracellular sensing of pH values

Science.gov (United States)

Shi, Wen; Li, Xiaohua; Ma, Huimin

2014-12-01

Intracellular pH regulates a number of cell metabolism processes and its sensing is thus of great importance for cell studies. Among various methods, fluorescent probes have been widely used for sensing intracellular pH values because of their high sensitivity and spatiotemporal resolution capability. In this article, the development of fluorescent probes with good practicability in sensing intracellular pH values and pH variation during 2009 - 2014 is reviewed. These fluorescence probes are divided into two kinds: small molecules and nanoparticles. Photophysical properties, advantages/disadvantages and applications of the two kinds of probes are discussed in detail.

Fluorescence background subtraction technique for hybrid fluorescence molecular tomography/x-ray computed tomography imaging of a mouse model of early stage lung cancer.

Science.gov (United States)

Ale, Angelique; Ermolayev, Vladimir; Deliolanis, Nikolaos C; Ntziachristos, Vasilis

2013-05-01

The ability to visualize early stage lung cancer is important in the study of biomarkers and targeting agents that could lead to earlier diagnosis. The recent development of hybrid free-space 360-deg fluorescence molecular tomography (FMT) and x-ray computed tomography (XCT) imaging yields a superior optical imaging modality for three-dimensional small animal fluorescence imaging over stand-alone optical systems. Imaging accuracy was improved by using XCT information in the fluorescence reconstruction method. Despite this progress, the detection sensitivity of targeted fluorescence agents remains limited by nonspecific background accumulation of the fluorochrome employed, which complicates early detection of murine cancers. Therefore we examine whether x-ray CT information and bulk fluorescence detection can be combined to increase detection sensitivity. Correspondingly, we research the performance of a data-driven fluorescence background estimator employed for subtraction of background fluorescence from acquisition data. Using mice containing known fluorochromes ex vivo, we demonstrate the reduction of background signals from reconstructed images and sensitivity improvements. Finally, by applying the method to in vivo data from K-ras transgenic mice developing lung cancer, we find small tumors at an early stage compared with reconstructions performed using raw data. We conclude with the benefits of employing fluorescence subtraction in hybrid FMT-XCT for early detection studies.

Native fluorescence detection of biomolecular and pharmaceutical compounds in capillary electrophoresis: detector designs, performance and applications: A review

NARCIS (Netherlands)

de Kort, B.J.; de Jong, G.J.; Somsen, G.W.

2013-01-01

This review treats the coupling of capillary electrophoresis (CE) with fluorescence detection (Flu) for the analysis of natively fluorescent biomolecular and pharmaceutical compounds. CE-Flu combines the excellent separation efficiency of CE with the high selectivity and sensitivity of Flu. In

[A cell-based detection of ciguatoxin using sodium fluorescence probe].

Science.gov (United States)

Yuan, Jian-hui; Yang, Hui; Tang, Huan-wen; Huang, Wei; Xu, Xin-yun; Liu, Jian-jun; Ke, Yue-bin; Cheng, Jin-quan; Zhuang, Zhi-xiong

2011-04-01

To establish a cell-based detection method of ciguatoxin using fluorescence assay. Mouse neuroblastoma N-2A cells were exposed to ouabain and veratridine and different concentrations of standard ciguatoxin samples (P-CTX-1) to establish the curvilinear relationship between the toxin dosage and fluorescence intensity using the sodium fluorescence probe CoroNaTM Green. The toxicity curvilinear relationship was also generated between the toxin dosage and cell survival using CCK-8 method. Based on these standard curves, the presence of ciguatoxin was detected in 33 samples of deep-sea coral fish. A correlation was found between the detection results of cell-based fluorescence assay and cytotoxicity assay, whose detection limit reached 103 g/ml and 1012 g/ml, respectively. The cell-based fluorescent assay sensitivity showed a higher sensitivity than cytotoxicity assay with a 2-4 h reduction of the detection time. The cell-based fluorescent assay can quickly and sensitively detect ciguatoxin and may serve as a good option for preliminary screening of the toxin.

A high resolution solar atlas for fluorescence calculations

Science.gov (United States)

Hearn, M. F.; Ohlmacher, J. T.; Schleicher, D. G.

1983-01-01

The characteristics required of a solar atlas to be used for studying the fluorescence process in comets are examined. Several sources of low resolution data were combined to provide an absolutely calibrated spectrum from 2250 A to 7000A. Three different sources of high resolution data were also used to cover this same spectral range. The low resolution data were then used to put each high resolution spectrum on an absolute scale. The three high resolution spectra were then combined in their overlap regions to produce a single, absolutely calibrated high resolution spectrum over the entire spectral range.

Green synthesis of highly fluorescent carbon quantum dots from sugarcane bagasse pulp

Energy Technology Data Exchange (ETDEWEB)

Thambiraj, S. [Nano-Bio Materials and Sensors Laboratory, PSG Institute of Advanced Studies, Coimbatore, 641 004, Tamil Nadu (India); Ravi Shankaran, D., E-mail: dravishankaran@hotmail.com [Nano-Bio Materials and Sensors Laboratory, PSG Institute of Advanced Studies, Coimbatore, 641 004, Tamil Nadu (India); National Centre for Nanoscience and Nanotechnology, University of Madras, Guindy Campus, Chennai, 600 025, Tamil Nadu (India)

2016-12-30

Graphical abstract: Schematic representation of CQDs from sugarcane bagasse carbon. - Highlights: • CQDs were synthesised from sugarcane bagasse waste with top down approaches. • Synthesis method is green, simple and efficient process. • CQDs possess high quantum yield, good stability and highly fluorescent in nature. • The morphological and topographical study of CQDs was done by HR-TEM and AFM and was observed that the average size is 4.1 ± 0.17 nm and surface thickness is 5 nm. - Abstract: Carbon quantum dots (CQDs) have great potential due to its advantageous characteristics of highly fluorescent nature and good stability. In this study, we aimed to develop a simple and efficient method for the green synthesis of fluorescent CQDs from sugarcane bagasse, a renewable and sustainable resource. The process involves the top down approach of chemical oxidation followed by exfoliation of sugarcane carbon. The synthesized CQDs was characterized by UV–vis absorption spectroscopy, Spectrofluorophotometry, Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), Raman spectroscopy, X-ray photon spectroscopy (XPS), Atomic force microscopy (AFM) and High-resolution transmission electron microscopy (HR-TEM). The synthesized CQDs possess stable fluorescent properties, good bio-compatibility and high quantum yield. The CQDs are highly crystalline with longitudinal dimensions of 4.1 ± 0.17 nm with an average roughness of around 5 nm. The XRD and TEM analysis indicates that the synthesized CQDs possess face centred cubic crystal structure. The results suggest that the proposed CQDs could be utilized for bio-sensor, bio-imaging and drug delivery applications.

Super-nonlinear fluorescence microscopy for high-contrast deep tissue imaging

Science.gov (United States)

Wei, Lu; Zhu, Xinxin; Chen, Zhixing; Min, Wei

2014-02-01

Two-photon excited fluorescence microscopy (TPFM) offers the highest penetration depth with subcellular resolution in light microscopy, due to its unique advantage of nonlinear excitation. However, a fundamental imaging-depth limit, accompanied by a vanishing signal-to-background contrast, still exists for TPFM when imaging deep into scattering samples. Formally, the focusing depth, at which the in-focus signal and the out-of-focus background are equal to each other, is defined as the fundamental imaging-depth limit. To go beyond this imaging-depth limit of TPFM, we report a new class of super-nonlinear fluorescence microscopy for high-contrast deep tissue imaging, including multiphoton activation and imaging (MPAI) harnessing novel photo-activatable fluorophores, stimulated emission reduced fluorescence (SERF) microscopy by adding a weak laser beam for stimulated emission, and two-photon induced focal saturation imaging with preferential depletion of ground-state fluorophores at focus. The resulting image contrasts all exhibit a higher-order (third- or fourth- order) nonlinear signal dependence on laser intensity than that in the standard TPFM. Both the physical principles and the imaging demonstrations will be provided for each super-nonlinear microscopy. In all these techniques, the created super-nonlinearity significantly enhances the imaging contrast and concurrently extends the imaging depth-limit of TPFM. Conceptually different from conventional multiphoton processes mediated by virtual states, our strategy constitutes a new class of fluorescence microscopy where high-order nonlinearity is mediated by real population transfer.

Peptide-assembled graphene oxide as fluorescent turn-on sensor for ultrasensitive Lipopolysaccharide (Endotoxin detection

Directory of Open Access Journals (Sweden)

Seng Koon Lim

2014-06-01

Full Text Available Introduction: Lipopolysaccharide (LPS, or endotoxin, a major component in the outer cell membrane of Gram-negative bacteria is a very powerful and toxic inflammatory stimulator, resulting in sepsis or septic shock, a significant medical problem affecting about 700 000 patients and causing 250 000 casualties annually in the United States itself. The detection of LPS is highly importance. However, the currently used enzymatic limulus amebocyte lysate assay is highly susceptible to changes in temperature and pH, interference factors, and requires cumbersome sample preparation. A more cost-effective, sensitive and robust detection method is needed. Objective: To design and develop biosensor for LPS detection by assembling a LPS-binding peptide (as LPS receptor with graphene oxide (GO, as fluorescence quencher. Methods: GO was synthesized using a modified Hummer’s method. A synthetic LPS-binding peptide was designed, fluorescent labelled, and assembled with GO in PBS buffer solution. The fluorescence recovery of the peptide-GO was measured upon addition of LPS from Gram negative bacteria: E. coli, K. pneumoniae, Samonella Thyphosa, P. aeruginosa, as well as living pathogenic bacteria. Specificity tests were conducted with various biological molecules to evaluate the sensing performance. Results & Discussion: Specific binding of LPS with peptide release the peptides from GO, resulting in fluorescence recovery, allowing ultrasensitive detection of LPS with the limit of detection of 130 pM, the most sensitive synthetic LPS sensors to-date. The LPS sensor is highly selective to LPS than other biological species. Conclusion: We developed a peptide-GO assembled fluorescence sensor for ultrasensitive and specific LPS/endotoxin detection. This is the most sensitive synthetic LPS sensor reported in the world.

Bloch surface wave structures for high sensitivity detection and compact waveguiding

Science.gov (United States)

Khan, Muhammad Umar; Corbett, Brian

2016-01-01

Resonant propagating waves created on the surface of a dielectric multilayer stack, called Bloch surface waves (BSW), can be designed for high sensitivity monitoring of the adjacent refractive index as an alternative platform to the metal-based surface plasmon resonance (SPR) sensing. The resonant wavelength and polarization can be designed by engineering of the dielectric layers unlike the fixed resonance of SPR, while the wide bandwidth low loss of dielectrics permits sharper resonances, longer propagation lengths and thus their use in waveguiding devices. The transparency of the dielectrics allows the excitation and monitoring of surface-bound fluorescent molecules. We review the recent developments in this technology. We show the advantages that can be obtained by using high index contrast layered structures. Operating at 1550 nm wavelengths will allow the BSW sensors to be implemented in the silicon photonics platform where active waveguiding can be used in the realization of compact planar integrated circuits for multi-parameter sensing.

Optimization of tube parameters in a tube excited X-ray fluorescence (TEXRF) system using secondary fluorescers