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https://www.readbyqxmd.com/read/28545043/a-multiplex-taqman-qpcr-assay-for-sensitive-and-rapid-detection-of-phytoplasmas-infecting-rubus-species
#1
Holger Linck, Erika Krüger, Annette Reineke
Rubus stunt is an economically important disease in the production of raspberries, blackberries, and loganberries. A fast, sensitive, and reliable diagnosis of phytoplasmas, the causal agent of the disease, is of prime importance to stop its spread by vegetative propagation and by insect vectors. Therefore, multiplex qPCR assays using TaqMan probes with different kinds of fluorophores in one reaction were developed, allowing the detection of phytoplasmas in general as well as a more specific detection of phytoplasmas belonging to group 16SrV and host DNA (either plant or insect)...
2017: PloS One
https://www.readbyqxmd.com/read/28544971/development-of-photostable-fluorophores-for-molecular-imaging
#2
REVIEW
Qinsi Zheng, Luke D Lavis
Advances in fluorescence microscopy promise to unlock details of biological systems with high spatiotemporal precision. These new techniques also place a heavy demand on the 'photon budget'-the number of photons one can extract from a sample. Improving the photostability of small molecule fluorophores using chemistry is a straightforward method for increasing the photon budget. Here, we review the (sometimes sparse) efforts to understand the mechanism of fluorophore photobleaching and recent advances to improve photostability through reducing the propensity for oxidation or through intramolecular triplet-state quenching...
May 22, 2017: Current Opinion in Chemical Biology
https://www.readbyqxmd.com/read/28542243/covalent-dye-attachment-influences-the-dynamics-and-conformational-properties-of-flexible-peptides
#3
Manuel P Luitz, Anders Barth, Alvaro H Crevenna, Rainer Bomblies, Don C Lamb, Martin Zacharias
Fluorescence spectroscopy techniques like Förster resonance energy transfer (FRET) and fluorescence correlation spectroscopy (FCS) have become important tools for the in vitro and in vivo investigation of conformational dynamics in biomolecules. These methods rely on the distance-dependent quenching of the fluorescence signal of a donor fluorophore either by a fluorescent acceptor fluorophore (FRET) or a non-fluorescent quencher, as used in FCS with photoinduced electron transfer (PET). The attachment of fluorophores to the molecule of interest can potentially alter the molecular properties and may affect the relevant conformational states and dynamics especially of flexible biomolecules like intrinsically disordered proteins (IDP)...
2017: PloS One
https://www.readbyqxmd.com/read/28542211/f%C3%A3-rster-resonance-energy-transfer-role-of-diffusion-of-fluorophore-orientation-and-separation-in-observed-shifts-of-fret-efficiency
#4
Bram Wallace, Paul J Atzberger
Förster resonance energy transfer (FRET) is a widely used single-molecule technique for measuring nanoscale distances from changes in the non-radiative transfer of energy between donor and acceptor fluorophores. For macromolecules and complexes this observed transfer efficiency is used to infer changes in molecular conformation under differing experimental conditions. However, sometimes shifts are observed in the FRET efficiency even when there is strong experimental evidence that the molecular conformational state is unchanged...
2017: PloS One
https://www.readbyqxmd.com/read/28540954/hydrazone-switches-and-things-in-between
#5
Ivan Aprahamian
This feature article surveys the various ways by which a structurally simple hydrazone can be used in accessing different functional materials, mainly photo/chemically activated switches, fluorophores and sensors. The modularity of this functional group enabled us to develop new switching mechanisms, pathways, cascades and loops, while addressing challenges in the field, such as waste accumulation, how to control cross-talk between different types of switches, developing photochromic compounds with benign activation wavelengths, among others...
May 25, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28540652/development-of-low-cost-instrumentation-for-single-point-autofluorescence-lifetime-measurements
#6
João Lagarto, Jonathan D Hares, Christopher Dunsby, Paul M W French
Autofluorescence lifetime measurements, which can provide label-free readouts in biological tissues, contrasting e.g. different types and states of tissue matrix components and different cellular metabolites, may have significant clinical potential for diagnosis and to provide surgical guidance. However, the cost of the instrumentation typically used currently presents a barrier to wider implementation. We describe a low-cost single point time-resolved autofluorescence instrument, exploiting modulated laser diodes for excitation and FPGA-based circuitry for detection, together with a custom constant fraction discriminator...
May 25, 2017: Journal of Fluorescence
https://www.readbyqxmd.com/read/28537289/a-novel-single-fluorophore-based-ratiometric-fluorescent-probe-for-direct-detection-of-isocyanates-in-air
#7
Zhenzhong Gao, Baichuan Han, Kai Chen, Jin Sun, Xianfeng Hou
A ratiometric fluorescent probe based on anthracene carboximide was developed for direct detection of isocyanates in air. The sensing mechanism was illustrated to be variation in the photophysical properties caused by the nucleophilic reaction of the probe molecule. A number of isocyanates were conveniently detected by a test paper.
May 24, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28536662/identification-of-abnormal-biliary-anatomy-utilizing-real-time-near-infrared-cholangiography-a-report-of-two-cases
#8
Joseph Bozzay, Diego Vicente, Elliot M Jessie, Carlos J Rodriguez
Biliary duct anomalies are commonly encountered during laparoscopic cholecystectomy. Advancements in the field of surgery allow for enhanced intraoperative detection of these abnormalities. Fluorophore injection and near-infrared (NIR) imaging can provide real-time intraoperative anatomic feedback without intraoperative delays and ionizing radiation. This report details two cases where the PINPOINT Endoscopic Fluorescence Imaging System (NOVADAQ, Ontario, Canada) was used to identify anomalies of the biliary tree and guide operative decision-making...
2017: Case Reports in Gastrointestinal Medicine
https://www.readbyqxmd.com/read/28535451/a-novel-carbohydrate-labeling-method-utilizing-transfer-hydrogenation-mediated-reductive-amination
#9
Zsuzsanna Kovács, Gábor Papp, Henrietta Horváth, Ferenc Joó, András Guttman
One of the most frequently used high-resolution glycan analysis methods in the biopharmaceutical and biomedical fields is capillary electrophoresis with laser-induced fluorescence (CE-LIF) detection. Glycans are usually labeled by reductive amination with a charged fluorophore containing a primary amine, which reacts with the aldehyde group at the reducing end of the glycan structures. In this reaction, first a Schiff base is formed that is reduced to form a stable conjugate by a hydrogenation reagent, such as sodium cyanoborohydride...
May 13, 2017: Journal of Pharmaceutical and Biomedical Analysis
https://www.readbyqxmd.com/read/28535395/a-zinc-chloride-doped-adhesive-facilitates-sealing-at-the-dentin-interface-a-confocal-laser-microscopy-study
#10
Manuel Toledano, Raquel Osorio, Estrella Osorio, Inmaculada Cabello, Manuel Toledano-Osorio, Fátima S Aguilera
The aim of this study was to ascertain the effect of Zn-doping of dental adhesives and mechanical load cycling on the micromorphology of the resin-dentin interdiffusion zone (of sound and caries affected dentin). The investigation considered two different Zn-doped adhesive approaches and evaluated the interface using a doubled dye fluorescent technique and a calcium chelator fluorophore under a confocal laser scanning microscopy. Sound and carious dentin-resin interfaces of unloaded specimens were deficiently resin-hybridized, in general...
May 19, 2017: Journal of the Mechanical Behavior of Biomedical Materials
https://www.readbyqxmd.com/read/28533371/blue-fluorescent-amino-acid-for-biological-spectroscopy-and-microscopy
#11
Mary Rose Hilaire, Ismail A Ahmed, Chun-Wei Lin, Hyunil Jo, William F DeGrado, Feng Gai
Many fluorescent proteins are currently available for biological spectroscopy and imaging measurements, allowing a wide range of biochemical and biophysical processes and interactions to be studied at various length scales. However, in applications where a small fluorescence reporter is required or desirable, the choice of fluorophores is rather limited. As such, continued effort has been devoted to the development of amino acid-based fluorophores that do not require a specific environment and additional time to mature and have a large fluorescence quantum yield, long fluorescence lifetime, good photostability, and an emission spectrum in the visible region...
May 22, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28530289/ca-nir-a-ratiometric-near-infrared-calcium-probe-based-on-a-dihydroxanthene-hemicyanine-fluorophore
#12
Mayeul Collot, Flavien Ponsot, Andrey S Klymchenko
Fluorescent calcium probes are essential tools for studying the fluctuation of calcium ions in cells. Herein, we developed Ca-NIR, the first ratiometric calcium probe emitting in the near infrared region. This probe arose from the fusion of a BAPTA chelator and a dihydroxanthene-hemicyanine fluorophore. It is efficiently excited with common 630-640 nm lasers and displays two distinct emission bands depending on the calcium concentration (Kd = ∼8 μM). The physicochemical and spectroscopic properties of Ca-NIR allowed for ratiometric imaging of calcium distribution in live cells...
May 22, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28529973/high-affinity-mannotetraose-as-an-alternative-to-dextran-in-cona-based-fluorescent-affinity-glucose-assay-due-to-improved-fret-efficiency
#13
Andrea K Locke, Brian M Cummins, Gerard L Coté
Diabetes mellitus affects millions of people worldwide and requires that individuals tightly self-regulate their blood glucose levels to minimize the associated secondary complications. Continuous monitoring devices potentially offer patients a long-term means to tightly monitor their glucose levels. In recent years, fluorescent affinity sensors based on lectins (e.g., Concanavalin A (ConA)) have been implemented in such devices. Traditionally, these sensors pair the lectin with a multivalent ligand, like dextran, in order to develop a competitive binding assay that changes its fluorescent properties in response to the surrounding glucose concentrations...
May 27, 2016: ACS Sensors
https://www.readbyqxmd.com/read/28529382/reversibly-manipulating-the-surface-chemistry-of-polymeric-nanostructures-via-a-grafting-to-approach-mediated-by-nucleobase-interactions
#14
Zan Hua, Robert Keogh, Zhen Li, Thomas R Wilks, Guosong Chen, Rachel K O'Reilly
"Grafting to" polymeric nanostructures or surfaces is a simple and versatile approach to achieve functionalization. Herein, we describe the formation of mixed polymer-grafted nanoparticles through a supramolecular "grafting to" method that exploits multiple hydrogen-bonding interactions between the thymine (T)-containing cores of preformed micelles and the complementary nucleobase adenine (A) of added diblock copolymers. To demonstrate this new "grafting to" approach, mixed-corona polymeric nanoparticles with different sizes were prepared by the addition of a series of complementary diblock copolymers containing thermoresponsive poly(N-isopropylacrylamide) (PNIPAM) to a preformed micelle with a different coronal forming block, poly(4-acryloylmorpholine) (PNAM)...
May 9, 2017: Macromolecules
https://www.readbyqxmd.com/read/28528641/preserving-the-photoswitching-ability-of-standard-fluorescent-proteins-for-correlative-in-resin-super-resolution-and-electron-microscopy
#15
Errin Johnson, Rainer Kaufmann
There are many different correlative light and electron microscopy (CLEM) techniques available. The use of super-resolution microscopy in CLEM is an emerging application and while offering the obvious advantages of improved resolution in the fluorescence image, and therefore more precise correlation to electron microscopy (EM) ultrastructure, it also presents new challenges. Choice of fluorophore, method of fixation, and timing of the fluorescence imaging are critical to the success of super-resolution CLEM and the relative importance, and technical difficulty, of each of these factors depends on the type of super-resolution microscopy being employed...
2017: Methods in Cell Biology
https://www.readbyqxmd.com/read/28528634/3d-subcellular-localization-with-superresolution-array-tomography-on-ultrathin-sections-of-various-species
#16
Sebastian M Markert, Vivien Bauer, Thomas S Muenz, Nicola G Jones, Frederik Helmprobst, Sebastian Britz, Markus Sauer, Wolfgang Rössler, Markus Engstler, Christian Stigloher
Array Tomography (AT) is a relatively easy-to-use and yet powerful method to put molecular identity in its full ultrastructural context. Ultrathin sections are stained with fluorophores and then imaged by light and afterward by electron microscopy to obtain a correlated view of a region of interest: its ultrastructure and specific staining. By combining AT with high-pressure freezing for superior structural preservation and superresolution light microscopy, even small subcellular structures can be mapped in 3D...
2017: Methods in Cell Biology
https://www.readbyqxmd.com/read/28527910/radiative-decay-engineering-8-coupled-emission-microscopy-for-lens-free-high-throughput-fluorescence-detection
#17
Liangfu Zhu, Ramachandram Badugu, Douguo Zhang, Ruxue Wang, Emiliano Descrovi, Joseph R Lakowicz
Fluorescence spectroscopy and imaging are now used throughout the biosciences. Fluorescence microscopes, spectrofluorometers, microwell plate readers and microarray imagers all use multiple optical components to collect, redirect and focus the emission onto single point or array imaging detectors. For almost all biological samples, except those with regular nanoscale features, emission occurs in all directions. With the exception of complex microscope objectives with large collection angles (NA ≤ 0.5), all these instruments collect only a small fraction of the total emission...
May 17, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/28527000/fluorescent-coelenteramide-containing-protein-as-a-color-bioindicator-for-low-dose-radiation-effects
#18
Alena S Petrova, Anna A Lukonina, Gennadii A Badun, Nadezhda S Kudryasheva
The study addresses the application of fluorescent coelenteramide-containing proteins as color bioindicators for radiotoxicity evaluation. Biological effects of chronic low-dose radiation are under investigation. Tritiated water (200 MBq/L) was used as a model source of low-intensive ionizing radiation of beta type. 'Discharged obelin,' product of bioluminescent reaction of marine coelenterate Obelia longissimi, was used as a representative of the coelenteramide-containing proteins. Coelenteramide, fluorophore of discharged obelin, is a photochemically active molecule; it produces fluorescence forms of different color...
May 19, 2017: Analytical and Bioanalytical Chemistry
https://www.readbyqxmd.com/read/28526819/systematic-comparison-of-2a-peptides-for-cloning-multi-genes-in-a-polycistronic-vector
#19
Ziqing Liu, Olivia Chen, J Blake Joseph Wall, Michael Zheng, Yang Zhou, Li Wang, Haley Ruth Vaseghi, Li Qian, Jiandong Liu
Cloning of multiple genes in a single vector has greatly facilitated both basic and translational studies that require co-expression of multiple factors or multi-units of complex protein. Many strategies have been adopted, among which 2A "self-cleaving" peptides have garnered increased interest for their polycistronic nature, small size and high "cleavage" efficiency. However, broad application of 2 A peptides is limited by the lack of systematic comparison of different 2As alone or in combination. Here we characterized the effect of varying gene position and 2As on the expression of proteins encoded in bi-, tri-, or quad-cistronic constructs...
May 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28525664/near-infrared-fluorescence-with-indocyanine-green-for-diagnostics-in-urology-initial-experience
#20
Andrey O Morozov, Yuri G Alyaev, Leonid M Rapoport, Dmitrii G Tsarichenko, Eugene A Bezrukov, Denis V Butnaru, Eugene S Sirota
INTRODUCTION: Fluorescence imaging with indocyanine green is used in urology for the detection of sentinel lymph nodes and identification of prostate margins in radical prostatectomy for delineation of resection zone and selective clamping of vessels in partial nephrectomy; for identification and evaluation of length of ureteral strictures; for assessment of perfusion and viability of anastomoses during reconstructive stage of cystectomy. Safety of this technique is proven, while its diagnostic value and usefulness is still controversial...
May 16, 2017: Urologia
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