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TY - JOUR. T1 - Engineering ESPT pathways based on structural analysis of LSSmKate red fluorescent proteins with large stokes shift. AU - Piatkevich, Kiryl D.. AU - Malashkevich, Vladimir N.. AU - Almo, Steven C.. AU - Verkhusha, Vladislav. PY - 2010/8/11. Y1 - 2010/8/11. N2 - LSSmKate1 and LSSmKate2 are monomeric red fluorescent proteins (RFPs) with large Stokes shifts (LSSs), which allows for efficient separation of absorbance and emission maxima, as well as for excitation with conventional two-photon laser sources. These LSSmKates differ by a single amino acid substitution at position 160 and exhibit absorbance maxima around 460 nm, corresponding to a neutral DsRed-like chromophore. However, excitation at 460 nm leads to fluorescence emission above 600 nm. Structures of LSSmKate1 and LSSmKate2, determined at resolutions of 2.0 and 1.5, respectively, revealed that the predominant DsRed-chromophore configurations are cis for LSSmKate1 but trans for LSSmKate2. Crystallographic and mutagenesis ...
Nitric Oxide (NO) is a highly-reactive radical gas that can modify a variety of cellular targets in both eukaryotes and bacteria. NO is produced endogenously by a wide variety of organisms: For example, as a cell-signaling molecule in mammals and bacteria via nitric oxide synthase (NOS) enzymes, and as a product of denitrification. As such, it is of great benefit to NO researchers to be able to sensitively detect intracellular NO and stable reactive nitrogen species (RNS) derived from NO. To this end, a protocol for fluorescent detection of intracellular NO/RNS in biofilm cultures of the Gram-positive pathogen Staphylococcus aureus has been optimized using the commercially-available cell-permeable fluorescent stain 4-Amino-5-Methylamino-2,7-Difluorofluorescein Diacetate (DAF-FM diacetate). This compound diffuses into cells and intracellular cleavage by esterase enzymes liberates weakly-fluorescent DAF-FM, which reacts with NO or other specific RNS to become highly fluorescent (Kojima et al., 1999).
Fluorescent Molecular Rotors are sensors of microenvironmental restriction that become fluorescent only if their rotation is constrained. It has been suggested that the change of fluorescence intensity is caused by the restriction of intramolecular rotational relaxation about the donor-acceptor bond of the fluorophores.Examples of molecular constraint include increased dye (aggregation)1, binding to antibodies2, or being trapped in the polymerization of actin3. There are also studies of membrane fluidity in endothelial cells under fluid shear stress4 where fluorescent molecular rotors are used.1. Bhattacharyya, A. et al., Indian J. Biochem. Biophys., 32, 442-446 (1995).2. Iwaki, T. et al., Biochem., 32, 7589-7592 (1993).3. Sawada, S. et al., Anal. Biochem., 204, 110-117 (1992).4. Farkas, D.L. and Leif R.C. (ed.), Optical diagnostics of living cells III, Proc SPIE 3291, 101-112 (2000
A red-emitting fluorescent probe was developed for the sensitive and selective detection of H2S. Upon treatment with H2S, this probe exhibited a remarkable fluorescence enhancement (10 fold) with a large Stokes shift (125 nm). The detection limit of this probe was as low as 5.7 nM based on S/N = 3. The appli
We present a technique for observing single fluorophore molecules in solution. A mode-locked laser beam is focused into the solution, thereby defining a two-photon excitation volume localized in three dimensions. Molecules diffusing into and out of this volume produce fluorescence bursts, which are detected with a high signal-to-background ratio. The theoretical foundations for the technique are laid out, including the attendant fluorescence rate distribution, and agree with experimental results obtained for Rhodamine B molecules in water.. © 1995 Optical Society of America. Full Article , PDF Article ...
Studying the implication of hydrogen peroxide in biological processes in plants remains a challenge due to the current shortcomings of H2O2-responsive probes. The use of ContPY1, a new fluorescent probe, which is highly selective and sensitive for H2O2, was investigated. To validate the use of ContPY1 on plants, we have generated protocols employing cells suspensions and leaves, and measured specifically H2O2 production by plants using spectrofluorometry and microscopy. © 2013 Landes Bioscience. ...
ortho-Aminobenzoic acid (Abz) has been used as a convenient fluorescent donor group in internally quenched fluorescent peptides, which are employed as substrates for several proteolytic enzymes. As Abz is usually bound to the N-amino terminal of these peptides, it is of interest to investigate the Abz group fluorescent properties bound to different amino acids. We report in this article the optical absorption and fluorescent properties, in aqueous media, of Abz bound to the alpha-amino group of Ala, Gly, Leu, ne, Val, Pro, Phe, Arg, Glu, Met, Asn, Tyr, and Trp, with monomethyl-amidated alpha-carboxyl group, in order to explore the origin of the drastic reduction of Abz attached to N-alpha amino group of prolyl-peptides, we also examined the fluorescence properties of Abz-NHCH3, Abz-N(CH3)(2), and Abz-pyrrolidine. Molecular dynamics simulation and NMR data indicated a lack of periplanarity of the Abz-dimethylamide, which could be the origin of low fluorescence quantum yield of ...
We have engineered a mutant of HIV Reverse Transcriptase that can be fluorescently labeled by covalent attachment of the environmentally sensitive fluorophore 7-diethylamino-3-((((2-maleimidyl)ethyl)amino)carbonyl)coumarin (MDCC). The result is a polymerase that is kinetically indistinguishable from the wild-type enzyme, but provides a signal to monitor changes in enzyme structure that result from conformational changes induced by substrate binding. Using this system, we have expanded the kinetic model governing nucleotide binding to include an enzymatic isomerization following initial nucleotide binding. In doing so, we define the role of induced-fit in nucleotide specificity and mismatch discrimination. Additionally, we have characterized the kinetics governing the specificity and discrimination of several widely administered Nucleotide Reverse Transcriptase Inhibitors (NRTIs) used to combat HIV infection including 3TC (Lamivudine), FTC (Emtricitabine), and AZT (Zidovudine) for the wild-type ...
Modular fluorescent tags: clickable BODIPYs (co-supervisor Dr. Christoph Nitsche). New dyes for electron and energy transfer. Lighting up sugars - fluorescent probes for mono-saccharides.
Fluorescent Dyes , Reactive Fluorescent Dyes , HiLyte Fluor 594 hydrazide - TFA Salt; HiLyte Fluor 594, an alternative to Alexa Fluor 594 and DyLight Fluor 594, has spectral characteristics similar to those of Texas Red. The labeling performance and stability are better than those of Texas Red. It has high extinction coefficient (80,000 M-1cm-1), high fluorescence quantum yield (0.9) and low correction factor (0.17). HiLyte FluorTM 594 based Bioconjugates exhibit little spectral overlap with green-fluorescent conjugates, and can be efficiently excited by 568 nm line of Ar-Kr laser and by the 594 nm line of orange He-Ne laser. Extinction Coefficient (M-1cm-1): 80,000 Fluorescence quantum yield: 0.90 Fluorescence Life Time (ns): 4.2; HiLyte Fluor594 hydrazide is a carbonyl-reactive fluorescent labeling dye. It can be used for labeling glycoproteins such as HRP.
ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT AUTHORS REQUEST.] The design, synthesis and spectroscopic properties of several long wavelength/NIR fluorescent sensors are discussed. Amongst them, the metal complex-indicator sensors are developed based on the indicator-displacement-assay (IDA) strategy; the other BODIPY derived fluorescent sensors are developed based on the indicator-spacer-receptor (ISR) strategy. For the IDA sensors, one [PBA-Zn receptor + ARS] sensor showed some unique dual wavelength patterns when different sugar analytes were introduced. The [Cu complex + Acid Blue 45] sensors have NIR fluorescent properties and displayed high affinity towards various amino acids and warfarin. For the ISR BODIPY sensors, two of the pH responsive sensors have fluorescent emission at 600nm and displayed appropriate pKas for a physiological environment. One piperazine-BODIPY sensor with a novel structure showed a turn-on NIR fluorescent emission when protonated. One ...
TY - GEN. T1 - Spray pyrolysis synthesis of particles possessing magnetic and fluorescent properties. Application of magnetic/fluorescent particles in immunoassays. AU - Dosev, D.. AU - Nichkova, M.. AU - Dumas, R.. AU - Liu, K.. AU - Kennedy, I. M.. PY - 2005. Y1 - 2005. N2 - Many types of fluorescent nanoparticles have been synthesized as alternatives to organic dyes in biochemistry. Magnetic beads also have a long history of biological applications. In this work we apply flame spray pyrolysis in order to engineer a novel type of particle that has both fluorescent and magnetic properties. The particles have magnetic cores of iron oxide and a fluorescent shell of europium - doped gadolinium oxide (Eu:Gd 2O 3). Measurements on a Vibrating Sample Magnetometer showed an overall paramagnetic response behavior of the composite particles. Fluorescence spectroscopy showed fluorescent spectra typical for the Eu ion in a Gd 2O 3 host with a narrow emission peak centered near 615 nm. Our synthesis method ...
Your German Partner for Transfection, Gene Expression, Fluorescent Probes, IVD Kits, Antibodies, Peptides, Lipidomics, Glycobiology, Lab Supplies, and more.
Your German Partner for Transfection, Gene Expression, Fluorescent Probes, IVD Kits, Antibodies, Peptides, Lipidomics, Glycobiology, Lab Supplies, and more.
Fluorescent molecular imaging helped surgeons visually identify lung adenocarcinomas during pulmonary resection, according to study results.
Optimized for western blotting with fluorescent-conjugated antibodies. Invitrogen™ iBind™ Fluorescent Detection (FD) Solution Kit is intended for use with the iBind™ Flex Western Device (SLF2000) for use in infrared detection and dual wavelength semi-quantitative analysis. iBind Fluorescent Detection Solution Kit,X1 bottle,X5 buffer, 2 vials X100 additive, 1 vial ...
We present LIVE-PAINT, a new approach to super-resolution fluorescent imaging inside live cells. In LIVE-PAINT only a short peptide sequence is fused to the protein being studied, unlike conventional super-resolution methods, which rely on directly fusing the biomolecule of interest to a large fluorescent protein, organic fluorophore, or oligonucleotide. LIVE-PAINT works by observing the blinking of localized fluorescence as this peptide is reversibly bound by a protein that is fused to a fluorescent protein. We have demonstrated the effectiveness of LIVE-PAINT by imaging a number of different proteins inside live S. cerevisiae. Not only is LIVE-PAINT widely applicable, easily implemented, and the modifications minimally perturbing, but we also anticipate it will extend data acquisition times compared to those previously possible with methods that involve direct fusion to a fluorescent protein. Oi et al. describe LIVE-PAINT, a new method to achieve superresolution fluorescent imaging within live cells.
There is no simple answer to this question as the quantum yield of a fluorescent dye can vary widely, depending on the dyes micro-environment. For example, the quantum yield of a dye attached to a protein may be very different from the quantum yield of the free dye. For dyes attached to a protein, the quantum yield is highly dependent on how many molecules of the dye are attached to the protein (i.e. degree of protein labeling). In general, a higher degree of protein labeling leads to a lower dye quantum yield due to fluorescence quenching via dye-to-dye interaction. For this reason, as the degree of labeling increases, fluorescence intensity of the labeled protein will eventually reach a maximum and start to decline thereafter. In fact, one of the best ways to compare the relative quantum yields of different dyes is to plot the total fluorescence of the labeled proteins as a function of degree of labeling by the dyes as we have done with CF® dyes and other commercial dyes. CF® dyes generally ...
Single-molecule detection has long relied on fluorescent labeling with high quantum-yield fluorophores. Plasmon-enhanced detection circumvents the need for labeling by allowing direct optical detection of weakly emitting and completely nonfluorescent species. This review focuses on recent advances in single molecule detection using plasmonic metal nanostructures as a sensing platform, particularly using a single particle-single molecule approach. In the past decade two mechanisms for plasmon-enhanced single-molecule detection have been demonstrated: (1) by plasmonically enhancing the emission of weakly fluorescent biomolecules, or (2) by monitoring shifts of the plasmon resonance induced by single-molecule interactions. We begin with a motivation regarding the importance of single molecule detection, and advantages plasmonic detection offers. We describe both detection mechanisms and discuss challenges and potential solutions. We finalize by highlighting the exciting possibilities in analytical ...
GelRed, GelGreen, EvaGreen - Fluorescence dyes, EtBr substitute Apoptosis and necrosis assay kitsCell biology accessory products Nucleic acid stains for microbiology Enzyme substrates Membrane and organelle stains Qubit® kits - DNA, RNA quantification Spectrophotometers a fluorometers Consumables GelRed Nucleic Acid Gel Stain, 10.000 x in water Biot41003 119 €144,30 € incl. Btw […]. ...
Our fluorophore conjugated secondary antibodies are available in a concentrated liquid formats and many are also offered in a convenient prediluted, ready-to-use format. ...
Real-time PCR was performed using the ABI 7700 Sequence detector (Applied Biosystems) as described previously (Biecker et al., 2004). A dual-labeled fluorogenic probe (labeled with a reporter dye at the 5′-end and a second dye, quenching the emission of the reporter dye, at the 3′-end) complementary to a sequence within each PCR product was added to the PCR reaction. Cleavage of the probe during elongation by the exonuclease activity of the TaqDNA polymerase separates the reporter from its quencher. Accumulation of PCR products is detected in real time by monitoring the increase in fluorescence of the reporter dye. The PCR reaction was performed in a volume of 25 μl containing 12.5 μl2× TaqMan PCR master mix (Applied Biosystems) as well as 1.2 μl (equivalent to 300 ng of total RNA) of cDNA. The concentrations of the primers and probes are given in Table 1. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) provided as ready-to-use primers (100 nM each) and probe (200 nM) by the ...
Naturally occurring lipid granules diffuse in the cytoplasm and can be used as tracers to map out the viscoelastic landscape inside living cells. Using optical trapping and single particle tracking we found that lipid granules exhibit anomalous diffusion inside human umbilical vein endothelial cells. For these cells the exact diffusional pattern of a particular granule depends on the physiological state of the cell and on the localization of the granule within the cytoplasm. Granules located close to the actin rich periphery of the cell move less than those located towards to the center of the cell or within the nucleus. Also, granules in cells which are stressed by intense laser illumination or which have attached to a surface for a long period of time move in a more restricted fashion than those within healthy cells. For granules diffusing in healthy cells, in regions away from the cell periphery, occurrences of weak ergodicity breaking are observed, similar to the recent observations inside living
A high dynamic range apparatus for separation and detection of polynucleotide fragments has a housing adapted to receive an electrophoresis gel holder containing an electrophoresis gel loaded with fluorophore-labeled samples; one or more laser diodes for providing radiation of a frequency suitable for excitation of the fluorophore which irradiates a an array of excitation/detection sites on the electrophoresis gel; an array of detectors aligned with the excitation/detection sites for collecting fluorescent emissions; and one or more components for increasing the dynamic range of the instrument by at least an order of magnitude. These components, which can be used individually or in combination include detectors that are connected to a signal processing system that modulates the period of signal integration employed so that large signals are totaled at short time intervals and smaller signals are totaled at longer time intervals; the use of a beam splitter to produces a high intensity beam of emitted
Quinoxalinones are widely exploited for their biological activities, but more rarely for their fluorescence behavior, partly due to a lack of data. Herein, we investigated the photophysical properties of selected 3-benzoylquinoxalin-2-ones and their NaBH4-reduced products, obtained by the rearrangement of be
MIDORIGreen Xtra is a new highly sensitive green fluorescent stain for a safe visualization of DNA and RNA in agarose gels. This DNA stain is a safe and better alternative to the traditional nucleic acid stain ethidium bromide (EtBr). Remarkably, agarose gels stained with MIDORIGreen Xtra have a very low background fluorescence, which makes the identification of low amounts of DNA very easy. ...
A unique DNA-binding dye with features ideal for both qPCR and High Resolution Melting® (HRM) analysis*. EvaGreen® dye binds to dsDNA via a novel release-on-demand mechanism, which permits the use of a relatively high dye concentration in qPCR without PCR inhibition (Ref. 1).. A unique feature of EvaGreen® dye is its safety. DNA-binding dyes are inherently dangerous due to their potential to cause mutation. With this in mind, Biotiums scientists designed EvaGreen® dye such that it cannot cross cell membranes, thus preventing the dye from being in contact with genomic DNA in live cells. All other commercial PCR dyes enter into cells in a matter of minutes. SYBR® Green I, for example, has been shown to be environmentally more toxic than ethidium bromide, a well-known mutagen (Ref. 2). Independent labs have confirmed that EvaGreen dye is nonmutagenic, noncytotoxic and safe to aquatic life for direct disposal in the drain. For details, download the EvaGreen® Dye Safety Report.. An added ...
To overcome this issue of reduced resolution in STED imaging due to photodegradation, ITbMs team led by Principal Investigators Shigehiro Yamaguchi, a synthetic chemist and Tetsuya Higashiyama, a plant biologist have developed a new fluorescent dye, C-Naphox that has enhanced photostability relative to conventional dyes. C-Naphox has demonstrated to be extremely photoresistant with almost no degradation of fluorescence even after prolonged STED imaging in live cells.. C-Naphox (diarylmethylene-bridged naphthophosphole P-oxide) consists of an aromatic framework with an amino moiety incorporated for its electron-donating properties and phosphorus oxide for its electron-accepting properties, leading to intense fluorescence emissions.. Although the previously synthesized molecules in the Yamaguchi group have also led to high fluorescence intensity, it is the carbon-bridged structure in C-Naphox that is the key to its extremely high resistance to high intensity light, says Aiko Fukazawa, an ...
PTI RatioMaster is a monochromator-based wide-field microscope system for measuring ratio-fluorescence or fluorescence dye intensity of labeled proteins in nanomolar concentrations. Ratio-fluorescence microscopy is the ultimate tool to study dynamic events in live cells and cellular structures. Unlike conventional intensity based fluorescence microscopy, ratio-fluorescence eliminates the effects of photo-bleaching, cell path length variation, non-uniformity of dye loading, non-uniform field of view, making ratio-fluorescence the most accurate method to determine intracellular ion concentrations. More importantly, PTI RatioMaster is fast enough to provide essential dynamic information about ion concentrations and interactions with other molecules, drugs or added reagents. The PTI RatioMaster is capable of dynamic ratio fluorescence measurements on a millisecond timescale. A xenon arc lamp provides high intensity, continuous broadband illumination. Alternating excitation wavelengths are selected ...
Description DNA Dye Non-Toxic products from Biocrede represent a new and safe class of nucleic acid stains for the visualization of double-stranded DNA, singl
Fluorescence microscopy is ideal for intracellular ion measurements and the most common of these measurements is intracellular calcium imaging measurements. Two of the most popular dyes for calcium imaging are Fura-2 and Indo-1. PTI EasyRatioPro is a monochromator-based wide-field microscope system for measuring ratio-fluorescence or fluorescence dye intensity of labeled proteins in nanomolar concentrations. Ratio-fluorescence microscopy is the ultimate tool to study dynamic events in live cells and cellular structures. Unlike conventional intensity based fluorescence microscopy, ratio-fluorescence eliminates the effects of photo-bleaching, cell path length variation, non-uniformity of dye loading, non-uniform field of view, making ratio-fluorescence the most accurate method to determine intracellular ion concentrations. More importantly, PTI EasyRatioPro is fast enough to provide essential dynamic information about ion concentrations and interactions with other molecules, drugs or added ...
IHC staining was visualized with a fluorescent microscope (Olympus BX51), and images were taken with a digital camera (Olympus DP30BW) using appropriate filters for different fluorophores or bright-field illumination for DAB stain. Identical images were taken for double IHC, overlaid, and pseudocolored using Olympus Software and Adobe Photoshop (Adobe Systems). Contrast and brightness were adjusted for fluorescent signals with Adobe Photoshop (Adobe Systems) for better visualization of neurons.. For Ad-iZ/EGFPf tracing from LepRb neurons in the DMH, we analyzed four mice with correct injections into the DMH and showed representative images of projection sites in the mPOA, DMH, PVN, and rRPa in Figure 4.. For FG tracing experiments, we focused our analysis on the mPOA and DMH/DHA to investigate colocalization of LepRb neurons with FG-traced neurons from the RMR/rRPa (n = 4), DMH/DHA (n = 3), or PVN (n = 2). Representative images were shown for the DMH/DHA and mPOA, and, in some cases, we ...
A robust lipophilic dye, based on the structures of the benzothiadiazole heterocycle, was shown to be a potent fluorescent stain for the selective imaging of lipid droplets (LDs) within both live and fixed human cells. Its small molecular framework, large Stokes shift, and vastly improved photostability over that of the current status quo, Nile Red, highlight its tremendous potential as a versatile chemical tool for facilitating LD imaging and research.
Dual-channel fluorometer for personal quantitation workflow. Designed to provide highly sensitive fluorescent detection when quantifying nucleic acids, the compact instrument is simple to operate. The Quantus Fluorometer is optimized with preprogrammed settings for Promega QuantiFluor™ Dye Systems to quantitate nucleic acids and offers the flexibility to create customized methods and quantitation settings for other fluorescent dyes. ...
Method for rapid-base sequencing in DNA and RNA with two-base labeling and employing fluorescent detection of single molecules at two wavelengths. Bases modified to accept fluorescent labels are used
Chromeo 505 is a bright green fluorescent dye that replaces fluorescein, FITC, Alexa 500, Dylight 505 or other dyes that are excitable at 488 nm. Chromeo 505 and its conjugates are bright, photostable and compatible with standard fluorescent microscopes.
Solon, Ohio (PRWEB) May 06, 2013 -- DNA quantitation is crucial to experimental design and interpretation of results, especially in sensitive molecular biology
Chromeo 642 is a bright dark-red fluorescent dye that replaces Cy5, Alexa 647, DyLight 649 or other dyes that are excitable from 630 nm-650 nm. Chromeo 642 and its conjugates are bright, photostable and compatible with standard fluorescent microscopes.
Contrast agents play an important role in the study of biological tissues and whole organisms, since they enable visualization of functional structures. Fluorescent contrast agents also enable specific targeting in therapeutic approaches. Developing optimized contrast agents is central to optimizing the performance of both imaging and therapy. The ideal contrast agent for optical microscopy combines high resolution, specific targeting of functional groups, 3D imaging (depth resolution), low toxicity, low bleaching (long observation time at high signal), and high signal to noise (low background). Traditional organic dyes and fluorescent proteins are excited in the ultraviolet (UV) or blue spectral region, and emit at a longer-wavelength that is Stokes-shifted. The use of the short-wavelength excitation leads to a short penetration depth of the excitation light and give rise to autofluorescence, photobleaching and photodamage to biological specimens. It is thus primarily suited to pathological and ...
A toxin is used to introduce an otherwise cell-impermeant fluorophore-antibody (or some thing which is equally specific) to bind to an intracellular protein which allows for super resolution imaging and single particle tracking inside the living cell.
Current techniques for observing the cytoskeleton can be difficult to get into living cells, can be toxic, and are usually limited in resolution and duration, since the signal wears off over time. A common technique is fluorescence microscopy, where fluorescent molecules (probes) are attached to cell structures and then lit up against a dark background.. The team of Kai Johnsson at EPFL has developed novel fluorescent probes that can easily enter live cells, are non-toxic, have long-lasting signals, and most importantly, offer unprecedented image resolution. In 2013, the researchers developed a fluorescent molecule called silicon-rhodamine (SiR), which switches on only when it binds to the charged surface of a protein like the ones found on the cytoskeleton. When SiR switches on, it emits light at far-red wavelengths.. The challenge was getting SiR to bind specifically to the cytoskeletons proteins, actin and tubulin. To achieve this, the scientists fused SiR molecules with compounds ...
In this study we employed a common high-throughput P450 enzyme assay kit to evaluate the three SCs against a specific P450 activity. The recombinant enzyme system may be more costly and less representative of physiologic conditions, but it is a more consistent platform that avoids the wide variability in enzyme expression and activity normally encountered in HLM (Snawder and Lipscomb, 2000) and hepatocytes (Rodriguez-Antona et al., 2002; Westerink and Schoonen, 2007). Also, the enzyme systems are highly specific and relatively stable, with no significant loss in activity noted after 7 hours at room temperature (Trubetskoy et al., 2005b).. Fluorescent high-throughput screening methods employ fluorescent P450 substrates that are efficiently metabolized by specific P450 isozymes to yield a product with altered fluorescent properties, usually increased fluorescent intensity (Trubetskoy et al., 2005b). The assay requires only low reactant volume to produce high signal-to-background ratio, which ...
MilliporeSigma offers a versatile range of dyes for microscopy, including high-quality Certistain® dyes, fluorescence dyes, dry dyes and dye mixtures.
Ratiometric reporters are tools to dynamically measure the relative abundance of a protein of interest. In these systems, a target protein fused to a fluorescent or bioluminescent reporter is expressed with fixed stoichiometry to a reference protein fused to a second reporter. Both fusion proteins are encoded on a single transcript but are separated during translation by a 2A
Abnormal concentrations of ATP are associated with many diseases and cancers, and quantitative detection of ATP is thus of great importance for disease diagnosis and prognosis. In the present work, we report a new dual recycling amplification sensor integrated with catalytic hairpin assembly (CHA) to achieve high sensitivity for fluorescent detection of ATP. The association of the target ATP with the aptamer beacons causes the allosteric structure switching of the aptamer beacons to expose the toehold regions, which hybridize with and unfold the fluorescently quenched hairpin signal probes (HP1) to recycle the target ATP and to trigger CHA between HP1 and the secondary hairpin probes (HP2) to form HP1/HP2 duplexes ...
We have synthesized a novel functionalized pillar[5]arene (PC5) and used it for fluorescent detection of iron ions (Fe³⁺). It displays a specificity response for iron ions over other common cations (Hg²⁺, Co²⁺, Ca²⁺, Ni²⁺, Pb²⁺, Cd²⁺, Zn²⁺, Cr³⁺, Cu²⁺, Mg²⁺ and Ag⁺) in a solution of DMSO/THF (1 : 4, v/v). Competitive cations did not show any significant changes in emission intensity and the fluore ...
Currently there are no approved biomarkers for the pre-symptomatic diagnosis of Alzheimers disease (AD). Cathepsin-D (Cat-D) is a lysosomal protease that is present at elevated levels in amyloid plaques and neurons in patients with AD and is also elevated in some cancers. We have developed a magnetic resonance imaging (MRI)/fluorescent contrast agent to detect Cat-D enzymatic activity. The purpose of this study was to investigate the cellular and tissue uptake of this MRI/fluorescent contrast agent. The agent consists of an MRI probe [DOTA-caged metal ion (Gd3+ or Tm3+)] and a fluorescent probe coupled to a cell-penetrating-peptide sequence by a Cat-D recognition site. The relaxivity of Gd3+-DOTA-CAT(cleaved) was measured in 10% heat-treated bovine serum albumin (BSA) phantoms to assess contrast efficacy at magnetic fields ranging from 0.24 mT to 9.4 T. In vitro, Tm3+-DOTA-CAT was added to neuronal SN56 cells over-expressing Cat-D and live-cell confocal microscropy was performed at 30 min. ...
TY - JOUR. T1 - Myosin V walks hand-over-hand. T2 - Single fluorophore imaging with 1.5-nm localization. AU - Yildiz, Ahmet. AU - Forkey, Joseph N.. AU - McKinney, Sean A.. AU - Ha, Taekjip. AU - Goldman, Yale E.. AU - Selvin, Paul R.. PY - 2003/6/27. Y1 - 2003/6/27. N2 - Myosin V is a dimeric molecular motor that moves processively on actin, with the center of mass moving ±37 nanometers for each adenosine triphosphate hydrolyzed. We have labeled myosin V with a single fluorophore at different positions in the light-chain domain and measured the step size with a standard deviation of ,1.5 nanometers, with 0.5-second temporal resolution, and observation times of minutes. The step size alternates between 37 + 2x nm and 37 - 2x, where x is the distance along the direction of motion between the dye and the midpoint between the two heads. These results strongly support a hand-over-hand model of motility, not an inchworm model.. AB - Myosin V is a dimeric molecular motor that moves processively on ...
Fluorescent probes are useful tools to investigate specific subcellular components in cells, tissues and organisms. A classic application of fluorescent ligands of a specific GPCR (G-protein coupled receptors) is the investigation of the receptor location, and, if their binding is reversible, it could provide pharmacological information such as affinity and proximity between interacting molecules. In cultured cell systems, methods based on fluorescence have permitted to observe the receptor cycling and the formation of oligomeric receptor complexes. In high throughput screening, fluorescent ligands represents a safer, more powerful and more versatile alternative to radioligands. [1] The introduction of the bulky fluorophore into a small molecule ligand could lead to deep modifications not only from a physicochemical point of view but also in its pharmacological properties. [2] In order to allow a correct interaction of the pharmacophore with its receptor, it is usually separated from the ...
Single molecule fluorescence measurements are used to probe the effects of GM1 in DPPC monolayers. Langmuir-Blodgett films of GM1 and DPPC were doped with ~10−8 mol% of the fluorescent lipid probe, BODIPY-PC, and transferred onto glass substrates at 23 mN/m. As shown previously, the individual orientation of each BODIPY-PC probe in the membrane can be measured using defocused polarized total internal reflection fluorescence microscopy, revealing changes in film properties at the molecular level. Here, BODIPY-PC tilt angle histograms are used to characterize the effects of GM1 in DPPC films from 0.05 mol% to 100 mol% GM1. At high GM1 levels (,5 mol% GM1), trends in the single molecule measurements agree with previous bulk measurements showing the turnover from condensing to expanding influence of GM1 at ~20 mol%, thus validating the single molecule approach. At biologically relevant, low concentrations of GM1 (,5 mol% GM1), where bulk fluorescence measurements are less informative, the single ...
TMA-DPH (1-(4-trimethylammonium)-6-phenyl-1,3,5-hexatriene), a hydrophobic fluorescent membrane probe, interacts with living cells by instantaneous incorporation into the plasma membrane, where it becomes fluorescent. It then follows the intracellular constitutive membrane traffic and acts as a bulk membrane marker of the endocytic pathway (Illinger, D., P. Poindron, P. Fonteneau, M. Modolell, and J. G. Kuhry. 1990. Biochim. Biophys. Acta. 1030:73-81; Illinger, D., P. Poindron, and J. G. Kuhry. 1991. Biol. Cell. 73:131-138). As such, TMA-DPH displays particular properties mainly due to partition between membranes and aqueous media. From these properties, original arguments can be inferred in favor of the maturation model for the endocytic pathway, against that of pre-existing compartments, in L929 cultured mouse fibroblasts. (a) TMA-DPH labeling is seen to progress from the cell periphery to perinuclear regions during endocytosis without any noticeable loss in fluorescence intensity; with a ...
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Spectral properties of carbocyanine dye 3-methyl-2-[3-methyl-2-(3-methyl-2,3-dihydro-1,3-benzothiazole-2-iliden)-1- butenyl]-1,3-benzothiazole-3-il iodide (Cyan betaiPr) in water solution, as well as in the presence of different types of double stranded DNA have been studied. While in water solution of free dye Cyan betaiPr stays mainly in monomeric form, in the presence of DNA the dye molecules form J-aggregates. The molecular structure of these J-aggregates causes the Davydov splitting of their absorption band, corresponding to the first electronic transition. A study of site-specificity showed that in the presence of poly (dA/dT) the majority of Cyan betaiPr molecules form J-aggregates, while in the presence of poly (dGC/dGC) dye molecules stay mainly in monomeric form and in presence of chicken erythrocytes DNA both J-aggregate and monomeric forms of dye are present. We suppose that Cyan betaiPr molecules aggregate in DNA groove, which serves as a template for J-aggregate forming. An ...
Development of Functional Fluorescent Molecular Probes for the Detection of Biological Substances. . Biblioteca virtual para leer y descargar libros, documentos, trabajos y tesis universitarias en PDF. Material universiario, documentación y tareas realizadas por universitarios en nuestra biblioteca. Para descargar gratis y para leer online.
Calcium ions play an important role in cell function, acting as effectors and/or signaling molecules for a variety of cellular biochemical and physiological processes. The development of a variety of Ca2+-sensitive fluorescent indicators and advancements in imaging technology have enhanced the ability to follow both global and localized changes in intracellular Ca2+ at ever improving temporal and spatial resolution. Ratiometric Ca2+ indicators like fura-2 permit the measurement of intracellular Ca2+ concentration (Grynkiewicz et al., 1985). But, they tend to have a limited dynamic range and a significant fluorescence background, which reduce their sensitivity to small local increases in Ca2+. On the other hand, certain nonratiometric Ca2+ indicators, such as fluo-3, although not able to give a dynamic direct read-out of the Ca2+ concentration, have very low fluorescence when not bound to Ca2+ and have a significant increase in fluorescence intensity upon binding Ca2+ (e.g., ∼200 times for ...
N-Alkylation of a novel pyridine sensor results in pyridinium salts whose conformations are stabilised by pyridinium cation-π interactions resulting in a fluorescent response that can be used to sense the presence of alkylating agents in solution at low concentration.. ...
Fluorescence Dye 620-M with Streptavidin conjugate. All the Fluorescent Dye M series products are designed to be maximally excited by one of the major light sources equipped in flow cytometers. Besides, in comparison with phycobiliprotein tandems, Fluorescent Dye M series possesses better photostability, higher conjugation yields, and little pH sensitivity making it an ideal choice in fluorescence imaging applications. (U0293) - Products - Abnova
FDSS is a kinetic plate reader in Fluorescence and Luminescence, and suitable for reading the kinetics of the living cells which those response, calcium flux, mobilization for example, is very fast and need to monitor its kinetic property. GPCR is one of the most highly used application field for FDSS kinetic reader monitoring the calcium transient of the living cells. From the basic fluorescence dye such like Fluo-4, Fura-2 and to more various type of fluorescence dye such like FRET, and Flash Luminescence (Aequorine etc.,) reading technology can be used in our Kinetic reading platform.. FDSS7000EX is a lamp&filter base High-end system, capable of 1536 cell based assay with multiple washing ability and 2 dispenser head setup for the sticky compounds. It has more flexibility in having multiple of reading settings. Ratiometric Fura-2, Single wavelength Fluo-3 type Calcium flux, mobilization reading are possible. Some dye which requires FRET can also be done by using the motorized emission filter ...
Fam Labeled Fluorogenic Probe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
We have examined the migration of murine macrophages from the vascular compartment to normal and inflammatory tissues by the adoptive transfer of resident peritoneal macrophages (RPM phi) fluorescently labeled with the hydrophobic dye 1,1-dioctadecyl 3,3,3,3-tetramethylindocarbocyanine perchlorate (DiI). After initial labeling of the plasma membrane of RPM phi, the dye accumulated stably in intracellular vesicles of low density (rho = 1.042-1.045 kg/l) and cells remained viable in culture for 4 weeks. Like the normal monocyte, DiI-RPM phi, but not exudate-derived or fixed cells, migrated to peritoneal exudates, following i.v. adoptive transfer, by a mechanism inhibitable by an antibody to the type 3 complement receptor. In the absence of an inflammatory stimulus there was no migration to the peritoneal cavity, and DiI-RPM phi accumulated within 4 h in the red pulp and marginal zone of the spleen. By day 6 these cells still formed a tight ring of fluorescence in the marginal zone alone, ...
노인성 치매에서 가장 많은 비율을 차지하는 알츠하이머병 (Alzheimers disease) 환자의 뇌에는 이 질병의 핵심 표지자로 알려져 있는 아밀로이드 단백질 (amyloid β protein)이 쌓여있으며, 이 단백질이 질병의 발달과 연관성에 대한 많은 연구가 있다. 아밀로이드 단백질 연구에 있어서 조직염색 (tissue staining)을 통한 공초점 (confocal microscope) 그리고 이광자 현미경 (two-photon microscope) 촬영이 큰 역할을 하고 있으며, 이에 아밀로이드 단백질 특이한 형광염료 (fluorescence dye)의 중요성이 부각되었다. 본 연구에서는 기존의 아밀로이드 단백질 형광염료와는 다른 새로운 형광염료를 개발하였다. 형광염료내의 전자기부자 (donor)와 수락자 (acceptor)사이의 종류 그리고 두 분자 사이의 거리를 조절함으로써 조직염색 시 더 적은 자가형광을 내며, 더 특이적으로 아밀로이드 ...
Emitted Light Filtering - We find that many researchers have optimized imaging/filtering conditions for EtBr, which is a red dye. Please pay close attention to the filters; though EtBr filters will work if there are no other options, they may absorb a lot of emitted shorter wavelength light and therefore lower the observed sensitivity. A GFP/Sybr Green filter is preferred for Midori Green dyes.. Excitation Light Source - Though transilluminators that emit shorter wavelengths (254 nm) are very good for EtBr, they are not good for Midori Green dyes. We recommend UV tables with emissions at 306 nm or higher-the longer the better! For optimal results, use blue LED and blue/green LED light sources for Midori Green Advance and Midori Green Direct. Midori Green Xtra is not compatible with UV; it should be used with blue LED or blue/green LED illumination only.. Natural Light Exposure - Midori Green is sensitive to photo-bleaching effects from long exposure to direct (or even indirect) sunlight. We ...
I used single molecule fluorescence and fluorescence lifetime measurements of fluorescent dyes conjugated to DNA to better understand the photophysical and photochemical interactions between organic dye molecules and DNA. This process is important, because fluorescent dyes are frequently used as labels for DNA, but interactions with specific DNA bases can significantly affect the fluorescent properties of the dyes, leading from chromatic shifts all the way to fluorescence quenching. Thus, it is very important to obtain a full understanding of the photophysics of these interactions if cDNA or siRNAs are used as molecular probes. I studied DNA hairpins that were labeled with different numbers of red-emitting Atto655 dyes. I investigated three different samples: a DNA hairpin labeled with three dyes, one with two dyes, and one with only one dye attached to specific bases and well-known spatial separations. Sample preparation consisted of denaturing and annealing the short synthesized single strands ...
The use of labelling or staining agents has greatly assisted the study of complex biological interactions in the field of biology. In particular, fluorescent labelling of biomolecules has been demonstrated as an indispensable tool in many biological studies. Types of fluorescent labelling agents that are commonly used include conventional classes of organic fluorophores such as fluorescein and cyanine dyes, as well as newer types of inorganic nanoparticles such as QDs, and novel fluorescent latex/silica nanobeads. The newer classes of fluorescent labels are gaining increasing popularity in place of their predecessors due to their better optical properties such as possessing an enhanced photostability and a larger Stokes shift over conventional organic fluorophores, for example. This paper gives an overview of the recent advances on these luminescent nanomaterials with emphases on their optical characteristics that are crucial in fluorescence microscopy, both advantages and limitations in their ...
The RNAscope® Multiplex Fluorescent assays provide the same exceptional sensitivity as our singleplex assays, allowing single-molecule detection of up to four RNA targets simultaneously. The RNAscope® Multiplex Fluorescent assays are ideal for co-localization studies of any genes in nearly any tissue type using fluorescent labels. Advanced Cell Diagnostics offers two types of multiplex fluorescent assays. RNAscope® Fluorescent Assay, our first generation assay, is an all in one kit, ideal for fresh or fixed frozen tissues.
This application note describes the use of the SPECTRAmax GEMINI XS fluorescence microplate reader, which helps in studying the changes in levels of intracellular cAMP.
Recombinant Proteins , Streptavidin and Labeled Streptavidin , Streptavidin, HiLyte Fluor 555 conjugated; HiLyte Fluor555-streptavidin conjugate has been optimized in fluorophore/protein labeling ratio to ensure high fluorescent signal and uncompromised streptavidin function. The spectrum of HiLyte Fluor555 is only slightly red-shifted compared to those of Cy3 dyes, resulting in an optimal match to filters designed for Cy3 dyes. The fluorescence of HiLyte Fluor555 can be observed at excitation/emission wavelength of 554 nm /570 nm. HiLyte Fluor555 is more photostable than Cy3 , providing researchers with additional time for capturing image. HiLyte Fluor555 - protein conjugates can sustain treatments during immunofluorescent staining, fluorescence in situ hybridization, flow cytometry and other biological applications without hydrolysis.
With respect to cellular analysis, the underlying principle of flow cytometry is that a cell suspension is focused into a single cell stream, which passes through a light source (typically a laser beam). The scattered and emitted fluorescent light (if the cells are fluorescently labeled) is subsequently measured using a range of detectors and these measurements are used to generate multi-parameter data sets that describe the physical characteristics of the cells and their fluorescent properties. The size and granularity of cells can be identified on the basis of their forward and side light scatter characteristics (FSc and SSc respectively). Their characteristics and/or expression of different proteins can be further defined by pre-staining with fluorescently-labeled antibodies or molecules that identify cellular components and / or integrity (viability) or, indeed, fluorescently-labeled proteins.. Flow cytometers can evaluate cells at an extremely rapid rate (up to several tens of thousands of ...
Fluorescence in situ hybridization (FISH) is a valuable cytogenetic technique for the detection and localization of specific DNA or RNA sequences on chromosomes. Typically, this technique can be used to define spatiotemporal expression patterns of target genes. Using fluorescent DNA probes with sequences complementary to the location of the chromosome of interest, gene expression patterns can be easily detected by fluorescence microscopy and further quantitatively analyzed. In addition, improved FISH analysis also allows simultaneous observation of multiple genes by labeling different sequence fragments with different fluorophores.. In plant systems, FISH analysis has been used in a variety of crops, including wheat, rye, cucumber and melon. Based on the information on Lifeasibles official website, Lifeasibles services cover every step of the FISH assay in the plant system:. • Probe designing and construction: aside from commercially available probes, Lifeasible also provides customized ...
Fluorescein is unionised at acidic pH and the fluorescence intensity changes with pH [19, 20]. As expected, our results clearly demonstrate this effect on fluorescence levels using a FRET pair and a quencher pair with FAM. By use of the ATTO495-ATTO647N FRET pair for Hoogsteen-based parallel triplex formation, a robust and reliably LightCycler method was established. This novel FRET pair is well-suited for Tm and ΔTm determinations over a broad pH range of parallel triplex formations.. Furthermore, this FRET pair clearly demonstrates the pH independence from pH 5.5 to 7.5 of antiparallel duplex Tm determinations in contrast to the pH dependent Tm determination of parallel triplex formation from pH 4.5 to 6.0. An interesting feature of this is the negative correlation between pH and fluorescence intensity as pH increases from 4.5 to 6.0 for parallel triplex formation (Figure 3c). This is in concordance with the expected lower efficacy of parallel triplex formation due to the lack of protonated ...
Widefield fluorescent image of epithelial cells (nuclei stained with DAPI, yellow; and filamentous actin stained with Alexa Fluor 488 phalloidin, magenta )
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On the substrate carrying a sub-wavelength grating covered with a thin metal layer, a fluorescent dye-labeled cell was observed by fluorescence microscope. The fluorescence intensity was more than 20 times greater than that on an optically flat glass substrate. Such a great fluorescence enhancement from labeled cells bound to the grating substrate was due to the excitation by grating coupled surface plasmon resonance. The application of a grating substrate to two-dimensional detection and fluorescence microscopy appears to offer a promising method of taking highly sensitive fluorescence images.. ©2008 Optical Society of America. Full Article , PDF Article ...
These can be very useful. Most allow you to plot out the excitation and emission curves of a number of fluorescent dyes. Some allow the user to add filter sets and excitation sources to see how well they work with the dyes being considered. Noteworthy examples include: an extensive database of single and 2-photon dye spectra assembled here at the University of Arizona (Utzinger & Boswell), Thermo-Fishers Fluorescence spectraviewer, Semrocks searchlight, Fluorophores.org, Chromas spectra viewer, Biolegends viewer, Leicas Fluoscout, Omega Opticals curvomatic, and Zeiss Fluorescence Dye and Filter Database ...
Coupling nanomaterials with biomolecular recognition events represents a new direction in nanotechnology toward the development of novel molecular diagnostic tools. Here a graphene oxide (GO)-based multicolor fluorescent DNA nanoprobe that allows rapid, sensitive, and selective detection of DNA targets in homogeneous solution by exploiting interactions between GO and DNA molecules is reported. Because of the extraordinarily high quenching efficiency of GO, the fluorescent ssDNA probe exhibits minimal background fluorescence, while strong emission is observed when it forms a double helix with the specific targets, leading to a high signal-to-background ratio. Importantly, the large planar surface of GO allows simultaneous quenching of multiple DNA probes labeled with different dyes, leading to a multicolor sensor for the detection of multiple DNA targets in the same solution. It is also demonstrated that this GO-based sensing platform is suitable for the detection of a range of analytes when ...
Single-molecule fluorescence spectroscopy and super-resolution microscopy are important elements of the ongoing technical revolution to reveal biochemical and cellular processes in unprecedented clarity and precision. Demands placed on the photophysical properties of the fluorophores are stringent a …
The rhodamine labeled f-actin did not appear in the composite image of the three different fluorophores due to the intensity of the fluorescence being too low to register. This could be due to photobleaching that had previously occurred in this specific area of the sample, or if the sample was not labeled adequately with enough fluorescent dye. Time-lapse data for all three fluorophores under the same condition revealed discrepancies in rate of decay and initial intensity for each fluorophore. A relative high initial intensity for DAPI labeled dsDNA can be explained by the relative high net local concentration of bright fluorophores. Each nucleus contains a high concentration of dsDNA, which when stained with DAPI, creates a large solid fluorescent region with overlapping fluorophores. This differs from both tubulin and f-actin, which are of tube like nature, and appear as porous regions of interest where background light can seep through and be analyzed, making the initial brightness in the ...
We recently finished our Ask the Expert discussion on Improving Live Cell Fluorescence Imaging. This week we had several interesting questions focused on combating the negative effects of imaging on cell health and viability including looking at media options as a possible solution.
Some APC-Cy7 conjugates show changes in their emission spectra with prolonged exposure to paraformaldehyde. For instruments equipped with a 561 nm Yellow-Green laser, a 585/15 nm filter is recommended for detection. When making decisions about which fluorochromes to use in your experiments, youll want to know their relative emission spectra. APC (Allophycocyanin) spectrum - APC (Allophycocyanin) is ... excitation and emission wavelengths using the interactive Spectrum Viewer - A web application for viewing and comparing spectra of various fluorescent compounds. BV605 is a tandem fluorochrome that combines BD Horizon BV421 and an acceptor dye with emission at 605nm. APC-H7 conjugates provide greater stability in light and paraformaldehyde fixatives and have less spillover into the APC channel than APC-Cy7 conjugates. However, because of its peak emission at 667 nm, Alexa Fluor® 647 cannot be seen well by eye, and it cannot be excited optimally with a mercury lamp. BV650 is a tandem fluorochrome ...
New techniques for biological optical imaging are of great interest for the detection and visualization of processes and disease in both clinical and research areas. One major advancement has been the use of far red and near infrared (NIR) light, as it has the ability to penetrate tissues deeper than other parts of the spectrum which are readily scatter and absorbed by the surroundings. In order to improve the signal to noise ratio and resolution of optical images, contrast agents are used. Fluorescent markers can be modified to attach to specific molecular targets, creating small molecular probes. These targets can be disease sites, or biological molecules which play a major role in processes such as tumor growth. It was our goal to create a new novel fluorescent probe, consisting of a cyanine based far red to NIR marker, and an n-hydroxysuccinimide (NHS) derivative to act as a linker, which could then bind with biological species containing primary amides such as proteins and antibodies, in this
The Quantus Fluorometer is a dual-channel fluorometer for your personal quantitation workflow. Designed to provide highly sensitive fluorescent detection when quantifying nucleic acids, the compact instrument is simple to operate.
A new fluorescent Zn2+ chemosensor (P1) based on a functionalized porphyrin was synthesized and characterized. P1 displayed dramatic ratiometric variations in absorption and fluorescent emission spectra upon exposure to Zn2+ due to the formation of a 1:1 Zn2+/P1 complex. The sensor also exhibited high selectivity and sensitivity toward Zn2+ over other common metal ions in the physiological pH range with a detection limit of 1.8 mM. The sensor showed fast response times and excellent reproducibility, thus confirming its potential applicability as a fluorescent sensor for Zn2+ sensing.
A fluorophore (or fluorochrome, similarly to a chromophore) is a fluorescent chemical compound that can re-emit light upon light excitation. Fluorophores typically contain several combined aromatic groups, or planar or cyclic molecules with several π bonds. Fluorophores are sometimes used alone, as a tracer in fluids, as a dye for staining of certain structures, as a substrate of enzymes, or as a probe or indicator (when its fluorescence is affected by environmental aspects such as polarity or ions). More generally they are covalently bonded to a macromolecule, serving as a marker (or dye, or tag, or reporter) for affine or bioactive reagents (antibodies, peptides, nucleic acids). Fluorophores are notably used to stain tissues, cells, or materials in a variety of analytical methods, i.e., fluorescent imaging and spectroscopy. Fluorescein, by its amine reactive isothiocyanate derivative FITC, has been one of the most popular fluorophores. From antibody labeling, the applications have spread to ...
SH2 protein microarray assays of HA4 specificity(a) Fluorescence images of SH2 chips following incubation with fluorophore-labeled HA4. Each SH2 domain is spott
TY - JOUR. T1 - Highly photostable ketopyrrolyl-BODIPYs with red aggregation-induced emission characteristics for ultrafast wash-free mitochondria-targeted bioimaging. AU - Wu, Hao. AU - Guo, Xing. AU - Yu, Changjiang. AU - Wong, Wai Yeung. AU - Hao, Erhong. AU - Jiao, Lijuan. PY - 2020/5. Y1 - 2020/5. N2 - Subcellular organelle-specific probes, including mitochondria-targeted fluorescent probes, have attracted enormous research interests because they can monitor or visualize the morphology or biological activities of specific organelles and play an indispensable role in disease diagnosis. To follow the process, highly specific and photostable fluorescent probes are in demand. However, commercially available mitochondria probes normally suffer from poor photostability under laser irradiation and aggregation caused quenching (ACQ) in the aggregate state. In this work, two simple aggregation-induced emission(AIE)-active meso-2-ketopyrrolyl BODIPYs were developed via a convenient one-pot synthetic ...
Phosphorescent molecules are attractive complements to fluorescent compounds for bioimaging. Time-gated acquisition of the long-lived phosphorescence signals provides an effective means to eliminate unwanted background noises due to short-lived autofluorescence. We have previously investigated the molecular principles governing modulation of photoinduced electron transfer in phosphorescence zinc probes that were based on biscyclometalated Ir(III) complexes (Woo, H. et al. J. Am. Chem. Soc. 2013, 135, 4771-4787). The studies established that phosphorescence turn-on responses would be attainable for Ir(III) complexes with high triplet-state energies. This sets an upper limit to an emission wavelength, restricting the development of red- or near-IR-phosphorescence turn-on probes. To address this challenge, we designed and synthesized a new phosphorescent probe having an electron-deficient 2-(2-pyridyl)pyrazine diimine ligand tethering a di(2-picolyl)amine (DPA) zinc receptor. This ligand control ...
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We offer Amine-Reactive Fluorophores; Bioconjugates; Biotin/Desthiobiotin; Buffers, Detergents; Caged Probes; Cell Analysis Kits; Cell stains; Click Chemistry; Enzyme Substrates; Fluorescent Mitochondrial Probes; Fluorescent Probes; Fluorescent/Non-Fluorescent Ion and pH Indicators; Fluorescent/Non-Fluorescent Lipids, Phospholipid Probes; Gel & Membrane Stains; Kits and Assays; Monomers; Natural Products and Their Fluorescent/None-Fluorescent Conjugates; Neurotransmitter receptors Probes; Nucleotides/Nucleosides; Protein Kinase Inhibitors (PKI); Quencher/Non-Fluorescent Probes; Reactive Probes and Related Products; Sensors; Steroids and Related Products
Nonlinear microscopy. Researcher using a two-photon excitation microscope to image living tissue (images on screen at left). Different cells within the tissue have been labelled with different fluorescent dyes. An infrared laser beam is focused through the objective lens. As it hits the fluorescent dyes it excites them, causing them to fluoresce. The laser emits pulses that last for only femtoseconds (one millionth of one billionth of a second). This imaging technique is able to penetrate deep into biological tissue and image entire networks of cells. - Stock Image C019/7668
Kinetics of B-fragment transport from EE/RE to the Golgi apparatus. (A) Confocal microscopy on living HeLa cells. Fluorophore-labeled B-fragment was internalize