for uses and theory of fluorescence Fluorescence in the life sciences Image FISH 13 21.jpg thumb A fluorophore labeled human cell . A fluorophore , in analogy to a chromophore , is a component of a molecule which causes a molecule to be Fluorescence in the life sciences fluorescent . It is a functional group in a molecule which will absorb energy of a specific wavelength and re emit energy at a different but equally specific wavelength. The amount and wavelength of the emitted energy depend on both the fluorophore and the chemical environment of the fluorophore. This technology has particular importance in the field of biochemistry and protein studies, e.g., in immunofluorescence and immunohistochemistry . ref name Pawley cite book author Tsien RY, Waggoner Aeditor Pawley JB chapter Fluorophores for confocal microscopy title Handbook of biological confocal microscopy publisher Plenum Press location New York year 1995 pages 267 74 isbn 0 306 44826 2 url http books.google.com books?id 16Ft5k8RC AC&pg PA267 accessdate 2008 12 13 ref Fluorescein isothiocyanate FITC , a reactive derivative of fluorescein , has been one of the most common fluorophores chemically attached to other, non fluorescent molecules to create new fluorescent molecules for a variety of applications. Other historically common fluorophores are derivatives of rhodamine TRITC , coumarin , and cyanine . ref cite book author Rietdorf J title Microscopic Techniques series Advances in Biochemical Engineering Biotechnology publisher Springer location Berlin year 2005 pages 246 9 isbn 3 540 23698 8 url http books.google.com books?id h9F RGrIoicC&pg PA247 accessdate 2008 12 13 ref Newer generations of fluorophores, many of which are proprietary, often perform better more photostable, brighter, and or less pH sensitive than traditional dyes with comparable excitation and emission. ref name Pawley ref Lakowicz, J.R. ... of the fluorophore might steric effects steric ally hinder the tagged molecule quantum dot 2 ... more details
Fluorophore assisted carbohydrate electrophoresis or FACE is a biochemical technology suited for detecting complex mixtures of high molecular weight N glycans. ref name Harish cite journal last Harish first P. M. Kumar date July 23, 1999 title Use of Fluorophore Assisted Carbohydrate Electrophoresis FACE in the Elucidation of N Linked Oligosaccharide Structures journal Methods in Biotechnology volume 10 pages 221 234 url http www.springerprotocols.com Abstract doi 10.1007 978 1 59259 261 6 18 doi 10.1007 978 1 59259 261 6 18 ref A specialized form of this technique is the DSA FACE, which is an acronym for DNA sequencer assisted flurophore assisted carbohydrate electrophoresis. DSA FACE has higher resolution and sensitivity than classical FACE. References reflist DEFAULTSORT Fluorophore Assisted Carbohydrate Electrophoresis Category Molecular biology techniques Category Molecular biology ... more details
Unsourced article date June 2010 Fluorescent labelling is the process of covalently attaching a fluorophore to another molecule, such as a protein or nucleic acid . This is generally accomplished using a reactive derivative of the fluorophore that selectively binds to a functional group contained in the target molecule. The most commonly labelled molecules are antibodies, proteins, amino acids and peptides which are then used as specific probes for detection of a particular target. ref http pharmaxchange.info press 2011 01 fluorescent labeling of biomolecules with organic probes Presentation on Fluorescent labelling of biomolecules with organic probes PharmaXChange.info ref Detection Fluorescent labels are generally used for detection of a protein or other labelled molecule via a fluorescence microscope , flow cytometer or some other fluorescence reading instrument. These can be useful in localization of a target within a cell, flow cytometry FACS analysis, western blot assays, and other immunoanalytical methods. Labelling techniques Fluorescent labelling is accomplished using a chemically reactive derivative of a fluorophore. Common reactive groups include amine reactive isothiocyanate derivatives such as Fluorescein isothiocyanate FITC and TRITC derivatives of fluorescein and rhodamine , amine reactive succinimidyl esters such as NHS fluorescein, and sulfhydryl reactive maleimide activated fluors such as fluorescein 5 maleimide. Reaction of any of these reactive dyes with another molecule results in a stable covalent bond formed between a fluorophore and a labelled molecule. Following a fluorescent labelling reaction, it is often necessary to remove any nonreacted fluorophore from the labelled target molecule. This is often accomplished by size exclusion chromatography , taking advantage of the size difference between fluorophore and labelled protein, nucleic acid, etc. Fluorophores may interact with the separation matrix and reduce the effiency of separation. For this reason ... more details
Unreferenced stub auto yes date December 2009 In molecular biology and biotechnology , a fluorescent tag is a part of a molecule that researchers have attached chemically to aid in detection of the molecule to which it has been attached. The tag is some kind of fluorescent molecule also known as fluorophore . Ethidium bromide , Fluorescein and Green fluorescent protein are common tags. See also Molecular tagging velocimetry DEFAULTSORT Fluorescent Tag Category Molecular biology Biochem stub ... more details
NBD can refer to Banking National Bank of Dubai National Bank of Detroit National Bank of Dominica Logistics Next Business Day Computing Network Based Defense The network block device of Linux Science The fluorophore Nitro benzoxadiazole Nucleotide binding domain Neurobiological brain disorder Internet Slang No Big Deal Other uses Negative binomial distribution New business development Slang abbreviation for Never Back Down from the movie, Never Back Down disambig it NBD ... more details
unreferenced date December 2010 File Molecular Beacons.jpg thumb Structure of molecular beacons in their native conformations top or hybridized with a DNA strand bottom Molecular beacons are oligonucleotide hybridization probe s that can report the presence of specific nucleic acid s in homogenous solutions. The terms more often used is molecular beacon probes . Molecular beacons are hairpin shaped molecules with an internally quenched fluorophore whose fluorescence is restored when they bind to a target nucleic acid sequence. This is a novel non radioactive method for detecting specific sequences of nucleic acids. They are useful in situations where it is either not possible or desirable to isolate the probe target hybrids from an excess of the hybridization probes. Molecular beacon probes Image SNP Molecularprobe 1.png right A typical molecular beacon probe is 25 nucleotides long. The middle 15 nucleotides are complementary to the target DNA and do not base pair with one another, and the five nucleotides at each end are complementary to each other and not to the target DNA. A typical molecular Beacon Structure can be divided in 4 parts Loop This is the 18 30 base pair region of the molecular beacon which is complementary to the target sequence. Stem The beacon stem sequence lies on both the ends of the loop. It is typically 5 7 bp long at the sequences at both the ends are complementary to each other. 5 fluorophore Towards the 5 end of the molecular beacon, is attached a dye that fluoresces in presence of a complementary target. 3 quencher non fluorescent The quencher dye is covalently attached to the 3 end of the molecular beacon and when the beacon is in closed loop shape, prevents the fluorophore from emitting light. If the mRNA detected is complementary to the strand in the loop, then the mRNA goes into the loop causing the separation of the fluorophore and the quencher therefore resulting in fluorescence. Applications of molecular beacons SNP detection Real t ... more details
one source date December 2010 In chemistry , fluorescence anisotropy assays the rotational diffusion of a molecule from the decorrelation of polarization in fluorescence , i.e., between the exciting and emitted fluorescent photon s. This decorrelation can measure the tumbling time of the molecule as a whole, or of a part of the molecule relative to the whole. From the rotational diffusion constants, one can estimate the rough shape of a macromolecule. ref http www.horiba.com fileadmin uploads Scientific Documents Fluorescence Anisotropy FL 3.pdf Fluorescence Anisotropy Studies ref Fluorescence anisotropy can be used for measuring the binding interaction between two molecules, to determine the binding constant or the inverse, the disassociation constant for the interaction. The basic idea is that a fluorophore excited by polarized light will also emit polarized light. However, if a molecule is moving, it will tend to scramble the polarization of the light by radiating at a different direction from the incident light. The scrambling effect is greatest with fluorophores freely tumbling in solution and decreases with decreased rates of tumbling. Protein interactions can be detected when one of the interacting partners is fused to a fluorophore upon binding of the partner molecule a larger, more stable complex is formed which will tumble more slowly thus, increasing the polarization of the emitted light and reducing the scrambling effect . This technique works best if a small molecule is fused to a fluorophore and binds to a larger partner this maximizes the difference in signal between bound and unbound states . If the fluorophore is attached to the larger protein in a binding pair, the difference in polarization between bound and unbound states will be smaller because the unbound protein will already be fairly stable and tumble slowly to begin with and the measurement will be less accurate. By titrating the amount of one of the proteins, a binding curve can be generate ... more details
between the observed fluorescence intensity physics intensity and the distance of the fluorophore ... an emitted photon per unit time, math P em math . Both probabilities are a function of the fluorophore height above the silicon surface, so the observed intensity will also be a function of the fluorophore height. The simplest arrangement to consider is a fluorophore embedded in silicon dioxide ... n 0 math . The fluorophore is excited by light of wavelength math lambda ex math and emits light ... dipole of excitation of the fluorophore. math P ex math is proportional to the squared projection ... e ex mid 2 math br The local electric field, math F in math , at the fluorophore is affected by interference ... be combined to give the probability of exciting the fluorophore per unit time math P ex math . br Many ... math gamma in math of the excitation light The angle of transition dipole math theta ex math of the fluorophore ... of a FLIC intensity plot showing the relative fluorescence intensity measured versus the distance of the fluorophore ... angle of the fluorophore dipoles. The polar coordinate system azimuthal angle math phi ex ... math I lambda ex math and the extinction coefficient of the fluorophore math epsilon lambda ex math ... and the distance of the fluorophore above the reflective surface. The fact that it is not an equality ... describes the relative fluorescence intensity measured versus the fluorophore height. The fluorophore ... depends on the sample being measured. For a sample with fluorophore height in the range of 10 ... and would produce the greatest contrast between fluorophore heights. Oxide thickness above a few hundred ... thicknesses is compared to the predicted ratio to calculate the fluorophore height above the oxide ... with the theoretical model allowing the distance of the fluorophore above the oxide surface math d textit f math to be a free parameter. The FLIC curves shift to the left as the distance of the fluorophore ... more details
the fluorescence emitted by the fluorophore when excited by the cycler s light source via F rster ... pmidlookup?view long&pmid 11013345 ref As long as the fluorophore and the quencher are in proximity ... releases the fluorophore from it and breaks the close proximity to the quencher, thus relieving the quenching effect and allowing fluorescence of the fluorophore. Hence, fluorescence detected in the real time PCR thermal cycler is directly proportional to the fluorophore released and the amount ... more details
fluorophore , a fluorescent dye which can be a small molecule, protein or quantum dot . Several techniques exist to exploit additional properties of Fluorophore fluorophores , such as F rster resonance ... levels. Main fluorescence The principle behind fluorescence is that the fluorophore fluorescent ... , and the extinction coefficient is the amount of light that can be absorbed by a fluorophore. Both the quantum yield and extinction coefficient are specific for each fluorophore and multiplied together ...?fldID 4DD9D52E 5056 8A76 4E6E E217FAD0D86B brightness ref Labelling Reactive dyes Main fluorophore ... of small molecule fluorophore is that of the transition metal ligand Coordination complex complexes ... and fluoresce longer and more stably. Other proteins are fluorescent but require a fluorophore cofactor ... quencher or another fluorophore, which has an excitation spectrum which overlaps with the emission ... at opposite ends of an aromatic ring system ref Evanko D. et al., A flaky but useful fluorophore ... dipole moment dipole moment when excited. When a fluorophore is excited, it generally has a larger dipole moment sub E sub than in the ground state sub G sub . Absorption of a photon by a fluorophore ... surrounding the fluorophore reorient 10 100 ps due to the change in polarity in the excited singlet ... state of the fluorophore is lowered longer wavelength , hence fluorophores that have a large ... however, in biochemistry environment sensitive fluorphore and solvatochromic fluorophore are used ..., cells or subcellular structures is accomplished by labeling an antibody with a fluorophore ... , which affords a quantitative, 3D view of the sample. See also Fluorophore Fluorescent microscopy ... more details
Unreferenced date December 2009 A luminophore is an atom or atomic grouping in a chemical compound that manifests luminescence . There exist organic and inorganic luminophores. It should be stressed that the correct, textbook terminology is luminophore , not lumophore , although the latter term has been frequently but erroneously used in the chemical literature. Luminophores can be divided into two subcategories fluorophore s and phosphor s. The difference between luminophores belonging to these two subcategories is derived from the nature of the excited state responsible for the emission of photon s. Some luminophores, however, cannot be classified as being exclusively fluorophore s or phosphors and exist in the gray area in between. Such cases include transition metal complexes such as ruthenium tris 2,2 bipyridine whose luminescence comes from an excited nominally triplet metal to ligand charge transfer MLCT state, but which is not a true triplet state in the strict sense of the definition and colloidal quantum dots , whose emissive state does not have either a purely singlet or triplet spin. Most luminophores consist of Conjugated system conjugated pi systems or transition metal complexes. There exist purely inorganic luminophores, such as zinc sulfide doped with rare earth metal ions, rare earth metal oxysulfides doped with other rare earth metal ions, yttrium oxide doped with rare earth metal ions, zinc orthosilicate doped with manganese ions, etc. Luminophores can be observed in action in fluorescent lights, TV screens, computer monitor screens, organic light emitting diode s and bioluminescence . Category Luminescence Category Chemical compounds be be x old cs Luminofor de Luminophor et Luminofoor fr Luminophore pl Luminofor ru sk Luminofor uk ... more details
A multiphoton fluorescence microscope MFM is a specialized optical microscope . Description The MFM uses pulsed long wavelength light to excite fluorophore s within the specimen being observed. The fluorophore absorbs the energy from two long wavelength photons which must arrive simultaneously in order to excite an electron into a higher energy state, from which it can decay, emitting a fluorescence signal. It differs from traditional fluorescence microscopy in which the excitation wavelength is shorter than the emission wavelength, as the summed energies of two long wavelength exciting photons will produce an emission wavelength shorter than the excitation wavelength. Multiphoton fluorescence microscopy has similarities to confocal laser scanning microscopy . Both use focused laser beams scanned in a raster pattern to generate images, and both have an optical sectioning effect. Unlike confocal microscopes, multiphoton microscopes do not contain pinhole apertures, which give confocal microscopes their optical sectioning quality. The optical sectioning produced by multiphoton microscopes is a result of the point spread function formed where the pulsed laser beams coincide. The multiphoton point spread function is typically dumbbell shaped longer in the x y plane , compared to the upright rugby ball shaped point spread function of confocal microscopes. The longer wavelength, low energy typically infra red excitation lasers of multiphoton microscopes are well suited to use in imaging live cells as they cause less damage than short wavelength lasers, so cells may be observed for longer periods with fewer toxic effects. Many researchers are currently working toward better and higher resolution multiphoton imaging developments. See also Two photon excitation microscopy References reflist External links Molecular Expressions http microscopy.fsu.edu primer techniques fluorescence multiphoton multiphotonhome.html introduction to multiphoton microscopy Optical microscopy Cate ... more details
unreferenced date November 2006 FTA Abs is a treponemal test for syphilis . Using antibodies specific for the Treponema pallidum species, such tests are more specific than non treponemal testing such as VDRL . In addition, FTA Abs turns positive earlier and remains positive longer than VDRL. Other treponemes, such as T. pertenue , may also produce a positive FTA Abs. FTA Fluorescent Treponemal Antibody Abs absorption should always be followed to confirm a positive Rapid plasma reagin RPR and or VDRL test for syphilis. The ABS suffix refers particularly to a processing step used to remove nonspecific antispirochetal antibodies present in normal serum. Procedure The antigen for the FTA Abs test is whole bacteria. The bacteria cannot be cultured on laboratory media, so the organisms used are a lyophilized suspension of T. pallidum extracted from rabbit testicular tissue. This is spread over and fixed to a slide. Patient serum is mixed with an absorbent the ABS part of the test containing an extract of a non pathogenic treponeme, Treponema phagedenis biotype Rieter. The purpose of the absorbent is to remove anti treponemal antibodies that are not specific for the syphilis bacteria. The pre adsorbed patient serum is then added to the slide if the patient has been infected by syphilis, their antibodies will bind to the bacteria. Fluorescein isothiocyanate FITC a fluorophore labeled anti treponeme antibody and TRITC another fluorophore labeled anti human antibodies are added as secondary antibodies. The spirochete location is identified using the FITC staining and the TRITC staining identifies whether the patient has anti T. pallidum antibodies binding to the same spirochete . Utility This test is not useful for following therapy, because it does not wane with successful treatment of the disease, and will continue to be positive for the life of the patient after primary exposure. ref cite journal last Singh first Ameeta E. coauthors Barbara Romanowski date 1 April 1999 tit ... more details
Orphan date February 2009 Image NBD TMA.png frame right NBD TMA NBD TMA 2 4 nitro compound nitro 2,1,3 azole benzoxadiazol 7 yl amine amino ethyl trimethylammonium is a small 139 Unified atomic mass unit u , positively charged 1 fluorescent dye. It was also known as EAM 1 N,N,N, Trimethyl 2 7 nitro 2,1,3 benzoxadiazol 4 yl amino ethanaminium iodide when it was briefly supplied by Macrocyclics Company as an iodide complex. NBD TMA has an excitation maximum at 458 nanometre nm and an emission maximum at 530 nm. It also has a smaller local excitation maximum around 343 nm. The molar extinction coefficient is about 13,000 cm sup 1 sup M sup 1 sup and its overall effective fluorescence is about 1 that of fluorescein . It is only mildly sensitive to halide ion collision quenching. NBD TMA was designed as a probe for monitoring kidney renal transport of organic ion cation s. As a small, positively charged fluorophore , it has also seen use as a tracer for measuring gap junction coupling in cases of cation selective connexin channels. Further reading PMID 10864014 PMID 15869481 Category Dyes ... more details
Essay like date February 2008 A dark quencher is a substance that absorbs excitation energy from a fluorophore and dissipates the energy as heat while a typical fluorescent Quenching fluorescence quencher re emits much of this energy as light ref Osterman, H., The Next Step in Near Infrared Fluorescence IRDye QC 1 Dark Quencher, 2009 Review Article. http biosupport.licor.com docs QC 1DarkQuencher v5.pdf Download PDF ref . Dark quenchers are used in molecular biology in conjunction with fluorophores. When the two are close together, such as in a molecule or protein, the fluorophore s emission is suppressed. This effect can be used to study molecular geometry and motion. An example of its use is in Taqman or invader assay, SNP genotyping methods. For instance, a hairpin loop with a fluorophore and quencher at the base of the stem is used. An unlabeled SNP specific PCR primer one of many with a specific 5 tail binds to the sequence to be probed, and the taq polymerase extends the sequence that will have a specific 5 end dependent on the SNP insensitive to polymorphisms upstream of the SNP in question . In the next run a primer molecular biology primer , complementary to that tail, with a hairpin loop is extended. In the next run the elongation of the complementary strand will linearise the hairpin separating the fluorophore and quencher . An alternative to using quenchers is to use F rster resonance energy transfer FRET where the combination of two dyes gives a signal ref Peng, X., Chen, H., Draney, D.R., Volcheck, W.M., A Non fluorescent, Broad Range Quencher Dye for FRET Assays, Analytical Biochemistry, 2009 Vol. 388 , pp. 220 228. http biosupport.licor.com docs NonfluorQuencherDyePaper09.pdf Download PDF ref . Mode of function Dark quenchers are dyes with no native fluorescence . Until the last few years, quenchers have typically been a second fluorescent dye, for example, fluorescein as the reporter and rhodamine as the quencher FAM TAM probes . However, quencher fluorescence ... more details
labeled ligands have to bear a bulky fluorophore that may cause it to hinder the ligand binding. Therefore, the fluorophore used, the length of the linker, and its position must be carefully selected. An example is by using FRET , where the ligand s fluorophore transfers its energy to the fluorophore ... more details
uses a single antibody that is chemically linked to a fluorophore . The antibody recognizes the target molecule and binds to it, and the fluorophore it carries can be detected via microscope ... conjugation of the antibody to the fluorophore. This reduces the number of steps in the staining ... molecule and binds to it, and the second the secondary antibody , which carries the fluorophore, recognises ... antibodies to directly carry a fluorophore. Different primary antibodies with different constant ... more details
also excite a fluorophore in one quantum event. Each photon carries approximately half the energy ... photon fluorescence lies in the 700&ndash 1000 nm infrared range. If the fluorophore absorbs two .... The fluorophore will then emit a single photon with a wavelength that depends on the type of fluorophore ... of the fluorophore, the probability for fluorescent emission from the fluorophores increases quadratically ... more details
Image Fp rhodamine.jpg frame right Fluorophosphonate rhodamine FP Rhodamine activity based probe for profiling of the serine hydrolase superfamily. In this probe the fluorophosphonate is the reactive group RG as it binds irreversibly to the active site serine nucleophile of serine hydrolase s and the tag is rhodamine , a fluorophore for in SDS PAGE gel visualization. Image Gel abpp eg.png frame left In SDS PAGE gel ABPP using probes with different fluorophores in the same lane to simultaneously profile differences in enzyme activities Activity based proteomics , or activity based protein profiling ABPP is a functional proteomics proteomic technology that uses specially designed chemical probes that react with mechanistically related classes of enzymes ref Berger AB, et al. Activity based protein profiling applications to biomarker discovery, in vivo imaging and drug discovery. American Journal of Pharmacogenomics 2004 http www.ncbi.nlm.nih.gov entrez query.fcgi?CMD search&DB pubmed Article ref . The basic unit of ABPP is the probe which typically consists of two elements a reactive group RG and a tag. Additionally, some probes may contain a binding group which enhances selectivity. The reactive group usually contains an electrophile that gets covalent bond covalently linked to a nucleophile nucleophilic residue in the active site of an active enzyme . An enzyme that is inhibited by enzyme inhibitor s or post translational modification s will not react with an activity based probe. The tag may be either a reporter such as a fluorophore or an affinity label such as biotin or an alkyne or azide for use with the Huisgen 1,3 dipolar cycloaddition also known as click chemistry ref Speers AE, et al. Activity Based Protein Profiling in Vivo Using a Copper I Cataqlyzed Azide Alkyne 3 2 Cycloaddition Journal of the American Chemical Society 2003 http www.scripps.edu cb cravatt pdf Speersetal2003.pdf Article ref . A major advantage of ABPP is the ability to monitor the availab ... more details
Citations missing date September 2009 Nick translation or Head Translation is a fluorescent tag tagging technique in molecular biology in which DNA Polymerase I is used to replace some of the nucleotides of a DNA sequence with their labeled analogues, creating a tagged DNA sequence which can be used as a probe in Fluorescent in situ hybridization or blot biology blotting techniques. This process is called nick translation because the DNA to be processed is treated with DNase to produce single stranded nicks. This is followed by replacement in nicked sites by DNA polymerase I , which elongates the 3 hydroxyl terminus, removing nucleotides by 5 3 exonuclease activity, replacing them with dNTPs. To radioactively label a DNA fragment for use as a probe in blotting procedures, one of the incorporated nucleotides provided in the reaction is radiolabeled in the alpha phosphate position. Similarly, a fluorophore can be attached instead for fluorescent labelling, or an antigen for immunodetection. When DNA polymerase I eventually detaches from the DNA, it leaves another nick in the phosphate backbone. The nick has translated some distance depending on the processivity of the polymerase. This nick could be sealed by DNA ligase , or its 3 hydroxyl group could serve as the template for further DNA polymerase I activity. Proprietary enzyme mixes are available commercially to perform all steps in the procedure in a single incubation. Nick translation could cause double stranded DNA breaks, if DNA polymerase I encounters another nick on the opposite strand, resulting in two shorter fragments. This does not influence the performance of the labelled probe in in situ hybridization. DEFAULTSORT Nick Translation Category Genetics Category Molecular biology Category Laboratory techniques Genetics stub ca Nick translation de Nick translation it Nick translation ... more details
Unreferenced date December 2009 Orphan date January 2009 Chembox verifiedrevid 400113589 ImageFile IAEDANS.svg ImageSize 200px IUPACName 5 2 2 Iodo 1 oxoethyl amino ethylamino 1 naphthalenesulfonic acid OtherNames Section1 Chembox Identifiers ChemSpiderID Ref chemspidercite correct chemspider ChemSpiderID 82727 InChI 1 C14H15IN2O4S c15 9 14 18 17 8 7 16 12 5 1 4 11 10 12 3 2 6 13 11 22 19,20 21 h1 6,16H,7 9H2, H,17,18 H,19,20,21 InChIKey ZMERMCRYYFRELX UHFFFAOYAI StdInChI Ref stdinchicite correct chemspider StdInChI 1S C14H15IN2O4S c15 9 14 18 17 8 7 16 12 5 1 4 11 10 12 3 2 6 13 11 22 19,20 21 h1 6,16H,7 9H2, H,17,18 H,19,20,21 StdInChIKey Ref stdinchicite correct chemspider StdInChIKey ZMERMCRYYFRELX UHFFFAOYSA N CASNo 36930 63 9 PubChem 91621 SMILES O S O O c1cccc2c1cccc2NCCNC O CI Section2 Chembox Properties Formula C sub 14 sub H sub 15 sub IN sub 2 sub O sub 4 sub S MolarMass 434.25 g mol Appearance Density MeltingPt BoilingPt Solubility Section3 Chembox Hazards MainHazards FlashPt Autoignition IAEDANS is an organic fluorophore fluorescent molecule . It stands for 5 2 iodoacetyl amino ethyl amino naphthalene 1 sulfonic acid. It is widely used as a marker in fluorescence spectroscopy. 1,5 IAEDANS has a peak excitation wavelength of 336 nm and a peak emission wavelength of 490 nm. The extinction coefficient of the dye is 5700. It is soluble in dimethylformamide DMF or buffer above pH 6 and reacts primarily with thiol s. The absorption spectrum IAEDANS overlaps well with the emission spectrum of tryptophan , making it useful as an acceptor in FRET experiments. It can also be used as a resonance energy donor to fluorophores such as fluorescein , Alexa Fluor 488, Oregon Green, and BODIPY FL. DEFAULTSORT Iaedans Category Sulfonic acids Category Luminescence Category Organoiodides de IAEDANS ... more details
Fluorescence Loss in Photobleaching , or FLIP , is a technique in fluorescence microscopy which can be used to examine the movement or diffusion of molecules inside cell biology cell s or cell membrane membrane s. Typically a cell membrane is labelled with a fluorescent fluorophore dye , and a specific area of the labeled membrane is photobleaching bleached using the beam from a confocal laser scanning microscope . The fluorescence intensity from that region of the membrane is measured over time. Motion of fluorescent molecules into and along the membrane slowly restores the fluorescence in the bleached region, while depleting the fluorescence in other regions by exchange of bleached for unbleached fluorophores . Measurement of the rate of this recovery provides an estimate of the lateral membrane fluidity. Changes in the size and shape of the bleached region can also indicate directional flow along the cell membrane. FLIP is also useful in verifying the continuity of membranous organelles e.g., the Golgi apparatus . A small circumscribed region of the organelle is continuously bleached. As fluorophores diffuse along the membrane into the illuminated spot, they are bleached eventually, the fluorescence of the entire organelle is depleted. It is closely related to another technique, Fluorescence recovery after photobleaching FRAP . The difference between FLIP and FRAP is that FLIP follows the path of the bleached fluorophores, while FRAP follows the recovery of the bleached region. ref name nature review cite journal last1 Lippincott Schwartzl first1 Jennifer last2 Snappl first2 Erik last3 Kenworthy first3 Anne title Studying protein dynamics in living cells journal Nature Reviews Molecular Cell Biology volume 2 pages 444 456 date June 2001 doi 10.1038 35073068 accessdate 2008 06 05 pmid 11389468 issue 6 ref References references See also Fluorescence microscope Category Microscopy Category Fluorescence category Cell imaging category Biochemistry methods Cell biology ... more details
Chembox ImageFile TCPO.png ImageSize 200px IUPACName Oxalic acid bis 2,4,6 trichloro phenyl ester OtherNames Bis 2,4,6 trichlorophenyl oxalate, bis 2,4,6 trichlorophenyl ethanedioate Section1 Chembox Identifiers CASNo 1165 91 9 PubChem ChemSpiderID 141097 SMILES O C C OC1 C Cl C C Cl C C1Cl O OC2 C Cl C C Cl C C2Cl InChI InChI 1 C14H4Cl6O4 c15 5 1 7 17 11 8 18 2 5 23 13 21 14 22 24 12 9 19 3 6 16 4 10 12 20 h1 4H Section2 Chembox Properties C 14 H 4 Cl 6 O 4 Appearance white crystalline powder Density 1.698 g cm sup 3 sup MeltingPt 188 192 C BoilingPt 500.9 C Solubility 0.01962 mg L Section3 Chembox Hazards MainHazards FlashPt 190.6 C Autoignition TCPO , or bis 2,4,6 trichlorophenyl oxalate, is the main chemical in glow stick s. Uses TCPO is one of the major ingredients in glow stick glow sticks . When combined with a fluorescent dye like 9,10 Bis phenylethynyl anthracene 9,10 bis phenylethynyl anthracene , a solvent such as diethyl phthalate , and a weak base usually sodium acetate or sodium salicylate , and hydrogen peroxide , the mixture will start a Chemiluminescence reaction to glow an fluorescent green color. ref http sites.google.com site nurdrage chemistry experiments how to make a glow stick with real chemicals How to Make a Glow Stick with Real Chemicals ref Red, yellow and blue colours can be made by replacing the 9,10 bis phenylethynyl anthracene with Rhodamine B, Rubrene and 9,10 diphenylanthracene respectively. In a glowstick a Fluorophore fluorescent dye converts much of the light energy produced into the visible spectrum producing a brighter glow. Preparation TCPO can be prepared from a solution of 2,4,6 trichlorophenol in a solution of dry toluene by reaction with oxalyl chloride in the presence of a base such as triethylamine . This method produces crude TCPO with a by product of triethylamine hydrochloride . References reflist Category Organochlorides Category Oxalates organic compound stub fr Oxalate de di 2,4,6 trichloroph nyle ... more details
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