Fluorophore
Encyclopedia
Chromophore
A chromophore is the part of a molecule responsible for its color. The color arises when a molecule absorbs certain wavelengths of visible light and transmits or reflects others. The chromophore is a region in the molecule where the energy difference between two different molecular orbitals falls...
, is a component of a molecule which causes a molecule to be fluorescent
Fluorescence in the life sciences
Fluorescence is used in the life sciences generally as a non-destructive way of tracking or analysing biological molecules by means of fluorescence....
. It is a functional group
Functional group
In organic chemistry, functional groups are specific groups of atoms within molecules that are responsible for the characteristic chemical reactions of those molecules. The same functional group will undergo the same or similar chemical reaction regardless of the size of the molecule it is a part of...
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
Biochemistry
Biochemistry, sometimes called biological chemistry, is the study of chemical processes in living organisms, including, but not limited to, living matter. Biochemistry governs all living organisms and living processes...
and protein
Protein
Proteins are biochemical compounds consisting of one or more polypeptides typically folded into a globular or fibrous form, facilitating a biological function. A polypeptide is a single linear polymer chain of amino acids bonded together by peptide bonds between the carboxyl and amino groups of...
studies, e.g., in immunofluorescence
Immunofluorescence
Immunofluorescence is a technique used for light microscopy with a fluorescence microscope and is used primarily on biological samples. This technique uses the specificity of antibodies to their antigen to target fluorescent dyes to specific biomolecule targets within a cell, and therefore allows...
and immunohistochemistry
Immunohistochemistry
Immunohistochemistry or IHC refers to the process of detecting antigens in cells of a tissue section by exploiting the principle of antibodies binding specifically to antigens in biological tissues. IHC takes its name from the roots "immuno," in reference to antibodies used in the procedure, and...
.
Fluorescein isothiocyanate
Fluorescein isothiocyanate
Fluorescein isothiocyanate is a derivative of fluorescein used in wide-ranging applications including flow cytometry. FITC is the original fluorescein molecule functionalized with an isothiocyanate reactive group , replacing a hydrogen atom on the bottom ring of the structure...
(FITC), a reactive derivative of fluorescein
Fluorescein
Fluorescein is a synthetic organic compound available as a dark orange/red powder soluble in water and alcohol. It is widely used as a fluorescent tracer for many applications....
, 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
Rhodamine
Rhodamine is a family of related chemical compounds, fluorone dyes. Examples are Rhodamine 6G and Rhodamine B. They are used as a dye and as a dye laser gain medium. They are often used as a tracer dye within water to determine the rate and direction of flow and transport...
(TRITC), coumarin
Coumarin
Coumarin is a fragrant chemical compound in the benzopyrone chemical class, found in many plants, notably in high concentration in the tonka bean , vanilla grass , sweet woodruff , mullein , sweet grass , cassia cinnamon and sweet clover...
, and cyanine
Cyanine
Cyanine is a non-systematic name of a synthetic dye family belonging to polymethine group. Cyanines have many uses as fluorescent dyes, particularly in biomedical imaging...
. Newer generations of fluorophores, many of which are proprietary, often perform better (more photostable, brighter, and/or less pH
PH
In chemistry, pH is a measure of the acidity or basicity of an aqueous solution. Pure water is said to be neutral, with a pH close to 7.0 at . Solutions with a pH less than 7 are said to be acidic and solutions with a pH greater than 7 are basic or alkaline...
-sensitive) than traditional dyes with comparable excitation and emission.
Size
The size of the fluorophore might stericSteric effects
Steric effects arise from the fact that each atom within a molecule occupies a certain amount of space. If atoms are brought too close together, there is an associated cost in energy due to overlapping electron clouds , and this may affect the molecule's preferred shape and reactivity.-Steric...
ally hinder the tagged molecule:
- quantum dotQuantum dotA quantum dot is a portion of matter whose excitons are confined in all three spatial dimensions. Consequently, such materials have electronic properties intermediate between those of bulk semiconductors and those of discrete molecules. They were discovered at the beginning of the 1980s by Alexei...
: 2-10 nm (diameter), 100-100,000 atoms - proteinProteinProteins are biochemical compounds consisting of one or more polypeptides typically folded into a globular or fibrous form, facilitating a biological function. A polypeptide is a single linear polymer chain of amino acids bonded together by peptide bonds between the carboxyl and amino groups of...
: Green fluorescent proteinGreen fluorescent proteinThe green fluorescent protein is a protein composed of 238 amino acid residues that exhibits bright green fluorescence when exposed to blue light. Although many other marine organisms have similar green fluorescent proteins, GFP traditionally refers to the protein first isolated from the...
(GFP) 26 kDaAtomic mass unitThe unified atomic mass unit or dalton is a unit that is used for indicating mass on an atomic or molecular scale. It is defined as one twelfth of the rest mass of an unbound neutral atom of carbon-12 in its nuclear and electronic ground state, and has a value of... - small moleculeSmall moleculeIn the fields of pharmacology and biochemistry, a small molecule is a low molecular weight organic compound which is by definition not a polymer...
: fluoresceinFluoresceinFluorescein is a synthetic organic compound available as a dark orange/red powder soluble in water and alcohol. It is widely used as a fluorescent tracer for many applications....
: about 20 atomAtomThe atom is a basic unit of matter that consists of a dense central nucleus surrounded by a cloud of negatively charged electrons. The atomic nucleus contains a mix of positively charged protons and electrically neutral neutrons...
s
Families
Several fluorescent proteinGreen fluorescent protein
The green fluorescent protein is a protein composed of 238 amino acid residues that exhibits bright green fluorescence when exposed to blue light. Although many other marine organisms have similar green fluorescent proteins, GFP traditionally refers to the protein first isolated from the...
s exist and are attached to a specific protein via a plasmid
Plasmid
In microbiology and genetics, a plasmid is a DNA molecule that is separate from, and can replicate independently of, the chromosomal DNA. They are double-stranded and, in many cases, circular...
constructor to form a fusion protein
Fusion protein
Fusion proteins or chimeric proteins are proteins created through the joining of two or more genes which originally coded for separate proteins. Translation of this fusion gene results in a single polypeptide with functional properties derived from each of the original proteins...
.
Fluorophores can be attached to protein to specific functional groups, such as
- amino groups (Active ester, Carboxylate, IsothiocyanateIsothiocyanateIsothiocyanate is the chemical group –N=C=S, formed by substituting sulfur for oxygen in the isocyanate group. Many natural isothiocyanates from plants are produced by enzymatic conversion of metabolites called glucosinolates. These natural isothiocyanates, such as allyl isothiocyanate, are also...
, hydrazineHydrazineHydrazine is an inorganic compound with the formula N2H4. It is a colourless flammable liquid with an ammonia-like odor. Hydrazine is highly toxic and dangerously unstable unless handled in solution. Approximately 260,000 tons are manufactured annually...
) - carboxyl groups (carbodiimideCarbodiimideA carbodiimide or a methanediimine is a functional group consisting of the formula RN=C=NR. Carbodiimides hydrolyze to form ureas, which makes them uncommon in nature.-Carbodiimide formation:...
) - thiolThiolIn organic chemistry, a thiol is an organosulfur compound that contains a carbon-bonded sulfhydryl group...
(maleimideMaleimideMaleimide is the chemical compound with the formula H2C22NH . This unsaturated imide is an important building block in organic synthesis. The name is a contraction of maleic acid and imide, the -CNHC- functional group...
, acetyl bromideAcetyl bromideAcetyl bromide is an acyl bromide compound. As is expected, it may be prepared by reaction between phosphorus tribromide and acetic acid:...
) - azideAzideAzide is the anion with the formula N3−. It is the conjugate base of hydrazoic acid. N3− is a linear anion that is isoelectronic with CO2 and N2O. Per valence bond theory, azide can be described by several resonance structures, an important one being N−=N+=N−...
(via click chemistryClick chemistryClick chemistry is a chemical philosophy introduced by K. Barry Sharpless of The Scripps Research Institute, in 2001 and describes chemistry tailored to generate substances quickly and reliably by joining small units together...
or non-specifically (glutaraldehydeGlutaraldehydeGlutaraldehyde is an organic compound with the formula CH22. A pungent colorless oily liquid, glutaraldehyde is used to disinfect medical and dental equipment...
).
These fluorophores are either quantum dots or small molecules. The former are fluorescent semiconductor nanoparticles. The latter molecules which fluoresce thanks to delocalized electrons which can jump a band and stabilize the energy absorbed, hence all fluorophores are aromatic or conjugated.
Benzene, one of the simplest aromatic hydrocarbons, for example, is excited at 254 nm and emits at 300 nm.
Additionally, various functional groups can be present to alter its proprieties, such as solubility, or confer special proprieties, such as boronic acid
Boronic acid
A boronic acid is an alkyl or aryl substituted boric acid containing a carbon–boron bond belonging to the larger class of organoboranes. Boronic acids act as Lewis acids. Their unique feature is that they are capable of forming reversible covalent complexes with sugars, amino acids, hydroxamic...
which binds to sugars or multiple carboxyl groups to bind to certain cations. When the dye contains an electron-donating and an electron-accepting group at opposite ends of the aromatic system, this dye will probably be sensitive to the environment's polarity (solvatochromic), hence called environment-sensitive. Often dyes are used inside cells, which are impermeable to charged molecules, as a result of this the carboxyl groups are converted into an ester, which is removed by esterases inside the cells, e.g., fura-2AM and fluorescein-diacetate.
Common dye families are:
- XantheneXantheneXanthene is a yellow organic heterocyclic compound. Its chemical formula is 1310. It is soluble in diethyl ether. Its melting point is 101-102 °C and its boiling point is 310-312 °C...
derivatives: fluoresceinFluoresceinFluorescein is a synthetic organic compound available as a dark orange/red powder soluble in water and alcohol. It is widely used as a fluorescent tracer for many applications....
, rhodamineRhodamineRhodamine is a family of related chemical compounds, fluorone dyes. Examples are Rhodamine 6G and Rhodamine B. They are used as a dye and as a dye laser gain medium. They are often used as a tracer dye within water to determine the rate and direction of flow and transport...
, Oregon green, eosin, Texas red, and Cal Fluor dyesBiosearch TechnologiesBiosearch Technologies, Inc. is a closely held biotechnology company based in Novato, California. This company specializes in custom synthesized oligonucleotides and DNA synthesis products.-Black Hole Quencher Dyes:... - CyanineCyanineCyanine is a non-systematic name of a synthetic dye family belonging to polymethine group. Cyanines have many uses as fluorescent dyes, particularly in biomedical imaging...
derivatives: cyanine, indocarbocyanineIndocyanine greenIndocyanine green is a cyanine dye used in medical diagnostics. It is used for determining cardiac output, hepatic function, and liver blood flow, and for ophthalmic angiography. It has a peak spectral absorption at about 800 nm. These infrared frequencies penetrate retinal layers, allowing ICG...
, oxacarbocyanine, thiacarbocyanine, merocyanineMerocyanineMerocyanines are a class of fluorescent dyes typified by merocycanine I.These dyes are usually intensely colored and have large extinction coefficients....
, and Quasar dyesBiosearch TechnologiesBiosearch Technologies, Inc. is a closely held biotechnology company based in Novato, California. This company specializes in custom synthesized oligonucleotides and DNA synthesis products.-Black Hole Quencher Dyes:... - NaphthaleneNaphthaleneNaphthalene is an organic compound with formula . It is a white crystalline solid with a characteristic odor that is detectable at concentrations as low as 0.08 ppm by mass. As an aromatic hydrocarbon, naphthalene's structure consists of a fused pair of benzene rings...
derivatives (dansylDansyl chlorideDansyl chloride or 5-naphthalene-1-sulfonyl chloride is a reagent that reacts with primary amino groups in both aliphatic and aromatic amines to produce stable blue- or blue-green–fluorescent sulfonamide adducts. Dansyl chloride is widely used to modify amino acids; specifically, protein sequencing...
and prodan derivatives) - CoumarinCoumarinCoumarin is a fragrant chemical compound in the benzopyrone chemical class, found in many plants, notably in high concentration in the tonka bean , vanilla grass , sweet woodruff , mullein , sweet grass , cassia cinnamon and sweet clover...
derivatives - oxadiazole derivatives: pyridyloxazole, nitrobenzoxadiazole and benzoxadiazole
- PyrenePyrenePyrene is a polycyclic aromatic hydrocarbon consisting of four fused benzene rings, resulting in a flat aromatic system. The chemical formula is . This colourless solid is the smallest peri-fused PAH...
derivatives: cascade blue etc. - Oxazine derivatives: Nile redNile redNile red is a lipophilic stain. It is produced by boiling a solution of Nile blue with sulfuric acid. As can be seen from the structural formulae, this process replaces an amino group with a carbonyl group. Nile red stains intracellular lipid droplets red...
, Nile blueNile blueNile blue is a stain used in biology and histology. It may be used with live or fixed cells, and imparts a blue colour to cell nuclei....
, cresyl violetCresyl violetCresyl violet is an organic compound with the chemical formula C19H18ClN3O. It is used in biology and medicine as a histological stain. Cresyl violet is an effective and reliable stain used for light microscopy sections. Initially, tissue sections are ‘defatted’ by passing through graded dilutions...
, oxazine 170 etc. - AcridineAcridineAcridine, C13H9N, is an organic compound and a nitrogen heterocycle. Acridine is also used to describe compounds containing the C13N tricycle....
derivatives: proflavin, acridine orange, acridine yellow etc. - Arylmethine derivatives: auramine, crystal violet, malachite green
- TetrapyrroleTetrapyrroleTetrapyrroles are compounds containing four pyrrole rings. With the exception of corrin, the four pyrrole rings are interconnected through one-carbon bridges, in either a linear or a cyclic fashion...
derivatives: porphinPorphinPorphin, sometimes spelled porphine, is the parent chemical compound for types of biochemically significant compounds called porphyrins. The chemical formula of porphin is C20H14N4...
, phtalocyanine, bilirubinBilirubinBilirubin is the yellow breakdown product of normal heme catabolism. Heme is found in hemoglobin, a principal component of red blood cells. Bilirubin is excreted in bile and urine, and elevated levels may indicate certain diseases...
The following dye families are trademark groups, and do not necessarily share structural similarities.
- CF dye (Biotium)
- BODIPYBODIPYBODIPY, short for boron-dipyrromethene, is a class of fluorescent dyes. It is composed of dipyrromethene complexed with a disubstituted boron atom, typically a BF2 unit. The IUPAC name for the BODIPY core is 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene...
(InvitrogenInvitrogenInvitrogen Corporation was a large, multinational biotechnology company headquartered in Carlsbad, California. In November 2008, a merger between Applied Biosystems and Invitrogen was finalized...
) - Alexa Fluor (Invitrogen)
- DyLight FluorDyLight FluorThe DyLight Fluor family of fluorescent dyes are produced by Dyomics in collaboration with Thermo Fisher Scientific. DyLight dyes are typically used in biotechnology and research applications as biomolecule, cell and tissue labels for fluorescence microscopy, cell biology or molecular...
(Thermo Scientific, Pierce) - AttoATTO dyesATTO Dyes are a series of fluorescent labels and dyes manufactured by ATTO-TEC GmbH in Siegen, Germany. The dyes have been designed for application in the area of life science, e.g...
and Tracy (Sigma Aldrich) - FluoProbesFluoprobesThe FluoProbes series of fluorescent dyes were developed by Interchim to improve performances of standard fluorophores. They are designed for labeling biomolecules, cells, tissues or beads in advanced fluorescent detection techniques....
(InterchimInterchimInterchim is a privately owned French company specialized in manufacturing and distribution of reagents, consumables and dedicated instruments for the R&D and industry laboratory in the fields of fine chemistry, chromatography and bio-analysis....
) - MegaStokes Dyes (Dyomics)
A few examples of commonly used dyes:
Reactive and conjugated probes
| Probe | Ex (nm) | Em (nm) | MW | Notes |
|---|---|---|---|---|
| Hydroxycoumarin | 325 | 386 | 331 | Succinimidyl ester |
| Aminocoumarin | 350 | 445 | 330 | Succinimidyl ester |
| Methoxycoumarin | 360 | 410 | 317 | Succinimidyl ester |
| Cascade Blue | (375);401 | 423 | 596 | Hydrazide |
| Pacific Blue | 403 | 455 | 406 | Maleimide |
| Pacific Orange | 403 | 551 | ||
| Lucifer yellow Lucifer yellow Lucifer yellow is a fluorescent dye used in cell biology. The key property of Lucifer yellow is that it can readily visualized in both living and fixed cells using a fluorescence microscope. Lucifer yellow was engineered by Walter W. Stewart at NIH and patented in 1978.- Preparations :For common... |
425 | 528 | ||
| NBD | 466 | 539 | 294 | NBD-X |
| R-Phycoerythrin (PE) | 480;565 | 578 | 240 k | |
| PE-Cy5 conjugates | 480;565;650 | 670 | aka Cychrome, R670, Tri-Color, Quantum Red | |
| PE-Cy7 conjugates | 480;565;743 | 767 | ||
| Red 613 | 480;565 | 613 | PE-Texas Red | |
| PerCP | 490 | 675 | Peridinin chlorphyll protein | |
| TruRed | 490,675 | 695 | PerCP-Cy5.5 conjugate | |
| FluorX | 494 | 520 | 587 | (GE Healthcare) |
| Fluorescein Fluorescein Fluorescein is a synthetic organic compound available as a dark orange/red powder soluble in water and alcohol. It is widely used as a fluorescent tracer for many applications.... |
495 | 519 | 389 | FITC; pH sensitive |
| BODIPY-FL | 503 | 512 | ||
| TRITC | 547 | 572 | 444 | TRITC |
| X-Rhodamine | 570 | 576 | 548 | XRITC |
| Lissamine Rhodamine B | 570 | 590 | ||
| Texas Red Texas Red Texas Red or sulforhodamine 101 acid chloride is a red fluorescent dye, used in histology for staining cell specimens, for sorting cells with fluorescent-activated cell sorting machines, in fluorescence microscopy applications, and in immunohistochemistry.Texas Red fluoresces at about 615 nm,... |
589 | 615 | 625 | Sulfonyl chloride |
| Allophycocyanin (APC) | 650 | 660 | 104 k | |
| APC-Cy7 conjugates | 650;755 | 767 | PharRed |
Abbreviations:
Ex (nm): Excitation wavelength in nanometers
Em (nm): Emission wavelength in nanometers
MW: Molecular weight
CyDyes
| Probe | Ex (nm) | Em (nm) | MW | Quantum yield |
|---|---|---|---|---|
| Cy2 | 489 | 506 | 714 | QY 0.12 |
| Cy3 | (512);550 | 570;(615) | 767 | QY 0.15 |
| Cy3B | 558 | 572;(620) | 658 | QY 0.67 |
| Cy3.5 | 581 | 594;(640) | 1102 | QY 0.15 |
| Cy5 | (625);650 | 670 | 792 | QY 0.28 |
| Cy5.5 | 675 | 694 | 1128 | QY 0.23 |
| Cy7 | 743 | 767 | 818 | QY 0.28 |
Nucleic acid probes
| Probe | Ex (nm) | Em (nm) | MW | Notes |
|---|---|---|---|---|
| Hoechst Hoechst stain Hoechst stains are part of a family of blue fluorescent dyes used to stain DNA. These Bis-benzimides were originally developed by the Hoechst AG, which numbered all their compounds so that the dye Hoechst 33342 is the 33342nd compound made by the company. There are three related Hoechst stains:... 33342 |
343 | 483 | 616 | AT-selective |
| DAPI DAPI DAPI or 4',6-diamidino-2-phenylindole is a fluorescent stain that binds strongly to A-T rich regions in DNA. It is used extensively in fluorescence microscopy... |
345 | 455 | AT-selective | |
| Hoechst Hoechst stain Hoechst stains are part of a family of blue fluorescent dyes used to stain DNA. These Bis-benzimides were originally developed by the Hoechst AG, which numbered all their compounds so that the dye Hoechst 33342 is the 33342nd compound made by the company. There are three related Hoechst stains:... 33258 |
345 | 478 | 624 | AT-selective |
| SYTOX Blue | 431 | 480 | ~400 | DNA |
| Chromomycin A3 | 445 | 575 | CG-selective | |
| Mithramycin | 445 | 575 | ||
| YOYO-1 | 491 | 509 | 1271 | |
| Ethidium Bromide | 493 | 620 | 394 | |
| Acridine Orange Acridine orange Acridine orange is a nucleic acid selective fluorescent cationic dye useful for cell cycle determination. It is cell-permeable, and interacts with DNA and RNA by intercalation or electrostatic attractions respectively. When bound to DNA, it is very similar spectrally to fluorescein, with an... |
503 | 530/640 | DNA/RNA | |
| SYTOX Green | 504 | 523 | ~600 | DNA |
| TOTO-1, TO-PRO-1 | 509 | 533 | Vital stain, TOTO: Cyanine Dimer | |
| TO-PRO: Cyanine Monomer | ||||
| Thiazole Orange | 510 | 530 | ||
| Propidium Iodide (PI) | 536 | 617 | 668.4 | |
| LDS 751 | 543;590 | 712;607 | 472 | DNA (543ex/712em), RNA (590ex/607em) |
| 7-AAD | 546 | 647 | 7-aminoactinomycin D, CG-selective | |
| SYTOX Orange | 547 | 570 | ~500 | DNA |
| TOTO-3, TO-PRO-3 | 642 | 661 | ||
| DRAQ5 | 647 | 681/697 | 413 | (Biostatus) (usable excitation down to 488) |
Cell function probes
| Probe | Ex (nm) | Em (nm) | MW | Notes |
|---|---|---|---|---|
| Indo-1 | 361/330 | 490/405 | 1010 | AM ester, low/high calcium (Ca2+) |
| Fluo-3 | 506 | 526 | 855 | AM ester. pH > 6 |
| Fluo-4 | 491/494 | 516 | 1097 | AM ester. pH 7.2 |
| DCFH | 505 | 535 | 529 | 2'7'Dichorodihydrofluorescein, oxidized form |
| DHR | 505 | 534 | 346 | Dihydrorhodamine 123, oxidized form, light catalyzes oxidation |
| SNARF | 548/579 | 587/635 | pH 6/9 |
Fluorescent proteins
| Probe | Ex (nm) | Em (nm) | MW | QY | BR | PS | Notes |
|---|---|---|---|---|---|---|---|
| Y66H | 360 | 442 | |||||
| Y66F | 360 | 508 | |||||
| EBFP | 380 | 440 | 0.18 | 0.27 | monomer | ||
| EBFP2 | 383 | 448 | 20 | monomer | |||
| Azurite | 383 | 447 | 15 | monomer | |||
| GFPuv | 385 | 508 | |||||
| T-Sapphire | 399 | 511 | 0.60 | 26 | 25 | weak dimer | |
| Cerulean | 433 | 475 | 0.62 | 27 | 36 | weak dimer | |
| mCFP | 433 | 475 | 0.40 | 13 | 64 | monomer | |
| ECFP | 434 | 477 | 0.15 | 3 | |||
| CyPet | 435 | 477 | 0.51 | 18 | 59 | weak dimer | |
| Y66W | 436 | 485 | |||||
| mKeima-Red | 440 | 620 | 0.24 | 3 | monomer (MBL) | ||
| TagCFP | 458 | 480 | 29 | dimer (Evrogen) | |||
| AmCyan1 | 458 | 489 | 0.75 | 29 | tetramer, (Clontech) | ||
| mTFP1 | 462 | 492 | 54 | dimer | |||
| S65A | 471 | 504 | |||||
| Midoriishi Cyan | 472 | 495 | 0.9 | 25 | dimer (MBL) | ||
| Wild Type GFP | 396,475 | 508 | 26k | 0.77 | |||
| S65C | 479 | 507 | |||||
| TurboGFP | 482 | 502 | 26 k | 0.53 | 37 | dimer, (Evrogen) | |
| TagGFP | 482 | 505 | 34 | monomer (Evrogen) | |||
| S65L | 484 | 510 | |||||
| Emerald | 487 | 509 | 0.68 | 39 | 0.69 | weak dimer, (Invitrogen) | |
| S65T | 488 | 511 | |||||
| EGFP | 488 | 507 | 26k | 0.60 | 34 | 174 | weak dimer, (Clontech) |
| Azami Green | 492 | 505 | 0.74 | 41 | monomer (MBL) | ||
| ZsGreen1 | 493 | 505 | 105k | 0.91 | 40 | tetramer, (Clontech) | |
| TagYFP | 508 | 524 | 47 | monomer (Evrogen) | |||
| EYFP | 514 | 527 | 26k | 0.61 | 51 | 60 | weak dimer, (Clontech) |
| Topaz | 514 | 527 | 57 | monomer | |||
| Venus | 515 | 528 | 0.57 | 53 | 15 | weak dimer | |
| mCitrine | 516 | 529 | 0.76 | 59 | 49 | monomer | |
| YPet | 517 | 530 | 0.77 | 80 | 49 | weak dimer | |
| TurboYFP | 525 | 538 | 26 k | 0.53 | 55.7 | dimer, (Evrogen) | |
| ZsYellow1 | 529 | 539 | 0.65 | 13 | tetramer, (Clontech) | ||
| Kusabira Orange | 548 | 559 | 0.60 | 31 | monomer (MBL) | ||
| mOrange | 548 | 562 | 0.69 | 49 | 9 | monomer | |
| mKO | 548 | 559 | 0.60 | 31 | 122 | monomer | |
| TurboRFP | 553 | 574 | 26 k | 0.67 | 62 | dimer, (Evrogen) | |
| tdTomato | 554 | 581 | 0.69 | 95 | 98 | tandem dimer | |
| TagRFP | 555 | 584 | 50 | monomer (Evrogen) | |||
| DsRed monomer | 556 | 586 | ~28k | 0.1 | 3.5 | 16 | monomer, (Clontech) |
| DsRed2 ("RFP") | 563 | 582 | ~110k | 0.55 | 24 | (Clontech) | |
| mStrawberry | 574 | 596 | 0.29 | 26 | 15 | monomer | |
| TurboFP602 | 574 | 602 | 26 k | 0.35 | 26 | dimer, (Evrogen) | |
| AsRed2 | 576 | 592 | ~110k | 0.21 | 13 | tetramer, (Clontech) | |
| mRFP1 | 584 | 607 | ~30k | 0.25 | monomer, (Tsien lab) | ||
| J-Red | 584 | 610 | 0.20 | 8.8 | 13 | dimer | |
| mCherry | 587 | 610 | 0.22 | 16 | 96 | monomer | |
| HcRed1 | 588 | 618 | ~52k | 0.03 | 0.6 | dimer, (Clontech) | |
| Katusha | 588 | 635 | 23 | dimer | |||
| mKate (TagFP635) | 588 | 635 | 15 | monomer (Evrogen) | |||
| TurboFP635 | 588 | 635 | 26 k | 0.34 | 22 | dimer, (Evrogen) | |
| mPlum | 590 | 649 | 51.4 k | 0.10 | 4.1 | 53 | |
| mRaspberry | 598 | 625 | 0.15 | 13 | monomer, faster photobleach than mPlum |
Abbreviations:
Ex (nm): Excitation wavelength in nanometers
Em (nm): Emission wavelength in nanometers
MW: Molecular weight
QY: Quantum yield
Quantum yield
The quantum yield of a radiation-induced process is the number of times that a defined event occurs per photon absorbed by the system. The "event" may represent a chemical reaction, for example the decomposition of a reactant molecule:...
BR: Brightness: Extinction coefficient * quantum yield / 1000
PS: Photostability: time [sec] to reduce brightness by 50%
Uses outside the life sciences
Additionally fluorescent dyes find a wide use in industry, going under the name of "neon colours", such as- multi-ton scale usages in textile dyeing and optical brighteners in laundry detergents
- advanced cosmetic formulations; safety equipment and clothing
- organic light-emitting diodes (OLED)
- fine arts and design (posters and paintings)
- synergists for insecticides and experimental drugs
- used as a dye in highlighters to give off a glow-like effect
See also
- Dark quencherDark quencherA dark quencher is a substance that absorbs excitation energy from a fluorophore and dissipates the energy as heat; while a typical quencher re-emits much of this energy as light . Dark quenchers are used in molecular biology in conjunction with fluorophores...
:Category:Fluorescent dyes
- Fluorescence recovery after photobleachingFluorescence recovery after photobleachingFluorescence recovery after photobleaching denotes an optical technique capable of quantifying the two dimensional lateral diffusion of a molecularly thin film containing fluorescently labeled probes, or to examine single cells. This technique is very useful in biological studies of cell membrane...
(FRAP) - an application for quantifying mobility of molecules in lipid bilayerLipid bilayerThe lipid bilayer is a thin membrane made of two layers of lipid molecules. These membranes are flat sheets that form a continuous barrier around cells. The cell membrane of almost all living organisms and many viruses are made of a lipid bilayer, as are the membranes surrounding the cell nucleus...
s. - Fluorescence in the life sciencesFluorescence in the life sciencesFluorescence is used in the life sciences generally as a non-destructive way of tracking or analysing biological molecules by means of fluorescence....
- InvitrogenInvitrogenInvitrogen Corporation was a large, multinational biotechnology company headquartered in Carlsbad, California. In November 2008, a merger between Applied Biosystems and Invitrogen was finalized...
External links
- The Database of fluorescent dyes
- Table of fluorochromes
- Fluorescence Tutorials
- Fluorescence SpectraViewer - check the compatibility of your fluorophores when designing multicolor experiments.
- The Molecular Probes Handbook - a comprehensive resource for fluorescence technology and its applications.