Fluoprobes
Encyclopedia
| Fluorescent dye | Color | mass (g/mol) | Absorb (nm) | Emit (nm) | ε (M−1cm−1) |
|---|---|---|---|---|---|
| FluoProbes 390 | violet | 343 | 390 | 479 | 24 000 |
| FluoProbes 488 | green | 804 | 493 | 519 | 85 000 |
| FluoProbes 532 | yellow | 765 | 532 | 553 | 117 000 |
| FluoProbes547H | orange | 736 | 557 | 574 | 150 000 |
| FluoProbes 594 | red | 1137 | 601 | 627 | 120 000 |
| FluoProbes647H | far-red | 761 | 653 | 674 | 250 000 |
| FluoProbes 682 | far-red | 853 | 690 | 709 | 140 000 |
| FluoProbes 752 | near-IR | 879 | 748 | 772 | 270 000 |
| FluoProbes 782 | near-IR | 976 | 783 | 800 | 170 000 |
| Abs = absorption maximum#, Em = emission maximum# .................................. ε = molar extinction coefficient |
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The FluoProbes series of fluorescent
Fluorescence
Fluorescence is the emission of light by a substance that has absorbed light or other electromagnetic radiation of a different wavelength. It is a form of luminescence. In most cases, emitted light has a longer wavelength, and therefore lower energy, than the absorbed radiation...
dyes were developed by Interchim
Interchim
Interchim 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....
to improve performances of standard fluorophore
Fluorophore
A fluorophore, in analogy to a chromophore, is a component of a molecule which causes a molecule to be fluorescent. It is a functional group in a molecule which will absorb energy of a specific wavelength and re-emit energy at a different wavelength...
s. They are designed for labeling biomolecule
Biomolecule
A biomolecule is any molecule that is produced by a living organism, including large polymeric molecules such as proteins, polysaccharides, lipids, and nucleic acids as well as small molecules such as primary metabolites, secondary metabolites, and natural products...
s, cells, tissues or beads in advanced fluorescent detection techniques.
- FluoProbes dyes are typically used to label proteins or nucleic acids (ultrafast -3 minutes- labeling of antibodiesAntibodyAn antibody, also known as an immunoglobulin, is a large Y-shaped protein used by the immune system to identify and neutralize foreign objects such as bacteria and viruses. The antibody recognizes a unique part of the foreign target, termed an antigen...
employs Lightning technology). Labeled products can be used for multiparameter detections, life time resolved fluorescence (TRF), polarisation anisotropy fluorescence, FRET, Quenching, FRAPFluorescence 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...
. They are typically used in biotechnology and research applications as fluorescence microscopyFluorescence microscopeA fluorescence microscope is an optical microscope used to study properties of organic or inorganic substances using the phenomena of fluorescence and phosphorescence instead of, or in addition to, reflection and absorption...
, cell biologyCell biologyCell biology is a scientific discipline that studies cells – their physiological properties, their structure, the organelles they contain, interactions with their environment, their life cycle, division and death. This is done both on a microscopic and molecular level...
or molecular biologyMolecular biologyMolecular biology is the branch of biology that deals with the molecular basis of biological activity. This field overlaps with other areas of biology and chemistry, particularly genetics and biochemistry...
, as well as infrared imaging. Derivatives with AmineAmineAmines are organic compounds and functional groups that contain a basic nitrogen atom with a lone pair. Amines are derivatives of ammonia, wherein one or more hydrogen atoms have been replaced by a substituent such as an alkyl or aryl group. Important amines include amino acids, biogenic amines,...
and Carboxyl suit peptide and nucleic acid synthesis, while reactive ones (with succinimidylN-HydroxysuccinimideN-Hydroxysuccinimide is a compound with a molecular weight of 115.09 and a melting point of 95 °C.As it is slightly acidic, it is an irritant to skin, eyes and mucous membranes....
, 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...
and hydrazideHydrazideHydrazides in organic chemistry are a class of organic compounds sharing a common functional group characterized by a nitrogen to nitrogen covalent bond with 4 substituents with at least one of them being an acyl group. The general structure for an hydrazide is R2-N-N-R3R4. A related class of...
) suit conjugation by conventional chemistry, while FluoProbes labeled antibodies and cellular probes (i.e. PhalloidinPhalloidinPhalloidin is one of a group of toxins from the death cap known as phallotoxins.-Background:Pioneering work on this toxin was done by the Nobel laureate Heinrich Wieland in the 1930s...
) suit direct use in immunoassays or cell assays.
- FluoProbes dyes that have comparable excitation and emission spectra to standard fluorophores such as 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....
s, 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...
s, cyanines Cy2/3/5/5.5/7CyanineCyanine 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...
, are claimed to solve limiting issues observed in some applications such as too high background, insufficient polarity, photobleachingPhotobleachingPhotobleaching is the photochemical destruction of a fluorophore. In microscopy, photobleaching may complicate the observation of fluorescent molecules, since they will eventually be destroyed by the light exposure necessary to stimulate them into fluorescing...
, insufficient brightness, or pHPHIn 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...
-sensitivity. I.e., FluoProbes488 reduces the backgroung in Flow CytometryFlow cytometryFlow cytometry is a technique for counting and examining microscopic particles, such as cells and chromosomes, by suspending them in a stream of fluid and passing them by an electronic detection apparatus. It allows simultaneous multiparametric analysis of the physical and/or chemical...
and in slide microscopy allowing sharper and brighter images. FluoProbes 488, 547H and 647H are found more photostable that is taken to good account in applications using long illuminations periods (i.e. scanning such as in confocal microscopyConfocal microscopyConfocal microscopy is an optical imaging technique used to increase optical resolution and contrast of a micrograph by using point illumination and a spatial pinhole to eliminate out-of-focus light in specimens that are thicker than the focal plane. It enables the reconstruction of...
), or for longer shelf-life of reagents (i.e. manufacturing diagnostics).
| FluoProbes dye | Color | Light sources (spectral line) |
|---|---|---|
| FluoProbes 390 | violet | Diode laser Laser diode The laser diode is a laser where the active medium is a semiconductor similar to that found in a light-emitting diode. The most common type of laser diode is formed from a p-n junction and powered by injected electric current... |
| FluoProbes 488 Fluorescein(FITC) 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... /Cy2 |
cyan | Argon laser (488.0nm) Ion laser An ion laser is a gas laser which uses an ionized gas as its lasing medium.Like other gas lasers, ion lasers feature a sealed cavity containing the laser medium and mirrors forming a Fabry–Pérot resonator. Unlike HeNe lasers, the energy level transitions that contribute to laser action come from ions... , Krypton laser (482.5nm) Ion laser An ion laser is a gas laser which uses an ionized gas as its lasing medium.Like other gas lasers, ion lasers feature a sealed cavity containing the laser medium and mirrors forming a Fabry–Pérot resonator. Unlike HeNe lasers, the energy level transitions that contribute to laser action come from ions... |
| FluoProbes 532 | yellow | Helium-neon laser (632.8nm) Helium-neon laser A helium–neon laser or HeNe laser, is a type of gas laser whose gain medium consists of a mixture of helium and neon inside of a small bore capillary tube, usually excited by a DC electrical discharge.- History of HeNe laser development:... |
| FluoProbes 547H TRITC/Cy3 |
orange | Argon laser (528.7nm) Ion laser An ion laser is a gas laser which uses an ionized gas as its lasing medium.Like other gas lasers, ion lasers feature a sealed cavity containing the laser medium and mirrors forming a Fabry–Pérot resonator. Unlike HeNe lasers, the energy level transitions that contribute to laser action come from ions... |
| FluoProbes 594 SR101 Sulforhodamine 101 Sulforhodamine 101 is a red fluorescent dye. In neurophysiological experiments which comprise calcium imaging methods, it can be used for a counterstaining of astrocytes to be able to analyze data from neurons separately.... /TR |
red | Argon laser (528.7nm) Ion laser An ion laser is a gas laser which uses an ionized gas as its lasing medium.Like other gas lasers, ion lasers feature a sealed cavity containing the laser medium and mirrors forming a Fabry–Pérot resonator. Unlike HeNe lasers, the energy level transitions that contribute to laser action come from ions... |
| FluoProbes 647H Cy5 |
far red | Krypton laser (647.1nm) Ion laser An ion laser is a gas laser which uses an ionized gas as its lasing medium.Like other gas lasers, ion lasers feature a sealed cavity containing the laser medium and mirrors forming a Fabry–Pérot resonator. Unlike HeNe lasers, the energy level transitions that contribute to laser action come from ions... , Laser (633nm) Laser A laser is a device that emits light through a process of optical amplification based on the stimulated emission of photons. The term "laser" originated as an acronym for Light Amplification by Stimulated Emission of Radiation... |
- The excitationExcited stateExcitation is an elevation in energy level above an arbitrary baseline energy state. In physics there is a specific technical definition for energy level which is often associated with an atom being excited to an excited state....
and emission spectra of the FluoProbes series covers much of the visible spectrumVisible spectrumThe visible spectrum is the portion of the electromagnetic spectrum that is visible to the human eye. Electromagnetic radiation in this range of wavelengths is called visible light or simply light. A typical human eye will respond to wavelengths from about 390 to 750 nm. In terms of...
up to the infraredInfraredInfrared light is electromagnetic radiation with a wavelength longer than that of visible light, measured from the nominal edge of visible red light at 0.74 micrometres , and extending conventionally to 300 µm...
region, matching the commonly used light sources and filters.
Similar lines of fluorescent dyes
Fluorophore
A fluorophore, in analogy to a chromophore, is a component of a molecule which causes a molecule to be fluorescent. It is a functional group in a molecule which will absorb energy of a specific wavelength and re-emit energy at a different wavelength...
provide an alternative to the FluoProbes Dyes.