Fluorescence Correlation Spectroscopy - Common Fluorescent Probes

Common Fluorescent Probes

The fluorescent species used in FCS is typically a biomolecule of interest that has been tagged with a fluorophore (using immunohistochemistry for instance), or is a naked fluorophore that is used to probe some environment of interest (e.g. the cytoskeleton of a cell). The following table gives diffusion coefficients of some common fluorophores in water at room temperature, and their excitation wavelengths.

Fluorescent dye (×10−10 m2 s−1) Excitation wavelength (nm) Reference
Rhodamine 6G 2.8, 3.0, 4.14 ± 0.05 @ 25.00 °C 514 ,
Rhodamine 110 2.7 488
Tetramethyl rhodamine 2.6 543
Cy3 2.8 543
Cy5 2.5, 3.7 ± 0.15 @ 25.00 °C 633 ,
carboxyfluorescein 3.2 488
Alexa-488 1.96,4.35 @ 22.5±0.5 °C 488

Atto655-maleimide 4.07 ± 0.1 @ 25.00 °C 663
Atto655-carboxylicacid 4.26 ± 0.08 @ 25.00 °C 663
2′, 7′-difluorofluorescein (Oregon Green488) 4.11 ± 0.06 @ 25.00 °C 498

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