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Why does trypan blue quench FITC or GFP in flow cytometry?
Posted May 18, 2020
Chemical ReagentsCell ViabilityCell Viability AssaysCellular ProcessesFlow Cytometry ReagentsSpectral Flow Cytometryflow cytometry
Answer
Trypan blue absorbs light in the green-yellow region with a maximum absorbance wavelength around 590 nm in water. It can absorb the fluorescence emitted by green-emission fluorochromes or fluorescent proteins, thus quenching FITC or GRP in flow cytometry.
Additional resources
Trypan Blue, sodium salt *10 mM aqueous solution*Graham, J. P., Rauf, M. A., Hisaindee, S., & Nawaz, M. (2013). Experimental and theoretical study of the spectral behavior of Trypan Blue in various solvents. Journal of Molecular Structure, 1040, 1–8. doi:10.1016/j.molstruc.2013.02.015 Shilova, O. N., Shilov, E. S., & Deyev, S. M. (2017). The effect of trypan blue treatment on autofluorescence of fixed cells. Cytometry Part A, 91(9), 917–925. doi:10.1002/cyto.a.23199