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Dasgupta A, Koerfer A, Kokot B, Urbančič I, Eggeling C, Carravilla P
Nat. Methods 21 (7) 1171-1174 [2024-07-00; online 2024-06-04]
Fluorescence microscopy is limited by photoconversion due to continuous illumination, which results in not only photobleaching but also conversion of fluorescent molecules into species of different spectral properties through photoblueing. Here, we determined different fluorescence parameters of photoconverted products for various fluorophores under standard confocal and stimulated emission depletion (STED) microscopy conditions. We observed changes in both fluorescence spectra and lifetimes that can cause artifacts in quantitative measurements, which can be avoided by using exchangeable dyes.
PubMed 38834747
DOI 10.1038/s41592-024-02297-4
Crossref 10.1038/s41592-024-02297-4
pmc: PMC11543600
pii: 10.1038/s41592-024-02297-4