Effects and avoidance of photoconversion-induced artifacts in confocal and STED microscopy
Anindita Dasgupta, Agnes Koerfer, Boštjan Kokot, Iztok Urbančič, Christian Eggeling, Pablo CarravillaNature methods, June 2024; doi: 10.1038/s41592-024-02297-4 (read more here).
In brief
High laser powers in fluorescence microscopy should be used with care; besides the well-known photobleaching, the dye’s emission spectrum can unexpectedly shift towards lower wavelengths (“photoblueing”). The fluorescence signal is consequently detected in another spectral channel attributed to another dye, possibly leading to erroneous experimental conclusions. Detailed characterization of this phenomenon was done by our unique combination of a STED microscope equipped with a spectral detector.
