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Article in Nature Methods
Iztok Urbančič and Boštjan Kokot (Laboratory of Biophysics, Condensed Matter Physics Department, F5) have, together with colleagues from the Institute for Applied Optics and Biophysics (Jena, Germany), published a paper titled Effects and avoidance of photoconversion-induced artefacts in confocal and STED microscopy in Nature Methods. Authors have shown that high laser powers in fluorescence microscopy should be used with care; besides the well-known photobleaching, the dye’s emission spectrum can 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. The characterization of this phenomenon was made possible by a unique combination of a STED microscope equipped with a spectral detector, implemented at JSI.
