Effects and avoidance of photoconversion-induced artifacts in confocal and STED microscopy.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 38834747.
- Also identified by DOI 10.1038/s41592-024-02297-4 and PMC identifier 11543600.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
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.
Medical subject headings
- Artifacts
- Microscopy, Fluorescence
- Microscopy, Confocal