Photobleaching is the permanent loss of fluorescence after extended exposure to light and is a major limiting factor in super-resolution microscopy (SRM) that restricts spatiotemporal resolution and observation time. Strategies for preventing or overcoming photobleaching in SRM are reviewed developing new probes and chemical environments. Photostabilization strategies are introduced first, which are borrowed from conventional fluorescence microscopy, that are employed in SRM. SRM-specific strategies are then highlighted that exploit the on–off transitions of fluorescence, which is the key mechanism for achieving super-resolution, which are becoming new routes to address photobleaching in SRM. Off states can serve as a shelter from excitation by light or an exit to release a damaged probe and replace it with a fresh one. Such efforts in overcoming the photobleaching limits are anticipated to enhance resolution to molecular scales and to extend the observation time to physiological lifespans.
Bibliographical noteFunding Information:
This work was supported by the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT (Grant No. 2021R1A2C2010792 and 2021R1A4A1032114). M.S.E. was supported by Korea Research Fellowship Program through NRF funded by the Ministry of Science and ICT (Grant No. 2019H1D3A1A01711).
© 2022 The Authors. Advanced Science published by Wiley-VCH GmbH.
- super-resolution fluorescence microscopy
ASJC Scopus subject areas
- Medicine (miscellaneous)
- General Chemical Engineering
- General Materials Science
- Biochemistry, Genetics and Molecular Biology (miscellaneous)
- General Engineering
- General Physics and Astronomy