Photochemical Mechanisms of Fluorophores Employed in Single‐Molecule Localization Microscopy

• Published in: Angewandte Chemie International Edition Vol. 62; no. 1; pp. e202204745 - n/a
• Main Authors: Kikuchi, Kai, Adair, Liam D., Lin, Jiarun, New, Elizabeth J., Kaur, Amandeep
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 02.01.2023; John Wiley and Sons Inc
• Edition: International ed. in English
• Subjects: Atmospheric chemistry; Biomolecules; Chemical compounds; Fluorescence; Fluorescent Dyes - chemistry; Fluorescent indicators; Fluorophores; Imaging; Localization; Medical research; Microscopy; Microscopy, Fluorescence - methods; Photochemicals; Photochemistry; Photoswitching; Probes; Review; Reviews; Sensors; Single Molecule Imaging - methods; Spatial distribution; Super-Resolution; Super-Resolution; Fluorescence; Photoswitching; Sensors; Photochemistry
• ISSN: 1433-7851; 1521-3773
• DOI: 10.1002/anie.202204745

Decoding cellular processes requires visualization of the spatial distribution and dynamic interactions of biomolecules. It is therefore not surprising that innovations in imaging technologies have facilitated advances in biomedical research. The advent of super‐resolution imaging technologies has empowered biomedical researchers with the ability to answer long‐standing questions about cellular processes at an entirely new level. Fluorescent probes greatly enhance the specificity and resolution of super‐resolution imaging experiments. Here, we introduce key super‐resolution imaging technologies, with a brief discussion on single‐molecule localization microscopy (SMLM). We evaluate the chemistry and photochemical mechanisms of fluorescent probes employed in SMLM. This Review provides guidance on the identification and adoption of fluorescent probes in single molecule localization microscopy to inspire the design of next‐generation fluorescent probes amenable to single‐molecule imaging. Single molecule localization microscopy has revolutionized our ability to investigate cellular structures and processes at the nanometer scale. This Review discusses the photochemical mechanisms harnessed to develop small‐molecule probes that exhibit photoswitching properties necessary for application in imaging methods based on single‐molecule localization. Strategies to develop improved photoswitchable probes and future directions are also discussed.