Correlative super-resolution microscopy with deep UV reactivation.

Autor: Prakash K; Division of Molecular Pathology, The Institute of Cancer Research, London, UK.; The Royal Marsden NHS Foundation Trust, London, UK.
Jazyk: angličtina
Zdroj: Journal of microscopy [J Microsc] 2024 Nov; Vol. 296 (2), pp. 133-138. Date of Electronic Publication: 2024 Jan 03.
DOI: 10.1111/jmi.13258
Abstrakt: Correlative super-resolution microscopy has the potential to accurately visualize and validate new biological structures past the diffraction limit. However, combining different super-resolution modalities, such as deterministic stimulated emission depletion (STED) and stochastic single-molecule localization microscopy (SMLM), is a challenging endeavour. For correlative STED and SMLM, the following poses a significant challenge: (1) the photobleaching of the fluorophores in STED; (2) the subsequent reactivation of the fluorophores for SMLM and (3) finding the right fluorochrome and imaging buffer for both imaging modalities. Here, we highlight how the deep ultraviolet (DBUE) wavelengths of the Mercury (Hg) arc lamp can help recover STED bleaching and allow for the reactivation of single molecules for SMLM imaging. We also show that Alexa Fluor 594 and the commercially available Prolong Diamond to be excellent fluorophores and imaging media for correlative STED and SMLM.
(© 2023 The Author(s). Journal of Microscopy published by John Wiley & Sons Ltd on behalf of Royal Microscopical Society.)
Databáze: MEDLINE
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