Journal
MICROSCOPY AND MICROANALYSIS
Volume 18, Issue 6, Pages 1419-1429Publisher
CAMBRIDGE UNIV PRESS
DOI: 10.1017/S1431927612013347
Keywords
single molecule; super-resolution; fluorescence; microscopy; PALM; STORM
Categories
Funding
- Institut Pasteur
- Fondation pour la Recherche Medicale
- EMBO
- financement jeune chercheur from Sidaction
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For over a decade fluorescence microscopy has demonstrated the capacity to achieve single-molecule localization accuracies of a few nanometers, well below the similar to 200 nm lateral and similar to 500 nm axial resolution limit of conventional microscopy. Yet, only the recent development of new fluorescence labeling modalities, the increase in sensitivity of imaging hardware, and the creation of novel image analysis tools allow for the emergence of single-molecule-based super-resolution imaging techniques. Novel methods such as photoactivated localization microscopy and stochastic optical reconstruction microscopy can typically reach a tenfold increase in resolution compared to standard microscopy methods. Their implementation is relatively easy only requiring minimal changes to a conventional wide-field or total internal reflection fluorescence microscope. The recent translation of these two methods into commercial imaging systems has made them further accessible to researchers in biology. However, these methods are still evolving rapidly toward imaging live samples with high temporal resolution and depth. In this review, we recall the roots of single-molecule localization microscopy, summarize major recent developments, and offer perspective on potential applications.
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