Journal
INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY
Volume 101, Issue -, Pages 74-79Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.biocel.2018.05.014
Keywords
Super-resolution microscopy; Fluorescence; Image processing; Live-cell imaging
Categories
Funding
- UK Biotechnology and Biological Sciences Research Council [BB/M022374/1, BB/P027431/1, BB/R000697/1]
- Wellcome Trust [203276/Z/16/2]
- 4-year MRC Research Studentship
- BBSRC [BB/M022374/1, BB/R000697/1, BB/P027431/1] Funding Source: UKRI
- MRC [MR/K015826/1] Funding Source: UKRI
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Super-resolution microscopy techniques break the diffraction limit of conventional optical microscopy to achieve resolutions approaching tens of nanometres. The major advantage of such techniques is that they provide resolutions close to those obtainable with electron microscopy while maintaining the benefits of light microscopy such as a wide palette of high specificity molecular labels, straightforward sample preparation and live-cell compatibility. Despite this, the application of super-resolution microscopy to dynamic, living samples has thus far been limited and often requires specialised, complex hardware. Here we demonstrate how a novel analytical approach, Super-Resolution Radial Fluctuations (SRRF), is able to make live-cell super-resolution microscopy accessible to a wider range of researchers. We show its applicability to live samples expressing GFP using commercial confocal as well as laser- and LED-based widefield microscopes, with the latter achieving long-term timelapse imaging with minimal photobleaching.
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