TY - JOUR
T1 - SRRF
T2 - Universal live-cell super-resolution microscopy
AU - Culley, Siân
AU - Tosheva, Kalina L.
AU - Matos Pereira, Pedro
AU - Henriques, Ricardo
N1 - Funding Information:
We thank Romain Laine (University College London) for critical reading of the manuscript. This work was funded by grants from the UK Biotechnology and Biological Sciences Research Council ( BB/M022374/1 ; BB/P027431/1 ; BB/R000697/1 ) (R.H. and P.M.P.) and the Wellcome Trust ( 203276/Z/16/Z ) (S.C. and R.H.). K.L.T. is supported by a 4-year MRC Research Studentship .
Publisher Copyright:
© 2018 The Authors
PY - 2018/8
Y1 - 2018/8
N2 - 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.
AB - 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.
KW - Fluorescence
KW - Image processing
KW - Live-cell imaging
KW - Super-resolution microscopy
UR - http://www.scopus.com/inward/record.url?scp=85047783919&partnerID=8YFLogxK
U2 - 10.1016/j.biocel.2018.05.014
DO - 10.1016/j.biocel.2018.05.014
M3 - Short survey
C2 - 29852248
AN - SCOPUS:85047783919
SN - 1357-2725
VL - 101
SP - 74
EP - 79
JO - International Journal of Biochemistry and Cell Biology
JF - International Journal of Biochemistry and Cell Biology
ER -