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journal article

Multiplane 3D superresolution optical fluctuation imaging

Geissbuehler, Stefan
•
Sharipov, Azat  
•
Godinat, Aurélien  
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2013
arXiv

By switching fluorophores on and off in either a deterministic or a stochastic manner, superresolution microscopy has enabled the imaging of biological structures at resolutions well beyond the diffraction limit. Superresolution optical fluctuation imaging (SOFI) provides an elegant way of overcoming the diffraction limit in all three spatial dimensions by computing higher-order cumulants of image sequences of blinking fluorophores acquired with a conventional widefield microscope. So far, three-dimensional (3D) SOFI has only been demonstrated by sequential imaging of multiple depth positions. Here we introduce a versatile imaging scheme which allows for the simultaneous acquisition of multiple focal planes. Using 3D cross-cumulants, we show that the depth sampling can be increased. Consequently, the simultaneous acquisition of multiple focal planes reduces the acquisition time and hence the photo-bleaching of fluorescent markers. We demonstrate multiplane 3D SOFI by imaging the mitochondria network in fixed C2C12 cells over a total volume of 65x65x3.5µm3 without depth scanning.

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Type
journal article
ArXiv ID

1310.5969

Author(s)
Geissbuehler, Stefan
Sharipov, Azat  
Godinat, Aurélien  
Bocchio, Noelia  
Dubikovskaya, Elena  
Lasser, Theo  
Leutenegger, Marcel  
Date Issued

2013

Published in
arXiv
Subjects

3D imaging

•

superresolution

•

stochastic switching

•

cross-cumulants

•

molecular statistics

Note

arXiv:1310.5969 [physics.optics]

Editorial or Peer reviewed

NON-REVIEWED

Written at

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Available on Infoscience
March 12, 2014
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/101744
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