Three-dimensional Super-resolution Optical Fluctuation Imaging

Super-resolution optical fluctuation imaging (SOFI) achieves three-dimensional super-resolution by computing higher-order spatio-temporal cross-cumulants of stochastically blink-ing fluorophores. In contrast to localization microscopy, SOFI is compatible with weakly emitting fluorophores and a wider range of blinking conditions. The main drawback of SOFI is the nonlinear response to brightness and blinking heterogeneities in the sample, which limits the use of higher cumulant orders. We present a balanced SOFI algorithm for mapping molecular parameters and for linearizing the brightness response and we outline a MATLAB toolbox for two- and three-dimensional SOFI analysis. We show super-resolved three-dimensional cell structures imaged with a multi-plane wide-field microscope. The simultaneous acqui-sition of several focal planes significantly reduces the acquisition time and helps limiting the photo-bleaching of the marker fluorophores.


Presented at:
Focus on Microscopy - Superresolution Microscopy III: Pointillism, Image Processing, Maastricht, The Netherlands, March 24-27, 2013
Year:
2013
Keywords:
Laboratories:


Note: The status of this file is: Involved Laboratories Only


 Record created 2013-04-08, last modified 2018-03-17

Abstract:
Download fulltextPDF
Presentation:
Download fulltextPPT
Video:
Download fulltextAVI
Rate this document:

Rate this document:
1
2
3
 
(Not yet reviewed)