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  4. Balanced Super-resolution Optical Fluctuation Imaging (bSOFI)
 
conference presentation

Balanced Super-resolution Optical Fluctuation Imaging (bSOFI)

Geissbühler, Stefan  
•
Bocchio, Noelia  
•
Dellagiacoma, Claudio  
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2012
18th International Workshop on "Single Molecule Spectroscopy and Ultra Sensitive Analysis in the Life Sciences"

Super-resolution optical fluctuation imaging (SOFI) achieves 3D super-resolution by computing higher-order cumulants of stochastically blinking fluorophores. In contrast to localization microscopy, SOFI is compatible with weakly emitting fluorophores and a wide 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. Balanced super-resolution optical fluctuation imaging (bSOFI), extends SOFI by the combination of several cumulant orders to map fluorescence-related molecular statistics, such as molecular state lifetimes, concentrations and brightness distributions with super-resolution. Since these parameters are often linked to the chemical microenvironment of the fluorophores, they report on static differences and/or dynamic changes within cells and thus add a “functional” dimension to super-resolution microscopy based on stochastic switching. Furthermore, the information obtained can be used to correct for the nonlinear brightness and blinking response of cumulants. We show experimental results of Alexa647-labeled microtubules in fixed HeLa cells with an up to five-fold resolution improvement compared to diffraction-limited widefield microscopy. Using a total-internal-reflection illumination scheme, we obtain depth information through the estimation of the spatial distributions of the molecular brightness as well as the blinking on-ratio.

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Type
conference presentation
Author(s)
Geissbühler, Stefan  
Bocchio, Noelia  
Dellagiacoma, Claudio  
Geissbühler, Matthias  
Berclaz, Corinne  
Leutenegger, Marcel  
Lasser, Theo  
Date Issued

2012

Subjects

Fluorescence microscopy

•

Super-resolution

•

Stochastic switching

•

Sofi

•

Balanced cumulants

•

Molecular statistics

•

Functional imaging

URL

URL

http://www.picoquant.com/
Written at

OTHER

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Event nameEvent placeEvent date
18th International Workshop on "Single Molecule Spectroscopy and Ultra Sensitive Analysis in the Life Sciences"

Berlin, Germany

September 5-7, 2012

Available on Infoscience
September 18, 2012
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/85493
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