Document Detail


High frame-rate fluorescence confocal angle-resolved linear dichroism microscopy.
MedLine Citation:
PMID:  23742559     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Angle-resolved linear dichroism is a recent technique that exploits images recorded using an illumination field whose polarization angle is sequentially rotated during acquisition. It allows to retrieve orientation information of the fluorescent molecules, namely the average orientation angle and the amplitude of the fluctuations around this average. In order to boost up the acquisition speed without sacrificing the axial sectioning, we propose to combine a spinning disk confocal excitation scheme together with an electrooptical polarization switching and a camera acquisition. The polarization distortions induced when passing through the spinning disk system have been quantified and effectively compensated. The signal to noise features of the camera have been analyzed in detail so that the precision of the method can be quantified. The technique has been successfully tested on giant unilamellar vesicles and on living cells labeled with different fluorescent lipid probes, DiIC18 and di-8-ANEPPQ. It was able to acquire precise orientation images at full frame rates in the range of a second, ultimately limited by the fluorophore brightness and the camera sensitivity.
Authors:
Xiao Wang; Alla Kress; Sophie Brasselet; Patrick Ferrand
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  The Review of scientific instruments     Volume:  84     ISSN:  1089-7623     ISO Abbreviation:  Rev Sci Instrum     Publication Date:  2013 May 
Date Detail:
Created Date:  2013-06-07     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0405571     Medline TA:  Rev Sci Instrum     Country:  United States    
Other Details:
Languages:  eng     Pagination:  053708     Citation Subset:  IM    
Affiliation:
Aix-Marseille Université, Institut Fresnel, F-13013 Marseille, France.
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