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Axial Resolution Enhancement by 4Pi Confocal Fluorescence Microscopy with Two-Photon Excitation.
MedLine Citation:
PMID:  19669529     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Confocal fluorescence microscopy and two-photon microscopy have become important techniques for the three-dimensional imaging of intact cells. Their lateral resolution is about 200-300 nm for visible light, whereas their axial resolution is significantly worse. By superimposing the spherical wave fronts from two opposing objective lenses in a coherent fashion in 4Pi microscopy, the axial resolution is greatly improved to approximately 100 nm. In combination with specific tagging of proteins or other cellular structures, 4Pi microscopy enables a multitude of molecular interactions in cell biology to be studied. Here, we discuss the choice of appropriate fluorescent tags for dual-color 4Pi microscopy and present applications of this technique in cellular biophysics. We employ two-color fluorescence detection of actin and tubulin networks stained with fluorescent organic dyes; mitochondrial networks are imaged using the photoactivatable fluorescent protein EosFP. A further example concerns the interaction of nanoparticles with mammalian cells.
Authors:
Sylvia Glaschick; Carlheinz Röcker; Karen Deuschle; Jörg Wiedenmann; Franz Oswald; Volker Mailänder; G Ulrich Nienhaus
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Publication Detail:
Type:  Journal Article     Date:  2008-06-19
Journal Detail:
Title:  Journal of biological physics     Volume:  33     ISSN:  0092-0606     ISO Abbreviation:  J Biol Phys     Publication Date:  2007 Dec 
Date Detail:
Created Date:  2009-08-11     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0417731     Medline TA:  J Biol Phys     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  433-43     Citation Subset:  -    
Affiliation:
Institute of Biophysics, University of Ulm, 89069, Ulm, Germany.
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