Document Detail

Measurement of rapid changes in cell volume by forward light scattering.
MedLine Citation:
PMID:  12937987     Owner:  NLM     Status:  MEDLINE    
Light scattering is an empirical technique employed to measure rapid changes in cell volume. This study describes a new configuration for the method of light scattering and its corroboration by measurements of cell height (as a measure of cell volume). Corneal endothelial cells cultured on glass cover-slips were mounted in a perfusion chamber on the stage of an inverted microscope. A beam of light was focused on the cells from above the stage at an angle of 40 degrees to the plane of the stage. The scattered light intensity (SLI), captured by the objective and referred to as forward light scatter (FLS), increased and decreased in response to hyposmotic and hyperosmotic shocks, respectively. The rapid increase and decrease in SLI corresponded to cell swelling and shrinkage, respectively. Subsequently, SLI decreased and increased as expected for a regulatory volume decrease (RVD) and increase (RVI), respectively. These data are in agreement with measurements of cell height, demonstrating that the method of light scatter in FLS mode is useful for monitoring rapid changes in cell volume of cultured cells. Changes in SLI caused by gramicidin were consistent with cell volume changes induced by equilibration of NaCl and KCl concentrations across the cell membrane. Similarly, an additional decrease in SLI was recorded during RVD upon increasing K+ conductance by valinomycin. Decreasing K+ conductance of the cell membrane with Ba2+ changed the time course of SLI consistent with the effect of the K+ channel blocker on RVD. Bumetanide and dihydro-ouabain inhibited increases in SLI during RVI. In conclusion, FLS is a valid method for qualitative analysis of cell volume changes with a high time resolution.
S P Srinivas; Joseph A Bonanno; Els Larivière; Danny Jans; Willy Van Driessche
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.     Date:  2003-08-21
Journal Detail:
Title:  Pflügers Archiv : European journal of physiology     Volume:  447     ISSN:  0031-6768     ISO Abbreviation:  Pflugers Arch.     Publication Date:  2003 Oct 
Date Detail:
Created Date:  2004-02-20     Completed Date:  2004-09-14     Revised Date:  2014-09-15    
Medline Journal Info:
Nlm Unique ID:  0154720     Medline TA:  Pflugers Arch     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  97-108     Citation Subset:  IM    
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MeSH Terms
Cell Size / physiology
Endothelial Cells / cytology*,  physiology*
Research Design / standards
Scattering, Radiation
Grant Support
E11107//PHS HHS; EY00834/EY/NEI NIH HHS; R01 EY008834/EY/NEI NIH HHS; R01 EY008834-11/EY/NEI NIH HHS

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