Document Detail


Evaluation of rapid volume changes of substrate-adherent cells by conventional microscopy 3D imaging.
MedLine Citation:
PMID:  15312195     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Precise measurement of rapid volume changes of substrate-adherent cells is essential to understand many aspects of cell physiology, yet techniques to evaluate volume changes with sufficient precision and high temporal resolution are limited. Here, we describe a novel imaging method that surveys the rapid morphology modifications of living, substrate-adherent cells based on phase-contrast, digital video microscopy. Cells grown on a glass substrate are mounted in a custom-designed, side-viewing chamber and subjected to hypotonic swelling. Side-view images of the rapidly swelling cell, and at the end of the assay, an image of the same cell viewed from a perpendicular direction through the substrate, are acquired. Based on these images, off-line reconstruction of 3D cell morphology is performed, which precisely measures cell volume, height and surface at different points during cell volume changes. Volume evaluations are comparable to those obtained by confocal laser scanning microscopy (DeltaVolume < or = 14%), but our method has superior temporal resolution limited only by the time of single-image acquisition, typically approximately 100 ms. The advantages of using standard phase-contrast microscopy without the need for cell staining or intense illumination to monitor cell volume make this system a promising new tool to investigate the fundamentals of cell volume physiology.
Authors:
F Boudreault; R Grygorczyk
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of microscopy     Volume:  215     ISSN:  0022-2720     ISO Abbreviation:  J Microsc     Publication Date:  2004 Sep 
Date Detail:
Created Date:  2004-08-17     Completed Date:  2004-10-28     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  0204522     Medline TA:  J Microsc     Country:  England    
Other Details:
Languages:  eng     Pagination:  302-12     Citation Subset:  IM    
Affiliation:
Research Centre, Centre hospitalier de l'Université de Montréal (CHUM) -- Hôtel-Dieu, 3850 Saint-Urbain, Montréal, Québec, H2W 1T7, Canada.
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MeSH Terms
Descriptor/Qualifier:
Cell Adhesion
Cell Line, Tumor
Cell Size
Humans
Hypotonic Solutions
Image Processing, Computer-Assisted*
Imaging, Three-Dimensional
Lung / cytology*,  pathology*
Microscopy, Phase-Contrast / instrumentation,  methods*
Chemical
Reg. No./Substance:
0/Hypotonic Solutions

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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