Quantification of cytoskeletal deformation in living cells based on hierarchical feature vector matching

Departments of 1  Physiology, 2  Pediatrics, and 3  Molecular Cell Biology, Maastricht University, 6200 MD Maastricht; and 4  Faculty of Biomedical Engineering, Eindhoven University of Technology, 5512 AZ Eindhoven, The Netherlands The cytoskeleton is a dynamic scaffold in living cells even in the a...

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Bibliographic Details
Published in:American Journal of Physiology: Cell Physiology 2002-08, Vol.283 (2), p.C639-C645
Main Authors: Delhaas, Tammo, van Engeland, Saskia, Broers, Jos, Bouten, Carlijn, Kuijpers, Nico, Ramaekers, Frans, Snoeckx, Luc H. E. H
Format: Article
Language:English
Subjects:
Algorithms
Animals
Cell Movement
CHO Cells
Cricetinae
Cytoskeleton - physiology
Cytoskeleton - ultrastructure
Green Fluorescent Proteins
Image Processing, Computer-Assisted
Indicators and Reagents
Luminescent Proteins - genetics
Microscopy, Confocal
Recombinant Fusion Proteins
Software
Vimentin - genetics
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Summary:Departments of 1  Physiology, 2  Pediatrics, and 3  Molecular Cell Biology, Maastricht University, 6200 MD Maastricht; and 4  Faculty of Biomedical Engineering, Eindhoven University of Technology, 5512 AZ Eindhoven, The Netherlands The cytoskeleton is a dynamic scaffold in living cells even in the absence of externally imposed forces. In this study on cytoskeletal deformation, the applicability of hierarchical feature vector matching (HFVM), a new matching method, currently applied in space research and three-dimensional surface reconstruction, was investigated. Stably transfected CHO-K1 cells expressing green fluorescent protein (GFP) coupled to vimentin were used to visualize spontaneous movement of the vimentin cytoskeleton of individual cells using a confocal laser scanning system. We showed that, with proper parameter and configuration settings, HFVM could recognize and trace 60-70% of all image points in artificially translated, rotated, or deformed images. If only points belonging to the cytoskeleton were selected for matching purposes, the percentage of matched points increased to 98%. This high percentage of recognition also could be reached in a time series of images, in which a certain degree of bleaching of the fluorescence over the recording time of 30 min was inevitable. In these images, HFVM allowed the detection as well as the quantification of spontaneous cytoskeletal movements of up to 10% of the cell width. Therefore, HFVM appears to be a reliable method of quantifying dynamic cytoskeletal behavior in living cells. vimentin; fibroblast; green fluorescent protein; confocal laser scanning
ISSN:0363-6143
1522-1563
DOI:10.1152/ajpcell.00535.2001