Vol. 283, Issue 2, C639-C645, August 2002
SPECIAL COMMUNICATIONS
Quantification of cytoskeletal deformation in living cells
based on hierarchical feature vector matching
Tammo
Delhaas1,2,
Saskia
van Engeland4,
Jos
Broers4,3,
Carlijn
Bouten4,
Nico
Kuijpers4,
Frans
Ramaekers3, and
Luc H. E. H.
Snoeckx1,4
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
