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A quantitative measure for alterations in the actin cytoskeleton investigated with automated high‐throughput microscopy
Author(s) -
Weichsel Julian,
Herold Nikolas,
Lehmann Maik J.,
Kräusslich HansGeorg,
Schwarz Ulrich S.
Publication year - 2010
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20818
Subject(s) - cytoskeleton , actin cytoskeleton , cytochalasin d , actin , microbiology and biotechnology , biology , cytometry , fluorescence microscope , flow cytometry , cell , immunology , physics , biochemistry , fluorescence , optics
The actin cytoskeleton modulates a large variety of physiological and disease‐related processes in the cell. For example, actin has been shown to be a crucial host factor for successful infection by HIV‐1, but the underlying mechanistic details are still unknown. Automated approaches open up the perspective to clarify such an issue by processing many samples in a high‐throughput manner. To analyze the alterations in the actin cytoskeleton within an automated setting, large‐scale image acquisition and analysis were established for JC‐53 cells stained for actin. As a quantitative measure in such an automated approach, we suggest a parameter called image coherency. We successfully benchmarked our analysis by calculating coherency for both a biophysical model of the actin cytoskeleton and for cells whose actin architecture had been disturbed pharmacologically by latrunculin B or cytochalasin D. We then tested the influence of HIV‐1 infection on actin coherency, but observed no significant differences between uninfected and infected cells. © 2009 International Society for Advancement of Cytometry