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A quantitative measure for alterations in the actin cytoskeleton investigated with automated high-throughput microscopy.

Cytometry. Part A : the journal of the International Society for Analytical Cytology (2009-11-10)
Julian Weichsel, Nikolas Herold, Maik J Lehmann, Hans-Georg Kräusslich, Ulrich S Schwarz
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Atto 633, BioReagent, suitable for fluorescence
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Atto 633 iodoacetamide, suitable for fluorescence
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Atto 633 NHS ester, BioReagent, suitable for fluorescence, ≥90% (HPLC)
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Atto 633 azide, suitable for fluorescence, ≥90% (HPLC)