A quantitative measure for alterations in the actin cytoskeleton investigated with automated high-throughput microscopy.
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.
Weichsel
Julian
J
BioQuant, University of Heidelberg, Heidelberg, Germany.
Herold
Nikolas
N
Lehmann
Maik J
MJ
Kräusslich
Hans-Georg
HG
Schwarz
Ulrich S
US
D000199:Actins / Q000648:ultrastructure*; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D002478:Cells, Cultured; D015638:Cytochalasin D / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure*; D015658:HIV Infections; D015497:HIV-1 / Q000187:drug effects; D006367:HeLa Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D008954:Models, Biological; D053778:Thiazolidines / Q000494:pharmacology
D000199:Actins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D053778:Thiazolidines; D015638:Cytochalasin D; C037068:latrunculin B