Microscopy-based HTS examines the mechanism of stress F-actin fiber disruption by cytochalasin D: orientation texture data collated with quantitative kinetic modeling.
We report a drug dose-response, end-point study of intracellular filamentous actin (F-actin) by automated fluorescence microscopy, complemented with theoretical kinetic simulation of drug action. We highlight the use of an advanced orientation-sensitive image processing procedure (<cos(2)theta> transform), specially tailored for the detection of ordered filamentous "patches" in cell images. To examine the extent of stress F-actin disruption caused by the drug, we compare the measured response based on the above transformation with the theoretical data obtained from a quantitative model. We show that the assay data are consistent with the first-order mass action kinetics predicted by a basic reaction model. As a concluding remark, we briefly discuss advantages, perspectives, and challenges of conventional fluorescent microscopy within the context of the quantitative high-throughput screening paradigm.
Alexandrov
Yuriy
Y
GE Healthcare, Cardiff, Wales, United Kingdom. yuriy.alexandrov@ge.com
Santos
Albert Francis
AF
Hather
Cath
C
Zaltsman
Alla Borisovna
AB
D000199:Actins / Q000187:drug effects* / Q000648:ultrastructure*; D000465:Algorithms; D001331:Automation; D015638:Cytochalasin D / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007700:Kinetics; D008853:Microscopy; D015233:Models, Statistical; D022502:Stress Fibers / Q000187:drug effects* / Q000648:ultrastructure*
D000199:Actins; D015638:Cytochalasin D