Alexandrov Yuriy, Santos Albert Francis, Hather Cath, Zaltsman Alla Borisovna
GE Healthcare, Cardiff, Wales, United Kingdom.
Assay Drug Dev Technol. 2009 Aug;7(4):366-73. doi: 10.1089/adt.2008.0170.
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.
我们报告了一项通过自动荧光显微镜对细胞内丝状肌动蛋白（F-肌动蛋白）进行药物剂量反应终点研究，并辅以药物作用的理论动力学模拟。我们重点介绍了一种先进的方向敏感图像处理程序（<cos(2)θ>变换）的使用，该程序是专门为检测细胞图像中有序的丝状“斑块”而定制的。为了检查药物引起的应激F-肌动蛋白破坏程度，我们将基于上述变换测得的反应与从定量模型获得的理论数据进行比较。我们表明，测定数据与基本反应模型预测的一级质量作用动力学一致。作为结束语，我们在定量高通量筛选范式的背景下简要讨论了传统荧光显微镜的优点、前景和挑战。
