Bosgraaf Leonard, van Haastert Peter J M, Bretschneider Till
University of Groningen, Cell Biochemistry Department, Haren, The Netherlands.
Cell Motil Cytoskeleton. 2009 Mar;66(3):156-65. doi: 10.1002/cm.20338.
The use of fluorescent markers in living cells has increased dramatically in the recent years. The quantitative analysis of the images requires specific analysis software. Previously, the program Quimp was launched for quantitating fluorescent intensities at the membrane or the cortex of the cell. However, Quimp is not well suited to quantitate local membrane displacement. Here we present Quimp2 that is capable of tracking membrane subregions in time, which enables the simultaneous quantification of fluorescent intensities and membrane movement. Quimp2 has two new tools, (i) conversion filters to analyze movies obtained with fluorescent, DIC and phase contrast different microscopes, and (ii) a macro that calculates the local membrane displacement and provides various options to display the results. Quimp2 is used here to investigate the molecular mechanism of cell movement by correlating the dynamics of local membrane movement with the local concentration of myosin and F-actin.
近年来，荧光标记物在活细胞中的应用急剧增加。图像的定量分析需要特定的分析软件。此前，Quimp程序被开发用于定量细胞的膜或皮质处的荧光强度。然而，Quimp不太适合定量局部膜位移。在此，我们展示了Quimp2，它能够实时跟踪膜亚区域，从而能够同时定量荧光强度和膜运动。Quimp2有两个新工具，（i）转换过滤器，用于分析通过荧光、微分干涉差（DIC）和相差不同显微镜获得的视频，以及（ii）一个宏，用于计算局部膜位移并提供多种显示结果的选项。在此使用Quimp2通过将局部膜运动的动力学与肌球蛋白和F-肌动蛋白的局部浓度相关联来研究细胞运动的分子机制。
