Quantifying cell-matrix adhesion dynamics in living cells using interference reflection microscopy
Quantifying cell-matrix adhesion dynamics in living cells using interference reflection microscopy
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DOI：
10.1111/j.1365-2818.2008.02069.x
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发表时间：
2008-10-01
影响因子：
2
通讯作者：
Dunn, G. A.
中科院分区：
文献类型：
--
作者：
Holt, M. R.;Calle, Y.;Dunn, G. A.
局灶性粘连和podosomes是整合素介导的细胞基质接触，可以使用干涉反射显微镜（OCT）可视化。在这里，我们已经开发了自动化的图像处理程序，以量化粘附营业额从活细胞的荧光显微镜图像。使用时间序列的图像，我们产生的粘连地图，揭示粘连的空间变化，并包含这些变化的时间序列的额外信息。这种地图被用来表征局灶性粘附动力学在小鼠胚胎成纤维细胞缺乏一个或两个等位基因的黏着斑蛋白基因。黏着斑蛋白表达的缺失导致粘着斑的组装、分解和/或移位增加，表明黏着斑蛋白对于稳定粘着斑是重要的。该方法也可用于研究在原代小鼠树突状细胞中观察到的足体的快速动力学。
Focal adhesions and podosomes are integrin-mediated cell-substratum contacts that can be visualized using interference reflection microscopy (IRM). Here, we have developed automated image-processing procedures to quantify adhesion turnover from IRM images of live cells. Using time sequences of images, we produce adhesion maps that reveal the spatial changes of adhesions and contain additional information on the time sequence of these changes. Such maps were used to characterize focal adhesion dynamics in mouse embryo fibroblasts lacking one or both alleles of the vinculin gene. Loss of vinculin expression resulted in increased assembly, disassembly and/or in increased translocation of focal adhesions, suggesting that vinculin is important for stabilizing focal adhesions. This method is also useful for studying the rapid dynamics of podosomes as observed in primary mouse dendritic cells.