Step-by-step quantitative analysis of focal adhesions.
Step-by-step quantitative analysis of focal adhesions.
复制标题
DOI：
10.1016/j.mex.2014.06.004
复制
发表时间：
2014
期刊：
影响因子：
1.9
通讯作者：
Pesen-Okvur, Devrim
中科院分区：
文献类型：
--
作者：
Horzum, Utku;Ozdil, Berrin;Pesen-Okvur, Devrim
通过结合ImageJ程序的不同插件简化了分析步骤。由于荧光图像的优化处理，可以更好地检测FA，并具有最小的假阴性。这种方法可以应用于量化各种荧光图像，包括高背景内的焦点和/或局部信号，如FA，细胞中许多复杂的信号结构之一。局灶性粘连（FA）是一种特殊的粘附结构，作为细胞与周围细胞外基质之间的细胞通讯单位。FA参与信号转导和肌动蛋白细胞骨架组织。脂肪酸介导的细胞粘附，这是一个关键的现象，在癌症的研究。由于细胞可以形成许多和微米级的FA，其定量分析需要良好优化的图像分析方法。在这里，我们优化了MDA-MB-231乳腺癌细胞的FA分析。优化是基于黏着斑蛋白的免疫荧光图像的适当处理，黏着斑蛋白是脂肪酸的标志物之一。所有图像处理步骤都使用ImageJ软件进行，该软件可免费获得并处于公共领域。我们的方法的优点是：
The analysis steps are simplified by combining different plugins of the ImageJ program. FAs are better detected with minimal false negatives due to optimized processing of fluorescent images. This approach can be applied to quantify a variety of fluorescent images comprising focal and/or localized signals within a high background such as FAs, one of the many complex signaling structures in a cell. Focal adhesions (FAs) are specialized adhesive structures which serve as cellular communication units between cells and the surrounding extracellular matrix. FAs are involved in signal transduction and actin cytoskeleton organization. FAs mediate cell adhesion, which is a critical phenomenon in cancer research. Since cells can form many and micrometer scale FAs, their quantitative analysis demands well-optimized image analysis approaches. Here, we have optimized the analysis of FAs of MDA-MB-231 breast cancer cells. The optimization is based on proper processing of immunofluorescence images of vinculin, which is one of the markers of FAs. All image processing steps are carried out using the ImageJ software, which is freely available and in the public domain. The advantages of our method are: