一种用于细胞迁移的快速且稳健的定量延时检测方法。

A fast and robust quantitative time-lapse assay for cell migration.

作者信息

Dai Lingsong, Alt Wolfgang, Schilling Karl, Retzlik Jens, Gieselmann Volkmar, Magin Thomas M, Kappler Joachim

机构信息

Institut für Physiologische Chemie, Nussallee 11, D-53115 Bonn, Germany.

出版信息

Exp Cell Res. 2005 Dec 10;311(2):272-80. doi: 10.1016/j.yexcr.2005.09.013. Epub 2005 Oct 24.

DOI:10.1016/j.yexcr.2005.09.013

PMID:16248998

Abstract

We describe a simple and widely applicable method to measure cell migration in time-lapse sequences of fluorescently labeled cells in culture. Briefly, binarized cell images obtained after thresholding were cumulatively projected, and the covered areas were measured. This procedure determines the time course of the track area successively covered by the cell population. Under conditions where cell growth is negligible, a robust index of cell motility is derived from normalized plots for the displacement of cells over time. We applied this method to quantitatively examine the migration of B35 neuroblastoma cells transiently expressing GFP and to C6 glioma cells after staining with Hoechst 33258. This sensitive assay detected the influence of agents which inhibit actin polymerization (cytochalasin B) or interfere with the maintenance of cell polarity (methyl-beta-cyclodextrin) on cell migration. Thus, this assay is a versatile tool to measure quickly the migration of different cell types using different labeling strategies.

摘要

我们描述了一种简单且广泛适用的方法，用于在培养的荧光标记细胞的延时序列中测量细胞迁移。简而言之，对阈值化后获得的二值化细胞图像进行累积投影，并测量覆盖面积。该过程确定了细胞群体连续覆盖的轨迹区域的时间进程。在细胞生长可忽略不计的条件下，从细胞随时间位移的归一化图中得出细胞运动性的稳健指标。我们将此方法应用于定量检测瞬时表达绿色荧光蛋白（GFP）的B35神经母细胞瘤细胞以及用Hoechst 33258染色后的C6胶质瘤细胞的迁移。这种灵敏的检测方法检测到了抑制肌动蛋白聚合的试剂（细胞松弛素B）或干扰细胞极性维持的试剂（甲基-β-环糊精）对细胞迁移的影响。因此，该检测方法是一种通用工具，可使用不同的标记策略快速测量不同细胞类型的迁移。