Gau David M, Roy Partha
Dept of Bioengineering, University of Pittsburgh, Pittsburgh, USA.
Bio Protoc. 2020 Apr 20;10(8). doi: 10.21769/bioprotoc.3586.
Cell migration is a fundamental cellular process that plays a crucial role in many physioglogical and pathological processes such as wound healing or cancer metastasis. Many assays have been developed to examine cell migration, such as the wound healing or scratch assay, Boyden Chamber or transwell assay, and the method we will describe here, single cell migration assay. In this assay, cells are plated sparsely on a collagen coated plate and live cell imaging is performed over a period of 2 h at 1 frame per minute. After imaging is completed, cells are tracked manually using ImageJ by tracking movement of the centroid of the cell. These data points are then exported and overall distance travelled from frame to frame is determined and divided by total time imaged to determine speed of the cell. This method provides a quick way to examine effect of cellular manipulation on cell migration before proceeding to perform more complex assays.
细胞迁移是一个基本的细胞过程，在许多生理和病理过程中发挥着关键作用，如伤口愈合或癌症转移。已经开发了许多检测细胞迁移的方法，如伤口愈合或划痕试验、Boyden小室或Transwell试验，以及我们在此将描述的方法——单细胞迁移试验。在该试验中，将细胞稀疏地接种在胶原包被的培养板上，并以每分钟1帧的速度进行2小时的活细胞成像。成像完成后，使用ImageJ通过跟踪细胞质心的移动来手动追踪细胞。然后导出这些数据点，并确定相邻帧之间移动的总距离，再除以成像的总时间以确定细胞的速度。这种方法提供了一种在进行更复杂的试验之前快速检测细胞操作对细胞迁移影响的方法。
