Description

Cell migration assays are an important technique in analyzing how altering different variables can affect cell motility. In the laboratory, cells are grown in a confluent layer on a 35mm dish then, a “wound” or “scratch” is introduced across this layer of cells by drawing a pipet across the field of cells to create an open space. The movement of cells into this open area is monitored for ~24 h by microscopy, most often brightfield images taken every 2-10 minutes. To quantify the movement of cells as a function of time and the overall closure of the open area, the researcher must define the open area on each image. This constitutes a major difficulty with these assays as several software programs are available for this analysis, but their reliability and efficiency vary. In this study, three different software packages were used to assess cell migration assays: MiToBo(an ImageJ plugin), SketchAndCalc(an iPad app), and ibidi’sAutomated Cellular Analysis System (ACAS). Migration assays for Hap1 cells, WT or having a single protein knockout, were completed in triplicate. Data, images taken every 10 min for a 24 h time period (149 images) were analyzed by each of the three programs. MiToBowas a very fast image analysis program but was not able to consistently distinguish between the cells and the open area. This led to over-or under-estimation of the open area. The SketchAndCalcapproach was an extremely accurate image analysis program but required the user to trace the edges of the open area on each image, requiring several hours to completely analyze each data set. The ACAS was also a very fast automated program like MiToBobut was much more consistent in distinguishing the cells from the open area. In directly comparing the SketchAndCalcwith the ACAS analysis, the WT cells had a 25% closure for the ACAS and a 26% closure for the SketchAndCalcand for the knocked-out cell line it was reported to have a 15% closure by the ACAS system and 13% by the SketchAndCalcprogram. These results support that the efficient analysis by the ACAS software provides reliable analysis of cell migration assay data.

COinS
 

Identifying Accurate and Efficient Data Analysis Tools for Wound Healing Assays

Cell migration assays are an important technique in analyzing how altering different variables can affect cell motility. In the laboratory, cells are grown in a confluent layer on a 35mm dish then, a “wound” or “scratch” is introduced across this layer of cells by drawing a pipet across the field of cells to create an open space. The movement of cells into this open area is monitored for ~24 h by microscopy, most often brightfield images taken every 2-10 minutes. To quantify the movement of cells as a function of time and the overall closure of the open area, the researcher must define the open area on each image. This constitutes a major difficulty with these assays as several software programs are available for this analysis, but their reliability and efficiency vary. In this study, three different software packages were used to assess cell migration assays: MiToBo(an ImageJ plugin), SketchAndCalc(an iPad app), and ibidi’sAutomated Cellular Analysis System (ACAS). Migration assays for Hap1 cells, WT or having a single protein knockout, were completed in triplicate. Data, images taken every 10 min for a 24 h time period (149 images) were analyzed by each of the three programs. MiToBowas a very fast image analysis program but was not able to consistently distinguish between the cells and the open area. This led to over-or under-estimation of the open area. The SketchAndCalcapproach was an extremely accurate image analysis program but required the user to trace the edges of the open area on each image, requiring several hours to completely analyze each data set. The ACAS was also a very fast automated program like MiToBobut was much more consistent in distinguishing the cells from the open area. In directly comparing the SketchAndCalcwith the ACAS analysis, the WT cells had a 25% closure for the ACAS and a 26% closure for the SketchAndCalcand for the knocked-out cell line it was reported to have a 15% closure by the ACAS system and 13% by the SketchAndCalcprogram. These results support that the efficient analysis by the ACAS software provides reliable analysis of cell migration assay data.

 

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