Wound healing assays, or scratch assays are used to investigate area infiltration by cells. Several methods to set up such an experiment are known. In the example shown here the scratch assay starts with the culture of a confluent monolayer of cells. Thereafter, a scratch is made in the monolayer to create a “wound”, this is generally done with a pipette tip. An image of the scratch is taken with a microscope at the start and sequentially at several other time points to analyze the closure of the scratch over time. Generally, the scratch width, the percentage of closure and/or the migration speed are parameters derived from this assay.
Scratch assay analysis
Using the imaging capabilities of the CytoSMART Omni users can investigate the migration patterns of cells in whole well comparison.
In the figure in the above a wound healing assays was performed on C6 rat glioma cells. The experiment was performed in a 48-well plate. Users can visually inspect the progression of the gap closure from a whole well overview and to cell level detail. The image analysis algorithms in the CytoSMART Cloud provide information on the gap surface closure and closure speed over time. The highlighted blue areas in the image are masks indicating what pixels are selected by the algorithm. The numerical data can easily be extracted after analysis. The two graphs shown here display the numerical data of the gap closure of the four wells shown in the example.
High throughput imaging allows you to easily compare samples
In the video below a single 96-well plate was imaged using the CytoSMART Omni. Time-lapse videos show that increasing concentrations of blebbistatin and cytochalasin-d leads to inhibition of cell migration.
NIH-3T3 mouse fibroblasts were used and imaged for 48 hours and a 1 hour time interval.
Scratch closure - video example CytoSMART Omni
whole well view, 48 well plate, c6 rat glioma cells - 23 hours, 1h imaging interval
whole well view, 48 well plate, normal Human Epidermal Keratinocytes 48 h, 1h imaging interval
Scratch closure - video example CytoSMART Lux2
Follow the migration of C6 rat glioma cells over 29 hours, 1 image/10 mins.