Counting Cell Detachment from Timelapse

Hello, I’m measuring the adhesion strength of epithelial cells with a flow chamber. I will take an image every second and I need to count the number of cells in each image as they detach due to shear stress. I’ll then make a graph of Number of Cells vs Time

I’m looking for any software out there that can do this, any ImageJ plugins that can do this, or any general advice on an method for which I can write my own macro to do it

The images look something like this, although in this image I just scratched the top left of the cells with a needle to view the background. In the experiment single cells will detach randomly from all over the surface


Made me think of this:
If you only have a few videos, it might be easiest to define the scratch manually. If you have many, it should be possible to use intensity and blur to get a good automatically thresholded area of interest, but that could be dangerous if you define the area by the first frame and that frame has several cells in the process of detatching.

Not sure about the details, but maybe it will get you started.

Hi Owen,

I know you say you want to count the number of cells.
This sounds like a standard cell-segmentation task and CellProfiler (CP) may be a good place to start - you may want to search using keywords 'segmentation, phase-contrast, bright-field, monolayer". For CP the first step is likely for you to invert your images so that the dark cells appear bright, since CP was developed to detect fluorescent cells.

Alternatively, as a proxy for cell count you may be interested in simply measuring the temporal development of the cell-free/covered area. This takes you to the more general are of scratch assays for studying wound healing, using particle image velocimetry (PIV). Here are some tools for that task (ImageJ and MATLAB):

Hope that helps.