Intensity Measurement

I am trying to measure the movement of fluorescence in cells over a time lapsed experiment. In my pipeline I first use a mask to remove saturated cells. I then identify the cells of interest which have a fluorescent membrane that moves into the cytoplasm over time. I am trying to find a way to measure the intensity of the cytoplasm over the time lapsed experiment but I can’t seem to find a way to reliably identify objects that change in fluorescence over time.

I have attached two sample images which show the movement of the fluorescence and my pipeline.
pipeline.cpproj (582 KB)



It appears as though your pipeline works well at detecting the relevant cells. Also, they don’t appear to move much over time, tracking using the overlap method seems like a good approach.

To track the fluorescent movement, I think you may be best served by using the MeasureObjectRadialDistribution module. This module divides the cell into concentric bins and measures intensity-based statistics from each. As the fluorescence shifts from the cell boundary to the interior, we would expect the intensity distribution to shift accordingly.

Regards,
-Mark