Hello! I’m trying to assay for a marker expressed in cell-cell junctions (let’s call it CB) from cells grown to various degrees of confluency, using fluorescent immunostaining. I’m subtracting the CB image from an actin stain and then calling IdentifySecondary against it, which yields surprisingly good segmentation but draws the cell boundary just inside the bright region of the CB stain, meaning that I can’t measure the CB-bright region as part of the cell.
I was tempted to dilate the identified cells a handful of pixels with ExpandOrShrink and then stamp out the cell body with IdentifyTertiaryRegion to isolate a CB-rich (or not) “border” region, but noticed to my dismay that when expanded cell boundaries overlapped, they merged into a single object.
My new strategy is to develop a module that will look at each identified cell, “expand” the border n pixels, and measure the intensity of that region without actually performing the cell-merging expand operation. My intuition suggests that this shouldn’t be hard but I’m not sure where to begin – I’m looking through modules now and we’ll see. I wanted to see if anyone had any tactical suggestions or had already done this, or if CP already supports what I’m trying to do somehow.