Hello,
I am currently working with a ZR-75-1 cell line that was treated for long time with fulvestrant to acquire resistance. The parental cell line behave normally while I believe that the resistant cells lowered the expression of some receptor needed to attach to the wells. They detach especially when treated with a fixative. I need to perform growth assays with these cells and I already tried many protocols (crystal violet, sulforhodamine B). I only manage to get a growth curve using cell titer blue, but I need a cell-number-based assay for confirmation. I am looking for a staining protocol that does not require fixation. Does anyone have a suggestion? Any suggestion is welcome to solve the problem! Thank you.
You may consider doing vital staining using propedium iodide or trypan blue for all cells and take count using hemocytometer or flowcytometry.
All the best
Dear Martina
If you wish to count the cells in the without fixation, you can stain them with Calcein-AM which is taken up by live cells (perhaps in combination with PI for dead cells) and use s fluorescence microscope
Best
Andreas
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