I have been trying to label somatic cells isolated from milk (resuspended in PBS) using fluorescein diacetate (conc. of 8micrograms/mL) and view it under fluorescence microscope (Ex/Em used is ~460/530). I can see the cells in brightfield but the fluorescence signal is almost non-existent. On increasing the exposure time to 100 or 200ms, I am able to see hardly one or two cells in a frame (where actually there are hundreds of cells in a frame) and there too, the signal intensity is extremely low. I tried increasing the incubation time and dye concentration but doesn't really help. Also tried adding a detergent like Triton X because I thought that perhaps some of the proteins or fats from the milk were somehow contaminating my sample and their hydrophobic nature was interfering with the labelling but that didn't change anything.
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