I have 50 mL of volume at the beginning of the culture and I just can take samples of 1 mL per day, so there are very few cells to count them using trypan blue (after washing with PBS and detaching with Tryple).
I've tried using Violet Crystal, but it gives me inconsistent results along the days that I've followed the culture.
Does anyone a better protocol, idea or suggestion?
One way is to label the cells within the scaffold with a nuclear stain such as DAPI and then do optical sectioning of the scaffold using confocal microscopy.
what is your initial cell density and dilution rate for counting? Maybe you can not count in first days with high dilution rates like 10-fold. But if you try 2-fold dilution for counting, maybe you can count with this way.
Dear Fauzi Mb Could you please share your protocol for counting cells with presto Blue assay? How do you do the standard curve to move from fluorescence to a cell number? Thank you in advance!
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