Im currently using a commercial TUNEL assay kit with TdT enzyme for flow cytometry analysis. However despite starting with half a million cells, and maintaining a pellet throughout the steps, after the enzymymatix labelling mix step, I seem to have a signficant cell loss and even after centrifuging I can see floating cells but a significantly reduced cell pellet making subsequent analysis difficult. The kit recommends starting with 1 million cells but I dont have that number available and I would have though 500,000 is sufficient. Im not resuspending the pellet during the washes too vigourously but I just have no idea why im losing all the cells. Any ideas?