Hi, I am Nao in Terry Fox Laboratory, BC Cancer Agency. I have a question on SP (side population) cell sorting using FACS Vantage. Small population of BM cells which have Hoechst33342-low phenotype are called side population cells and are enriched in hematopoietic stem cells. We used to work with FACStar Plus for this sorting and recently swiched to FACS Vantage. But since then, we have not got proper profile. It looks to me that bigger population which usually locates right next to SP cells (cells in G0/G1) moves toward Hoechst-low position and overlaps with SP. We could not figure out what was going on, and somehow the profile changed into normal all of a sudden. So we know that our machine is capable of visualizing SP cells, but we are still not confident in performing this sorting properly for next experiment. If there are somebodies who can think of what was the reason why it did not work in the first place and what procedure to take when it happens, please let me know. Thank you Nao
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