Dear flowers, In addition to Dr. Waddick's questions concerning CFSE, we too have some problems with it. I have been using CFSE to track human cord blood HSC in vivo and check for cell divisions. - I cannot use frozen material, because then I get clump forming, which results in loss of 75% of my cells. Is this normal? - It is extremely difficult to compensate the overflow from CFSE towards the FL-3 and the FL-2 channel, making it difficult top combine CFSE with PE or Cy labeled antibodies. Gating out of dead cells using propidium iodide also becomes very difficult. Do others also have the same problems, or do you have the magical settings? - How long can you keep CFSE in good shape frozen in DMSO (-20°C)? Kind regards, Tessa
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