When I use Ficoll-paque PLUS to isolate PBMC part, the plasma fraction sometimes becomes jelly-like. It is hard to remove it with a pipette. I even have to use forceps to help remove the "Jello". This doesn't always happen, but it does happen. Anybody has an idea about this?
Additional information about the peripheral blood sample that you are using would be helpful. Was an anticoagulant used? If so, which one? Also, storage of the sample and sedimentation/centrifuge conditions might be beneficial in discovering what might be causing problems.
i have the same comment as Jared. More info about the patient would be good as i believe you get your samples randomly or from the same donor as always. is it EDTA blood or Heparin blood. do you always dilute your blood or use it as it is without dilution. all these things can influence the consistency of the plamsa fraction.
I used BD vacutainer K2 EDTA tube. The samples is fresh, the centrifuge is 400g for 30 min. The only thing I should mention is that my colleague use 1000 rpm to centrifuge the blood samples to collect platelets before I use ficoll to get PBMC. But even though, most time I have no problem with isolating PBMC.
you should dilute the blood with pbs containing edta, the plasma is clotting
Yes, I always dilute the blood 1:1 with MACS Buffer from MACS (Miltenyi Bioscience) before I continue with ficolling. This buffer is actually PBS with EDTA. Again I need to emphasize I have no problem with blood from most of the donors. This only happens may be 3 times in about 60 donors samples. Thanks
RBCs might have already lysed due to which it becomes jelly-like. I happened to see this 3-4 times in my lab while performing ficoll. This usually happens when the blood was over 48-72 hours.
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