I will get a sample of bone marrow cells, containing appr. 1000 cells in 200 µl 1xPBS. For my downstream application, I would need to concentrate them down to 30-40 µl volume. Does anyone have experience with such procedure? I would centrifuge them 5 min at 800 g, followed by 5 min @ 1500 g, followed by gentle aspiration of 160 µl of the supernatant. Is it a good idea?
Hi
This will be good enough to concentrate by centrifugation if cell can bear little shear stress at less rpm. I think you can use one more way by using ultracentrifuge tube with filter membrane where you can collect the cell pallet which you can resuspend in your desired volume and concentration
I agree with Dominic, 800 and 1500g were high speed. In my culture centrifuge, 1500rpm means 300g. Often, we use speed around 300g and below. If you do not need to change buffer, just one is perfect.
If you cells are in a microfuge tube simply place the tube in a microcentrifuge press the button used for a quick spin. for ~10 seconds and you cells will be pelleted, just remove the PBS and their ready to go. If your microcentifuge lacks this option (quick spin) set the RPMs to lowest setting. Turn it on for 10-20 seconds and turn off and proceed with your protocol. It works every time.
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