Hi,
I would like to know if some of you had experience with the difference between fixed angled rotors versus swing-out buckets for the centrifuge steps in Ficoll/Percoll protocols for isolation of monocytes. We use swing-out buckets, but I read in a Percoll manual that it is not recommended to use swing-out buckets. For Ficoll, I cannot find any recommendation. So I would like to hear some experiences from others, what are you using and why?
So in short. Should you use fixed-angle rotors when working with Ficoll and Percoll?
Aren't most centrifuge swing out buckets? The only one I've seen with the fixed buckets are the large centrifuges that we use to spin down liters of virus/bacteria, and whatnot.
I'm not sure why they won't recommend the swing out buckets, because that's what I've always used for the gradient separations. Only thing...don't forget to turn off the break.
In our institute everyone that i know, including myself, use swing-bucket centrifuges for ficoll/ histopaque.
Swing buckets were always preferred as they give a separation that will remain when you transfer your tubes to an upright position.
Fixed buckets will centrifuge your contents at an angle, and subsequently it is possible to lose the clarity of your spin down layers when you place these tubes in an upright position.
The deciding factor may be how fast you need to spin your samples. Ultimately fixed buckets are more stable and can spin faster. However, you are looking isolate PBMC/ monocytes, as was I, which require much lower speeds.
Agreed, best separation with swing out rotors and make sure the brake is off to maintain a clean separation.
Can we separate PBMC without using a swinging bucket centrifuge? or with a centrifuge with the fixed rotor?
When you use swing out rotor what angle do you prefer for good separation of cells by ficoll/ histopaque - 90 or 45o ?
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