cell cetrifugation_how fast can you go? - (Jun/07/2006 )

Hi!

I was wondering how fast can you centrifuge cells without damage (so that you can still resuspend them and do a trypan blue count) in a table top centrifuge?

thanks for help

j

i usually spin mine at 1500rpm without any hassle and normally with more than enough pellet formation

but i have spun cell succesfully up to 3000rpm

i routinely pin at 200g.
If needed, 300g is ok.

I use 700-800 RPM for spinning down cells, either after thawing from Liquid nitrogen or trypsinisation. Remember that centrifugation KILLS/DAMAGES cells, the lower the spin, the less the damage.
Also Trypan Blue is a very INSENSITIVE method for looking at cell damage. You may get a 98% viability with trypan, but with the same sample do FACS Analysis looking at Annexin V and find vaibility down at 50%. Look at Ctyochrome C levels in the cells for an even more sensitive method of seeing if the cells are happy.

please, RPM doesn't mean anything if you don't precise with which rotor.
expressing centrifugation by g or rcf is better.
Cheers

How can I check this cytochrome c?

Dear Missele,

5 minutes at 100g or 800rpm on my sigma centrifuge is easily fast enough to pellet cells without causing major damage.

Dear Josephine,

We have an in house method for looking at whole cells and their Cytochrome C levels. When cells are stressed they release cytochrome C.
We also look at Oxygen Consumption, Complex I, II, III and IV oxidation/reduction states. All are very very sensitive methods of looking at the vaibility and well being of whole cells.
When we see major changes in the function of the cells, Trypan Blue measurement would still be 98%, therefore too insensitive to pick up these changes.