We have power trips going on quite frequently in our lab and to say the least this is a nightmare for my PBMC isolations. I was wondering if it would ruin the layer separation permanently or is there still a chance to get a nice layer by resuming the spin after electricity restoration? I wanted an opinion from someone who has experienced such an event.
Hi Saira,
you should not have any problems. We had power cuts in the past here and the separation worked just fine.
Keep up with your work and do not get discouraged.
Best wishes,
Nikos
Hi Saira..
I have had this problem. If the speed is decreasing and in between your power is back and speed increases, then you may have a problem. You should switch off power supply to centrifuge when power failure is there and allow it to come to rest. When power is back, you can spin again and layer will be fine.
I agree with Arya. The centrifugation, the speed and the brake are delicate moments in buffy coat formation. Hope you resolve the electricity problems, maybe you have a short circuit of an instrument in your lab.
Agree with all foreposters. Important IMHO is that
i) the centrifuge has no defects in bearing(s)
ii) if you still are in the process of centrifugation and you'll face a power cut, disconnect from power supply and let swing out the rotor without using break function.
iii) when electric power is back, start the centrifuge again (ensuring all electronically set parameters e.g. time& g-force have been saved, otherwise you have to program once again).
iv) Dear Saira, would you mind to communicate the parameters of your buffy coat preparation in this forum?? (fixed angle vs. swing out rotor?)
From my experience the biggest problem will be if the brake acts due to a power failure as this will ruin your layer on the Ficoll (or whatever you use). But how your centrifuge reacts to the power cut (break on or off) could depend on the type of centrifuge you use.
It should not hamper. If the machine stops with a jerk, then the buffy coat may be disturbed.Otherwise the there is normal spin, the layer will not be disturbed,Moreover, if the layer is disturbed spmehow,you would see it from outside in the glass tube.
Hello
i agree with Christa Walther .
I often use ficoll separations and have found smeared gradients when there is something wrong with the brake settings of the centrifuge. Is it possible that someone adjusted your brake settings?
Good Luck
You can still recover the buffy coat. Just gently mix the blood again and spin one more time at the same time and speed. Then you should be able to get a nice buffy coat again.
Best!!
Switch off the brake of the centrifuge. Then you don´t loose anything.
I agree with Prasanta K Ray, we have also power cut issues but there is always a nice layer.
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