cell loss during antibody staining for flow cytometry - FACS staining protocol (Jan/11/2011 )

Hello,

I have used 1.6 ml eppendorf vials to stain cells for FACS and many cells are lost after spinning and aspirating the supernatant during washes. Should I use 12x75 mm polystyrene tubes instead? Are there adapters to centrifuge these tubes?

Thanks!

Vic

victorius on Wed Jan 12 02:38:54 2011 said:

I usually use 15ml Falcon tubes, depends on the number of cells you are using. 5ml (12x75) tubes work as well, there are also adapters around. Maybe you should ask in your department if anyone has them. Polystyrene works better for me than Polypropylene.
Anyway, you have to expect a certain cell loss during staining and washing. You can try if changing the tubes helps to minimize this.

rsm

I use 1.5MCT and never face such problem becuase I align the tubes so that the tapering (opening) end of cap is facing inside the axis of the eppenndof minicentrifuge, and the pellet formed is always on the out base of the tube, i hope you might have got it (1500rpm for 5 mins is my stnd centrifuging conditions)

I would recommend 4000rpm for 2mins if you want to use 1.5ml tubes...

Thank you all!

Anyway, you have to keep in mind that you loose at least 5% of cells every centrifugation. It's true.
As for FACS tubes, almost all cell culture-size centrifuges have an appropriate adapter for that stuff.