Centrifuged cells stick to wall of reaction tube - (Jan/07/2013 )
Tabaluga on Tue Jan 8 20:44:13 2013 said:
Thanks everyone for sharing your ideas !
@pito: That's true, but I see this as my last resort. I don't think the lab will buy more expensive tubes because it's weird, I seem to be the only person having this problem (my cells perhaps ??)
@casandra: I already try to pipet the rest of the stuck cells down when resuspending, but somehow the cell number still diminishes greatly. Maybe due to shear forces when I remove the supernatant with the pump or something like that ?
@bob1: Good idea, I never tried that. I can try this out tomorrow and will certainly report back whether it worked.
But adding substances to prevent sticking, costs money and time too..
You could do what bob1 said, but I wonder if you could collect everything of your sample in an easy way like that.
Its always a question about whats best in the end: buying more expensive tubes or messing a long time trying to avoid buying them.
@ casandra: Well I wouldn't say it's fluffy but it can move around if you aspire the supernatant close to it. Also i always leave a tiny bit of supernatant as to avoid that, however I cannot avoid the cells on the wall getting under "vacuuming stress", at least I think that's where the problem may be. And I already have to pipet off manually a lot in the later parts of my procedures when I use formaldehyde or other fixatives under the fume hood... luckily no carpal tunnel syndrome yet though 
@ pito: I know. when other ways fail, I'll see to it that we can test these non-sticking tubes at least. Which ones do you use yourself ?
Unfortunately, bob1's suggestion didn't work for me. So I guess that means I'll have to really look for different tubes... Thanks everyone! If you happen to have any other ideas I'd be grateful to hear them of course !