Posted 13 March 2012 - 11:08 AM
I'm a relatively new FACS user, as is everyone in my lab. We've had a basic introduction to flow cytometry and the machine is running relatively well, however, we frequently have problems with clogs. If the nozzle isn't clogged, then there is something blocking (or at least causing problems) with the sample line. We work with adherent cells, and unfortunately everyone has just sort of been using whatever buffer they want when they do FACS. I assume this is part (sample preparation being the obvious other half) of the problem. Could I get some suggestions from experienced FACS users for what a good FACS buffer is and why you add what you add. I have one guy sorting with his regular media, one guy using 10% FCS in his buffer and when I tell them it's not good, they just tell me: It makes my cells happy and I don't have any problems with clogs! Of course, the next person using the FACS more likely than not will have a clog, so I would like to have some feedback from more experienced users. Are they right and anything can be used as a buffer? How do I convince them if they need to use less FBS? I am under the impression that a max of 2%FBS in PBS with 1mM EDTA is the basic FACS buffer. Any help so that we can reduce the amount of time wasted on troubleshotting due to clogs would be helpful!
Sorry, I'm just a lowly tech and it's hard to get some PhDs to listen, ergo I need some solid reasoning behind my argument. Thank you in advance.
Posted 13 March 2012 - 12:42 PM
Posted 13 March 2012 - 07:55 PM
I'm not saying for definite that the FBS concentration can't be a problem, but it is routinely used in FACs buffers so it can't be that bad. And I've used as high as 5% before with no problems.
The facility where I do my flow is VERY strict about cleaning the machine between runs, so perhaps you could introduce that to your lab?
Edited by leelee, 13 March 2012 - 07:58 PM.
Posted 13 March 2012 - 07:57 PM
Posted 13 March 2012 - 08:02 PM
Posted 14 March 2012 - 01:03 AM