Tritiated thymidine incorporation assay and biosafety cabinet - (May/15/2007 )

Dear all,

Recently I got some worries about 3H works. I would like to know that during the tritiated thymidine incorporation assay to assess the proliferation of PBMC, would you add the tritated thymidine within a biosafety cabinet? If you do, which type of biosafety hood will you use? ClassIIA, B1, B2, III? If you don't, would you concern about contamination problem as you got to incubate the plate for 6 hrs after thymidine addition?

Actually, after searching for a long time over the internet, I've found that triated thymidine may generate [3H]2O waste and which can be relatively, volatile. Is that true? Well, I know that it can penetrate gloves though (thus some people suggested that we should wear 2 gloves).

Sorry if those questions seemed stupid.

Many thanks,
Kallichroma

I also wish to know about the proper way of handling 3H.

In my lab, one PhD student add 3H to the tissue culture plates outside the hood.



Thank in advance.