RE: [Histonet] H.Pylori

From:"Monfils, Paul"

I haven't done this with H. pylori, but I have done it with other organisms, including gram+ and gram- bacteria, acid-fast organisms, and yeast as a fungus control. I prefer lung as the tissue because there are so many natural spaces to receive the organisms. If you inject into something like liver, the tissue will have to separate to receive the injection, and there will be little to hold the injected culture in place.  My preferred method is to take a block of fresh tissue, push the needle almost all the way through, then inject as I withdraw the needle. Do this three or four times, then leave the tissue in a covered petrie dish with a moist piece of paper towel to maintain humidity, at room temperature for a couple of hours. This allows the organisms to settle and adhere to the tissue. Then just drop the tissue into formalin, let it fix, and process as usual.  I have also injected into pre-fixed lung and gotten usable results, but not as good as with the above method. In this case there is no need to wait a couple of hours because the organisms will die as soon as they are injected, so just inject and drop into formalin.

For gram+ controls I use a mix of Bacillus subtilis and Staphylococcus epidermidis.  For gram- a mix of Escherichia coli and Micrococcus luteus. I purchase the cultures from a biological supply company - - for less than $10 each, in liquid medium, not on plates.  I spin them down to concentrate the organisms, pour off most of the medium, resuspend by vortexing, mix the cultures if I want a mix, then draw into a syringe and inject as above.

> ----------
> From: on behalf of Vacca Jessica
> Sent: 	Thursday, January 25, 2007 10:05 AM
> To:
> Subject: 	[Histonet] H.Pylori
> Does anyone that works in a Hospital have their Micro department grow H.
> pylori and create their own controls by injecting into the tissue? If
> you do, do you mind getting me their procedure for this? Thanks in
> advance for your help.
> Jessica Vacca
> Histology Supervisor
> Brandon Regional Hospital
> 119 Oakfield Dr.
> Brandon Fl 33511
> (813) 571-5193 or (813) 681-5551 ext 2454
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