I have been having some trouble maintaining cultures of candida which I have aquired from ATCC. My first plate, made from the rehydrated powder they send always looks fine, giving reasonable numbers of good sized, unifrom colonies. But when I make a culture from that plate, spin down the cells, and then dilute and make new plates for counting I always end up with plates overgrown with tiny colonies which look quite different from the colonies yielded from the original rehydration. The numbers of these small colonies gives improbably high counts given my dilutions. I feel quite confident that I am using good sterile technique and that my problem is not due to contamination. Additionally I can isolate any of these colonies, stain them, and they appear as candida when viewed microscopically.
Does anyone know what I am experiencing? Is this due to spore formation? Any help would be really appreciated.
Plate your diluent. You probably have a contaminant.
Also, when setting up from ATCC, it's best to make a couple of passes to
"wake up" your organism before working with it. Sub a colony from your
original plate to a new SABS (or similar medium) and let grow. Do this
again. Put your bug onto a slant to save it (or freeze it using beads, which
I don't know how to do but someone else may). You can also do it the "old
way" we used to do - put on a slant and overlay with sterile mineral oil.
Keeps for about a year at room temperature that way.
Hope this helps.
Judy Dilworth, M.T. (ASCP)
"Christopher M Havens" <[Only registered users see links. ]> wrote in message
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I have been having some trouble maintaining cultures of candida which I
have aquired from ATCC.