We are currently having problems with our TB cultures. They are very clumpy. Does anyone have suggestions on how to solve this problem?
A possibility is to draw them up into a syringe using a non beveled wide gauge needle a few times. It needs to be fairly gentle though so as not to shear the cells. Another possibility is to triturate them by drawing them up very slowly using either fire polished glass pasteur pipettes or a 1 ml gilson pipette.
A possibility is to draw them up into a syringe using a non beveled wide gauge needle a few times. It needs to be fairly gentle though so as not to shear the cells. Another possibility is to triturate them by drawing them up very slowly using either fire polished glass pasteur pipettes or a 1 ml gilson pipette.
Li Li Have you tried coating the culture flask with something that they might adhere to when they are first plated down in preference to each other?
I have been told that growing mycobacteria in Middlebrook medium in liquid culture gives cells that clump much less than growing cells on agar medium. Have you tried that? Also, if you have a mixture of suspended cells and clumps, you can let this stand in a test tube for a while and clumps will settle to the bottom. You can then take off the top layer containing suspended single cells. Have you tried that?
HI, I use a magnetic stirrer on a very low setting while culturing this prevents them from clumping to a large degree, but clumping is unavoidable once they reach around 10-8/ml. Also If you have the stirrer speed set too fast you will end up with a ring around the flask.
I don't use the shaker anymore, I use a stirrer with a small magnetic flea, a lot less clumping.
Paul Wheeler at the VLA in Weybridge, UK tells me that 0.02% tyloxapol can be used in the culture medum. Another option is adding D-arabinose according to a publication by M. Daffe in FEMS Microbiol Letters 1996 v.144 p.167-70.
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