Anybody able to help with the following? ------- Forwarded Message Follows ------- From: Dorothy Reimer <reimed@genemedicine.com> To: cyto-inbox Date sent: Wed, 23 Jul 1997 17:49:35 -0500 Subject: some flow questions Dear Terry: I am interested in following some internal fluorescence through tumour cells during gene transfection procedures. I have been able to follow it by flow to some extent, but I find that there is some autofluorescence associated with these cells (directly dispersed from tumours which were injected with some formulations). I have some information to believe that trypan blue can be used to decrease to autofluorescence thereby enhancing the signal to noise ratio. I have tried this with my cells, but fail to see a shift to the left indicating a decrease in autofluorescence. I have tried several different concentrations of TB in addition to several other things. But, after talking with Mark, I am wondering about the mechanics of how TB works. Do you as an experienced flow person have any suggestions or comments to me about the use of TB as an inhibitor for autofluorescence. Or is there something else that I could try. I really need to find a way to enhance the separation on flow, since the population of cells that I have that are positive are potentially quite low. I would sure appreciate any ideas you can pass my way. Thanks in advance for your help. Dody Reimer, Senior Scientist, Gene Medicine. Dr. Terry Hoy Principal Research Officer Department of Haematology Telephone 44 1222 743458 University of Wales College of Medicine FAX 44 1222 744655 Heath Park E-mail hoy@cardiff.ac.uk CARDIFF CF4 4XN http://www.cf.ac.uk/uwcm/hg/hoy/index.html U.K.
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