Could anyone advise me on fixatives for cells to be flow sorted and subjected to RNA extraction for RT-PCR? Normally, we run the cells, unfixed, on the same day, on an analytical flow cytometer. If we can't run them the same day, we fix in 2% paraformaldehyde. This sort will involve a drive to Toronto, so doing it the same day is not practical. Based on an abstract in Cytometry (full paper ordered but still not received!), 95% ethanol + 5% acetic acid + RNase inhibitors should be OK from the RNA standpoint, but we were concerned about the effect on cell morphology. (Large size and granularity are key features of our cells.) We tested the ethanol fixation today, and the scatter plots are a mess. It would be great if formalin were OK for RNA recovery. We have another animal scheduled for Monday to test fixation on, so we are really keen for input! Heidi Dr. Heidi Engelhardt, Adjunct Professor Department of Animal and Poultry Science University of Guelph Guelph, Ontario CANADA N1G 2W1 phone: 519-824-4120, ext. 3658 or 8355 fax: 519-767-0573 email: hengel@aps.uoguelph.ca www: http://www.aps.uoguelph.ca/faculty/heidi.html
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