Hello everyone, I need help in analyzing samples that require logarithmic pulse area vs. width. I have tried unsuccessfully to reproduce results from a method described in a journal article to evaluate micronuclei in V79 cells (D. Roman, et al, "Evaluation of a New Procedure for the Flow Cytometric Analysis of In Vitro, Chemically Induced Micronuclei in V79 Cells", Environ. & Mol. Mutagen. 32:387-396, 1998). The method uses a two-step technique to destroy the cellular membrane and cytoplasm, and release and stain the nuclei and micronuclei with ethidium bromide. It requires analysis of ethidium bromide width versus area using a logarithmic scale to separate debris from micronuclei. The analysis was done using a BD FACSVantage flow cytometer. However, we have a BD FACSCalibur which can only analyze width vs. area using a linear scale. Using the linear scale does not sufficiently separate the populations. Has anyone successfully accomplished this procedure without using the logarithmic scale? Any and all comments and suggestions are appreciated. Many thanks. Anne Illi-Love Department of Microscopy and Microanalysis DR45M, Bldg. AP31 Abbott Laboratories, Abbott Park, IL 60064 (847) 937-0991 anne.h.illi-love@abbott.com
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