Extension Scientist, School of Forest Resources, Purdue Univ., W. Lafayette, IN
Professor, Purdue Univ., W. Lafayette, IN
Associate Professor, College of Pharmacy, Univ. Georgia, Athens, GA
Associate Research Scientist, Univ. Georgia, Savannah River Ecology Laboratory, Aiken, SC
Pages: 12 Published: Jan 1999
Flow cytometry (FCM) has been used to demonstrate altered DNA content in fish, reptiles, birds and mammals exposed to radionuclides, PAHs and other contaminants. However, artifacts resulting from sample preparation, handling, variations in instrument parameters or other factors may confound such measurements. Some artifacts resemble genotoxic responses and so could lead to erroneous positive conclusions. As part of ongoing studies of effects of various pollutants on DNA content in fishes, we tested sample handling and preparation methods for the induction of artifacts. We describe QA/QC methods, including control of staining conditions, doublet discrimination by comparison of peak versus integral fluorescence, internal DNA standards, and the use of time versus fluorescence plots. Consistent application of these practices is essential to obtain valid measurements of DNA content in environmental samples, and neglect of these can result in poor quality data and the acceptance of incorrect hypotheses.
quality control, quality assurance, flow cytometry, DNA content, aneuploidy, light scatter, fluorescent staining, artifacts
Paper ID: STP15818S