SYMPOSIA PAPER Published: 01 January 1999
STP15818S

Quality Control in the Application of Flow Cytometry to Studies of Environmentally-Induced Genetic Damage

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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.

Author Information

McCreedy, CD
School of Forest Resources, Purdue Univ., W. Lafayette, IN
Robinson, JP
Purdue Univ., W. Lafayette, IN
Dallas, CE
College of Pharmacy, Univ. Georgia, Athens, GA
Jagoe, CH
Univ. Georgia, Savannah River Ecology Laboratory, Aiken, SC
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Details
Developed by Committee: E47
Pages: 401–412
DOI: 10.1520/STP15818S
ISBN-EB: 978-0-8031-5408-7
ISBN-13: 978-0-8031-2618-3