Improved Sensitivity and Resolution in the Flow Cytometric DNA Analysis of Human Solid Tumor SpecimensUse ofIn VitroFine-Needle Aspiration and Uniform Staining Reagents

Bruce Allen Bach, William A. Knape, Mark G. Edinger, Raymond R. Tubbs
1991 American Journal of Clinical Pathology  
Twenty-five unselected fresh colon or breast tumors were studied to identify specific components of sample preparation, sample staining, and flow cytometer operation affecting the sensitivity of DNA stemline analysis. Solid tumors were disaggregated using both a published method for mechanical/enzymatic whole cell (M/EWC) isolation and a fine-needle aspiration (FNA) technique. Staining FNA samples with CycleTEST™ propidium iodide reagents demonstrated improved sensitivity in the recognition of
more » ... ear diploid and near tetraploid aneuploid populations: 9 of 20 resolvable aneuploid DNA stemlines identified in FNA suspensions were not detected or clearly resolved in M/EWC There is considerable interest, by pathologists and clinicians, in the use of flow cytometric DNA stemline analysis to characterize human neoplasms for clonal diversity and potential for clinical progression as measured by proliferation fraction. 1 " 15 Propidium iodide staining of representative samples of disaggregated solid tumor specimens, immediately followed by flow cytometric DNA stemline analysis, permits both objective and quantitative measurements of DNA aneuploidy, 616 " 18 and, under appropriate conditions when mathematical models are applied, simultaneous estimates of cell-cycle fraction (S-phase estimation). These cell-cycle fractions, as a measure of cellular proliferation, may be correlated to the biological behavior of the tumor.
doi:10.1093/ajcp/96.5.615 fatcat:btkl3s3wobfodefzkaupqafoou