The use of trout erythrocytes and human lymphocytes for standardization in flow cytometry

A method of standardization of flow cytometric ploidy measurements using trout erythrocytes and human lymphocytes is described. The sources of errors of the ratio between the modal channel number of erythrocytes and lymphocytes were investigated. The sample standard deviation was 0.3%, and the variation between persons of the same sex was approximately 0.5%. A distinct difference (1.82%) between the two sexes indicated that small deviations of DNA content can be detected. The standardization method was applied to the analysis of biopsy specimens from bladder tumors, normal bladder mucosa, and lymphocytes from the same patients. In diploid populations the standard deviation of the DNA indices was 1.5-2% for normal bladder mucosa but approximately 2.5% for tumors. These values indicate that deviations amounting to 4-5% from the diploidy are detectable by a single analysis of one sample.