Flow cytometric detection of chromosome abnormalities by measuring centromeric index, DNA content, and DNA base...

This paper highlights two improvements of the on-line centromeric index (CI) analysis for the detection of chromosome abnormalities. On-line CI versus DNA content analysis of an EBV-transformed cell line, with a deletion (11)(p13p15.1), of a patient with aniridia and Wilms' tumour demonstrates the first improvement of the method of on-line CI analysis for flow karyotyping and sorting; a reciprocal translocation, insertion, or deletion can, when the cell type contains not more than a few of these types of abnormalities, be traced to the p-arm or q-arm of the relevant chromosome. On-line CI analysis was also performed with chromosomes isolated from a transitional cell carcinoma of the bladder. Cytogenetic analysis of this cell line showed numerous chromosomal abnormalities. Chromosomes of this cell line were also karyotyped by bivariate flow cytometry using a different set of parameters: Hoechst 33,258 fluorescence intensity (HOfl) versus chromomycin A3 fluorescence intensity (CAfl). A comparison of these results reveals the second improvement of the CI method for the detection of chromosome abnormalities; bivariate analysis of CI versus propidium fluorescence (PIfl) are complementary to bivariate analysis of HOfl versus CAfl. Chromosomes with distributions that fuse together in the HO/CA flow karyotype may be distinguished as individual peaks on the basis of their CI values.