Slit-scanning technique using standard cell sorter instruments for analyzing and sorting nonacrocentric human chromosomes, including small ones

We have investigated the performance of two types of standard flow cell sorter instruments, a System 50 Cytofluorograph and a FACSTar PLUS cell sorter, for the on-line centromeric index (CI) analysis of human chromosomes. To optimize the results, we improved the detection efficiency for centromeres in two ways. A higher efficiency was obtained first by elongation of the chromosomes and second by introducing a high resolution lens system for laser beam focusing. In the two-parameter flow karyotype of CI and DNA content of human chromosomes, distinct peaks are produced not only by the larger chromosomes 1-8 and X, but by the smaller nonacrocentric chromosomes 9-12 and 16-20 as well. As the chromosomes 9-12 cannot be distinguished by other flow karyotyping methods, we discriminated and sorted chromosomes 12 and 10 from 9 and 11 to investigate the capacity for the separation of chromosomes in this group. A purity of at least 90% was achieved; in the isolated population the fraction chromosomes 12 was 55%; the remaining 45% were chromosomes 10 (40%) and unidentifiable chromosomes (5%)