The suitability of air cooled helium cadmium (HeCd) lasers for two color analysis and sorting of human chromosomes

Commonly available HeCd lasers operate at wavelengths of 325 and 442 nm, close to the published absorption maxima of the Hoechst and chromomycin dyes used for vibariate flow karyotype analyses. Although bivariate chromosome analysis has heretofore required the use of water cooled argon ion lasers operating at hundreds of milliwatts, the HeCd lasers usually produce about one tenth this power (30 mW at 325 nm and 50 mW at 442 nm). Recently a sorting quartz flow cell design has been introduced which allows improved sensitivity compared to jet-in-air operation. The goal of this study was to determine if such an improvement in sensitivity would allow the use of HeCd air cooled lasers for bivariate flow karyotype analysis. They examined the resolution of chromosomes isolated from the human fibroblast cell line GM130 as well as a hamster-human somatic cell hybrid (Q826-20) containing a single human chromosome (No. 5). Resolution with the HeCd lasers was comparable to that obtained using water cooled argon ion lasers. Data are shown illustrating the results obtained with the two different excitation configurations. The advantages of using 442 nm exciation for chromomycin, compared to 457 nm are illustrated. In conclusion, small air cooled HeCd lasers are capablemore » of providing adequate resolution for vibariate chromosome analysis and sorting for many flow cytogenetic applications.« less