NASA/American Cancer Society high-resolution flow cytometry project-I

Background: The NASA/American Cancer Society (ACS) now cytometer can simult aneously analyze the electronic nuclear volume (ENV) and DNA content of cel ls. This study describes the schematics, resolution, reproducibility, and s ensitivity of biological standards analyzed on this unit. Methods: Calibrated beads and biological standards (lymphocytes, trout eryt hrocytes [TRBC], calf thymocytes, and tumor cells) were analyzed for ENV ve rsus DNA content. Parallel data (forward scatter versus DNA) from a convent ional now cytometer were obtained. Results: ENV linearity studies yielded an R value of 0.999. TRBC had a coef ficient of variation (CV) of 1.18 +/- 0.13. DNA indexes as low as 1.02 were detectable. DNA con tent of lymphocytes from 42 females was 1.9% greater t han that for 60 males, with a noninstrumental variability in total DNA cont ent of 0.5%. The ENV/DNA ratio was constant in 15 normal human tissue sampl es, but differed in the four animal species tested. The ENV/DNA ratio for a hypodiploid breast carcinoma was 2.3 times greater than that for normal br east tissue. Conclusions: The high-resolution ENV versus DNA analyses are highly reliabl e, sensitive, and can be used for the detection of near-diploid tumor cells that are difficult to identify with conventional cytometers. ENV/DNA ratio may be a useful parameter for detection of aneuploid populations. Cytometr y 43:2-11, 2001. Published 2001 Wiley-Liss, Inc.dagger