Chromosomal DNA measurements represent the most accurate methodology for the quantitative analysis of human karyotype. A modified FACS 440 signal collection is described here. The FSC signal voltage coming from the optical bench has been split in two equal parts by a T-connector placed at the preamplifier entrance. The side scatter channel that is not employed during chromosome analysis has been used for the half-FSC signal. This modification allowed the contemporary use of FSC signal both for gating and analysis leading to a better discrimination of background, and therefore to a more accurate definition of sorting windows.