Chromosome sorting was key for sequencing of the human genome and is still used today in fields as diverse as plant biology and cancer. The technique has remained largely unchanged since inception and involves staining chromosomes released from metaphase cells with the dyes Hoechst (AT base pair specific) and Chromomycin A3 (GC base pair specific). The respective excitation of these dyes relies on a UV and mid blue laser (such as 445nm or 457.9nm), neither of which are common features of most flow cytometers. To increase the accessibility of this technique, we have explored an alternative dye to Chromomycin A3 - 7-Aminoactinomycin D (7AAD). 7AAD has a similar GC base pair specificity as Chromomycin A3 but is excitable by a 561 nm laser (and to a lesser extent a 488nm laser), both relatively common on modern flow cytometers. Chromosome suspensions were stained with 7AAD and Hoechst or CA3 and Hoechst and flow karyotypes were generated from three instruments, a MoFlo Legacy, LSR Fortessa X-20 and an ARIA Fusion. 561 nm excitation of 7AAD yielded flow karyotypes with clearly resolvable chromosome clusters. Cluster resolution was decreased especially for larger chromosomes relative to samples stained with CA3. Resolution of flow karyotypes from 7AAD stained chromosomes excited with a 488 nm laser was reduced relative to 561 nm excitation but still surprisingly useable. Use of the 405 nm laser for excitation of Hoechst did not yield flow karyotypes with sufficient resolution to identify chromosome clusters. In conclusion 7AAD can act as a subpar substitute to Chromomycin A3 if lacking a mid 400 nm laser, yielding flow karyotypes with sufficient resolution for many purposes.