I am looking to isolate spermatid cells, as they are haploid cells that do not posses highly condensed chromatin of mature sperm cells. It is paramount that cells are minimally disturbed, and all FACS/MACS methods I've come across employ intercalating DNA dyes (like Hoechst Blue/Red) that might interfere with DNA analysis (single cell next generation genomics). STA-PUT methods, although very simple and inexpensive, are based on potentially damaging enzyme digestion and subsequent time-consuming gradient separation. The best idea is to conjugate spermatid cells with fluorophore (or magnetic bead) conjugated antibody for spermatid-specific cell-membrane protein and employ cell sorting.
To that result, have there been works (or protocols, etc.) that employ high throughput cell sorting based on (mouse) spermatid-specific cell-membrane proteins? The best target for spermatid-specific cell-membrane protein would be nectin-3 - are there any better spermatid-specific cell-membrane proteins?
Furtheremore, it is especially important that our haploid fraction is not contaminated with diploid faction, and to that result STA-PUT and other gradient based methods do not achieve high enough purity to be the sole method for isolation.
Thank you for your time!