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Although a number of more contemporary techniques have been developed, velocity sedimentation using the STAPUT method remains as a gold standard for this purpose. The gentle nature of the technique, wherein germ cell subpopulations are separated by sedimentation at unit gravity, results in the isolation of viable and high-purity cells. We provide an updated and simplified step-by-step version of the STAPUT protocol for the purification of mouse male germ cells. As per the original method, the protocol described herein allows for the purification of mouse spermatocyte and round spermatids, however it also allows for successful purification of elongating, and elongated spermatid populations, and is optimised for the preservation of cellular ultrastructure. This method yields sufficient numbers of high-purity cells from one adult mouse for RNA or protein extraction or for immunolocalisation studies.<\/jats:p>","DOI":"10.1093\/molehr\/gaz056","type":"journal-article","created":{"date-parts":[[2019,10,2]],"date-time":"2019-10-02T11:14:32Z","timestamp":1570014872000},"page":"675-683","source":"Crossref","is-referenced-by-count":23,"title":["An optimised STAPUT method for the purification of mouse spermatocyte and spermatid populations"],"prefix":"10.1093","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2009-0165","authenticated-orcid":false,"given":"Jessica E M","family":"Dunleavy","sequence":"first","affiliation":[{"name":"School of Biological Sciences, Faculty of Science, Monash University, Melbourne, 3800, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne E","family":"O\u2019Connor","sequence":"additional","affiliation":[{"name":"School of Biological Sciences, Faculty of Science, Monash University, Melbourne, 3800, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Moira K","family":"O\u2019Bryan","sequence":"additional","affiliation":[{"name":"School of Biological Sciences, Faculty of Science, Monash University, Melbourne, 3800, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2019,10,22]]},"reference":[{"key":"2019112917103804200_ref1","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1002\/cyto.a.20129","article-title":"Flow cytometric characterization of viable meiotic and postmeiotic cells by Hoechst 33342 in mouse spermatogenesis","volume":"65","author":"Bastos","year":"2005","journal-title":"Cytometry A"},{"key":"2019112917103804200_ref2","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1083\/jcb.74.1.68","article-title":"Spermatogenic cells of the prepuberal mouse: isolation and morphological characterization","volume":"74","author":"Bellve","year":"1977","journal-title":"J Cell Biol"},{"key":"2019112917103804200_ref3","first-page":"50648","article-title":"Separation of spermatogenic cell types using STA-PUT velocity sedimentation","author":"Bryant","year":"2013","journal-title":"J Visualized Exp. 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