title: Flow Cytometry for the Isolation and Characterization of Rodent Meiocytes.
abstract: Molecular analyses in mammalian meiotic cells have been hindered by the difficulty in isolating stage-specific cell populations, and this is especially true for early meiotic prophase stages (leptotene and zygotene). Here, we describe a method for obtaining cells in different spermatogenic stages from rodents including lepto-zygotene meiocytes at very high purity levels. The procedure includes an approach for the mechanical disaggregation of the testicular tissue, staining with a vital, noncytotoxic dye that is excitable with a blue laser, isolation of the cell populations by flow sorting, and different alternative protocols for the collection of the sorted cells.
authors.affiliation: Sección Bioquímica/Biología Molecular, Facultad de Ciencias, Universidad de la República (UDELAR), 11400, Montevideo, Uruguay. adriana.geisinger@gmail.com.|Departamento de Biología Molecular, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Avda. Italia 3318, 11600, Montevideo, Uruguay. adriana.geisinger@gmail.com.
authors.forename: Adriana
authors.initials: A
authors.lastname: Geisinger
authors.affiliation: Departamento de Biología Molecular, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Avda. Italia 3318, 11600, Montevideo, Uruguay.
authors.forename: Rosana
authors.initials: R
authors.lastname: Rodríguez-Casuriaga
mesh_terms: D000818:Animals; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods; D005434:Flow Cytometry / Q000379:methods*; D008297:Male; D049469:Meiotic Prophase I; D051379:Mice; D051381:Rats; D012377:Rodentia; D013091:Spermatogenesis; D013737:Testis / Q000166:cytology*
keywords: Flow cytometry; Flow sorting; Meiosis; Spermatogenesis; Testicular cell suspension