Chapter title |
Flow Cytometry for the Isolation and Characterization of Rodent Meiocytes
|
---|---|
Chapter number | 11 |
Book title |
Meiosis
|
Published in |
Methods in molecular biology, March 2017
|
DOI | 10.1007/978-1-4939-6340-9_11 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6338-6, 978-1-4939-6340-9
|
Authors |
Geisinger, Adriana, Rodríguez-Casuriaga, Rosana, Adriana Geisinger, Rosana Rodríguez-Casuriaga |
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. |
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