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Meiosis pp 217-230 | Cite as

Flow Cytometry for the Isolation and Characterization of Rodent Meiocytes

  • Adriana GeisingerEmail author
  • Rosana Rodríguez-Casuriaga
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1471)

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.

Key words

Spermatogenesis Testicular cell suspension Flow cytometry Meiosis Flow sorting 

Notes

Acknowledgments

This work was supported by Agencia Nacional de Investigación e Innovación (ANII, Uruguay) (Project N° FCE-1-2011-1-6742 to A.G.). The authors thank Federico Santiñaque for his invaluable expertise in the FCM analysis and sorting procedures and for critical reading of the manuscript.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Adriana Geisinger
    • 1
    • 2
    Email author
  • Rosana Rodríguez-Casuriaga
    • 2
  1. 1.Sección Bioquímica/Biología Molecular, Facultad de CienciasUniversidad de la República (UDELAR)MontevideoUruguay
  2. 2.Departamento de Biología MolecularInstituto de Investigaciones Biológicas Clemente Estable (IIBCE)MontevideoUruguay

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