Germline Stem Cells

Volume 450 of the series Methods in Molecular Biology™ pp 95-107

High-Resolution Light Microscopic Characterization of Spermatogonia

  • Hélio Chiarini-GarciaAffiliated withDepartment of Morphology—ICB Federal University of Minas Gerais, Laboratory of Structural Biology and Reproduction
  • , Marvin L. MeistrichAffiliated withDepartment of Experimental Radiation Oncology, The University of Texas, M.D. Anderson Cancer Center

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It is possible to distinguish the morphological features of the spermatogonial nuclei and nucleoli and to further identify their distinct generations using an appropriate method to fix whole testes via vascular perfusion with glutaraldehyde, postfixation by immersion in reduced osmium, embedding in araldite, and staining of semithin tissue sections. A well-trained individual can distinguish each of the spermatogonial types in rodents (Aundiferentiated, A1, A2, A3, A4, In, and B) using this tissue preparation technique based on their morphological details and without correlation with the stages of the epithelium cycle or other parameters. The possibility of distinguishing each spermatogonial type by their morphological characteristics allows a more accurate evaluation of their kinetics during the spermatogenic cycle. Moreover, the understanding of spermatogonial behavior is a means to elucidate the functional control of the spermatogenesis, which consequently allows the determination of their effects on the fertility of humans and other animals.


High-resolution light microscopy morphology perfusion spermatogonia stem cell