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Cytological Analysis of Spermatogenesis: Live and Fixed Preparations of Drosophila Testes
Authors:Poojitha Sitaram  Sarah Grace Hainline  Laura Anne Lee
Institution:1.Department of Cell and Developmental Biology, Vanderbilt University Medical Center
Abstract:Drosophila melanogaster is a powerful model system that has been widely used to elucidate a variety of biological processes. For example, studies of both the female and male germ lines of Drosophila have contributed greatly to the current understanding of meiosis as well as stem cell biology. Excellent protocols are available in the literature for the isolation and imaging of Drosophila ovaries and testes3-12. Herein, methods for the dissection and preparation of Drosophila testes for microscopic analysis are described with an accompanying video demonstration. A protocol for isolating testes from the abdomen of adult males and preparing slides of live tissue for analysis by phase-contrast microscopy as well as a protocol for fixing and immunostaining testes for analysis by fluorescence microscopy are presented. These techniques can be applied in the characterization of Drosophila mutants that exhibit defects in spermatogenesis as well as in the visualization of subcellular localizations of proteins.
Keywords:Basic Protocol  Issue 83  Drosophila melanogaster  dissection  testes  spermatogenesis  meiosis  germ cells  phase-contrast microscopy  immunofluorescence
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