Male meiotic chromosome preparation

Several techniques have been developed for preparation of male meiotic chromosomes (see, e.g.. Cooper 1965 Ashbumer 1989 Miyazaki and Orr-Weaver 1992). All of these techniques allow a clear visualization of chromosomes, but not of other cellular structures such as the cytoskeleton and the spindle. We describe here three of these techniques that in our hands give good reproducible results. The first technique, developed by Lifschytz and Hareven (1977) and described below in Protocol 5.9, is particularly usefid for the analysis of adult testes. The other two techniques (Ripoll et al. 1985 Cenci et al. 1997) are routinely used in our laboratory for larval and pupal testis preparations. The technique described in Protocol 5.10 is a very simple method for preparing aceto-orcein-stained chromosomes. The other technique outlined in Protocol 5.11 allows preparation of unstained chromosomes that can be subsequently stained with various dyes such as Giemsa, Hoechst 33258, and DAPl, or processed for in situ hybridization. 

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