The Acrylamide FISH Protocol for Pollen Mother Cells (etc) Note: 3D Acryl FISH method to be published in 2012, ES Howe et al. The 1 page short protocol is here: FISH_meth_short.html The long, detailed protocol from anther fixation to scope is here. The latest version, AcrFISH4.doc, is here (12/99). Web page version AcrFISH4.html or text file version AcrFISH4.txt I developed this procedure by adapting the FISH and embedding protocols used by Abby Dernburg and Sue Parmelee in the Sedat lab. This protocol was used to analyze the telomere bouquet stage of meiotic prophase. The Bass et al. (1997) contains the the original complete description of this method for use with higher plant meiocytes. See also methods chapter due out some time in 2012, Methods in Mol Biol, ES Howe, SP Murphy, & HW Bass, in Plant Meiosis Methods). In addition to maize meiotic cells, we have successfully used this protocol to obtain 3-D FISH images from root tips, shoot meristems, or vibratome-sections from maize, arabidopsis, tobacco, oat, and lily. This method provides good preservation of the nucleus, even during FISH. Bass H.W., Marshall, W.F., Sedat J.W., Agard D.A., and Cande W.Z. (1997) Telomeres cluster de novo before the initiation of synapsis; a 3-dimensional spatial analysis of telomere positions before and during meiotic prophase J Cell Biol 137(1):5-18 Thanks to Lisa Harper for help in initial writing of the long protocol.