The Acrylamide FISH Protocol for Pollen Mother Cells (etc)

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 publication (below) contains the the original complete
description of this method for use with higher plant meiocytes, and is 
the appropriate source (not this web page) for methodology or citation.

Although the bouquet paper was limited to meiocytes, I have also 
successfully used this protocol to obtain 3-D FISH images from root 
tips, shoot meristems, and vibratome-sectioned leaf material.  
It is a very nice way to preserve the integrety of the nucleus, 
cell, and even tissue 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 writing up much of the long protocol.

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