[Histonet] Hoescht and Giemsa

Xenophanes _ xenophanes__ <@t> hotmail.com
Tue Feb 3 10:52:23 CST 2009

Hello all.  I am excited to be a new participant of the Histonet list serve.  I hope that you can provide some assistance in a little problem I am having.Simply, I wish to photograph blood smear slides (methanol fixed) with both Hoescht and Giemsa.  I want to find items via normal light microscopy from the Giemsa stain and visualize the Hoescht via our UV imager on the same scope.  The problem I am having is that performing the Giemsa stain first doesn't work out well because the Hoescht staining solution leeches out the Giemsa and produces washed out samples with little purple staining.  I am trying to do the Hoescht stain first, but I am afraid that the Giemsa staining conditions might reduce the ability to resolve the fluorescent signal from the Hoescht dye.Furthermore, Hoescht binds DNA in the minor groove, while the Giemsa binds DNA on the phosphate groups.  Does any one know about possible inhibition of binding between these two compounds since they are physically close on the DNA strand itself?Thank you again for your time and assistance!-Blood Smear-O-Rama
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