[Histonet] Hoescht and Giemsa

[Lewin, Anne]

I have never tried that type of double staining, but DAPI also stains DNA in the minor groove and may not interfere with your Giemsa stain.  You can get DAPI already diluted in mounting media - Vector Labs makes a nice aqueous mount one that hardens so you don't have to use fingernail polish to seal the edges.

>-----Original Message-----
>From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-
>bounces <@t> lists.utsouthwestern.edu] On Behalf Of Xenophanes _
>Sent: Tuesday, February 03, 2009 11:52 AM
>To: histonet <@t> lists.utsouthwestern.edu
>Subject: [Histonet] Hoescht and Giemsa
>
>
>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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