[Histonet] is Methyl Green suitable for water-based mounting media?

[andres]

Hi everybody,

I'm trying to counterstain my in situ hybridizations with methyl green and 
want to mount them in glycerol or similar.  I'd like to ask if any of you 
have done (or if it washes away with time...since all the protocols I've 
seen are followed by dehydration prior to mounting).

Also, since I can't get here the already prepared solution and I've seen 
several ways to prepare the staining solution from powder(Aldrich) any 
suggestion about how to do it??


Thanks a lot !



Andrés Kamaid

Dpto. Histología
Facultad de Medicina
Universidad de la República
Montevideo - Uruguay




At 17:04 01/02/2006, James Watson wrote:
>Steve,
>
>I routinely use 5% glycerin in the absolute alcohols for my mouse 
>tissues.  this has shortened our soaking times and eliminated the drying 
>artifact.  I started using this years ago on whole cross sections of dog 
>hearts that were processed at a 2.5 cm thickness and on frog leg muscle 
>that was very dry.  I am in the process of completeing some comparative 
>studies and hope to get this published soon.  if you want my processing 
>schedules let me know.
>
>James Watson
>GNF San Diego
>jwatson <@t> gnf.org
>
>________________________________
>
>From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Steven Coakley
>Sent: Tue 1/31/2006 10:55 AM
>To: Histonet <@t> lists.utsouthwestern.edu
>Subject: [Histonet] processing
>
>
>
>Good afternoon,
>
>   I'm working with mouse/rat muscle and trying to fine tune my processing 
> schecdule.
>   My last run of muscle were very dry.  I have a copy of the NSH Animal 
> Processing Manual and trying to strike a balance between those listed on 
> page 5-6.  I've noticed most of the protocols do not call for heat or 
> P/V.  The scedule I currently use is as follows: 1-70% (hold), 1-80%, 
> 2-95%, 3-100%, 3-xylene all for 30 minutes each and at 38C with P/V.  3 
> paraffins, 45 minutes each at 55C with P/V.  I'm fairly new to research 
> and still trying to make the "mental" switch in handling the different tissue.
>
>   Thanks everyone,
>
>   Steve
>
>
>
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