[Histonet] RE: ISH preparation

[Joana Moreira]

Leica's Thermobrite Elite?
http://www.leicabiosystems.com/pathology-imaging/cytogenetics/details/product/leica-thermobrite-elite/

Joana Moreira
Supervisor - Anatomical Pathology
Department of Pathology

Sidra Medical & Research Center
PO Box 26999 | Doha, Qatar
Direct Line  +974-4404-2036
jmoreira <@t> sidra.org | www.sidra.org




Message: 6
Date: Wed, 25 Jun 2014 08:38:29 -0400
From: Emily Brown <talulahgosh <@t> gmail.com>
Subject: Re: [Histonet] ISH preparation
To: Jennifer.Johnson <@t> genzyme.com
Cc: "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
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That may be what you have to do, as the first day of ISH needs to be RNase free!
When I do them, it is 10 slides at a time and we have dishes and a lab bench set aside for RNase free procedures only.  Obviously this is not feasible in all workplaces!
I would just think about it logically.  Use gloves when you can, always.
 Can you keep the hybridizing part RNase free? I don't know what machines you use for this--we do process the slides manually in glass dishes and then hybridize in a slide container and oven kept for RNase free purposes.
 After hybridization, you don't really need to be too careful.

Now I'm wondering if there's some sort of magical machine that does ISH!

Emily

"By bitching and bitching and bitching, they could exhaust the drama of their own horror stories. Grow bored. Only then could they accept a new story for their lives. Move forward."

-Chuck Palahniuk, "Haunted"


On Mon, Jun 23, 2014 at 2:06 PM, <Jennifer.Johnson <@t> genzyme.com> wrote:

> Hi Folks!
> I was hoping to get a little input about preparing samples for ISH.
> We currently RNAse-away our tools at the time of collection, toss the
> tissues into formalin, process normally and then cut them using RNAse
> away cleaned microtomes/forceps on a bath of DEPC treated water (in a
> cleaned water bath).
>
> How do other labs prepare paraffin embedded samples?
>
> Do you use special reagents - formalin?  Do you clean the processor
> and use treated reagents there too?  Do you wash the slides in DEPC
> treated water?
>
> I am trying to figure out how far we need to go here.  We are a high
> throughput lab and we deal with all kinds of tissues and I don't want
> to have to take down a processor and keep it RNAse free if I don't have to!
> I will run the experiments and test different methods, but I was
> hoping some of you could share your wisdom and give me a place to start.
> Thanks,
> JJ
>
> Jennifer Johnson
> Staff Scientist
> Genzyme Corp.
> Department of Pathology
> 5 Mountain Road
> Framingham, MA 01701-9322
> Phn - 508-271-3610
> Fax - 508-872-9080
>
>
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