[Histonet] uh...

[William Chappell]

Oh yes, how to fix it.

Hydrate through clearite, 100% ETOH, 95% ETOH, and water.  Spend an extra 5 mins in running tap water -- that should wash out most of the eosin. Then counterstain again with eosin.

Dehydrate with the clean ETOHs and Clearite.

That should do it.

Another possibility I just thought of, I never used clearite for coverslipping, even in labs that primarily used clearite -- I always finished up in Xylene.

Will



--- On Thu, 4/7/11, William Chappell <chapcl <@t> yahoo.com> wrote:

> From: William Chappell <chapcl <@t> yahoo.com>
> Subject: Re: [Histonet] uh...
> To: "<sgoebel <@t> mirnarx.com>" <sgoebel <@t> mirnarx.com>
> Cc: "<histonet <@t> lists.utsouthwestern.edu>" <histonet <@t> lists.utsouthwestern.edu>
> Date: Thursday, April 7, 2011, 2:11 PM
> Sounds like your dehydrating alcohol
> is not dehydrating completely. I would switch out the 100%'s
> and the clearite. Either they got "wet" during the course of
> the day or someone made an oops when changing the stain
> line. 
> 
> Hope that helps. 
> 
> Will
> 
> Sent from my iPhone
> 
> On Apr 7, 2011, at 2:06 PM, <sgoebel <@t> mirnarx.com>
> wrote:
> 
> > So I just stained a group of slides all at the same
> time with the same
> > conditions.  About 40 of 150 the eosin looks like
> it is bleeding out of
> > the sections...this has never happened before? 
> What could be the cause
> > and how do I fix it?  Everything is as normal,
> but again I am being
> > forced to use the crappy stuff called clear
> rite.  Could this be the
> > cause of the bleeding?
> > 
> > Thanks
> > 
> > 
> > 
> > Sarah Goebel, BA, HT(ASCP)
> > 
> > Histotechnologist
> > 
> > Mirna Therapeutics
> > 
> > 2150 Woodward Street
> > 
> > Suite 100
> > 
> > Austin, Texas  78744
> > 
> > (512)901-0900 ext. 6912
> > 
> > 
> > 
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