[Histonet] coverslipping question
b427297 <@t> aol.com
b427297 <@t> aol.com
Thu Nov 14 09:07:26 CST 2013
Are you using tape or glass coverslips? We have found that when using tape coverslips, at least two very clean 100% alcohol steps must be added before slides go into the clearing reagent, be it xylene or Histoclear. If not, brown spots appear on some tissues, usually liver and bone. If there is any TRACE of water retained in the tissue, it will show up as a brown artifact on the tissue. This doesn't happen with conventional glass coverslipping and mounting medium - I think the mounting medium compensates for the retained moisture in the tissues.
From: Kim Donadio <one_angel_secret <@t> yahoo.com>
To: Gautier, Nicole M. <ngauti <@t> lsuhsc.edu>
Cc: histonet <histonet <@t> lists.utsouthwestern.edu>
Sent: Wed, Nov 13, 2013 1:37 pm
Subject: Re: [Histonet] coverslipping question
I would try adding 1 or 2 more 100% alcohols before the histoclear.
Sent from my iPhone
On Nov 12, 2013, at 9:32 AM, "Gautier, Nicole M." <ngauti <@t> lsuhsc.edu> wrote:
> My lab has been having a problem with "specks" appearing on our slides after
they come out of Histoclear. Our protocol is to dehydrate in 2 minutes each of
70%, 80%, 95%, and 100% ethanol before 2 minutes 2 times in Histoclear. The
slides are perfectly clear when they come out of the ethanols, but not when they
come out of the Histoclear.
> Since it's not a problem we had when we first started, I tried changing out
the Histoclear, but the problem remained. At the end of last week, I changed
out everything - all glassware, all ethanols, all Histoclear. The problem went
away and the 24 slides looked fine. Today, the slides are looking speckled
> So I guess my questions are: What could be causing the speckled appearance of
the slides? and How often should I have to change the ethanols and Histoclear?
I seem to remember years ago when I did this that we only changed the solutions
> Nicky Gautier
> Research Associate
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
More information about the Histonet