[Histonet] Waste drum labeling

[Cynthia Pyse]

I would also be interested in the answers to this.
Cindy 

Cindy Pyse, CLT, HT (ASCP)
Laboratory Manager
X-Cell Laboratories
20 Northpointe Parkway Suite 100
Amherst, NY 14228
716-250-9235 etx. 232
e-mail cpyse <@t> x-celllab.com

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Amber
McKenzie
Sent: Wednesday, November 28, 2012 1:16 AM
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Waste drum labeling

For EPA standards, how do you label your waste drums?  I have one 55 lb drum
that we only pour ventana waste in.  What numbers do you write in the
diamond to cover all the kits that you stain with?  2 reps from EPA came by
my lab yesterday and said i needed an emergency phone list by our phone??
Does anyone have the checklist for EPA that small labs should follow?

________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu
[histonet-bounces <@t> lists.utsouthwestern.edu] on behalf of Cristi Rigazio
[cls71877 <@t> gmail.com]
Sent: Tuesday, November 27, 2012 3:01 PM
To: Rene J Buesa
Cc: histonet <@t> lists.utsouthwestern.edu; Elizabeth Cameron
Subject: Re: [Histonet] Uneven Staining

As most other bases have been covered, do you leave space between the slides
in the rack?  Once in a blue moon we had stains from the same rack that
differed in intensity, after much troubleshooting we started putting space
between the slides and we haven't had the problem since.  Just an idea.
Cristi

Sent from my iPhone

On Nov 27, 2012, at 12:24 PM, Rene J Buesa <rjbuesa <@t> yahoo.com> wrote:

> This is quite a puzzling situation.
> You have ruled out dewaxing (that was my first idea of cause) but I do not
think that soaking will be the problem because the sections are fixed and
infiltrated with paraffin, so there is no possibility of "diluting" in
water.
> Thick/thin sections in a series is a very rare occurrence so I am also at
a loss with this problem. Weak staining solutions could cause it but all
sections in a series will be equally weak.
> Just in case, since you have covered the dewaxing, issue do the following:
do not leave the sections too long in the water bath and dry them in the
oven as soon as they are drained.
> René J.
>
> From: Elizabeth Cameron <Elizabeth.Cameron <@t> jax.org>
> To: "histonet <@t> lists.utsouthwestern.edu" 
> <histonet <@t> lists.utsouthwestern.edu>
> Sent: Tuesday, November 27, 2012 1:35 PM
> Subject: [Histonet] Uneven Staining
>
> About a year ago, I posted about issues with staining mouse kidney 
> sections (see below for original post).  We realized that part of the
problem was the sections drying during staining, which has been resolved,
but we are still having some issues with some slides staining well and
others being very pale.  This time it seems to be consistent across the
slide, so I know it is not a drying issue.  They are serial sections, so
they are all cut together and stained together, and they are from the same
animal.  I don't believe deparaffinization is an issue - we have tried
longer times with fresh solutions and it doesn't make a difference.  We have
also tried re-staining the slides after letting them sit in xylene
overnight, and there is no difference in the staining.  I am now thinking
that it may be related to how long the sections soak or how long they sit on
the waterbath.  They are NBF fixed, and there is a fine line between soaking
well for hydration and oversoaking, resulting in swelling.  Has anyone
experienced anything like this?  Any other ideas?  I am at a loss, and I'd
really like for this to work!
> Thanks,
> Liz
>
> We are doing a Hale's colloidal iron stain (no counterstain) on serial
sections of mouse kidneys.  We are staining an entire kidney at a time
(about 90-150 slides), and after many successful runs, we are now finding
some slides in each batch with very uneven staining.  Half of a section will
stain as it should, and the rest of the section is very pale.  It seems to
be in a similar area from one section to the next, but not exactly the same
area.  It does not look like a deparaffinization issue.  There may be two or
three slides in a row like this, or just one section on a slide, followed by
30-40 that look fine.  The sections are 6 microns. Any ideas on why this
might be happening?  Thanks!
>
>
>
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