I am looking for a reliable method for staining mouse muscle with ATP'ase pH
4.3, 4.6 and 9.4.
Any help will be greatly appreciated.
-----Original Message-----
From: [email protected]
[mailto:[email protected]] On Behalf Of Tom McNemar
Sent: Wednesday, February 09, 2011 9:05 AM
To: 'Erin Sarricks'; [email protected]
Subject: RE: [Histonet] PAS with Diastase Digestion
Spit for 5 minutes.... old school....
Tom McNemar, HT(ASCP)
Histology Co-ordinator
Licking Memorial Health Systems
(740) 348-4163
(740) 348-4166
[email protected]
www.LMHealth.org
-----Original Message-----
From: [email protected]
[mailto:[email protected]] On Behalf Of Erin Sarricks
Sent: Wednesday, February 09, 2011 7:38 AM
To: [email protected]
Subject: [Histonet] PAS with Diastase Digestion
Hi Histonet-
I recently ran a PAS/D stain and had some issues with it. Both "with" and
"without" slides came out looking the same so I'm guessing my digestion step
didn't work! I used a Malt Diastase solution (0.5g to 500mL water) for my
digestion. This is the procedure I used:
1. Deparaffinize and hydrate to water.
2. Place the sections labeled “with” in diastase solution preheated to
37˚C for 1 hour. Hold the sections labeled “without” in distilled water.
3. Wash in running water for 5 minutes
4. Place all section (with and without) in 0.5% periodic acid solution
for 5 minutes
5. Wash in 3 changes of distilled water
6. Place in Schiff reagent for 15 minutes
7. Wash in lukewarm tap water for 5 minutes (immediately sections turn
dark pink color).
8. Counterstain in Mayer’s Hematoxylin for 3 minutes.
9. Wash in tap water for 10 minutes
10. Dehydrate starting with 95% ETOH, clear, and coverslip.
I am wondering if my solution possibly got too warm in the oven and hindered
the enzyme activity, or is it possible I left it in too long? Any tips would
be much appreciated! Oh, and I have about 300 slides to stain, so spitting on
them is my last last last resort! Haha! Thanks in advance for all your help!
Erin Sarricks, HT (ASCP)
Histology Laboratory Technician
USAMRICD Comparative Pathology Branch
Office: Bldg E-3081 Room 178
E-mail: [email protected]
_______________________________________________
Histonet mailing list
[email protected]
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
This e-mail, including attachments, is intended for the sole use of the
individual and/or entity to whom it is addressed, and contains information from
Licking Memorial Health Systems which is confidential or privileged. If you are
not the intended recipient, nor authorized to receive for the intended
recipient, be aware that any disclosure, copying, distribution or use of the
contents of this e-mail and attachments is prohibited. If you have received
this in error, please advise the sender by reply e-mail and delete the message
immediately. You may also contact the LMH Process Improvement Center at
740-348-4641. E-mail transmissions cannot be guaranteed to be secure or
error-free as information could be intercepted, corrupted, lost, destroyed,
arrive late or incomplete, or contain viruses. The sender therefore does not
accept liability for any errors or omissions in the contents of this message,
which arise as a result of e-mail transmission. Thank you.
------------------------------------------------------------
CONFIDENTIALITY NOTICE:
The information in this e-mail may be confidential and/or
privileged. If you are not the intended recipient or an
authorized representative of the intended recipient, you
are hereby notified that any review, dissemination, or
copying of this e-mail and its attachments, if any, or
the information contained herein is prohibited. If you
have received this e-mail in error, please immediately
notify the sender by return e-mail and delete this e-mail
from your computer system. Thank you.
============================================================
_______________________________________________
Histonet mailing list
[email protected]
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
