If we don't give it a go, how would we know?
Now I know!!
Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA
-----Original Message-----
From: Jacqui Detmar [mailto:[EMAIL PROTECTED]
Sent: Thursday, 16 October 2008 12:52 AM
To: Tony Henwood; [EMAIL PROTECTED];
histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] restarting DAB reaction
Hey there. Actually, I recently tried to re-do a DAB reaction
on two slides (mouse placental sections) that I had cover-slipped about
a year ago, thinking it would work. Nothing happened. Even the RBCs
just sat there and laughed at my clumsy attempt. I slunk away in shame.
Jacqui
Jacqui Detmar, Post-doctoral Fellow
Samuel Lunenfeld Research Institute,
Mount Sinai Hospital,
25 Orde Street, room 6-1001 AJ
Toronto, ON, Canada
M5T 3H7
Tel: 416-586-4800 x5607
Fax: 416-586-8588
email: [EMAIL PROTECTED]
________________________________
From: [EMAIL PROTECTED] on behalf of
Tony Henwood
Sent: Tue 10/14/2008 11:36 PM
To: [EMAIL PROTECTED]; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] restarting DAB reaction
Yes,
I would also agree that this would probably be the case.
I have not tried to redo the DAB step after counterstaining,
dehydrating, clearing and mounting.
I would expect it not to work.
Though I could easily be corrected.
Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA
-----Original Message-----
From: [EMAIL PROTECTED]
[mailto:[EMAIL PROTECTED] On Behalf Of
[EMAIL PROTECTED]
Sent: Wednesday, 15 October 2008 12:58 PM
To: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] restarting DAB reaction
Emily:
Although it might seem that the reapplication of a
substrate-chromogen
solution?after removal of the coverslip/mounting media and
rehydration?should work, it?very rarely does.? The scientific
reason is
that, after the specimen has been dehydrated and
coverslipped,?the
enzyme label (e.g. polymer-HRP) applied in the original staining
procedure is 'dead' -- meaning that it lacks the?power to drive
another
chromogen-polymerization reaction.? Your best bet is start over,
'from
scratch'... Good Luck,
J.D.?Myers, M.S., CT(ASCP)
***************************************
Message: 1
Date: Mon, 13 Oct 2008 13:17:49 -0400
From: "Emily Sours" <[EMAIL PROTECTED]>
Subject: [Histonet] restarting DAB reaction
To: histonet@lists.utsouthwestern.edu
Has anyone ever tried to continue a DAB reaction after the
slides have
been washed and coverslipped? I can't think of a reason why this
wouldn't work, since we just wash the slides in PBS and water
after the
DAB step. We use a vectastain kit for this--would we need to add
more of
the ABC reagent (after removing the coverslips and washing in
PBS) and
then do DAB?
Emily
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