[Histonet] restarting DAB reaction

[Tony Henwood]

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:detmar <@t> lunenfeld.ca] 
	Sent: Thursday, 16 October 2008 12:52 AM
	To: Tony Henwood; jmyers1 <@t> aol.com;
histonet <@t> 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:   detmar <@t> lunenfeld.ca
	 

________________________________

	From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of
Tony Henwood
	Sent: Tue 10/14/2008 11:36 PM
	To: jmyers1 <@t> aol.com; histonet <@t> 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: histonet-bounces <@t> lists.utsouthwestern.edu
	[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
	jmyers1 <@t> aol.com
	Sent: Wednesday, 15 October 2008 12:58 PM
	To: histonet <@t> 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" <talulahgosh <@t> gmail.com>
	Subject: [Histonet] restarting DAB reaction
	To: histonet <@t> 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
	
	_______________________________________________
	Histonet mailing list
	Histonet <@t> lists.utsouthwestern.edu
	http://lists.utsouthwestern.edu/mailman/listinfo/histonet
	
	
*********************************************************************
	This email and any files transmitted with it are confidential
and intended solely for the use of the individual or entity to whom they
are addressed. If you are not the intended recipient, please delete it
and notify the sender.
	
	Views expressed in this message and any attachments are those of
the individual sender, and are not necessarily the views of The
Children's Hospital at Westmead
	
	This note also confirms that this email message has been
	virus scanned and although no computer viruses were detected,
The Childrens Hospital at Westmead accepts no liability for any
consequential damage resulting from email containing computer viruses.
	
**********************************************************************
	
	
	_______________________________________________
	Histonet mailing list
	Histonet <@t> lists.utsouthwestern.edu
	http://lists.utsouthwestern.edu/mailman/listinfo/histonet
	


*********************************************************************
This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender.

Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead

This note also confirms that this email message has been
virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses.
**********************************************************************