[Histonet] restarting DAB reaction

Tony Henwood AnthonyH <@t> chw.edu.au
Wed Oct 15 16:52:38 CDT 2008


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
	
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