[Histonet] 20% NaOH Nail pre-Treatment... Questions

[Due, Brice]

Hello Rene, thank you very much for the paper you sent me! I will read it
tomorrow.
   
> 20% NaOH is too strong.
 
Actually, floating 4um nail sections on 20% NaOH does work. Very well. We just
validated it today and the attending/staff derm path people are very happy with
the results. We will continue to "float" difficult nails until we implement and
validate a pre-soak protocol in the grossing lab. The key to making foatation
work well is to re-float the sections on a regular water bath for several
minutes to rinse out the NaOH. Residual NaOH will prevent adhesion to even the
stickiest slides.
 
My guess is the keratin network is crosslinked in many ways, and the NaOH
treatment is degrading only some of them. What I have observed with 4um sections
is they flatten out and "spread" while floating on NaOH -- very similar to how
nail clippings swell up when soaked in 20% NaOH. Of course this can go too far
and leave you with mush, but 4um sections can certainly withstand 20% NaOH (warm
~40C, not RT) for at least 15min. The derm pathologist said the morphology
looked good.
 
There's always more than one way... :)
 
Thanks again for the paper!
-brice

________________________________

From: Rene J Buesa [mailto:rjbuesa <@t> yahoo.com] 
Sent: Wednesday, January 27, 2010 3:49 PM
To: Joe The Toe Nocito; Due, Brice
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] 20% NaOH Nail pre-Treatment... Questions


20% NaOH is too strong.
Under separate cover I am sending another procedure.
René J.

--- On Tue, 1/26/10, Due, Brice <BDUE <@t> PARTNERS.ORG> wrote:



	From: Due, Brice <BDUE <@t> PARTNERS.ORG>
	Subject: [Histonet] 20% NaOH Nail pre-Treatment... Questions
	To: "Joe "The Toe" Nocito" <jnocito <@t> satx.rr.com>
	Cc: histonet <@t> lists.utsouthwestern.edu
	Date: Tuesday, January 26, 2010, 6:05 PM
	
	
	Hello Joe, first thanks so much for sharing your expertise here on
Histonet.
	
	I work in the main histo lab at Mass General Hosp. in Boston and I have
	interested the powers that be in your NaOH protocol. I initially used
your NaOH
	idea to attach an "impossible" nail to slides by floating 4um section on
20%
	NaOH for 10-15min. (The transfers between regular water bath and NaOH
bath (and
	back to rinse) were surprisingly easy because the concentrated NaOH
prevents
	adhesion to even the stickiest adhesive slides.)
	
	Anyhow, I have some questions for you about your lab's implementation. I
found
	your most complete post at
	
http://lists.utsouthwestern.edu/pipermail/histonet/2008-October/040078.html
	
	But I am concerned about the safety of 20% NaOH in routine use. Would
you mind
	posting or emailing me any SOPs you have that address the safety issues?

	
	Specifically both the histology lab and the "grossing" labs here use RDO
decal
	solutions at every bench. So there is/will be concern about the proper
handling
	of 20% NaOH around RDO... by overworked PAs and residents, etc. etc.
Feel free
	to tell me your safety almost-horror stories so we can (try to) prevent
them
	from occuring here.
	
	I've seen that some people use 10% NaOH instead. Have you experimented
with
	this? Any reasons you stick with 20% beyond the usual "it ain't broke,
so..."?
	Have you (or anyone on Histonet) ever tried to find the minimum
effective
	concentration? Increasing the pre-soak time isn't an issue here since
I've been
	assured that nail turnaround times are non-critical.
	
	Along these lines, do you or anyone have any histochemical references
for the
	reactions that are taking place? With this info it might be possible to
predict
	the minimum effective pH. It would also be nice to have references for
the SOP.
	(I know about the coverslip "crushes" some derm clinicians do with fresh
	scrapings. Alternately, do you know of any histochemical references for
this?)
	
	Thank you so much for sharing your time and info! And if I happen to
answer any
	of my own questions I will be sure to post what I learn.
	
	Sincerely,
	-brice
	Massachusetts General Hospital 
	Surgical Pathology, Histology
	
	
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