RE: [Histonet] Destaing H&E

From:Gary Gill

After removing the cover glass, proceed as follows:

No.	Solution		Time		Note

1-2	Xylene		1 min/each	Removes traces of mountant.  Dip
repeatedly; inspect surface.  A wavy rather than a
smoothly glistening surface denotes incomplete rinsing and indicates further

3-5	Abs. EtOH		1 min/each	Prepares slides for next

6	1.0% HCl*		min-1 hr	Removes hematoxylin.
Monitor the progress of decolorization by
periodic microscopic inspection.
7	1.5% NH4OH
	in 70% ethanol**	1 min		Removes eosin.  Time
required may vary and should be adjusted as needed.  Repeated
dipping aids uniform decolorization.

8-9	Tap water		1 min/each	To H&E stain

*	0.23 mL HCl concentrated (i.e., 36.5-38% w/w, S.G. 1.1854-1.1923, or
5.5 mL N/2 HCl) in water q.s. 100 mL
**	5.7 mL NH4OH concentrated, 29.2% w/w, S.G. 0.900, in 75 mL 95%
ethanol, and water q.s. 100 mL

Gary Gill

-----Original Message-----
From: [] 
Sent: Monday, February 23, 2004 9:51 AM
Subject: [Histonet] Destaing H&E

What is the simple method of destaining H&E slides?
Most of the precious slides are too dark..

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