RE: [Histonet] Re: H&E staining of frozen section

From:"Gladney, Diane C Ms MACH"


I do H&E staining on frozen sections using sections that are 6 - 8 microns
thick. We pick the sections up with a room temp. slide which is then placed
in a coplin jar containing 95% Ethanol (ethyl alcohol). This technique
requires some practice so I suggest practicing with a block of OCT compound
without any tissue to perfect your technique before you attempt this with
any tissue.

 The following staining protocol is used to stain the slides and they turn
out beautiful (I have used this protocol for more years than I care to think

1.  95% Ethanol (brought from Cryostat) - Dip until clear
2.  Running tap water (gentle stream of water running into a bowl or
appropriate container) - dip until clear
3.  Distilled water - dip until clear
4.  Hematoxylin 2 (Richard Allen Scientific) - 30 seconds
5.  Running tap water - until clear
6.  Bluing Reagent (Richard Allen Scientific) - dip 3 - 4 times
7.  Running tap water - 30 seconds
8.  95% Ethanol - 3 - 4 dips
9.  Eosin - Y (Richard Allen Scientific) - 1 - 2 dips
10.  95% Ethanol - 2 changes - 5 - 10 dips
11.  100% Ethanol - 3 changes - 5 - 10 dips
12.  Xylene - 2 changes - 5 - 10 dips
13.  Mount with resinous medium (coverslip)

I have a staining series set-up for doing frozen sections. The tissues are
already mounted on the slide before you do the staining series, therefore
you just need to coverslip when you finish staining the slides. I hope that
this has helped you and meets your needs.

Happy Cutting,

Diane C. Gladney, HT (ASCP)
Histology /Cytology Supervisor
Moncrief Army Community Hospital
P.O. BOX 484
4500 Stuart Ave.
FT. Jackson, SC 29207

(803) 751-2530
DSN 734-2530


-----Original Message-----
[]On Behalf Of Lori
Sent: Monday, April 26, 2004 9:18 PM
To: ''
Subject: [Histonet] Re: H&E staining of frozen section 

Does anyone have a protocol for H&E staining of frozen sections?  I am
particularly interested in breast, but any tissue type would be useful.  The
specimens were prepared in a cryomold using OCT.    



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