[Histonet] PTAH Procedure

Tony Henwood AnthonyH <@t> chw.edu.au
Mon Apr 26 19:24:26 CDT 2004


Ruth, we use the Cherukian Method:

Regards

Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318

http://www.histosearch.com/homepages/TonyHenwood/default.html
http://us.geocities.com/tonyhenwoodau/index.html

Document Procedure:	
Phosphotungstic Acid Haematoxylin
Principle:	
Cherukian's modification employs an eosin solution that stains the
erythrocytes red and differentiates them from the blue fibrin.

Fixation:      10% buffered formalin.
Microtomy:     paraffin sections at 5um.

Controls:	
Use brain sections and section of muscle.  A good stain will demonstrate the
dendrites as blue where as in a bad stain they appear light grey to salmon
in colour.  Nuclei, fibrin, platelets and muscle will be blue, red cells and
collagen appear red. Muscle striations should be well defined.

Reagents: 

1.   Eosin:   - Warning: Flammable - see MSDS
           Eosin Y, water soluble (CI 145380)       0.5g
           Distilled Water                         	  10ml
           80% Ethanol                             	190ml

           Working:   10ml stock and
                   Before use add 50ul glacial acetic acid.

2.   1% Periodic Acid

3.   PTAH solution
            Haematoxylin (CI 75290)       	   0.5g
            Phosphotungstic Acid           		    10g
            Distilled water               		500ml

Dissolve solid ingredients in separate portions of the water.  Use gentle
heat for Haematoxylin.  Combine solutions when cool.  Add 0.088g potassium
permanganate to ripen.  The stain is ready to use.

 

Procedure:

1.	Dewax and hydrate sections to 80% alcohol.
2.	Place slides in eosin for 30 seconds.
3.	Wash slides in distilled water for a few seconds.
4.	Place slides in 1% periodic acid for 20 minutes.
5.	Wash slides in water for 3 minutes.
6.	Place slides in PTAH for 30-90 minutes in 60oC oven.  Check from 30
minutes on.
7.	Dehydrate, clear and mount.



Results: 
Dendrites, nuclei, fibrin, platelets and muscle - 	blue
Red blood cells and collagen - 			red.


Notes: 

Reference:	 

	1.	Cherukian, C.J., Histologic. 8(4); 105, (1977).
	2.	Luna, L., Histologic. 5(2); 66, (1975).

-----Original Message-----
From: Cazares, Ruth [mailto:RCazares <@t> schosp.org] 
Sent: Saturday, 24 April 2004 5:31 AM
To: histonet <@t> pathology.swmed.edu
Subject: [Histonet] PTAH Procedure


Hello again Histonetters,

 

I was wondering if anyone out there knows if there is a procedure for PTAH
that does not require the tissue to be treated in Zenker's?  Any and all
information will be greatly appreciated.  Thanks.

 

Ruth Cazares

 




***  Confidentiality Statement ***
This e-mail is intended only for the use of the individual or entity to
which it is addressed and may contain information that is privileged and
confidential. If the reader of this message is not the intended recipient,
please notify the sender immediately by replying to this message and then
delete it from your system. Any review, dissemination, distribution, or
reproduction of this message by unintended recipients is strictly prohibited
and may be subject to legal restriction.
Thank you for your cooperation.
*************************



_______________________________________________
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 footnote 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.
**********************************************************************





More information about the Histonet mailing list