Hi Catherine,
I have staining protocol for alcian blue and nuclear fast red, see bellow. I
looked in Conn's biological stains book (9th edition,p.188)and saw that
chlorantine
fast red 5B is soluble in water. I think you can substitute Nuclear fast red
with chlorantine
fast red 5B and try and see the results. Connective tissue will be deeper red.
Use controls and try it.
Iliana Dimitrova, MSc, RT
Histology Supervisor
Medical Education and Laboratory Support Services (MELSS)
Faculty of Medicine
Memorial University of Newfoundland
St. John's, NL Canada
Alcian Blue Staining Protocol
NovaUltra Special Stain Kits
Description: Alcian blue stains acid mucosubstances and acetic mucins.
Excessive amounts of non-sulfated acidic mucosubstances are seen in
mesotheliomas, certain amounts occur normally in blood vessel walls but
increase in early lesions of atherosclerosis. Strongly acidic mucosubstances
will be stained blue, nuclei will be stained pink to red, and cytoplasm will be
stained pale pink.
Fixation: formalin fixed, paraffin embedded tissue sections.
Solutions and Reagents:
3% Acetic Acid Solution:
Glacial acetic acid ----------------- 3 ml
Distilled water --------------------- 97 ml
Alcian Blue Solution (pH 2.5):
Alcian blue, 8GX -------------------- 1 g
Acetic acid, 3% solution ----------- 100 ml
Mix well and adjust pH to 2.5 using acetic acid.
0.1% Nuclear Fast Red Solution:
Nuclear fast red ------------------- 0.1 g
Aluminum sulfate------------------ 5 g
Distilled water ---------------------100 ml
Dissolve aluminum sulfate in water. Add nuclear fast red and slowly heat
to boil and cool.
Filter and add a grain of thymol as a preservative.
Procedure:
1. Deparaffinize slides and hydrate to distilled water.
2. Stain in alcian blue solution for 30 minutes.
3. Wash in running tap water for 2 minutes.
4. Rinse in distilled water.
5. Counterstain in nuclear fast red solution for 5 minutes.
6. Wash in running tap water for 1 minute.
7. Dehydrate and through 95% alcohol, 2 changes of absolute alcohol, 3
minutes each.
8. Clear in xylene or xylene substitute.
9. Mount with resinous mounting medium.
Results:
Strongly acidic sulfated mucosubstances -------------- blue
Nuclei ------------------------------------------------------ pink to red
Cytoplasm ------------------------------------------------- pale pink
Positive Controls:
Small intestine, appendix, or colon.
-----Original Message-----
From: [email protected]
[mailto:[email protected]] On Behalf Of Ross, Catherine
Sent: December 13, 2011 5:45 AM
To: [email protected]
Cc: Grainger, Shirley
Subject: [Histonet] Stain recipe
All,
Greetings from across the pond!
One of our histology technicians is looking for a recipe for an alcian
blue/sirius red stain, otherwise known as "Alcian blue 8G with chlorantine
fast red 5B". It is not one I have used but I suggested tapping into the
wealth of knowledge on this forum - can anyone help, please?
Thank you in advance,
Catherine
Dr. Catherine L. Ross DVM MSc(VetPath) MRCVS
Covance Laboratories (UK) Ltd.
Otley Road,
Harrogate, N. Yorks.
HG3 1PY
T: +44 (0) 1423 848759
E: [email protected] <mailto:[email protected]>
-----------------------------------------------------
Confidentiality Notice: This e-mail transmission
may contain confidential or legally privileged
information that is intended only for the individual
or entity named in the e-mail address. If you are not
the intended recipient, you are hereby notified that
any disclosure, copying, distribution, or reliance
upon the contents of this e-mail is strictly prohibited.
If you have received this e-mail transmission in error,
please reply to the sender, so that we can arrange
for proper delivery, and then please delete the message
from your inbox. Thank you.
_______________________________________________
Histonet mailing list
[email protected]
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
This electronic communication is governed by the terms and conditions at
http://www.mun.ca/cc/policies/electronic_communications_disclaimer_2011.php
_______________________________________________
Histonet mailing list
[email protected]
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
