Double Staining Epoxy Sections

4/18/97

Dear List

Is there a reference or does someone have experience with double staining of

Epoxy semi-thin sections. I would like to use Basic Fuchsin for red and

Toluidine Blue for the blue but I haven't been able to find out any

information. Anyone willing to share first hand experience would be

appreciated. By the way I am looking for areas with Epstein-Barr viral

inclusions in the nucleus.

TIA

Jack Megill

BMS

Princeton, NJ

Megill_John_R.PRILVMS3@msmail.bms.com

References as follows:

Aparicio, SR, and Marsden, P: Methylene blue-basic fuchsin. Journal of

Microsc. (Eng.) 89:139-141, 1969.

Huber, JD, Parker, F, and Odland, GF; Basic fuchsin and alkalinized

methylene blue. Stain Technol. 43:83-87, 1968.

Humphrey, CD, and Pittman, Fe: Methylene blue-azure II and basic fuchsin.

Stain Technol 42:9-14, 1974.











Nancy A. Monteiro-Riviere, Ph.D.,BCFE,BCFM

Professor of Investigative Dermatology/Toxicology

North Carolina State University

College of Veterinary Medicine

Cutaneous Pharmacology and Toxicology Center

4700 Hillsborough Street

Raleigh, NC 27606

Telephone: 919-829-4426

FAX: 919-829-4358

email: Nancy_Monteiro@ncsu.edu

CTPC Homepage: http://cptc.ncsu.edu

This is a protocol we have used that gives wonderful results:

Polychrome stain

1. Blue Stain: Methylene blue .13gm , Azure II .02gm, Glycerol

10ml, Methyl alcohol 10ml, D.H2O 80ml

(stir and filter, keeps 6 mo.)

2. Red stain: STOCK SOL: Basic fuchsin .2gm, DH2O 100ml (stir and

filter, keeps 6mo.)

WORKING SOL: dilute stock 1:4 in DH2O (fresh daily)

3. Sodium Hydroxide 1% fresh daily

Procedure:

1. Flood slide with blue stain 15-60 seconds, depending on

temperature and material.

2. Add 4-6 drops NaOH to the stain and mix by tilting the slide,

about 10 seconds total time.

3. Wash in running water and dry on hotplate. Blue stain can be

destained by heating.

4. Add red stain for 15-30 seconds on hotplate. (Red stain cannot

be destained)

5. Rinse with running water and dry.

References: Mackay and Mead. 1970. 28th EMSA meetings pp.296-297.

Modified by Griffin and Fahrenbach, Oregon Regional Primate Research

Center

Hope this is of some help

Linda M. Fox

Dept. of CBN and Anatomy

Loyola University Medical School

2160 S. First Ave.

Maywood, Illinois 60153

lfox1@wpo.it.luc.edu

I have copies of the LKB handbook, given to me by LKB in

1977(April/August)

Title: Stains for Plastic Embedded Tissue Sections.

1. comparasion of three different methylene blue-basic fuchsin

stains.(Aparicio,Huber,Jha.) August 1977

2. Stainning of sections from different animal, human, and plant tissues

with a methylene blue-azure ll-basic fuchsin stain.(Humphrey) April

1977.

Should you require a copy, please contact me so that I can send you a

photocopy.

All the best

Vijay H Bandu

bandu@emu.unp.ac.za

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