RE: Microwave mucin stain
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From: | RGreen@SARCOM.COM (by way of Marvin Hanna) |
To: | histonet@histosearch.com |
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Here is a great microwave procedure...
MUCIN-MUCICARMINE (Southgate's Method)
FIXATIVE
Any well-fixed tissue will stain with this method.
SPECIMEN TYPE
Cut paraffin sections at 3-4 microns.
EQUIPMENT AND SUPPLIES
Equipment Vendor Manufacturer
Cat. No.
Coplin jars, plastic Allegiance Bel-Art
S7666
Graduated cylinders Allegiance Corning
3662-0050
Pipettes Allegiance
P4650-15
Microwave oven (700 W) Grant Campus General Electric
JE1445G
Microwave oven (700W) Riverside Campus Quasar
MQS0841E
Supplies Vendor Manufacturer
Cat. No.
Hydrochloric Acid Allegiance Mallinckrodt
2611-2.5(NY)
Mucicarmine
Stock Solution Sigma
HT30-1-8
Tartrazine, 0.25% Sigma
T0388
Hematoxylin C.I.75290 Allegiance J.T. Baker
M906-03(BC)
Ferric Chloride Allegiance Mallinckrodt
5029-500(NY)
95% Alcohol Pharmco Pharmco
1200040
CONTROLS
A mucin positive section of small intestine.
REAGENT PREPARATION (Time Required: 1-2 minutes)
CAUTION: AVOID EYE AND SKIN CONTACT
WEAR GLOVES AND PROTECTIVE CLOTHING
DO NOT INHALE OR INGEST
WEAR GOGGLES OR SAFETY GLASSES
A. 29% Ferric Chloride Solution
CAUTION: CORROSIVE, IRRITANT
Combine the following:
Distilled Water 100 ml
Ferric Chloride 29 gram
B. Weigert's Iron Hematoxylin Solution A
CAUTION: TOXIC, FLAMMABLE
Combine the following:
95% Alcohol 400 ml
Hematoxylin 4.0 g
C. Weigert's Iron Hematoxylin Solution B
CAUTION: CORROSIVE, IRRITANT
Combine the following:
Distilled Water 380 ml
29% Ferric Chloride 16 ml
Hydrochloric Acid 4 ml
D. Weigert's Iron Hematoxylin Working Solution
CAUTION: CARCINOGENIC, ACID, IRRITANT
Combine the following:
Weigert's Iron Hematoxylin Solution A 25 ml
Weigert's Iron Hematoxylin Solution B 25 ml
E. Mucicarmine Stock Solution
CAUTION: IRRITANT, FLAMMABLE
F. Working Mucicarmine Stain
CAUTION: IRRITANT, FLAMMABLE
Combine the following:
Tap Water 35 mls
Mucicarmine Stock Solution 5 mls
G. 0.25% Tartrazine
CAUTION: IRRITANT
PROCEDURE NOTES
A. Process a control slide with the sections to be stained.
B. The rose color attributable to carmine staining will be obscured if
sections are over counter-stained with tartrazine.
C. Always use plastic labware in the microwave.
PROCEDURE (Time Required: 30-45 minutes)
1. Decerate sections to distilled water.
2. Stain in Weigert's Iron Hematoxylin for 10 minutes.
3. Wash in running water for 5-7 minutes.
4. Stain in Working Mucicarmine using the microwave.
A. Place slides in mucicarmine stain and microwave on high
power for 30 seconds.
B. Remove plastic coplin jar with slides from microwave and
allow to incubate for 15 minutes or longer.
5. When staining is complete, discard solution and rinse slides in
distilled water for 5 seconds.
6. Stain with tartrazine for 5 seconds.
7. Rinse quickly in distilled water, 1 dip.
8. Dehydrate, clear, and coverslip.
RESULTS
Mucin - deep rose to red
Capsule of Cryptococcus - deep rose to red
Nuclei - black
Other tissue elements - yellow
REFERENCES
1. Biological Stain Commission, University of Rochester Medical Center,
Rochester, New York, Staining Procedures Used by the Biological Stain
Commission. The Williams and Wilkins Co., Baltimore, MD, 1060.
2. Green RM, Minturn T: Grant/Riverside Methodist Hospitals, Grant
Histology Reference Manual, 1996.
3. Kolmer JA, Spaulding EH, Robinson HW: Approved Laboratory
Technique, Appleton, Century Crofts, Inc., 1951.
4. Mallory FB: Pathological Technique, Hafner Publishing Co., 1968.
5. Theory and Practice of Histotechnology, 2nd edition,
Sheehan-Hrapchak, pages 163, 168, 169.
6. Mayer, P (1896) Uber Schleimf#228#rbung. Mitteilungen qus der
Zoologischen Station zu Neapel, 12, 203.
7. Southgate HW. (1927) Note on preparing mucicarmine, Journal of
Pathology and Bacteriology, 30, 729.
8. Green RM, Harrison L: Grant/Riverside Methodist Hospitals Histology
Special Stain Procedure Manual, Columbus, OH, 1998. Ph (614)566-9177.
9. Ross J: Riverside Methodist Hospitals Histology Procedure Manual,
1980.
Best of luck,
Rod Green
Sarcom Enterprise Education
8405 Pulsar Place
Columbus, Ohio 43240
Ph: 614.854.1450 x6611
E-mail: rgreen@sarcom.com
-----Original Message-----
From: Goodwin, Diana [mailto:DGoodwin@CHSNJ.org]
Sent: Tuesday, January 18, 2000 11:18 AM
To: 'Histonet'
Subject: Microwave mucin stain
Hello, Histonet.
Does anyone have a good, reliable microwave method for mucin?
Thanks in advance.
Diana Goodwin, HT
Trenton, NJ
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