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Caution: Avoid Contact and Inhalation

This document provides instructions for staining mucin using Southgate's mucicarmine stain. The stain uses a solution of carmine, aluminum hydroxide, and aluminum chloride to bind to the acid substrates in mucins, staining them a deep rose color. The procedure involves deparaffinizing tissue sections, staining with Mayer's hematoxylin and the mucicarmine solution, then differentially staining with metanil yellow before mounting. The stain specifically identifies mucin deposits in tissue samples and can determine the origin of tumor cells. Safety precautions include working in a hood and wearing proper PPE when preparing and working with the acidic and flammable reagents.

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75 views5 pages

Caution: Avoid Contact and Inhalation

This document provides instructions for staining mucin using Southgate's mucicarmine stain. The stain uses a solution of carmine, aluminum hydroxide, and aluminum chloride to bind to the acid substrates in mucins, staining them a deep rose color. The procedure involves deparaffinizing tissue sections, staining with Mayer's hematoxylin and the mucicarmine solution, then differentially staining with metanil yellow before mounting. The stain specifically identifies mucin deposits in tissue samples and can determine the origin of tumor cells. Safety precautions include working in a hood and wearing proper PPE when preparing and working with the acidic and flammable reagents.

Uploaded by

AneemA
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
You are on page 1/ 5

SURGICAL PATHOLOGY - HISTOLOGY Date:

STAINING MANUAL - CARBOHYDRATES Page: 1 of 2


MUCICARMINE STAIN - SOUTHGATE’S - MUCIN
PURPOSE: To stain mucin which is a secretion produced by a variety of
epithelial cells and connective tissue cells. Excess mucin is secreted by
epithelial cells in certain inflammations and in certain intestinal
carcinomas. The mucicarmine technique is also useful in determining the
site of a primary tumor in that finding mucin positive tumor cells in an
area that does not contain mucin producing cells would indicate the tumor
did not arise from that area. It is also useful in staining encapsulated
fungi; cryptococcus.

PRINCIPLE: aluminum is believed to form a chelation complex with the


carmine, changing the molecule to a positive charge allowing it to bind
with the acid substrates of low density such as mucins.

CONTROL: Small intestine

FIXATIVE: 10% buffered formalin.

TECHNIQUE: Cut paraffin sections 4-5 m.

EQUIPMENT: Rinse glassware in DI water. Stirring hot plate, magnetic


stir bars, 500 ml beaker, coplin jars, microwave oven.

REAGENTS:
Southgate's Mucicarmine Mayer's Hematoxylin:
Solution: See Congo Red Procedure
Carmine, alum lake 1.0 gm
Metanil Yellow Solution:
Aluminum hydroxide 1.0 gm
Metanil yellow 0.25 gm
50% alcohol 100.0 ml
Distilled water 100.0 ml
Mix well, add: Glacial acetic acid 0.25 ml

Aluminum chloride, Mix well, label and date, good for 1


anhydrous 0.5 gm year.

Boil gently for 21/ 2 minutes. Cool, Caution: avoid contact and inhalation.
filter, refrigerate. Good for 6
months.
CAUTION: FLAMMABLE, avoid contact
and inhalation.
CARBOHYDRATES
MUCICARMINE Page: 2 of 2
SAFETY: Use the hood when preparing stains and microwaving solutions.
Wear gloves and lab coat.

Add aluminum chloride slowly, it is water reactive. Avoid breathing dust.

Metanil Yellow; target organ effects on reproductive system after chronic


feeding studies in male rats. Avoid contact and inhalation of dyes.

PROCEDURE:
1. Deparaffinize and hydrate to distilled water.
2. Mayer's hematoxylin for 10 minutes.
3. Wash in running tap water, 5 minutes.
4. *Mucicarmine solution, microwave HI power, 45 seconds.
5. Rinse quickly in distilled water.
6. Metanil yellow, 30 seconds to 1 minute.
7. Dehydrate quickly in three changes of absolute alcohol, clear and
coverslip in Permount.
*Conventional Method: Mucicarmine solution at room temperature for
1 hour.

RESULTS:
Mucin deep rose
Nuclei black
Other tissue elements yellow

REFERENCES:
Bancroft J, Stevens A, Theory and Practice of Histological Techniques, 2nd
Ed, 1982, pp201-202, Churchill-Livingstone, NY
Carson F, Histotechnology A Self-Instructional Text, 1990, 1st Ed, pp124-
125, ASCP, Ill
Crookham,J, Dapson,R, Hazardous Chemic ls in the Histopathology
Laboratory, 2nd ED, 1991, Anatech

Prepared: By:

Approved: By:
Downloaded from WebPath: Internet Pathology Laboratory
http://www-medlib.med.utah.edu/WebPath/webpath.html
PROCEDURE CARD
MUCIN - SOUTHGATE'S MUCICARMINE STAIN
CONTROL: Small intestine

PROCEDURE:
1. Deparaffinize and hydrate to distilled water.
2. Mayer's hematoxylin for 10 minutes.
3. Wash in running tap water, 5 minutes.
4. *Mucicarmine solution, microwave HI power, 45 seconds.
5. Rinse quickly in distilled water.
6. Metanil yellow, 30 seconds to 1 minute.
7. Dehydrate quickly in three changes of absolute alcohol, clear and coverslip in Premount.
*Conventional Method: Mucicarmine solution at room temperature for 1 hour.

RESULTS:
Mucin deep rose
Nuclei black
Other tissue elements yellow

SAFETY/PPE: Use the hood when preparing mucicarmine stain and microwaving solutions.
Wear gloves and lab coat. All aluminum chloride slowly; water reactive.

Southgate's Mucicarmine Solution: Metanil Yellow Solution:


Carmine, alum lake 1.0 gm Metanil yellow 0.25 gm
Aluminum hydroxide 1.0 gm Distilled water 100.0 ml
50% alcohol 100.0 ml Glacial acetic acid 0.25 ml
Mix well, add: Mix well, label and date, good for 6 months.

Aluminum chloride, anhydrous 0.5 gm


Mayer's Hematoxylin:
Boil gently for 21/2 minutes. Cool, filter and See Congo Red
store in the refrigerator. Label with date and
initals, good for 6 months.

CAUTION: Flammable, corrosive.


SOUTHGATE'S MUCICARMINE: METANIL YELLOW SOLUTION:
Carmine, alum lake 1.0 gm Metanil yellow 0.25 gm
Aluminum hydroxide 1.0 gm Distilled water 100.0 ml
50% alcohol 100.0 ml Glacial acetic acid 0.25 ml

Mix well, add: Mix well, label and date, good for
6 months.
Aluminum chloride, anhydrous
0.5 gm
Caution: avoid contact and inhalation.
Boil gently for 21/2 minutes.
Cool, filter and store in the DATE:
refrigerator. Label with date and
initals, good for 6 months. TECH:
CAUTION: Flammable, corrosive.
Aluminum chloride is water EXPIRATION:
reactive.

SAFETY/PPE: Wear gloves, METANIL YELLOW


goggels and lab coat. Work under
hood. Add aluminum chloride DATE:
slowly.
TECH:
DATE:

TECH:

EXPIRATION:

MUCICARMINE STAIN

DATE:

TECH:
Mayer's Hematoxylin: MAYER'S HEMATOXYLIN
Hematoxylin 1.0 gm
DATE:
Distilled water 1000.0 ml
Sodium iodate 0.2 gm TECH:
Ammonium or Potassium
aluminum sulfate 50.0 gm
Citric acid 1.0 gm
Chloral hydrate 50.0 gm

Dissolve the hematoxylin in


water, heat to boiling, boil for 5
minutes. Remove from heat, add
sodium iodate, wait 10 minutes to
ripen. Add the remainder of the
chemicals in the order given,
dissolving each one before adding
the next. Label, initial, stable for
1 year.

SAFETY/PPE: Wear gloves,


goggles and lab coat. Work in well
ventilated area or under hood.

DATE:

TECH:

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