RE: Microwave mucin stain

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From:RGreen@SARCOM.COM (by way of Marvin Hanna)
To:histonet@histosearch.com
Reply-To:
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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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