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A modification of the mallory connective tissue stain as a stain for keratin
ORAL PATHOLOGY General Section .
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A MODIFICATION AS A STAIN Patricia Department
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.
OF TEE
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MALLORY
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.
.
CONNECTIVE
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TISSUE
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.
.
STAIN
FOR KERATIN
Ryoub of
.
a,nd Gerald Shklar,
Oral Pathology,
Tufts
Boston, ~llass.
L'nixcrsity
School of Dmtal
Medicine
a series of investigations over the past year concerning the histochemistry of oral carcinoma and the oral lesions of dermatologic disease it was felt that a differential staining technique for keratin and prekeratin would be of considerable value. A modification of the Mallory connective tissue The staining stain was found to be very helpful in meeting this requirement technique is as follows: Fixation: Formalin-fixed tissue is preferred. Technique: Cut paraffin sections at 6 microns.
D
URING
Solutions:
f uchsin solutio?l 5.0 (:nl. Acid fuchsin ---_----__--------Distilled water __----_---------100.0 C.C. Aniline blue-Orange G solution 0.5 Chl. Aniline blue, water-soluble _---_2.0 Gm. Orange G -----__------------___ 1.0 Gm. Phosphotungstic acid -- ______--_Distilled water ------__--__---- 100.0 cc. Staining procedure: 1. Deparaffinize sections through 2 changes of xylem, absolute alcohol, and 95 per cent alcohol, to distilled water as usual. 2. Stain sections in acid fuchsin solution for 3 minutes. 3. Transfer directly to aniline blue-orange G solution for 45 minutes. 4. Transfer directly to 95 per cent alcohol for several changes. 5. Dehydrate with 2 changes of absolute alcohol, clear with I! to 3 changes of xylene, and mount in balsam. For consistent. results, use fresh solutions.
Acid
MODIFICATION
Volume 16 Number 5
OF MALLORY
CONNECTIVE
TISSUE
STAIN
581
The keratin stains a brilliant red with this technique. The stratified squamous epithelium generally stains gray. Occasionally the upper areas of the stratum spinosum stain orange, and this appears to illustrate the formation of a prekeratin-like substance. The connective tissue stains the usual deep blue. Erythrocytes in small blood vessels stain deep red.
Fig.
l.-Medium-power intense stain
photomicrograph of epidermoid of the keratin as distinct from
carcinoma of buccal mucosa. the remainder of the epithelium.
Note
the
We have been using this stain routinely for the past year in all cases of oral tumors, hyperkeratotic lesions, and oral manifestations of dermatologic disease.l The stain is very helpful in its emphasis of keratin and prekeratin-like substance and is a useful auxiliary to the hematoxylin and eosin stain. This stain may prove helpful in the diagnosis of epidermoid carcinoma where the keratin formed is small in amount and abnormal. We are currently attempting to evaluate the significance of this stain and to clarify the chemical reactions involved. REFEREKCE
1. Shklar, G., and Meyer, I.: Lesions
in the
Mouth,
The Histopathology
and Histochemistry
of Dermatologic
ORAL SURG., ORAL MED. & ORAL PATH. 14: 1069, 1961.
.
.
.
.
.
A MODIFICATION AS A STAIN Patricia Department
.
.
OF TEE
.
.
.
MALLORY
.
.
.
.
.
CONNECTIVE
.
.
.
TISSUE
.
.
.
.
STAIN
FOR KERATIN
Ryoub of
.
a,nd Gerald Shklar,
Oral Pathology,
Tufts
Boston, ~llass.
L'nixcrsity
School of Dmtal
Medicine
a series of investigations over the past year concerning the histochemistry of oral carcinoma and the oral lesions of dermatologic disease it was felt that a differential staining technique for keratin and prekeratin would be of considerable value. A modification of the Mallory connective tissue The staining stain was found to be very helpful in meeting this requirement technique is as follows: Fixation: Formalin-fixed tissue is preferred. Technique: Cut paraffin sections at 6 microns.
D
URING
Solutions:
f uchsin solutio?l 5.0 (:nl. Acid fuchsin ---_----__--------Distilled water __----_---------100.0 C.C. Aniline blue-Orange G solution 0.5 Chl. Aniline blue, water-soluble _---_2.0 Gm. Orange G -----__------------___ 1.0 Gm. Phosphotungstic acid -- ______--_Distilled water ------__--__---- 100.0 cc. Staining procedure: 1. Deparaffinize sections through 2 changes of xylem, absolute alcohol, and 95 per cent alcohol, to distilled water as usual. 2. Stain sections in acid fuchsin solution for 3 minutes. 3. Transfer directly to aniline blue-orange G solution for 45 minutes. 4. Transfer directly to 95 per cent alcohol for several changes. 5. Dehydrate with 2 changes of absolute alcohol, clear with I! to 3 changes of xylene, and mount in balsam. For consistent. results, use fresh solutions.
Acid
MODIFICATION
Volume 16 Number 5
OF MALLORY
CONNECTIVE
TISSUE
STAIN
581
The keratin stains a brilliant red with this technique. The stratified squamous epithelium generally stains gray. Occasionally the upper areas of the stratum spinosum stain orange, and this appears to illustrate the formation of a prekeratin-like substance. The connective tissue stains the usual deep blue. Erythrocytes in small blood vessels stain deep red.
Fig.
l.-Medium-power intense stain
photomicrograph of epidermoid of the keratin as distinct from
carcinoma of buccal mucosa. the remainder of the epithelium.
Note
the
We have been using this stain routinely for the past year in all cases of oral tumors, hyperkeratotic lesions, and oral manifestations of dermatologic disease.l The stain is very helpful in its emphasis of keratin and prekeratin-like substance and is a useful auxiliary to the hematoxylin and eosin stain. This stain may prove helpful in the diagnosis of epidermoid carcinoma where the keratin formed is small in amount and abnormal. We are currently attempting to evaluate the significance of this stain and to clarify the chemical reactions involved. REFEREKCE
1. Shklar, G., and Meyer, I.: Lesions
in the
Mouth,
The Histopathology
and Histochemistry
of Dermatologic
ORAL SURG., ORAL MED. & ORAL PATH. 14: 1069, 1961.