Special staines used in oral pathology.

1. SPECIAL STAIN
BY
DR VINESH RAJ.S
MOSc(APPLIED ORAL PATHOLOGY CANDIDATE),
DEPARTMENT OF ORAL MEDICINE & ORAL PATHOLOGY
FACULTY OF DENTISTRY,UM

2. CONTENT
1) HEMATOXYLIN & EOSIN
2) SPECIAL STAINS ON :-
3) SPECIAL STAINS USED IN ORAL PATHOLOGY
4) REFERENCES
1.CARBOHYDRATES
2.AMYLOID
3.LIPIDS
4.NUCLEIC ACID
5.PIGMENTS & MINERALS
6.MICROORGANISM
7.CONNECTIVE TISSUE

3. HEMATOXYLIN
HEMATOXYLIN
•Hematoxylin is an organic compound having the chemical formula C16H14O6. This
compound is extracted from the heartwood of the logwood tree. This is a natural dye
that has been long used as a histologic stain, ink and as a dye in the textile and leather
industry. As a dye, this material has several names such as Palo De Campeche,
longwood extract, bluewood and blackwood.
NATURE
•BASIC
•Haematoxylin can be considered as a basic dye. It is used to stain acidic structures a
purplish blue. DNA in the nucleus, and RNA in ribosomes and in the rough
endoplasmic reticulum are both acidic, and so haemotoxylin binds to them and stains
them purple.
STAINING
•BLUE/BLACK
•NUCLEUS
• Helmenstine, Anne Marie, Ph.D. (2020, August 27). What Histology Is and How It's Used.
Retrieved from https://www.thoughtco.com/histology-definition-and-introduction-4150176

4. EOSIN
• Eosin is a group of fluorescent acidic compounds having bonds with basic or eosinophilic
compounds to form salts. The salts are formed with compounds such as proteins which
contain amino acid residue including arginine, and lysine. Eosin can stain protein residues
dark red or pink colour as a result of the action of bromine on fluorescein. Additionally, we
can use this dye to stain proteins in the cytoplasm and to stain collagen and muscle fibres
for studying under the microscope.
NATURE
• ACIDIC
• Eosin is anionic and acts as an acidic dye. It is negatively charged and stains basic (or
acidophilic) structures red or pink. Most proteins in the cytoplasm are basic, and so eosin
binds to these proteins and stains them pink. This includes cytoplasmic filaments in muscle
cells, intracellular membranes, and extracellular fibres.
• PINK
• CYTOPLASM, MUSCLE CELLS,COLLAGEN
EOSIN
• Helmenstine, Anne Marie, Ph.D. (2020, August 27). What Histology Is and How It's Used.
Retrieved from https://www.thoughtco.com/histology-definition-and-introduction-4150176

5. SPECIAL
STAINS
1)IMPORTANT TOOLS FOR PATHOLOGIST TO VISUALIZ
AND IDENTIFY SPECIFIC CELLS,TISSUES AND
MICROORGANISM.
2)SPECIFIC STAINS ARE BASED ON CHEMICAL
REACTIONS THAT USES VARIOUS COLOUR DYES AND
REAGENTS.
3)THE STAINING TECHNIQUES ARE BASED ON
ACIDIC,BASIC AND BUFFERS THAT ARE SENSITIVE
AND TAKE UP SPECIAL DYES.
4)BASED ON BIOLOGICAL STAIN COMMISION THERE
ARE 64 CERTIFIED SPECIAL STAINS

6. SPECIAL
STAINS
1.CARBOHYDRATES
2.AMYLOID
3.LIPIDS
4.NUCLEIC ACID
5.PIGMENTS & MINERALS
6.MICROORGANISM
7.CONNECTIVE TISSUE

7. CARBOHYDRATE
CARBOHYDRATE
POLYSACCHARIDES
GLYCOGEN-PAS
DIASTASE DIGESTION OF
GLYCOGEN-PAS-D
NEUTRAL MUCOSUBSTANCE
FOUND IN STOMACH,INTESTINES
AND PROSTATE-PAS
ACID MUCOSUBSTANCE
EPITHELIAL-ALCIAN
BLUE/TOLUDINE BLUE
CONNECTIVE TISSUE-ALCIAN BLUE
MUCICARMINE-MUCIN
MUCOPROTIEN
GLYCOPROTEIN-PAS
MUCOLIPIDS
GLYCOLIPIDS-PAS

8. PERIODIC ACID SCHIFF
METHOD
REAGENTS 1.PERIODIC ACID
2.SCHIFF REAGENT
TECHNIQUE DEPARAFFINIZE AND HYDRATE TO DISTILLED WATER-OXIDISE IN
1% PERIODIC ACID FOR 5 MINS-RINSE-SCHIFF’S REAGENT FOR
30MINS-WASH-HARRIS HAEMATOXYLIN FOR 1 MIN-WASH-
DEHYDRATE,CLEAR AND COVERSLIP
RESULT GLYCOGEN AND MUCIN-MAGENTA
NUCLEI-BLUE
POSITIVE FOR GLYCOGEN,STARCH,MUCIN,BASEMENT MEMBRANE,ALPHNA
ANTI TRYPSIN,RETICULIN,FUNGI(CAPSULES),PANCREATIC
ZYMOGEN GRANULES,THYROID COLLOID,CORPORA
AMYLACEA,RUSSELS BODIES
MODIFICATION MILD PAS TECHNIGUE-FOR N-ACETYLSIALIC ACID CONTAINING
MUCIN
PAS-DIASTASE-FOR AMYLASE-DIGEST GYGOCEN-NEGATIVE
REACTION(DIFFERENTIATE GLYCOGEN TO MUCIN)
DIAGNOSIS 1)FUNGI-CANDIDAL HYPAE
2)MUCINS-FUNGUS IN BASEMENT MEMBRANE IN
ADENOCARCINOMA
3)MACROPHAGES-WHIPPLE’S DISEASE(BACTERIAL DISEASE IN
JOINT AND DIGESTIVE DISEASE)
4)STORAGE DISORDER-GLYCOGEN
5)GLYCOGEN-SEMIOMA,RHABDOMYOSARCOMA,EWING’S
SARCOMA
6)ROUTINE STAIN FOR KIDNEY AND LIVER BIOPSIES
Bancroft JD, Gamble M (2008).Theory and practice of histological techniques. 6th Edition. Churchill
Livingstone Elsevier. Chaps 4; 6-15; 17-19; 21 - 24; Pages 53 – 75; 83 – 261; 309 – 365; 433 - 517.

10. PAS-D PAS

11. ALCIAN BLUE
REAGENTS 1)ALCIAN BLUE
2)ALUMINIUM SULFATE
3)NUCLEAR FAST RED
TECHNIQUE DEPARAFFINIZE AND HYDRATE TO WATER-RINSE-ACETIC
ACID FOR 3 MINS-ALCIAN BLUE FOR 30 MINS-WASH-
HARRIS HAEMATOXYLIN FOR 30 SECS-WASH-
DEHYDRATE,CLEAR AND COVERSLIP
RESULT ACID MUCOPLYSACCHARIDES-BLUE
NUCLEUS-RED
POSITIVE FOR MUCIN-TO DIFFERENTIATE ACID MUCIN FROM NEUTRAL
MUCIN
MODIFICATION CRITICAL ELECTROLYTE CONCENTRATION(CEC)-
DIFFERENTIATE ACIDIC MUCIN AND PROTEOGLYCANS
DIAGNOSIS 1)GI TRACT DISEASE-BARRETS ESOPHAGUS
2)MESOTHELIOMA
3)ATHEROSCLEROSIS
4)DENOCARCINOMA
https://www.fixationonhistology.com/post/the-alcian-blue-stain-for-histology

13. MUCICARMINE
REAGENTS 1)CARMINE
2)ALUMINIUM HYDROXIDE
3)ALUMINIUM CHLORIDE,ANHYDROUS
4)50% ALCOHOL
TECHNIQUE DEPARAFFINIZE AND HYDRATE TO DISTALLED WATER-HARRIS
HAEMOTOXYLIN FOR 12 MINS-WASH-2 DIPS IN 0.5% ACID
ALCOHOL-WASH-MUCICARMINE FOR 30 MINS-RINSE-
DEHYDRATE,CLEAR AND MOUNT
RESULT ACID MUCIN-RED
NUCLEI-BLACK
OTHER TISSUE ELEMENTS-LIGHT YELLOW
POSITIVE FOR MUCIN
MODIFICATION -
DIAGNOSIS 1)MUCIN-EPITHELIAL AND CONNECTIVE CELLS
2)MUCIN-EPITHELIAL CELLS DURING INFLAMMATIONS
3)MUCIN-INTESTINAL CARCINOMA
4)MUCIN PRODUCING TUMOR-DETERMINE PRIMARY SITE OF
TUMOR
5)FUNGI CAPSULES-CRYPTOCOCCUS
Bancroft JD, Gamble M (2008).Theory and practice of
histological techniques. 6th Edition. Churchill Livingstone
Elsevier. Chaps 4; 6-15; 17-19; 21 - 24; Pages 53 – 75; 83 –
261; 309 – 365; 433 - 517.

15. AMYLOID
• EXTRACELLULAR,AMORPHOUS,EOSINOPHILIC
MATERIAL
• COMPOSED OF PROTEIN IN AN ANTIPARALLEL
BETA PLEATED SHEET CONFIGURATION
• IN H&E STAIN,CAN BE CONFUSED WITH
HYLALINE AND FIBRINOID SUBSTANCES
• EARLIEST SPECIAL STAIN USED FOR AMYLOID
WAS IODINE BY VIRCHOW
• MOST USEFUL METHODS ARE:-CONGO
RED,THIOFLAVINE T AND METHYL VIOLET,GREEN
METHODE
Bancroft JD, Gamble M (2008).Theory and practice
of histological techniques. 6th Edition. Churchill
Livingstone Elsevier. Chaps 4; 6-15; 17-19; 21 - 24;
Pages 53 – 75; 83 – 261; 309 – 365; 433 - 517.

16. CONGO RED STAIN
REAGENTS 1)80% ALCOHOL SATURATED WITH SODIUM CHLORIDE
2)1% AQUEOUS SOLUTION OF CONGO RED
3)1% AQUEOUS SOLUTION NaOH
TECHNIQUE TAKE SECTION TO WATER-STAIN NUCLEI IN HARRIS
HAEMATOXYLINE FOR 5 MIN-WASH-CONGO RED FOR 10
MINS-DEHYDRATE IN 2 CHANGES OF ABSOUTE
ALCOHOL-CLEAR IN XYELENE AND MOUNTH
RESULT AMYLOID-PINK
NUCLIE-BLUE
POSITIVE FOR AMYLOID DEPOSIT
MODIFICATION KMnO4/CR-TO DIFFERENTIATE PRIMARY AND
SECONDARY AMYLOIDOSIS
POLORIZED-CONGO RED STAINED EXHIBITS APPLE-
GREEN BIREFRINGENCE
DIAGNOSIS 1)NUEROFIBRILLARY TANGLES OF ALZHEIMER’S
2)INTRACELLULAR INCLUSIONS SEEN IN ADRENAL
CORTICAL CELLS
3)CELLULOSE AND CHITIN
4)DENSE COLLAGEN
Bancroft JD, Gamble M (2008).Theory and
practice of histological techniques. 6th Edition.
Churchill Livingstone Elsevier. Chaps 4; 6-15;
17-19; 21 - 24; Pages 53 – 75; 83 – 261; 309 –
365; 433 - 517.

18. THIOFLAVIN T
-FLOURESCENT METHOD
-TO DIFFERENTIATE AL(PRIMARY) AMYLOID AND AN(SECONDARY) AMYLOID
-AA AMYLOID LOSE THEIR AFFINITY FOR CONGO RED BUT AL AMYLOID IS RESISTANT
METHYL VIOLET / GREEN METHOD
-METACHROMIA STAIN
-MV-BLUISH STAIN AMYLOID AS PINK
-MG-GREEN STAIN AMYLOID PINK WITH GREEN NUCLEUS

19. LIPIDS
SIMPLE COMPOUND DERIVED
-FATS
-OIL
-WAXES
-TRIGLYCERIDES
-PHOSPOLIPIDS
-GLYCOLIPIDS
-DERIVED FROM
SOMPLE AND
COMPOUND LIPID
-CHOLESTROL
-BILE ACIDS
COMMONLY USED STAINS ARE OILD RED O AND SUDAN BLACK B
Bancroft JD, Gamble M (2008).Theory and practice of
histological techniques. 6th Edition. Churchill Livingstone
Elsevier. Chaps 4; 6-15; 17-19; 21 - 24; Pages 53 – 75; 83 – 261;
309 – 365; 433 - 517.

20. OIL RED O
REAGENTS 1)60% TRIETHYL PHOSPHATE
TECHNIQUE FROZEN SECTION-FREE FLOATING IN DISTILLED WATER-1
DIP IN 60%AQUEOUS TRIETHYL PHOSPHATE/70%
ALCOHOL-OIL O RED FOR 30 MIN-DISTILLED WATER FOR 1
MIN-HAEMATOXYLIN FOR 1 MIN-DISTLLIED WATER FOR 5
MIN-MOUNT SECTION TO SLIDE-DRY ON HOT PLATE-
MOUNT IN AQUAMOUNT AND RING WITH PARAFFIN
RESULT LIPID-RED
NUCLEI-BLUE
POSITIVE FOR LIPID
MODIFICATION -
DIAGNOSIS 1)FAT IN ABNORMAL PLACE
2)FATTY EMBOLI
3)LIPOSARCOMA
Bancroft JD, Gamble M (2008).Theory and practice of
histological techniques. 6th Edition. Churchill Livingstone
Elsevier. Chaps 4; 6-15; 17-19; 21 - 24; Pages 53 – 75; 83 – 261;
309 – 365; 433 - 517.

22. SUDAN BLACK B
REAGENTS 1)SUDAN BLACK B
TECHNIQUE WASH-SECTION IN 60% TRIETHYL PHOSPHATE-SUDAN
BLACK B FOR 10 MINS-SECTION IN 60% TRIETHYL
PHOSPHATE-SUDAN –WASH-IN MAYER’S CARMALUM
FOR 3 MINS-WASH-MOUNT IN GLYCERIN JELLY
RESULT LIPID-BLACK
NUCLEI-RED
POSITIVE FOR LIPID
MODIFICATION -
DIAGNOSIS 1)FAT IN ABNORMAL PLACE
2)FATTY EMBOLI
3)LIPOSARCOMA
Bancroft JD, Gamble M (2008).Theory and practice of
histological techniques. 6th Edition. Churchill Livingstone
Elsevier. Chaps 4; 6-15; 17-19; 21 - 24; Pages 53 – 75; 83 – 261;
309 – 365; 433 - 517.

24. NUCLEIC ACIDS
• NUCLEIC ACID HAVE 2 COMPONENT WHICH ARE
• 1)DNA
• 2)RNA
• THEY CONSIST OF SUGAR,PHOSPHATE AND
NITROGEN BASE
• DEMONSTRATION OF NUCLEIC ACIDS DEPENDS
UPON EITHER
• 1)REACTION OF DYES WITH PHOSPHATE GROUPS
• 2)PRODUCTION OF ALDEHYDES FROM SUGAR
• NO HISTOCHEMICAL METHODS ARE AVAILABLE
TO DEMONSTRATE THE NITROGENOUS BASE

25. • EXTRACTION TECHNIQUE
1)DIGESTION METHODS
PURE DEOXYRIBONUCLEASE WILL
DIGEST DNA AND PURE
RIBONUCLEASE WILL DIGEST RNA
2)CHEMICAL METHODS
• BY PERCHLORIC ACID
• TRICHLOROACETIC ACID
• HYDROCHLORIC ACID
STAINS FOR DNA STAINS FOR RNA
1)FEULGEN
TECHNIQUE(DEMONSTRATE
SUGAR)
2)METHYL GREEN PYRONIN
TECHNIQUE(DEMONSTRATE
PHOSPHATE)
3)ACRIDINE
ORANGE(FLOURESCENT
METHOD)
4)GALLOCYNANIN-CHROME
ALUM METHOD
1)GALLOCYNANIN-CHROME
ALUM METHOD

26. FEULGEN
TECHNIQUE(
DEMONSTRA
TE SUGAR)

27. PIGMENTS & MINERALS
ENDOGENOUS
PIGMENTS
EXOGENOUS
PIGMENTS
ARTIFACT PIGMENTS
1)HEMATOGENOUS
2)NON-
HEMATOGENOUS
1)ASBESTOS
2)SILICA
3)LEAD
4)CARBON
1)FORMALIN
2)MALARIA
3)MERCURY
4)SCHISTOSOME
Bancroft JD, Gamble M (2008).Theory and practice of
histological techniques. 6th Edition. Churchill Livingstone
Elsevier. Chaps 4; 6-15; 17-19; 21 - 24; Pages 53 – 75; 83 – 261;
309 – 365; 433 - 517.

28. DEFINITION STAINS
2 COMPONENT HAEM-IRON AND
OXYGEN AND GLOBIN-COLOURLESS
PROTIEN
1)LEUCO-PATENT BLUE
2)DUNN-THOMPSON
METHOD(MODIFIED VAN GIESON
3)BENZIDINE METHOD
HAEMOGLOBIN
DEFINITION STAINS
IRON STORED IN BONE MARROW IN THE
FORM OF THE GOLDEN-BROWN
PIGMENT.
1)PERL’S PRUSSIAN BLUE
2)TIRMANN-SCHMELZER TURNBULL BLUE
REACTION
HEMOSIDERIN

29. DEFINITION STAINS
THESE CONSIST OF A NUMBER OF
DIFFERENT PIGMENTS,THE TWO MAIN
CONSTITUENTS BEING BILIRUBIN AND
HAEMATOIDIN.
1)GMELIN TECHNIQUE
2)STEIN’S METHOD
3)FOUCHET’S METHOD
4)VAN GIESON STAIN
BILE PIGMENTS
DEFINITION STAINS
THIS TERM COVERS A NUMBER OF
PIGMENTS FOUND THROUGHOUT THE
BODY. IT IS SEEN AS GOLDEN-BROWN
DROPLETS THAT HAVE A VARIABLE
COMPOSITION
CEROID TYPE OF LIPOFUSCIN SEEN IN
LIVER
1)SUDAN BLACK
2)ZIEHL-NEELSEN
3)MASSON FONTANA
4)PAS
5)TOLUDINE BLUE
1)PAS
2)PERL’S METHOD
3)SCHMORL’S REACTION
LIPOFUSCIN

30. MELANIN
DEFINITION STAINS
MELANIN IS A NORMAL TISSUE PIGMENT
AND IS SEEN IN THE MELANOCYTES OF
THE SKIN
1)MASSON FONTANA
2)SCHMORL REACTION
3)DOPA OXIDASE REACTION
REAGENTS 1)FONTANA SILVER
2)UNDERGOES ARGENTAFFIN
REACTION
RESULT 1)MELANIN-BLACK
2)NUCLEI-RED
1) MASSON FONTANA STAINS 2)SCHMORL REACTION
REAGENTS 1)PRUSSIAN BLUE
2)FERRIC SALT
RESULT 1)MELANIN-DARK BLUE
2)NUCLEI-RED
Bancroft JD, Gamble M (2008).Theory and practice of histological techniques. 6th Edition. Churchill
Livingstone Elsevier. Chaps 4; 6-15; 17-19; 21 - 24; Pages 53 – 75; 83 – 261; 309 – 365; 433 - 517.

33. MINERALS
TYPE OF MINERALS STAINS
IRON
CALCIUM
COPPER
LEAD
ALUMINIUM & BERYLLIUM
SILVER
PERL’S REACTION
ALIZARIN RED S
VON KOSSA SILVER METHOD
RUBEANIC ACID METHOD
RHODIZONATE METHOD
SOLOCHROME METHOD
PARADIMETHYLAMINOBENZYLIDENE-
RHODANINE

34. MICROORGANISM
REAGENTS 1)CRYSTAL VIOLET STAIN
2)GRAMS IODINE SOLUTION
3)ETHYL ALCOHOL
4)ACETONE-XYLENE SOLUTION
5)BASIC FUCHINS
6)PICRIC ACID
RESULT 1)GRAM POSITIVE BACTERIA-BLUE
2)GRAM NEGATIVE BACTERIA-RED
3)NUCLEI-RED
4)OTHER TISSUES ELEMENTS-YELLOW
GRAM STAINING FOR BACTERIA-MODIFIED BROWN-BRENN METHOD
• http://stmichaelshospitalresearch.ca/staff-services/research-facilities/facilities/histology-core/routine-he
stain/#:~:text=Haematoxylin%20can%20be%20considered%20as,them%20and%20stains%20them%20purple.

35. ACID FAST STAINING FOR BACTERIA(TB)-NIEHL NEELSON(ZN) STAIN
REAGENTS 1)CARBOL FUCHSIN SOLUTION
2)1% ACID ALCOHOL
3)0.1% METHYL BLUE SOLUTION
RESULT 1)ACID FAST BACILLI-BRIGHT RED
2)OTHER TISSUE-PALE BLUE
3)CASEOUS MATERIAL-VERY PALE GRAYISH BLUE
REAGENTS 1)CARBOL FUCHSIN SOLUTION
2)5% SULPHURIC ACID ALCOHOL
3) METHYL BLUE SOLUTION
RESULT 1)M.LEPRAE-BRIGHT RED
2)NUCLEI AND OTHER TISSUE-PALE BLUE
MYCOBACTERIUM LEPRAE-MODIFIED FITE TECHNIQUE
REAGENTS 1)ACETATE BUFFER pH-3.6
2)1% SILVER NITRATE
RESULT 1)SPIROCHETES-BLACK
2)BACKGROUND-GOLDEN-YELLOW
SPIROCHETES-WARTHIN STARRY METHOD

36. FUNGAL STAINS
• FUNGAL CELL WALLS ARE RICH IN
POLYSACCHARIDES WHICH CAN BE
CONVERTED BY OXIDATION TO
DIALDEHYDES.DIALDEHYDES ARE THEN
DETECTED BY SILVER SOLUTION
• (GOMORII METHEAMINE SILVER
NITRATE (GMS) TECHNIQUE)
REAGENTS 1)4% CHROMIC ACID
2)1% SODIUM BISULFITE
3)5% SODIUM THIOSULFATE
4)0.21% SILVER NITRATE
5)GOLD CHLORIDE 0.1%
6)LIGHT GREEN SOLUTION
RESULT 1)FUNGI,PNEUMOCYSTIS.MELANIN-BLACK
2)MUCIN & GLYCOGEN-DARK GREY
3)BACGROUND-PALE GREEN
4)HYPHAE & YEAST FORM-SHARPLY DELINEATED IN BLACK
AGAINTS GREEN BACKGROUND
OTHER STAIN
USED FOR
FUNGAL
PAS/PAS-D
• http://stmichaelshospitalresearch.ca/staff-services/research-facilities/facilities/histology-core/routine-he
stain/#:~:text=Haematoxylin%20can%20be%20considered%20as,them%20and%20stains%20them%20purple.

38. OTHER
MICROORGANISM
MICROORGANISM STAINS
HELICOBACTER PYLORI CRESYL VIOLET ACETATE
METHOD
RICKETTSIA & VIRAL
INCLUSION
MACCHIAVELLO’S STAIN
VIRAL INCLUSION LENDRUM’S PHLOXINE-
TARTRAZINE
PARASITES GIEMSA STAIN

39. CONNECTIVE TISSUE
BLOOD
SPECIALIZED
PROPER
CARTILAGE
BONE
DENSE
LOOSE
ELASTIC
FIBROCARTILAGE
HYALINE
SPONGY
COMPACT
REGULAR
IRREGULAR
DENSE
ALVEOLAR
RETICULAR
ADIPOSE

40. COLLAGEN FIBRES STAINS
1)MASSON’S TRICHROME TECHNIQUE
2)VAN GIESON’S STAIN
3)MALLORY’S PHOSPHOTUNGSTIC ACID
HEMATOXYLIN
4)MSB TECNIQUE
5)PAS
6)HEIDENHAIN’S AZAN STAIN
7)LILLIE’S ALLOCHROME METHOD
8)LUXOL FAST BLUE G

41. MASSON’S
TRICHROME
TECHNIQUE
• -TRICHROME STAINS DEPENDS ON THE TISSUE PERMEABILITY
AND DYE MOLECULAR SIZE
• -ERYTHOCYTE PROTEIN HAS DENSE NETWORK WITH SMALL
PORES
• MUSCLE CELLS-LARGER PORES
• COLLAGEN-LEAST DENSE NETWORK AND QUITE POROUS
• HEAT INCREASES RATE OF STAINING AND PENETRATION
• LOW PH (1.5-3) ALSO HELPS WITH BETTER PENETRATION
• NUCLEAR STAIN FOR TRICHROME-IRON HEMTOCYLIN IS
PREFFERED AS ITS MORE RESISTANCE TO ACIDITY
-DYE SOLUTIONS,ALUM HEMATOXYLINS ARE DECOLORIZED
-CAN USE CELESTIN BLUE-ALUM HEMATOXYLIN SEQUENCE
-RECOMMENDED FIXATIVES ARE BOUIN’S,ZENKER’S.FORMAL-
MERCURY AND ZINC FORMALIN

42. REAGENTS 1)WEIGERT’S IRON HEMATOXYLIN
2)ACID FUCHSIN
3)GLACIA ACETIC ACID
4)PHOSPHOMOLYBDIC ACID
5)METHYL BLUE
RESULT 1)NUCLEI-BLUE/BLACK
2)CYTOPLASM,MUSCLE,RBC-RED
3)COLLAGEN-BLUE
DIAGNOSIS 1)DEMONSTRATE COLLAGEN AND MUSCLE IN NORMAL TISSUE
2)DIFFERENTIATE COLLAGENA ND MUSCLE IN TUMORS
3)IDENTIFY AN INCREASE IN COLLAGENOUS TISSUE
4)INDICATE FIBROTIC CHANGE IN CIRRHOSIS OF LIVER
5)INDICATE FIBROTIC CHANGE IN PYELONEPHRITIS
6)DISTINGUISH TUMORS THAT HAVE ARISEN FROM MUSCLE CELL AND
FIBROBLASTS
Bancroft JD, Gamble M (2008).Theory and practice of histological techniques. 6th Edition. Churchill Livingstone
Elsevier. Chaps 4; 6-15; 17-19; 21 - 24; Pages 53 – 75; 83 – 261; 309 – 365; 433 - 517.

44. VAN GIESON
TECHNIQUE
REAGENTS 1)WEIGERT’S IRON HEMATOXYLIN
2)SATURATED PICRIC ACID
3)ACID FUCHSIN
RESULT 1)COLLAGEN-BRIGHT RED
2)NUCLEI-BLUE/BLACK
3)CYTOPLASM,MUSCLE,RBC,ELASTIN,RETICULIN-YELLOW
Bancroft JD, Gamble M (2008).Theory and practice of
histological techniques. 6th Edition. Churchill
Livingstone Elsevier. Chaps 4; 6-15; 17-19; 21 - 24;
Pages 53 – 75; 83 – 261; 309 – 365; 433 - 517.

46. SPECIAL STAIN ORAL DISEASES
PERIODIC ACID SCHIFF (PAS) STAIN / GOMORII
METHEAMINE SILVER NITRATE (GMS)
TECHNIQUE
FUNGI-CANDIDA ALBICAN HYPHAE
GLYCOGEN STORAGE DISORDER
EWING’S SARCOMA
RHABDOMYOSARCOMA
ALCIAN BLUE STAIN ADENOCARCINOMA
ADENOSQUAMOUS CARCINOMA
MAST CELL LEUKEMIA
MYXOMA
MAY BE USEFUL IN FNA DIAGNOSIS OF
SALIVARY GLAND PLEOMORPHIC ADENOMA
MUCICARMINE STAIN USED TO IDENTIFY CANCER THAT PRODUCES
MUCIN:-
EXAMPLE WOULD BE TO DISTINGUISH
BETWEEN HIGH GRADE MUCOEPIDERMOID
CARCINOMA OF THE PAROTID, WHICH STAINS
POSITIVE VS SQUAMOUS CELL CARCINOMA
OF THE PAROTID WHICH DOES NOT.
ORAL CRYPTOCOCCUS NEOFORMANS
SPECIAL STAINS USED IN ORAL PATHOLOGY

47. SPECIAL STAINS ORAL DISEASES
FEULGEN STAIN TO RESEARCH THE NUCLEAR
MORPHOLOGY AND PLOIDY OF CANCER
CELLS.
OIL O RED & SUDAN BLACK B LIPOMA AND ANOTHER LIPOMATOUS
LESION
CONGO RED, METHYL VIOLET / GREEN
METHOD
AMYLOIDOSIS
NIEHL NEELSON(ZN) STAIN MYCOBACTERIUM TUBERCULOSIS
WARTHIN STARRY SYPHILIS
MASSON’S TRICHROME TECHNIQUE USED TO DIFFERENTIATE TUMORS THAT
HAVE ORIGINATED FROM MUSCLE CELLS
FROM THOSE THAT HAVE DEVELOPED
FROM FIBROBLASTS.
LEIOMYOMA, ANGIOLEIOMYOMA, AND
SMOOTH MUS-CLE HAMARTOMA,
MYOFIBROMA
• Sow-Yeh Chen, John E. Fantasia, Arthur S. Miller,Myxoid lipoma of oral soft tissue: A clinical and ultrastructural
study,Oral Surgery, Oral Medicine, Oral Pathology,Volume 57, Issue 3,1984,Pages 300-307,ISSN 0030-
4220,https://doi.org/10.1016/0030-4220(84)90186-5.

48. SPECIAL STAINS ORAL DISEASES
PERL’S STAIN ORAL EXFOLIATIVE CYTOLOGY FOR SICKLE
CELL ANAEMIA AND THALASSAEMIA
MASSON FONTANA METHOD MELANOMA

49. REFERENCES
1. Helmenstine, Anne Marie, Ph.D. (2020, August 27). What Histology Is and How
It’s Used. Retrieved from https://www.thoughtco.com/histology-definition-
and-introduction-4150176
2. http://stmichaelshospitalresearch.ca/staff-services/research-
facilities/facilities/histology-core/routine-he
stain/#:~:text=Haematoxylin%20can%20be%20considered%20as,them%20and
%20stains%20them%20purple.
3. https://www.fixationonhistology.com/post/the-alcian-blue-stain-for-histology
4. Bancroft JD, Gamble M (2008).Theory and practice of histological techniques.
6th Edition. Churchill Livingstone Elsevier. Chaps 4; 6-15; 17-19; 21 - 24; Pages
53 – 75; 83 – 261; 309 – 365; 433 - 517.
5. https://www.pathologyoutlines.com/topic/stainsalcianblue.html
6. https://en.wikipedia.org/wiki/Mucicarmine_stain
7. Sow-Yeh Chen, John E. Fantasia, Arthur S. Miller,Myxoid lipoma of oral soft
tissue: A clinical and ultrastructural study,Oral Surgery, Oral Medicine, Oral
Pathology,Volume 57, Issue 3,1984,Pages 300-307,ISSN 0030-
4220,https://doi.org/10.1016/0030-4220(84)90186-5.
8. https://www.jstor.org/stable/30082447?seq=1
9. Masson Trichrome Stain Helps Differentiate Myofibroma from Smooth Muscle
Lesions in the Head and Neck Region November 2008 Journal of the Formosan
Medical Association 107(10):767-73DOI: 10.1016/S0929-6646(08)60189-8
10. A Non-Invasive Technique which Demonstrates the Iron in the Buccal Mucosa
of Sickle Cell Anaemia and Thalassaemia Patients who Undergo Repeated
Blood TransfusionsHarika Chittamsetty, M.S. Muni Sekhar, Syed Afroz Ahmed,
Charu Suri, Sridevi Palla, S. Muni Venkatesh, Shahela TanveerJ Clin Diagn
Res. 2013 Jun; 7(6): 1219–1222. Published online 2013 Apr
27. doi: 10.7860/JCDR/2013/5551.3084PMCID: PMC3708240
11. DiNardo, A. R., Schmidt, D., Mitchell, A., Kaufman, Y., & Tweardy, D. J. (2015).
First Description of Oral Cryptococcus neoformans Causing Osteomyelitis of the
Mandible, Manubrium and Third Rib with Associated Soft Tissue Abscesses in
an Immunocompetent Host. Clinical microbiology & case reports, 1(3), 017.

50. THANK YOU

51. SPECIAL THANKS TO MLT’S
FROM OMOP(UM)