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Special Stain.pptx

This document discusses special stains used in oral pathology. It begins by describing hematoxylin and eosin staining, which colors nuclei purple and cytoplasm pink. It then discusses several other special stains including: Periodic acid Schiff for glycogen and mucins. Alcian blue for acidic mucins. Mucicarmine specifically stains acid mucins red. Congo red and thioflavin T stain amyloid deposits. Oil red O and Sudan black B stain lipids. Feulgen staining demonstrates DNA sugars. Finally, pigments, minerals, and stains for microorganisms are briefly mentioned.

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SPECIAL STAIN BY DR VINESH RAJ.S MOSc(APPLIED ORAL PATHOLOGY CANDIDATE), DEPARTMENT OF ORAL MEDICINE & ORAL PATHOLOGY FACULTY OF DENTISTRY,UM
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
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
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
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
SPECIAL STAINS 1.CARBOHYDRATES 2.AMYLOID 3.LIPIDS 4.NUCLEIC ACID 5.PIGMENTS & MINERALS 6.MICROORGANISM 7.CONNECTIVE TISSUE
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
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.
PAS-D PAS
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
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.
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.
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.
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
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.
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.
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.
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
• 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
FEULGEN TECHNIQUE( DEMONSTRA TE SUGAR)
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.
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
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
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.
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
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.
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
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.
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
CONNECTIVE TISSUE BLOOD SPECIALIZED PROPER CARTILAGE BONE DENSE LOOSE ELASTIC FIBROCARTILAGE HYALINE SPONGY COMPACT REGULAR IRREGULAR DENSE ALVEOLAR RETICULAR ADIPOSE
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
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
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.
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.
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
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.
SPECIAL STAINS ORAL DISEASES PERL’S STAIN ORAL EXFOLIATIVE CYTOLOGY FOR SICKLE CELL ANAEMIA AND THALASSAEMIA MASSON FONTANA METHOD MELANOMA
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.
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Special Stain.pptx

  • 1.
    SPECIAL STAIN BY DR VINESHRAJ.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 anorganic 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 isa 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 FORPATHOLOGIST 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.
  • 7.
    CARBOHYDRATE CARBOHYDRATE POLYSACCHARIDES GLYCOGEN-PAS DIASTASE DIGESTION OF GLYCOGEN-PAS-D NEUTRALMUCOSUBSTANCE 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 REAGENTS1.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.
  • 11.
    ALCIAN BLUE REAGENTS 1)ALCIANBLUE 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)ALUMINIUMCHLORIDE,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 • COMPOSEDOF 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 REAGENTS1)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 -TODIFFERENTIATE 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 -DERIVEDFROM 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 REAGENTS1)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 REAGENTS1)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 • NUCLEICACID 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)DIGESTIONMETHODS 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.
  • 27.
    PIGMENTS & MINERALS ENDOGENOUS PIGMENTS EXOGENOUS PIGMENTS ARTIFACTPIGMENTS 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 COMPONENTHAEM-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 CONSISTOF 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 ISA 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 MINERALSSTAINS 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 VIOLETSTAIN 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 STAININGFOR 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 • FUNGALCELL 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 PYLORICRESYL VIOLET ACETATE METHOD RICKETTSIA & VIRAL INCLUSION MACCHIAVELLO’S STAIN VIRAL INCLUSION LENDRUM’S PHLOXINE- TARTRAZINE PARASITES GIEMSA STAIN
  • 39.
  • 40.
    COLLAGEN FIBRES STAINS 1)MASSON’STRICHROME 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 STAINSDEPENDS 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 IRONHEMATOXYLIN 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’SIRON 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 ORALDISEASES 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 ORALDISEASES 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 ORALDISEASES PERL’S STAIN ORAL EXFOLIATIVE CYTOLOGY FOR SICKLE CELL ANAEMIA AND THALASSAEMIA MASSON FONTANA METHOD MELANOMA
  • 49.
    REFERENCES 1. Helmenstine, AnneMarie, 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.
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  • 51.
    SPECIAL THANKS TOMLT’S FROM OMOP(UM)