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Stains in dermatology .pptx
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- 2. WHAT IS STAIN? Staining is an auxiliary technique used in microscopy to enhance contrast in the microscopic images. Stains may be used to define and examine the bulk tissue 1. Cell population 2. Muscle fibers 3. Connective tissues 4. Organelles
- 3. SPECIAL STAINS Usedin addition to H & E staining to selectively Stain cells & cellular components Gives information on: Presence of certain class of molecules Their localization Number of molecules
- 4. STAINS FOR MUCOPOLYSACCARIDES 1. PAS 2.ALCIAN BLUE 3. MUCICARMINE 4. COLLOIDAL IRON
- 5. PAS Most commonlyused special stain Stains neutral mucopolysaccharides (Glycogen) Basement membranes (e.g SLE) Fungi, Parasides Fibrin, Inclusion bodies
- 9. ALCIAN BLUE Beststain for mucin Alcian blue (pH 2.5) - Acid mucopolysaccharides (glycosaminoglycain’s) - Light blue Alcian blue (pH 0.5) - Sulfated mucopolysaccharides (heparin sulfate) - Blue Does NOT stain neutral mucins
- 10. RESULTS Acid mucin& proteoglycans : BLUE Nuclei : RED
- 12. MUCICARMINE To demonstrateacid & neutral mucopolysaccharides - Epithelial mucin - Capsule of cryptococcus neoformans Results Acid epithelial mucins : Deep rose to red Nuclei : Black Others tissue elements : Light yellow
- 14. COLLOIDAL IRON Amucin stain for dermal/connective tissue mucin Stains acid mucopolysaccharides RESULTS Proteoglycans , Acid mucins : BRIGHT BLUE Collagen : RED Muscle & cytoplasm : YELLOW
- 16. STAINS FOR PIGMENTS& MINERALS 1. Fontana-Masson 2. Von-kossa 3. Alzarin red S 4. Prussian blue stain
- 17. FONTANA-MASSON Stains melanin& argentaffin granules – BLACK (nuclei will be red) Useful for quantifying melanocytes (e.g. in vitiligo) & in cases of minocycline pigmentary alteration Also stains cryptococcus RESULTS Melanin , Argentaffin, Chromaffin : BLACK Nuclei : RED
- 18. USES To identifymelanin & argentaffin granules In diagnosis of malignant melanoma Argentaffin granules are found in carcinoid tumors
- 21. VON-KOSSA Stains calciumsalts – BLACK Calcification of vessel walls & elastic tissue ( Calcinonis cutis, pseudoxanthoma elasticum, calciphylaxis, elastosis & elastofibroma)
- 24. PRUSSIAN BLUE STAIN(PERLS IRON) Hemosiderin and ferric ions Useful for identifying iron as the source of pigment Stains iron – BLUE RESULT Ferric iron : Blue Nuclei : Red
- 26. ALIZARIN RED Stainfor calcium salts RESULTS Calcium : ORANGE RED Background : GREEN
- 28. STAINS FOR CONNECTIVE TISSUE Trichrome-Masson Verhoeff-Van Gieson
- 29. TRICHROME-MASSON Smooth muscle, cytoplasm, keratin – Red Useful for distinguishing leiomyoma’s from dermatofibromas & neural tumors Collagen – Blue/green Useful in evaluating the characteristics of dermal collagen Nucleus - Black
- 30. USES It isused to differentiate b/w collagen & smooth muscle in tumor To identified the increased collagen deposition : Keloid RESULT Nuclei : Blue/Black Cytoplasm, Muscle, & Erythrocytes : Red Collagen : Blue
- 32. VERHOEFF-VAN GIESON ORWEIGERT Used to differentiate collagen and smooth muscle Can be used to demonstrate the presence of collagen in pathological conditions( DLE, EN, RA, SLE, MORPHOEA) Stains nuclei : Blue Cytoplasm, muscle, fibrin : Yellow Collagen : Bright Elastic fibers : Black
- 34. STAINS FOR AMYLOID Congo red Thioflavin T Crystal violet
- 35. CONGO RED Stainsamyloid : PINKISH-RED Gives apple-green birefringence to amyloid The most specific method for amyloid
- 38. THIOFLAVIN T Amyloidshows yellow fluorescence RESULT UV light source : Silver-blue Blue light fluorescence : Yellow
- 40. CRYSTAL VIOLET • Stain Amyloidpurple-violet
- 41. STAINS FOR FAT Sudan black B Sudan orange Oil red O
- 42. SUDAN BLACK B Most sensitive of all fat dyes Stains neutral fats – Blue-Black Stains phospholipid – Grey
- 44. OIL RED O Stains fat red Frozen/Fresh tissue ( once tissue is fixed &processed into paraffin blocks, this method does not work) This may be very helpful in seeing the fat globules in sebaceous carcinoma
- 45. RESULT Fat : BrilliantRed Nuclei : Blue FAT NUCLEI
- 46. STAINS FOR MASTCELLS Giemsa Toluidine blue Chloroacetate esterase ( Leder stain)
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- 48. TOLUIDINE BLUE Mast cells stain– Purple (Metachromatic staining) Background stain – Blue (orthochromatic staining) Mastocytosis MAST CELL
- 49. STAINS FOR MICROORGANISMS H&E Gram Giemsa Gomori methenamine silver (GMS) PAS Fontana-Masson Warthin-Starry Ziehl-Neelson stain
- 50. H&E May demonstrate fungi,bacteria
- 51. GRAM STAINING Differentiatesbacteria by the chemical & physical properties of their cell well by detecting peptidoglycan Gram +ve bacteria retain the crystal violet dye & thus are stained violet : BLUE Gram –ve bacteria : PINK
- 53. GOMORI METHENAMINE SILVER(GMS) Fungal cell walls Stains fungi & parasites brown or black with a green background Pneumocystis carnii, histoplasma , leshmania
- 56. GIEMSA STAIN Usedto stain bacteria & protozoa H. pylori, rickettsia & chlamydia Bacteria stains : BLUE Cytoplasm stains from : Pink to rose Nuclei : BLUE Eosinophils are also easily detected
- 58. WARTHIN-STARRY A silvernitrate stain Spirochetes – BLACK Background – Golden yellow
- 60. ZIEHL-NEELSEN Acid fastrefers to cell walls containing high lipid content Used to stain Mycobacteria, oocysts of Cryptosporidium parvum, Cyclospora, Isospora Acid fast cells stain : RED Non acid fast cells stain : BLUE
- 62. ACID FAST STAINING TECHNIQUES Ziehl-Neelsen stain ( classic type) Kinyoun Auramine-rhodamine (fluorescent dyes) Fite : for leprosy bacillus which is less acid fast
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