STAINS FOR CONNECTIVE TISSUE

MALLORY’S PHOSPHOTUNGSTIC ACID HEMATIXYLIN STAIN VAN GIESON’S STAIN MASSON’S TRICHROME STAIN
FIXATIVE Zenker’s fixative (mercuric chloride fixative) Any good well-fixed tissue and 10% BNF Bouin’s fluid
TECHNIQUE 6u 6u 6u
SOLUTIONS Phosphotungstic acid hematoxylin Weigert’s Iron Hematoxylin – nuclear stain Bouin’s
Van Gieson’s Picric – Fuchsin solution - counterstain Weigert’s Iron Hematoxylin – nuclear stain
Bieberch Scarlet – Acid Fuchsin
Phosphomolybdic Acid -Phosphotungstic acid
Aniline Blue – counter stain (PAPA mordant)
Light green – counterstain (5% PTA mordant)
1% Acetic Water – removes excess countersatin
RESULTS Nuclei – blue Collagen – red Nuclei – black
Fibrin – blue Muscle, cornified epithelium – yellow Cytoplasm, keratin, muscle fiber – red
Fibroglia and microglia – blue Nuclei – blue to black Collagen, mucous - blue
Collagen fiber – yellowish to brown red
Coarse elastic fiber – purplish tint
STAINS FOR ELASTIC TISSUE FIBER STAINS FOR RETICULUM
VERHOEFF’S ELASTIC TISSUE STAIN WILDER’S RETICULUM STAIN LILLIE’S BIERBRICH SCARLET MALLORY’S ANILINE BLUE
PICROANILINE BLUE
FIXATIVE Any Formalin, zenker, or helly’s, 10% BNF
TECHNIQUE 6u 6-10u
SOLUTIONS 2% Ferric chloride – used for differentiation 10% Phosphomolybdic acid – for oxidation PTA
Van Gieson’s Stain Uranium nitrate – deep synthesizer Acid fuchsin
Sodium thiosulfate (hypo) Ammoniacal nitrate – colorless silver complex to dark Aniline blue
brown silver chloride
Reducing – dark brown silver oxide to black metallic
silver
1% Gold Chloride – toning, yellow color to lavander
Sodium Thiosulfate (hypo)
RESULTS Elastic fibers – blue to black Reticular fibers – black Nuclei – Red Nuclei & muscle tissue – red
Nuclei – blue to black Collagen – rose color Collagen fiber – blue Collagen fiber - blue
Collagen – red Other tissue elements – Depending on counterstain Muscle tissue - red
Other tissue elements - yellow
STAINS FOR CYTOPLASMIC GRANULES STAINS FOR MAST CELL GRANULES
FONTANA – MASSON SATIN FOR ARGENTAFFIN GRANULES DOMINICI’S METHOD MALLORY’S PHLOXINE – METHYLENE BLUE STAIN
FIXATIVE formalin Any and Zenker – gives better solution
TECHNIQUE 6–10 u 6u
SOLUTIONS Gold chloride Acid – fuchsin – orange g
Sodium thiosulfate Toluidine- metachromatic stain
Nuclear fast red (kernechrot) Nuclear fast red – (kernechrot)
RESULTS Argentaffin granules – black Granules of mast cells – purplish red Nuclei & bacteria – blue
Nuclei – pink Other tissue elements – pink to rose Mast cells granules – purple
Ricketssia – blue to violet
Collagen & other tissue elements – bright rose
 STAINS FOR CARBOHYDRATE, CONNECTIVE TISSUE, GROUND SUBSTANCE, AND AMYLOID
BEST’S CARMINE STAIN FOR PERIODIC ACID-SCHIFF (PAS) ALCIAN BLUE STAIN (SHORT METHOD) CRYSTAL VIOLET AMYLOID STAIN
GYCOGEN REACTION w/w/out HYALURONIDASE
FIXATIVE Absolute alcohol or carnoy’s 10% formalin & Zenker’s solution 10% Neutral Formalin formalin 10% or alcohol
fluid
TECHNIQUE 6u 6u 6u (avoid overheating) 6u
SOLUTIONS Carmine stock Coleman’s Feulgen reagent 3% Acetic-acid mordant to alcian blue Stock crystal violet
Working Schiff’s Leuco-fuchsin Alcian Blue Working
Differentiating Hyaluronidase Abopon Mounting Medium (water-soluble, temporary mount)
RESULTS Glycogen – pink to red +Reaction – rose to purplish-red Acid mucopolysaccharides – blue Amyloid – Purplish Violet
Nuclei - Blue Nuclei – blue Nuclei - Pink Other tissue elements - blue
Fungi – red
Background – pale green
STAINS FOR CARBOHYDRATE, CONNECTIVE TISSUE, GROUND SUBSTANCE, AND AMYLOID OTHERS
GRAM’S IODINE CONGO RED – METHOD BENNHOLD’S & METACHROMATIC STAIN MOWRY IRON STAIN METACHROMATIC STAINING
HIGHMAN’S
FIXATIVE
TECHNIQUE
SOLUTIONS Harris Hematoxylin & Congo red
RESULTS Amyloid - Red
STAINS FOR PIGMENTS AND MINERALS
GOMORI’S IRON REACTION TUMBULL’S BLUE REACTION MALLORY’S FUCHSIN STAIN VON KOSSA’S METHOD FOR STEIN’S BILE PIGMENT DAHL’S METHOD
(Fe pigments of hemosiderin) FOR FERROUS Fe(HEMOSIDERIN) FOR HEMOFUCHCIN STAIN DEMONSTRATING CALCIUM STAIN FOR CACLIUM
FIXATIVE Alcohol/ 10% Formalin BNF Zenkers, absolute OH, 10% BNF OH preferred and 10% formalin OH or 10% formalin 4-6 hrs
TECHNIQUE 5u 5u 6um 6u 6-7u
SOLUTIONS 20% HCl Neutral Red Stock Sol’n A Fresh hematoxylin - first is 5% Silver nitrate (stain) Stock tincture of iodine Alizarin red S –
10% Potassium ferrocyanide Stock Solution B unoxidized hematoxylin, 5% Sodium thiosulfate Stuck Lugol’s Staining sol’n
Nuclear Fast Red (Kernechtrot) Working Solution because the iron itself in the Nuclear Fast red (kernechtrot) Stein’s working iodine Light green -
Stain tissue will oxidize hematoxylin stain reagent counterstain
5% Soidum thiosulfate
Nuclear fast red
(Kernechrot) Stain
RESULTS Nuclei – red Iron Pigments – Bright blue (ferrous Nuclei – blue Calcium salts – black Nuclei – red
Cytoplasm – Pink to rose iron) Other- red (hemofuchsin) Nuclei – Red Cytoplasm – pink to rose
Iron pigments - blue Nuclei – red Unstained (hemosiderin) Cytoplasm – red to rose
Cytoplasm – pink to rose
STAINS FOR NERVOUS SYSTEM TISSUE
HOLZER’S STAIN FOR GLIA FIBERS BODIAN’S METHOD (nerve fibers and nerve endings) WEIL-WEIGERT STAIN for myelin sheath (Lillie’s variant)
FIXATIVE Formalin alcohol or 10% formalin Formalin, alcohol formalin, Bouin’s fluid 10% formalin or Orth’s fluid (chromate fixatives), chromic acid
TECHNIQUE 6-8u 6–8u Paraffin: 15u
Celloidin 25u
SOLUTIONS Phosphomolybdic alcohol Protargol (Silver protonate solution) Stock 1% Alcoholic hematoxylin
Absolute alcohol chloroform mixture 1% Gold Chloride Stock 4% Ferric ammonium sulfate
 Crystal violet stain (staining solution) 2% Oxalic Acid Working Hematoxylin sol’n – stainig sol’n
Potassium bromide Aniline Blue stain Borax – for glowing
Differentiating Differentiating Stock safranin
Workin safranin
RESULTS Glia Fibers – Deep violet Myelinated fibers, non-myelinated fibers of the central Nuclei – Red
Background – Pale violet peripheral nervous system & neurofibrils - black Cytoplasm – pink to rose
STAINS FOR MICROORGANISMS AND OTHER CELLS
KINYOUN’S ACID FAST ZIEHL – NELSEN STAIN (for FITE – FARACO MaCallum – GOODPASTURE BROWN & BRENN (B&B) METHOD SCHLEIF’S STAIN (for
STAIN acid fast bacteria) STAIN STAIN (for bacteria in tissue) (for bacteria, nocardia, & amiticytes negri bodies)
FIXATIVE Formalin-preferred (others Any 10% BNF Any Zenker’s
may be used sections to
water
TECHNIQUE Paraffin: 15u 4-6u 4-5um Paraffin and celloidin sections 6u
Celloidin: 25u are used
SOLUTIONS Kinyoun’s carbol fuchsin Carbol fuchsin Xylene peanut oil Goodpasture’s stain Gram’s iodine Stock Schleifstein’s Stain
1% acid alcohol 1% acid alcohol 1% Acid alcohol Gram’s iodine Saturated basic fuchsin Sol’n A
Stock methylene blue Working Methylene blue Zhiel-neelsen carbol Stirling’s gentian Violet stain Working crystal violet (fresh) Stock Schleifstein’s Stain
Working methylene blue fuchsin sol’n Saturated Picric Acid Working basic fuchsin Sol’n B
Working methylene Working Scheliefstein’s
blue 5% Iodine
5% Sodium thiosulfate
(Hypotonic)
RESULTS Acid fast bacteria – bright Acid fast bacteria – bright red M.Leprae & Other acid Gram (+) Organism – Blue Negri bodies – Deep
red Erythrocyte – yellowish orange – fast bacteria – bright Gram (-) Organism – Red magenta
Background – light blue Other tissue elements – pale red Nuclei – Red Cytoplasm – Blusih violet
blue Background – light blue Other tissue elements - yellow Erythrocytes - copper
STAINS FOR AMOEBA STAINS FOR SPIROCHETES STAINS FOR RICKETTSIA
1. Gridley’s Method LEVADITI’S METHOD WARTHIN-STARRY-FAULKNER 1. Pinkerton – citric acid for differentiation
for E. Histolytica METHOD (for spirochetes in tissue 2. Giemsa stain
• Results: section) 3. Mallory’s Phloxine-Methylene Blue stain
- Amoeba- Blue FIXATIVE OTHER STAINS
green TECHNIQUE 1. Feulgen stain – specific chemical test for DNA. Section of Lymph node is used as control
- Ingested RBC- SOLUTIONS Silver Nitrate sol’n – 2. Methylene Green Pyronin Stain – stains nuclear chromatin, DNA, RNA, and plasma cell
Deep rose fresh tissue:strong silver 3. Gomori’s Aldehyde Fuchsin Stain – Staining B cells of pancreas (deep blue to purple), use
2. Heidenheim’s Iron nitrate sol’n paraffin, since stain will not diffuse through celloidin.
Hematoxylin Reducing Sol’n 4. Mallory’s-Heidenhain’s Aniline Blue Stain (Azan stain) – stain for a cell of the pancreas
3. PTAH RESULTS Spirochetes – Black (red), collagen fibers (blue)
Background – Pale Yellow – Light 5. Gomori’s Chromium Hematoxylin-Phloxin-Stain (Chrome-alum hematoxylin stain) –
brown differential stain for islet cells of pancreas
• Fixative: Bouin’s sol’n
• A cells – red
• B cells – blue
• D cells – pink to red (indistinguishable from a-cells)
6. Masson’s Hematoxylin-Phloxine-Safron (HPS) Stain
 • Bone & cartilage – yellow
• Muscle tissue – red
• Nuclei – blue
• Red cells – bright pink
7. May Grunwalk Giemsa stain for bone marrow
• Nuclei – blue
• Cytoplasm – pink to rose
8. Pearse Trichrome PAS – differential stain for cells of the pituitary, (stain’s basement
membrane, glycogen, and fungi (purple red)
• Nuclei – blue to brown
• Acidophils – Orange to yellow
• Basophils – Deep purple to red
STAINS FOR FATS
OIL RED O FAT STAIN NILE BLUE SULFATE (FOR NEUTRAL FATS AND FATTY ACIDS)
FIXATIVE Formalin Formalin
TECHNIQUE Frozen sections – 10-15u, collect in distilled water Cut frozen sections in distilled water
SOLUTIONS Oil Red O Nile Blue sulfate stain
Glycerin jelly – temp. mount Abopon – water miscible mounting medium
RESULTS Fat – orange to bright red Neutral fats – Pink
Nuclei - blue Fatty acids – blue to violet
Nucleic & Elastic tissue – Dark blue