Histochemistry Practical Review Flashcards
Giemsa
Routine nuclear stain for bone marrow
Nuclei: blue
Bacteria: blue
Mast cells: dark blue with red granules
White blood cells: light blue
Rickettsias: intense reddish purple
Collagen and muscle: pale pink
Red blood cells: grey, yellow, or pink
PAS Periodic Acid Schiff (With and Without Diastase Digestion)
Demonstrates glycogen and neutral mucosubstances
Carbs WIthout digestion: Bright rose
Carbs With digestion: unstained/pale pink
Basement membranes and fungal cells walls also stain bright rose due to polysaccharides
Alcian Blue
pH 2.5 both sulfonated and carboxylated acid mucopolysaccharides
pH 1 only sulfonated acid mucopolysaccharides
Mucosubstances: Blue
Background: pink to red (NFR counterstain)
Nuclei: red
Alcian Blue with PAS
Acid mucopolysaccharides: blue
Neutral mucopolysaccharides: magenta
Other substances: purple
Mucicarmine
stains sulfonated and carboxylated epithelial (acid) mucins (like alcian blue)
also encapsulated yeast Cryptococcus neoformans which has a mucinous capsule
Mucin: deep rose to red (ex: bright red goblet cells)
Nuclei: black/grey
Background: metanil yellow counterstain
Congo Red
Demonstrates amyloid (most specific method)
Amyoid: deep pink to red, apple green birefringence under polarized light
Elastic tissue: pale pink
Nuclei: blue (Harris hematoxylin)
Crystal Violet
rapid screen for amyloid, less specific
Amyloid: purplish violet
Other tissue elements: blue
Colloidal Iron
Prussian blue reaction ddemonstrates carboxylated and sulfonated acid mucosubstances (like mucicarmine and alcian blue) but not as specific
acid mucopolysaccharides: deep prussian blue (esp catillage)
nuclei: red-pink (NFR counterstain)
cytoplasm: pink (NFR)
Van Gieson is an alternative counterstain that results in a yellow background
Masson’s Trichrome
Differentiate between collagen and smooth muscle
Nuclei: Black
Cytoplasm, keratin, and muscle: red
Collagen and mucin: blue (or light green if using alternate counterstain)
Verhoeff Elastic
demonstrate elastic fibers
Elastic fibers: blue-black to black
Nuclei: blue to black
Collagen: Red
Other tissue elements: yellow (due to Van Gieson counterstain)
Movat Pentachrome
Demonstrate the various connective tissues
Nuclei and elastic fibers: Black
Collagen: Yellow
Ground substance: Blue to green
Muscle: Red
Fibrinoid: Intense red
Gordon and Sweet’s Reticulum
argyrophil silver stain for reticular fibers (esp in liver)
Reticulin: Black (sharp lines/squiggles)
Other tissue elements: pink-red if NFR counterstain is used
Jones (methenamine silver)
PAMS (periodic acid methenamine silver)
methenamine silver stain for basement membranes in kidney
Basement membrane: Crisp black
Background: Green (or pink if you use NFR)
Oil Red O
for lipids in frozen sections
Fat: Red
Other tissues: Blue or technique dependant
Sudan Black B
for lipids in frozen sections
Fat: Blue-black
Nuclei: Red (NFR counterstain)
Bielschowsky
argyrophil silver stain for neuro tangles and plaques (for alzhiemers)
Tangles, plaques, axons: Black
Amyloid (plaque cores and vasculature): Magents
Lipofucsin (wear and tear pigment): Magenta
LFB Luxol Fast Blue
stains for myelin (white matter)
inside spinal cord, outside brain
White matter (myeline): blue to blue-green
Grey matter: unstained/colorless
Should have high contrast macroscopically
AFB Acid Fast Bacteria Kinyoun
AFB Fite
Kinyoun detects acid fast myco bacteria
So does Fite, but it also detects M. leprae (leprosy)
AFB: Bright red
Background: Light blue
Brown Hopps Gram Stain
Gram + and Gram - bacteria for classification
Gram + (Thick wall): Blue black
Gram - (Thin wall): Red
Background Tissue: Yellow
Nuclei: light red
GMS Grocott Methenamine Silver
Argyrophil silver stain that demonstrates oxidized polysaccharides in fungal cell walls
Fungi: crisp black cells wals with visible internal structures
Mucin: Taupe to dark grey
Background: Green
Steiner and Steiner
Demonstrate spirochetes, argyrophil silver stain
Spirochetes, H. pylori, and other non-fillamentous bacteria: Dark brown to black
Background: Light Yellow
Iron (Prussian Blue)
Bone marrow and spleen
Nuclei and hemofuchsin: bright red
Hemosiderin (stored iron): blue
Background: Pink (NFR counterstain)
Copper (Rhodanine)
esp for detecting Wilson disease in the liver
Copper: bright red to red-yellow
Nuclei: Bright Blue
Von Kossa
For Calcium (indirect detection method of ions in calcium salts) argyropil method reduced by sunlight
Calcium Salts: Black
Background: Red (NFR counterstain)
Bile (Hall)
demonstrates bilirubin which is oxidized to biliverdin
Bile or bilirubin: Emerald green to olive Drab
Background: Yellow (liver)
Fontana Masson
Reducers such as argentaffn granules, melanin, and formalin pigment
argentaffin silver stain, often use skin as ctrl
Melanin, argentaffin granules: Black
Nuclei: Pink (NFR counterstain)
