Histochemistry Practical Review

Question 1

Giemsa

Answer 1

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

Question 2

PAS Periodic Acid Schiff (With and Without Diastase Digestion)

Answer 2

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

Question 3

Alcian Blue

Answer 3

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

Question 4

Alcian Blue with PAS

Answer 4

Acid mucopolysaccharides: blue

Neutral mucopolysaccharides: magenta

Other substances: purple

Question 5

Mucicarmine

Answer 5

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

Question 6

Congo Red

Answer 6

Demonstrates amyloid (most specific method)

Amyoid: deep pink to red, apple green birefringence under polarized light
Elastic tissue: pale pink
Nuclei: blue (Harris hematoxylin)

Question 7

Crystal Violet

Answer 7

rapid screen for amyloid, less specific

Amyloid: purplish violet
Other tissue elements: blue

Question 8

Colloidal Iron

Answer 8

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

Question 9

Masson’s Trichrome

Answer 9

Differentiate between collagen and smooth muscle

Nuclei: Black
Cytoplasm, keratin, and muscle: red
Collagen and mucin: blue (or light green if using alternate counterstain)

Question 10

Verhoeff Elastic

Answer 10

demonstrate elastic fibers

Elastic fibers: blue-black to black
Nuclei: blue to black
Collagen: Red
Other tissue elements: yellow (due to Van Gieson counterstain)

Question 11

Movat Pentachrome

Answer 11

Demonstrate the various connective tissues

Nuclei and elastic fibers: Black
Collagen: Yellow
Ground substance: Blue to green
Muscle: Red
Fibrinoid: Intense red

Question 12

Gordon and Sweet’s Reticulum

Answer 12

argyrophil silver stain for reticular fibers (esp in liver)

Reticulin: Black (sharp lines/squiggles)
Other tissue elements: pink-red if NFR counterstain is used

Question 13

Jones (methenamine silver)
PAMS (periodic acid methenamine silver)

Answer 13

methenamine silver stain for basement membranes in kidney

Basement membrane: Crisp black
Background: Green (or pink if you use NFR)

Question 14

Oil Red O

Answer 14

for lipids in frozen sections

Fat: Red
Other tissues: Blue or technique dependant

Question 15

Sudan Black B

Answer 15

for lipids in frozen sections

Fat: Blue-black
Nuclei: Red (NFR counterstain)

Question 16

Bielschowsky

Answer 16

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

Question 17

LFB Luxol Fast Blue

Answer 17

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

Question 18

AFB Acid Fast Bacteria Kinyoun

AFB Fite

Answer 18

Kinyoun detects acid fast myco bacteria

So does Fite, but it also detects M. leprae (leprosy)

AFB: Bright red
Background: Light blue

Question 19

Brown Hopps Gram Stain

Answer 19

Gram + and Gram - bacteria for classification

Gram + (Thick wall): Blue black
Gram - (Thin wall): Red
Background Tissue: Yellow
Nuclei: light red

Question 20

GMS Grocott Methenamine Silver

Answer 20

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

Question 21

Steiner and Steiner

Answer 21

Demonstrate spirochetes, argyrophil silver stain

Spirochetes, H. pylori, and other non-fillamentous bacteria: Dark brown to black
Background: Light Yellow

Question 22

Iron (Prussian Blue)

Answer 22

Bone marrow and spleen

Nuclei and hemofuchsin: bright red
Hemosiderin (stored iron): blue
Background: Pink (NFR counterstain)

Question 23

Copper (Rhodanine)

Answer 23

esp for detecting Wilson disease in the liver

Copper: bright red to red-yellow
Nuclei: Bright Blue

Question 24

Von Kossa

Answer 24

For Calcium (indirect detection method of ions in calcium salts)
argyropil method reduced by sunlight

Calcium Salts: Black
Background: Red (NFR counterstain)

Question 25

Bile (Hall)

Answer 25

demonstrates bilirubin which is oxidized to biliverdin

Bile or bilirubin: Emerald green to olive Drab
Background: Yellow (liver)

Question 26

Fontana Masson

Answer 26

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)