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AP0509 Component 2 Exam > Common Stains in Cellular Pathology > Flashcards

Common Stains in Cellular Pathology Flashcards

1
Q

H&E gives diagnosis in how many histological clinical reports?

A
  • 70%
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2
Q

What colours does H&E stain?

A
  • nuclei - blue/black
  • collagen - pink
  • muscle - deep pink
  • RBCs - orange
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3
Q

What colour does elastic fibres stain from H&E?

A
  • red/pink
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4
Q

Common methods used for elastic fibres

A
  • Orcein
  • Verhoeff’s
  • Weigert’s
  • Aldehyde fuchsin
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5
Q

What structures do haematoxylin stain?

A
  • acidic or basophilic (basic dye)

- cell nucleus, ribosomes, rough ER

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6
Q

What structures do eosin stain?

A
  • basic or acidophilic (acidic dye)

- cytoplasm, cell walls, extracellular fibres

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7
Q

What types of mucins does AB/PAS stain?

A
  • PAS; neutral and acid

- AB; acid (blocks PAS from staining acid mucins)

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8
Q

What does increased mucin production indicate?

A
  • adenocarcinomas (breast, pancreas, bronchus, stomach, ovary and colon)
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9
Q

What is mucus composed of and what does it coat?

A
  • mucins and inorganic salts suspended in water

- epithelial surfaces (respiratory, GIT, genital tracts)

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10
Q

What does mucus act as?

A
  • diffusion barrier against contact w/noxious substances

- and as lubricant to minimise shear stresses

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11
Q

What are mucins?

A
  • family of large heavy glycosylated proteins

- some membrane bound, some secreted on mucosal surfaces and saliva

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12
Q

Schiff reagent in PAS staining

A
  • dialdehydes react with it

- chromophore restored as modified

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13
Q

Oxidising in 0.5% Periodic Acid

A
  • oxidation of carbohydrates to aldehydes
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14
Q

Washing well in running water

A
  • enhances colour
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15
Q

Three different ways of producing metallic silver

A
  • argentaffin
  • argyrophil
  • ion-exchange
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16
Q

Argentaffin reaction

A
  • stains enterochromaffin cells

- tissues contain reducing groups w/enough strength and quantity to give deposit w/o reducing agent

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17
Q

Argyrophil reaction

A
  • external reducer is used to enhance deposition of silver atoms from silver ions
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18
Q

Ion-exchange reaction

A
  • silver nitrate reacts w/carbonates and phosphates in mineralised bone forming insoluble silver salt
  • silver salts blackened by UV light or hydroquinone solution
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19
Q

Advantages of reticulin staining

A
  • stable (doesn’t fade)
  • densely dark deposits
  • slender objects are thickened
  • very sensitive method
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20
Q

Disadvantages of reticulin staining

A
  • may strip section of slide
  • can be unreliable and capricious
  • stains everything in contact w/ silver solution
  • silver solutions are expensive
  • heavy metal; needs special waste discharge
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21
Q

What is added as a reducing agent in silver impregnation?

A
  • formalin
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22
Q

Treating w/ Sodium Thiosulfate

A
  • excess silver in unprecipitated state is removed
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23
Q

Rinsing in distilled water then treat w/ 4% Iron Alum

A
  • a mordant (enhances silver salts
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24
Q

Reduce in 10% formalin in tap water w/agitation of slide

A
  • reducing agent, causes deposition of metallic silver
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25
Q

Treat w/Ammoniacal Silver Nitrate w/agitation of slide

A
  • silver in a solution which can be easily reduced
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26
Q

Washing off in water and bleaching w/1% Oxalic Acid

A
  • decolourises background stain
27
Q

Treat w/Acidified Potassium Permanganate solution

A
  • oxidising agent which produces aldehyde groups
28
Q

Paraffin wax treatment

A
  • necessary to remove wax embedding medium

- rehydrate sections prior to staining (wax embedded sections are impervious to aqueous and alcoholic dyes)

29
Q

Where are reticulin fibres found in?

A
  • bone marrow
  • lymph nodes
  • spleen
  • liver
  • kidney
30
Q

Von Kossa staining

A
  • used on undecalcified sections of bone tissue
  • none calcium specific stain (reacts w/phosphates/carbonates in mineralised bone/teeth)
  • allows for identification of lesions, metabolic bone disease, abnormal calcium deposition in tissues
31
Q

Martius yellow in MSB staining

A
  • quickly goes in, tightly held in RBCs, quickly washed out of collagen/muscle cytoplasm
32
Q

Brilliant crystal scarlet red in MSB staining

A
  • fills cytoplasm/muscle/collagen but can’t enter RBCs as they’re saturated w/ Martius yellow
33
Q

Aniline blue in MSB staining

A
  • collagen needs to be filled, any excess rinsed off
34
Q

Phosphotungistic acid in MSB staining

A
  • dye excluder/differentiator, competes w/red dye + takes it out of tissue
  • Reaction is stopped when collagen is only faintly stained
35
Q

What is Masson Trichrome stain used for?

A
  • detection of collagen fibres in tissues

- on formalin-fixed wax embedded sections

36
Q

Weigert iron haematoxylin solutions stain what?

A
  • nuclei (deep blue)
37
Q

Beibrich scarlet-acid fuchsin (red) in Masson Trichome staining

A
  • fills cytoplasm/muscle/collagen/RBCs
38
Q

Phosphomolybdic-phosphotungistic acid in Masson Trichrome staining

A
  • dye excluder/differentiator
  • competes w/red dye and takes it out of tissue
  • reaction is stopped when collagen is only faintly stained
39
Q

Aniline blue in Masson Trichrome stain

A
  • fills collagen and excess is rinsed off
40
Q

Results of Masson Trichrome staining

A
  • nuclei - black
  • muscle, cytoplasm and keratin - red
  • collagen - blue
41
Q

Who 2 acid dyes are used in Van Gieson’s stain?

A
  • acid fuchsin

- picric acid

42
Q

What does Van Gieson’s stain differentiate?

A
  • muscle from collagen (yellow/red)

- also identifies bile (bottle green)

43
Q

Acid fuchsin in Van Gieson’s stain

A
  • larger molecules
  • displaces picric acid from collagen fibres
  • has larger pores and allow larger molecules to enter
44
Q

Picric acid in Van Gieson’s stain

A
  • small molecules
  • penetrate all the tissue rapidly
  • only firmly retained in close textured RBCs and muscle
45
Q

Types of Trichrome stains

A
  • Picro Mallory
  • Azan
  • Masson Trichrome
  • Masson Goldner
  • MSB
46
Q

What principles do trichrome stains work on?

A
  • density/permeability of tissues
  • molecular size of dye
  • chemical attraction (acid/base)
47
Q

Stains and results for elastin

A

Stains: Miller’s elastin, van Gieson’s
Results: elastin (black), collagen (red), fibrin (yellow)

48
Q

Stain and result for collagen III

A
  • silver impregnation

- black

49
Q

Stain and results for collagen IV

A
  • PAS/haematoxylin

- Results: basement membrane (pink), nuclei (blue)

50
Q

Stains and results for amyloid

A
  • Crystal violet: amyloid (blue)
  • Sirius red: amyloid (red)
  • thioflavin T: fluorescent dye
  • Congo red: amyloid (red), also gives green birefringence
51
Q

Stains and results for fibrin/collagen

A
  • MSB: collagen (blue), fibrin (red), muscle (red), RBCs (yellow)
  • Masson Trichrome: collagen (blue), muscle and erythrocytes (red)
  • Phosphotungistic acid: fibrin (blue), collagen (orange/red), muscle (blue)
52
Q

Stain in haematology (blood smear)

A
  • Leishman, May Grunwald, Giemsa, Wright etc

- based on Romanowsky staining (combo of methylene blue/eosin)

53
Q

Stain in cytopathology (cervical smear)

A
  • papanicolaou stain
54
Q

Fungi stains in microbiology

A
  • Grocott, PAS - important when monitoring immunodeficiency
55
Q

Bacteria stains in microbiology

A
  • Gram stain
  • Ziehl Neelsen - mycobacterium (TB)
  • Giemsa, Warthin-Starry - helicobacter pylori in stomach (acute/chronic gastritis)
56
Q

Orthochromasia

A
  • regular blue staining of other structures
57
Q

Metachromasia

A
  • purple-red staining of most cell granules w/toluidine blue
58
Q

Colour shift

A
  • certain tissue structures (chromotropes) can stain a different colour to colour of dye solution
59
Q

Classic methods used for neutral lipids

A
  • Oil Red O

- Sudan dyes

60
Q

Neutral lipids

A
  • elective solubility when dye leaves solvent and stains lipid
  • aqueous mountant needed, as neutral lipid will dissolve in dehydrating alcohols
61
Q

Preferred method in most labs for amyloid

A
  • congo red

- allows ‘dichroism’ w/polarised light - apple green birefringence

62
Q

What makes amyloids difficult to demonstrate?

A
  • different amino acid compositions - may stain different

- many tinctorial stains lack specificity

63
Q

What is amyloid resistant to?

A
  • enzyme degradation

- accumulates between cells in many organs

AP0509 Component 2 Exam (21 decks)

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