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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
Treat w/Ammoniacal Silver Nitrate w/agitation of slide
- silver in a solution which can be easily reduced
26
Washing off in water and bleaching w/1% Oxalic Acid
- decolourises background stain
27
Treat w/Acidified Potassium Permanganate solution
- oxidising agent which produces aldehyde groups
28
Paraffin wax treatment
- necessary to remove wax embedding medium | - rehydrate sections prior to staining (wax embedded sections are impervious to aqueous and alcoholic dyes)
29
Where are reticulin fibres found in?
- bone marrow - lymph nodes - spleen - liver - kidney
30
Von Kossa staining
- 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
Martius yellow in MSB staining
- quickly goes in, tightly held in RBCs, quickly washed out of collagen/muscle cytoplasm
32
Brilliant crystal scarlet red in MSB staining
- fills cytoplasm/muscle/collagen but can't enter RBCs as they're saturated w/ Martius yellow
33
Aniline blue in MSB staining
- collagen needs to be filled, any excess rinsed off
34
Phosphotungistic acid in MSB staining
- dye excluder/differentiator, competes w/red dye + takes it out of tissue - Reaction is stopped when collagen is only faintly stained
35
What is Masson Trichrome stain used for?
- detection of collagen fibres in tissues | - on formalin-fixed wax embedded sections
36
Weigert iron haematoxylin solutions stain what?
- nuclei (deep blue)
37
Beibrich scarlet-acid fuchsin (red) in Masson Trichome staining
- fills cytoplasm/muscle/collagen/RBCs
38
Phosphomolybdic-phosphotungistic acid in Masson Trichrome staining
- dye excluder/differentiator - competes w/red dye and takes it out of tissue - reaction is stopped when collagen is only faintly stained
39
Aniline blue in Masson Trichrome stain
- fills collagen and excess is rinsed off
40
Results of Masson Trichrome staining
- nuclei - black - muscle, cytoplasm and keratin - red - collagen - blue
41
Who 2 acid dyes are used in Van Gieson's stain?
- acid fuchsin | - picric acid
42
What does Van Gieson's stain differentiate?
- muscle from collagen (yellow/red) | - also identifies bile (bottle green)
43
Acid fuchsin in Van Gieson's stain
- larger molecules - displaces picric acid from collagen fibres - has larger pores and allow larger molecules to enter
44
Picric acid in Van Gieson's stain
- small molecules - penetrate all the tissue rapidly - only firmly retained in close textured RBCs and muscle
45
Types of Trichrome stains
- Picro Mallory - Azan - Masson Trichrome - Masson Goldner - MSB
46
What principles do trichrome stains work on?
- density/permeability of tissues - molecular size of dye - chemical attraction (acid/base)
47
Stains and results for elastin
Stains: Miller's elastin, van Gieson's Results: elastin (black), collagen (red), fibrin (yellow)
48
Stain and result for collagen III
- silver impregnation | - black
49
Stain and results for collagen IV
- PAS/haematoxylin | - Results: basement membrane (pink), nuclei (blue)
50
Stains and results for amyloid
- Crystal violet: amyloid (blue) - Sirius red: amyloid (red) - thioflavin T: fluorescent dye - Congo red: amyloid (red), also gives green birefringence
51
Stains and results for fibrin/collagen
- 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
Stain in haematology (blood smear)
- Leishman, May Grunwald, Giemsa, Wright etc | - based on Romanowsky staining (combo of methylene blue/eosin)
53
Stain in cytopathology (cervical smear)
- papanicolaou stain
54
Fungi stains in microbiology
- Grocott, PAS - important when monitoring immunodeficiency
55
Bacteria stains in microbiology
- Gram stain - Ziehl Neelsen - mycobacterium (TB) - Giemsa, Warthin-Starry - helicobacter pylori in stomach (acute/chronic gastritis)
56
Orthochromasia
- regular blue staining of other structures
57
Metachromasia
- purple-red staining of most cell granules w/toluidine blue
58
Colour shift
- certain tissue structures (chromotropes) can stain a different colour to colour of dye solution
59
Classic methods used for neutral lipids
- Oil Red O | - Sudan dyes
60
Neutral lipids
- elective solubility when dye leaves solvent and stains lipid - aqueous mountant needed, as neutral lipid will dissolve in dehydrating alcohols
61
Preferred method in most labs for amyloid
- congo red | - allows 'dichroism' w/polarised light - apple green birefringence
62
What makes amyloids difficult to demonstrate?
- different amino acid compositions - may stain different | - many tinctorial stains lack specificity
63
What is amyloid resistant to?
- enzyme degradation | - accumulates between cells in many organs

AP0509 Component 2 Exam (21 decks)

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