Staining

Question 1

What is tinctorial staining?

Answer 1

Dyes added in turn, each bind to different components and stain a different colour (work like textile dyes)

Question 2

What is histochemical staining?

Answer 2

Stains that utilise specific chemical reactions b/w chemicals and tissue components, resulting in a coloured compound

Question 3

Perl’s Prussian Blue is an example of tinctorial or histochemical staining?

Answer 3

Histochemical

Question 4

Periodic Acid Schiff (PAS) staining is an example of tinctorial or histochemical staining?

Answer 4

Histochemical

Question 5

Martius Scarlett Blue (MSB) staining is an example of tinctorial or histochemical staining?

Answer 5

Tinctorial

Question 6

Give an example of tinctorial staining

Answer 6

Martius Scarlett Blue (MSB)

Question 7

Give an example of histochemical staining

Answer 7

Perl’s Prussian Blue

Periodic Acid Schiff reaction

Question 8

Why do we stain sections?

Answer 8

Tissue processed into wax = fairly colourless
Insufficient refractive index for brightfieqld light microscopy
Insufficient contrast for microscopy

Question 9

What are the three ways stains colour tissues?

Answer 9

Stain-Tissue interactions (electrostatic interactions, vDWs, H bonding, covalent bonding)
Solvent-Solvent interactions (hydrophobic effect)
Stain-Stain interactions (dye molecules attract each other, form aggregates)

Question 10

Why is stain retained in tissues?

Answer 10

Not always

Will only be retained if the stain has a higher affinity for the tissue than the surrounding subsequent solvents

Question 11

What are the 3 things that can be adjusted to control staining?

Answer 11

Rate of reagent uptake
Rate of reaction
Rate of reagent loss

Question 12

A mordant must be added to haematoxylin before it will stain tissues. Why?

Answer 12

Ensure it is sufficiently positively charged to bind to negative tissue components

Question 13

What are the two main things that need to be done to haematoxylin before it will stain tissues?

Answer 13

Oxidised and a mordant added

Question 14

Haematoxylins are classified according to their mordant. What are the most commonly used?

Answer 14

Alum haematoxylins (+iron haematoxylins)

Question 15

Haemtoxylin staining can be progressive or regressive. What does this mean?

Answer 15

Progressive - left on tissue for specific time period until staining is optimal
Regressive - overstrained then differentiated out using acid alcohol followed by a blue-ing step

Question 16

Eosin is commonly mixed with what to sharpen staining?

Answer 16

Weak acetic acid solution

Question 17

Eosin is differentiated using what?

Answer 17

Water

Question 18

What the advantages of the H&E?

Answer 18

Inexpensive
Readily available reagents
Easy to use
Easy to modify
Easy to automate
Provides good staining of the majority of tissue components

Question 19

What does haematoxylin stain and what colour?

Answer 19

Stains acidic structures purple/blue (eg. cell nuclei, organelles w/ RNA)

Question 20

What does eosin stain and what colour?

Answer 20

Stains basic eosinophilic structures pink/red (inc. cytoplasm, cell walls, extracellular fibres)

Question 21

What is Congo Red used to stain?

Answer 21

Amyloid (β amyloid fibrils)

Question 22

What is Alcian Blue/PAS used to stain?

Answer 22

(Carbohydrates)

Mixed mucins

Question 23

What is Periodic Acid Schiff (PAS) used to stain?

Answer 23

(Carbohydrates)
Neutral mucins
Glycogen
Glycoproteins

Question 24

What is Masson’s Trichrome used to stain?

Answer 24

Connective muscle tissue - collagen, fibrin & muscle, nuclei

Question 25

What is Perl's Prussian Blue used to stain?

Answer 25

Iron

Question 26

What is Oil Red O used to stain?

Answer 26

Triglycerides and lipids

Question 27

What is Reticulin used to stain?

Answer 27

Reticulin fibres

Question 28

What is Alcian Blue used to stain?

Answer 28

(Carbohydrates) | Acidic mucins

Question 29

Alcian Blue, PAS and a combination of both can all be used to stain what?

Answer 29

Carbohydrates (mucins and glycoproteins)

Question 30

Alcian Blue, PAS and a combination of both can all be used to stain carbohydrates (mucins and glycoproteins). What is a factor that may affect which one is used?

Answer 30

Where the section was located/Acidity of the mucins

Question 31

Where may acidic mucins be found and what can be used to stain them?

Answer 31

Colon and oesophagus | Alcian Blue

Question 32

Where may neutral mucins be found and what can be used to stain them?

Answer 32

Stomach, cervix/vagina, bronchus, prostate | PAS

Question 33

Where may mixed mucins be found and what can be used to stain them?

Answer 33

Salivary | Alcian Blue/PAS

Question 34

What is Rhodanine used to stain?

Answer 34

Copper

Question 35

What is Orcein used to stain?

Answer 35

Copper

Question 36

What is Masson Fontana used to stain?

Answer 36

Melanin

Question 37

What is used to stain amyloid?

Answer 37

Congo red

Question 38

What is used to stain carbohydrates (acidic mucins)?

Answer 38

Alcian blue

Question 39

What is used to stain carbohydrates (neutral mucins)?

Answer 39

PAS

Question 40

What is used to stain carbohydrates (mixed mucins)?

Answer 40

Alcian Blue/PAS

Question 41

What can be used to stain copper?

Answer 41

Rhodanine, Orcein

Question 42

What can be used to stain melanin?

Answer 42

Masson Fontana

Question 43

What can be used to stain glycogen?

Answer 43

PAS

Question 44

What is Cresyl Violet used to stain?

Answer 44

Neuronal tissue

Question 45

What can be used to stain neuronal tissue?

Answer 45

Cresyl Violet

Question 46

What can be used to stain reticulin fibres?

Answer 46

Reticulin

Question 47

What can be used to stain lipids?

Answer 47

Oil red O, Sudan black

Question 48

What can Sudan Black be used to stain?

Answer 48

Lipids

Question 49

Congo Red stains amyloid fibres what colour?

Answer 49

Red under Brightfieqld light microscopy | Apple green under polarised light

Question 50

What can be used to stain elastic fibres?

Answer 50

Miller's Verhoeff's IHC

Question 51

What can Miller's stain be used to identify?

Answer 51

Elastin/elastic fibres

Question 52

What can Verhoeff's be used to identify?

Answer 52

Elastin/elastic fibres

Question 53

What can be used to stain fibrin?

Answer 53

Phosphotungsten acid haematoxylin (PTAH) | Trichrome stains - Masson Trichrome, Martius Scarlett Blue (MSB)

Question 54

Give an example of a trichrome stain

Answer 54

Masson trichrome | Martius Scarlett Blue

Question 55

What does phosphotungsten acid haematoxylin (PTAH) stain?

Answer 55

Fibrin

Question 56

What colour does phosphotungsten acid haematoxylin (PTAH) stain?

Answer 56

Blue - fibrin | Orange - other connective tissue

Question 57

What colour does Masson Trichrome stain?

Answer 57

Red - fibrin and muscle Blue - collagen Brown/black - nuclei

Question 58

What colour does Martius Scarlett Blue stain?

Answer 58

Red - fibrin Blue - collagen Yellow - RBCs

Question 59

What does Masson Trichrome stain?

Answer 59

Fibrin and muscle (red) Collagen (blue) Nuclei (brown/black)

Question 60

What does Martius Scarlett Blue stain?

Answer 60

Fibrin (red) Collagen (blue) Yellow (RBCs)

Question 61

Describe the principles of trichrome staining

Answer 61

Three different stains are added to the section in a particular sequence, dependent on their molecular size/penetration rate

Question 62

Which are some stains that can be used to enhance visualisation of bacteria?

Answer 62

Gram stain Ziehl-Neelson (ZN) Giemsa

Question 63

Which stain can be used to stain Hepatitis B?

Answer 63

Orcein | IHC

Question 64

How are viruses best visualised in CellPath?

Answer 64

Electron microscopy | Can used some special stains to visualise viral inclusion but IHC best

Question 65

What are two histologically significant viruses?

Answer 65

HSV HPV Hep B CMV