Staining Techniques

Question 1

What is the most common stain used?

Answer 1

Haematoxylin & Eosin (H&E)

Question 2

What are the features of Haematoxylin? (3)

Answer 2

• Stains acidic structures
• Stains Nuclei, Ribosomes and Rough Endoplasmic Reticulum (sites where RNA + DNA are present)
• Stains structures Purplish Blue

Question 3

What are the features of Eosin? (3)

Answer 3

• Stains basic structures
• Stains cytoplasmic proteins
• Stains red/pink

Question 4

What does Periodic Acid-Schiff Reaction (PaS) Stain?

Answer 4

Stains Complex Carbohydrates Magenta

Question 5

Which stain stains complex Carbohydrates Magenta?

Answer 5

Periodic acid-Sciff Reaction (PaS)

Question 6

What feature of GI and Respiratory Tracts would Periodic acid-Schiff stain Magenta?

Answer 6

Mucin from goblet cells

Question 7

What 6 features are PaS positive?

Answer 7

• Basement membranes
• Brush borders
• Cartilage
• Collagen (to some extent)
• Glycogen
• Mucin

Question 8

Masson’s Trichrome is what type of technique?

Answer 8

A Connective Tissue Technique, for supportive tissue, especially collagen

Question 9

What is stained Blue by Masson’s Trichrome?

Answer 9

Nuclei and Basophilic Structures

Question 10

What is stained Green/Blue by Masson’s Trichrome?

Answer 10

Collagen (colour dependent on stain variant)

Question 11

What four structures are stained Bright Red by Masson’s Trichrome?

Answer 11

Cytoplasm, Muscles, Erythrocytes, Keratin

Question 12

What is stained by Alcian Blue? (2)

Answer 12

Mucin, Cartilage

Question 13

What stains are most often used along with Alcian Blue?

Answer 13

van Gieson (turns Blue-> Green) or Haematoxylin & Eosin

Question 14

What stains Mucin or Cartilage Blue?

Answer 14

Alcian Blue

Question 15

van Gieson is what type of method?

Answer 15

Connective Tissue Method

Question 16

What tissue is van Gieson especially used for? (2)

Answer 16

Blood vessels and Skin

Question 17

What is stained Red by van Gieson?

Answer 17

Collagen

Question 18

What is stained Blue by van Gieson?

Answer 18

Nuclei

Question 19

What is stained Yellow by van Gieson? (2)

Answer 19

Erythrocytes and Cytoplasm

Question 20

When van Gieson is combined with an elastin stain, what colour does Elastin stain?

Answer 20

Blue/Black

Question 21

What colour does a Reticulin stain Reticulin fibres?

Answer 21

Blue/Black

Question 22

Which stains are often used to counterstain Nuclei along with a Reticulin Stain?

Answer 22

Haematoxylin (Purple-Blue) or Neutral Red (Red)

Question 23

What is Azan used to stain? (2)

Answer 23

Connective Tissue or Fine Cytological Detail (especially Epithelium)

Question 24

What is stained Bright Red by Azan?

Answer 24

Nuclei

Question 25

What is stained Blue by Azan? (3)

Answer 25

Collagen, Basement Membrane, Mucin

Question 26

What is stained Orange/Red by Azan? (2)

Answer 26

Muscles, Erythrocytes

Question 27

What tissues is Giemsa used for?

Answer 27

Smears, including blood cells and bone marrow

Question 28

What is stained Dark Blue/Violet by Giemsa?

Answer 28

Nuclei

Question 29

What is stained Pale Blue by Giemsa?

Answer 29

Cytoplasm

Question 30

What is stained Pale Pink by Giemsa?

Answer 30

Erythrocytes

Question 31

What uncommon fixation medium can Toluidine Blue be used on?

Answer 31

Epoxy Resin sections

Question 32

What tissues is Toluidine Blue often used on?

Answer 32

Glomeruli and Nerves (on HR investigation)

Question 33

What does Toluidine Blue stain reddish purple?

Answer 33

Mast Cell Granules (in Paraffin) (called Metachromasia)

Question 34

What is Goldner's Trichrome used to stain, and in what fixation medium?

Answer 34

Acrylic Resin stains of undecalcified bone Distinguishes mineralised bone and unmineralised osteoid

Question 35

Which component of Goldner's Trichrome stains Nuclei

Answer 35

Haematoxylin

Question 36

Which alternative to Goldner's Trichrome for distinguishing minerality of bone does not show cellular detail?

Answer 36

von Kossa Stain (a silver method)

Question 37

When were Silver and Gold Methods most used?

Answer 37

End of the 19th Century

Question 38

What are Silver and Gold methods useful in identifying?

Answer 38

Fine structures, such as cell processes: - Neurones, Motor End Plates, Intercellular Junctions

Question 39

What colours do Silver and Gold Methods stain sections?

Answer 39

Black, Brown and Golden

Question 40

What are Nissl and methylene blue used to stain?

Answer 40

The Rough Endoplasmic Reticulum of Neurones

Question 41

What are clumps of Rough ER seen in Neurones called?

Answer 41

Nissl Substance

Question 42

What stains are used for the Rough ER of Neurones

Answer 42

Nissl and methylene blue

Question 43

What do Sudan Black and Osmium stain?

Answer 43

Lipid Containing Structures (e.g. Myelin)

Question 44

What colour do Sudan Black and Osmium stain lipid containing structures?

Answer 44

Brown/Black

Question 45

Which stain of lipid-containing structures also provides contrast in EM

Answer 45

Osmium

Question 46

How are immunohistology antibodies produced?

Answer 46

Antigen is injected into lab animal Antibodies are produced These are harvested and propagated using monoclonal technology

Question 47

How are immunohistology stains applied?

Answer 47

Antibody solution placed over a section on a glass slide Excess antibody solution is washed away

Question 48

What is the antibody prelinked to in immunohistology stains?

Answer 48

An indicator substance, often a fluorescent substance or an enzyme

Question 49

Which fluorescent substance is often used as an indicator in Immunohistology?

Answer 49

Fluorescein

Question 50

What is it called when fluorescein is used as the indicator substance in Immunohistology?

Answer 50

Immunofluorescence

Question 51

What is the constraint of Immunofluorescence?

Answer 51

It requires a fluorescence microscope

Question 52

How does the use of an enzyme indicator work in Immunohistology?

Answer 52

Section is bathed in Immunohistology marker solution, then excess is washed away Solution of substrate is laid over the tissue section The enzyme converts colourless substrate to coloured product After a period of incubation the product is visible using LM

Question 53

What enzyme is commonly used in Immunohistology?

Answer 53

Horseradish peroxidase

Question 54

What is it called when peroxidase is used as the indicator substance in Immunohistology?

Answer 54

Immunoperoxidase technique

Question 55

Which immunohistology technique is useful with electron microscopy?

Answer 55

Immunogold labelling