Connective Tissues - Trichrome, Reticulin, Elastin (Wk 8)

Question 1

What colour does COLLAGEN stain in Van Gieson’s

Answer 1

Red

Question 2

Removes red staining from collagen

Answer 2

Phosphomolybdic acid

Question 3

What colour does MUSCLE stain in Masson’s Trichrome

Answer 3

Red

Question 4

Main coloured staining component in Verhoeff’s

Answer 4

hematoxylin

Question 5

Melanin staining is an ___________ reaction (type of silver rxn)

Answer 5

argentaffin

Question 6

3 highly cellular tissues that reticulin would be found in

Answer 6

Liver
Lymph nodes
Spleen

Question 7

In trichrome, aniline blue would most likely stain what?

Answer 7

Collagen

Question 8

Type of hematoxylin required in the trichrome method

Answer 8

Iron

Question 9

Typical stain for unsaturated lipids?

Answer 9

Oil Red O

Question 10

Reticulin staining is an _________ reaction (type of silver rxn)

Answer 10

argyrophil

Question 11

Stroma consists of ______ and connective tissue fibers

Answer 11

ECM

Question 12

Why are only unsaturated lipids stainable w/ Oil Red O

Answer 12

Stain requires lipids to be liquid at room temp - only UNsaturated lipids can be liquid at RT (saturated lipids are solid)

Question 13

Reagent used as a final differentiating wash in trichrome methods

Answer 13

Acetic acid

Question 14

Chromophore in Oil Red O?

Answer 14

Diazo bonds

Question 15

What 2 reagents do you combine to make ammoniacal silver?

Answer 15

Silver nitrate + ammonium hydroxide

Question 16

This component has the larger pore size in the conventional theory of trichrome staining?

Answer 16

Collagen

Question 17

This is the smaller dye used in Van Gieson’s.

Answer 17

Picric acid

Question 18

This general type of hematoxylin is used when doing Van Gieson’s or Masson’s trichromes due to their acidic nature.

Answer 18

Iron mordanted

Question 19

Final differentiating solution in many trichrome methods

Answer 19

acetic acid

Question 20

Reticulin method - what reagent is used to detect the aldehydes created in the reticulin?

Answer 20

Ammoniacal silver

Question 21

Stain that demonstrates elastin

Answer 21

Verhoeff’s

Question 22

3 main components of Verhoeff’s

Answer 22

Hematoxylin
Ferric chloride
Iodine

Question 23

This elastin stain is derived from lichens

Answer 23

Orcein

Question 24

What colour does COLLAGEN stain in Masson’s trichrome?

Answer 24

Green or Blue

Question 25

Why are the coloured molecules used in lipid stains technically not dyes?

Answer 25

No auxochromes

Question 26

Cause of gray background in Verhoeff's?

Answer 26

Due to iodine which is later removed with alcohol

Question 27

Which component in Van Gieson's is considered to be the cytoplasmic, muscle stain?

Answer 27

Picric acid

Question 28

Routine stain that collagen stains with?

Answer 28

Eosin

Question 29

Which component in Van Gieson's will demonstrate collagen?

Answer 29

Acid Fuschin

Question 30

What component of Gomori's Reticulin stain removes unreduced silver and gold?

Answer 30

Sodium thiosulphate

Question 31

4 components of Movat's Pentachrome and what they demonstrate

Answer 31

Alcian blue - acid mucoproteins (light blue) Hematoxylin - nuclei, elastin (dark blue) Saffron - collagen (yellow) Acid fuchsin - muscle, fibrin (red)

Question 32

This chemical group is blocked by prolonged fixation in formalin and may result in poor trichrome staining.

Answer 32

Amino groups

Question 33

Charge on all tissue components in trichrome method is ___________

Answer 33

cationic

Question 34

Supportive structures of organs are called?

Answer 34

Stroma

Question 35

Why is IRON hematoxylin required to stain nuclei in trichrome stains?

Answer 35

Due to strong acidity of staining solutions (can't use reg hematoxylin)

Question 36

What is used to differentiate in Verhoeff's? What is the term for it?

Answer 36

Ferric chloride Mordant differentiation

Question 37

What colour does MUSCLE stain in Van Gieson's

Answer 37

Yellow

Question 38

Elastin is produced by __________ and ______________

Answer 38

fibroblasts, smooth muscle cells

Question 39

2 functions of ferric chloride in Verhoeff's

Answer 39

Oxidizing agent | Mordant

Question 40

Oxidizes vicinal diols to produce aldehydes in the reticulin stain

Answer 40

Potassium permanganate (could also use periodic acid)

Question 41

This is typically used as a secondary fixative when doing Masson's Trichrome

Answer 41

Bouin's

Question 42

Stain that involves "mordant differentiation"

Answer 42

Verhoeff's

Question 43

Why is Bouin's routinely used as a secondary fixative before trichrome staining?

Answer 43

Restores the lost positive charges from formalin fixation

Question 44

Intermediate reagent used in some trichrome methods (mordant/colourless intermediate dye)

Answer 44

phosphomolybdic acid

Question 45

Stain for saturated lipids?

Answer 45

Sudan Black

Question 46

What component in the reticulin stain reduces ammoniacal silver to metallic silver?

Answer 46

Formalin

Question 47

What is the charge of the dyes used in most trichrome methods?

Answer 47

Anionic

Question 48

Ratio of picric/acid fuchsin in Van Gieson's

Answer 48

50:1

Question 49

3 types of connective tissue fibers

Answer 49

- Reticulin - Elastin - Collagen

Question 50

Usual counterstain in Verhoeff's?

Answer 50

Van Gieson's

Question 51

2 different words that refer to the coloured molecules in lipid stains

Answer 51

Lysochromes | Lipochromes

Question 52

What is the general name for the stains used to differentiate between collagen and muscle.

Answer 52

Trichrome

Question 53

This stain is a simpler and older trichrome method

Answer 53

Van Gieson's

Question 54

In the Gomori's Reticulin silver method, what is used to detect aldehydes in the reticulin fibers?

Answer 54

ammoniacal silver

Question 55

What is the process called in Gomori's Reticulin method which uses gold chloride?

Answer 55

Toning

Question 56

What stains collagen yellow in Movat's pentachrome?

Answer 56

Saffron (not picric acid!)

Question 57

2 most common special stains to demonstrate collagen

Answer 57

Masson's trichrome stain | Van Gieson's stain

Question 58

2 examples of stroma?

Answer 58

Collagen, reticulin

Question 59

Precaution when performing Verhoeff's

Answer 59

Possible to over-differentiate

Question 60

How to properly dispose of silver reagents?

Answer 60

Neutralize with salt (otherwise can form explosive compounds)

Question 61

This reagent removes the connective tissue dye in Masson's Trichrome.

Answer 61

Acetic Acid

Question 62

Why must you avoid prolonged washing in water or dilute alcohol when doing Van Gieson's?

Answer 62

Picric acid is water-soluble

Question 63

What is the sensitizing reagent in reticulin stain?

Answer 63

Iron alum (follows oxidizing agent to enhance contrast)

Question 64

What does it mean when the stain is an argentaffin reaction?

Answer 64

External reducing agent is NOT required to produce a visible product

Question 65

This type of connective tissue fiber would be found in highly cellular tissues such as liver, lymph nodes, spleen

Answer 65

Reticulin

Question 66

Main functional cells of an organ are called?

Answer 66

Parenchyma e.g. hepatocytes

Question 67

This term applies to items which are capable of reducing and binding silver, but require an external reducing agent in order to be visible.

Answer 67

Argyrophil

Question 68

This is another name for Reticulin.

Answer 68

Type III Collagen

Question 69

Purpose of potassium metabisulfite in reticulin stain?

Answer 69

Removes brown discolouration from potassium permanganate (oxidizing agent)

Question 70

Why is using charged slides suggested for silver staining?

Answer 70

Ammoniacal silver is very alkaline which may cause the tissue to lift off the slide