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Histology Exam > Pigment > Flashcards

Pigment Flashcards

1
Q

What are the three groups of pigment?

A
  1. Artifact
  2. Endogenous
  3. Exogenous
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2
Q

What are the two subgroups of endogenous pigments?

A

a. Hematogenous
b. non-hematogenous

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3
Q

Non hematogenous pigments can be subdivided into two categories?

A
  • Lipidic
  • Non-lipidic
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4
Q

What are examples of artifact pigments?

A
  • Formalin pigments
  • Mercury pigments
  • Chrome pigments
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5
Q

What are examples of hematogenous pigments?

A
  • Hemoglobin
  • Hemosiderin
  • porphyrins
  • bile pigments
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6
Q

What are examples of non-hematogenous pigments?

A
  • Melanin (NL)
  • Lipofuchsin (L)
  • Ceroid (L)
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7
Q

What are examples of exogenous pigments?

A
  • Carbon
  • Asbestos fibers
  • Tattoo pigments
  • Metals
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8
Q

How are artifact pigments formed?

A

Usually from fixation

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9
Q

What are exogenous pigments?

A

Formed externally and taken into the body

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10
Q

How are formaldehyde pigments formed?

A

During fixation using acidic formaldehyde solutions.

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11
Q

How can formaldehyde pigments be removed?

A

With saturated alcoholic picric acid prior to staining

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12
Q

How are mercuric pigments formed?

A

While fixing with mercuric fixatives.

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13
Q

How can mercuric pigments be removed?

A

Iodine solution followed by sodium thiosulphate.

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14
Q

What do hemosiderin pigments look like?

A

yellow / brown

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15
Q

Where might hemoglobin be seen, and what stain does it have an affinity for?

A
  • renal tubules post HA and recent hemorrhages
  • stains vividly with acidic dyes
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16
Q

List two methods which will demonstrate iron.

A
  • Prussian Blue
  • Turnbull’s blue reaction
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17
Q

How are bile pigments demonstrated?

A

By oxidizing bilirubin (yellow/brown) to a greenish biliverdin

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18
Q

Where is Melanin found in the human body?

A
  • Hair
  • Skin
  • Retina
  • Iris
  • CNS
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19
Q

What is a malignancy related to melanin called?

A

Melanocarcinoma

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20
Q

What is the principle behind lipofuchsins.

A

Slow progressive oxidation process of lipids and lipoproteins, into a brown pigment.

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21
Q

Which demonstration methods are useful for lipofuchsins?

A
  • PAS
  • Oil Red O
  • SBB
  • Schmorl’s
  • Long Ziehl-Neelson
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22
Q

What are ceroids?

A

Yellow/brown pigments that are rarely seen in humans, and are positively stained by acid fast staining methods.

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23
Q

What are trophi?

A

Large amount of Uric acid and urate crystals which are deposited in soft tissue

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24
Q

How can sodium urates be fixed for demonstration?

A

Requires alcohol fixation

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25
Q

What principle can be employed to demonstrate urates?

A

Argentaffin - Methenamine silver method

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26
Q

What are neuroendocrine cell’s? and what might be demonstrated in these cells?

A
  • Adrenal chromaffin cells
  • Endocrine cells
  • Enterochromaffin cells (GI)
  • “C” cells of the thyroid
  • Pituitary cells
    —-> Cytoplasmic granules <—-
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27
Q

How can cells of the neuroendocrine be demonstrated?

A
  • Argentaffin / Argyrophilic reactions
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28
Q

Which minerals can be demonstrated by special stains?

A
  • Calcium
  • Ferric iron and Ferrous Iron
  • Copper
  • Phosphate
  • Carbon (trioxide??)
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29
Q

What is the purpose of Pearl’s Prussian blue technique?

A

Demonstrate ferric iron in tissue

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30
Q

What is the principle of Pearl’s Prussian blue technique?

A

Detect ferric iron in bound protein complex, endpoint reaction being insoluble coloured pigment.

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31
Q

Which fixative is ideal for Pearl’s Prussian blue technique?

A
  • Alcohol
  • NBF
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32
Q

How large should sections be for the purpose of demonstrating iron?

A

4-5 microns

33
Q

What reagents are implicated in the Pearl’s Prussian blue technique?

A
  • 2% potassium ferrocyanide
  • 2% HCL
  • Nuclear-fast red
34
Q

What will nuclei look like after being stained with the Pearl’s Prussian blue technique?

A

Bright red

35
Q

What will Hemosiderin look like after being stained with the Pearl’s Prussian blue technique?

36
Q

What will the background look like after being stained with the Pearl’s Prussian blue technique?

37
Q

If you are trying to demonstrate iron in section you suspect might have calcium deposits what must you avoid doing? and why?

A

Avoid decalcification because the process removes iron.

38
Q

What is the purpose of the Turnbull blue staining method?

A

Detect ferrous iron in tissue sections

39
Q

Why might a technologist have to use this technique?

A

Because iron is not normally stored as Fe2+, as it is toxic in this state.

40
Q

What is the principle of Turnbull blue staining method?

A

Ferrous iron is turned into an insoluble bright blue pigment by treating sections with potassium ferrocyanide.

41
Q

What are the ideal fixatives to use when performing Turnbull blue staining method?

A
  • Alcohol
  • NBF
42
Q

What reagents are used to make the potassium ferrocyanide solution?

A
  • Potassium ferrocyanide
  • HCL
43
Q

What reagents are used for the Turnbull blue staining method?

A
  • HCL
  • Potassium ferricyanide solution
  • Acetic acid
  • Nuclear-fast red
44
Q

What solutions are used to make the nuclear-fast red in the Turnbull blue staining method?

A
  • Nuclear-fast red
  • Aluminum sulfate
  • Distilled water
45
Q

What will ferrous iron look like after being stained with the Turnbull blue staining method?

46
Q

What is the purpose of the Masson-fontana technique?

A

Demonstration of Argentaffin substances such as melanin.

47
Q

What is the reducing substance used in the Masson-fontana technique?

A

Phenolic groups

48
Q

What is the purpose of using gold chloride in the Masson-fontana technique??

49
Q

What is the purpose of using hypo in the Masson-fontana technique?

A

Fixing agent

50
Q

What is the ideal fixative to use for the Masson-fontana technique?

A
  • NBF
  • Avoid Alcohol
51
Q

What are the reagents used in the Masson-fontana technique?

A
  • Silver nitrate
  • Concentrated ammonium hydroxide
  • Fontana silver solution
  • Gold chloride
  • Sodium thiosulphate
  • Nuclear-fast red
52
Q

What will melanin look like when stained with the Masson-fontana technique?

53
Q

What will Argentaffin granules look like when stained with the Masson-fontana technique?

54
Q

What will Nuclei look like when stained with the Masson-fontana technique?

55
Q

You have a tissue section you suspect has both melanin pigments and argentaffin granules, post staining with the Masson-fontana technique you only see black pigment what would you do?

A

Bleach with Potassium permanganate followed by oxalic acid or peroxide, do bleach out the melanin.

56
Q

What is the purpose of Gomori’s methenamine silver method?

A

Demonstrate urate in tissue

57
Q

What is the principle of Gomori’s methenamine silver method?

A

Urates react with silver to reduce silver to it’s metalic form.

58
Q

What fixative is ideal for Gomori’s methenamine silver method?

A

Absolute alcohol is required

59
Q

Which reagents are used in the Gomori’s methenamine silver method?

A
  • Silver nitrate
  • Methenamine solution
  • Stock methenamine silver nitrate solution
  • Sodium borate
  • working methenamine silver nitrate
  • sodium thiosulphate
  • Light green
60
Q

What will urates look like following staining with Gomori’s methenamine silver method?

61
Q

What will the background stain following Gomori’s methenamine silver method?

62
Q

What are the two main methods for demonstrating Calcium?

A
  • Von kossa technique
  • Alizarin Red S
63
Q

What is the purpose of the Von kossa technique?

A

Demonstrate calcium in tissue sections

64
Q

What is the principle of the Von kossa technique?

A
  • Indirectly stains calcium by reacting with the radicals attached or bound to calcium, as calcium can only be demonstrable when bound to carbon and phosphate
    —-> Argyrophilic reaction <—-
65
Q

What reduces the Silver salts into their metallic form in the Von Kossa technique?

A

Bright lights

66
Q

What removes and un reduced Silver salts from the tissue section in the Von Kossa technique?

67
Q

What reagents are used in the Von Kossa technique?

A
  • Silver nitrate solution
  • Sodium thiosulphate
  • Nuclear-fast red
68
Q

What will calcium salts look like following the Von kossa technique?

A

Black / Brown

69
Q

What will the background look like following the Von kossa technique?

70
Q

What will nuclei look like following the Von kossa technique?

71
Q

You stain a section of tissue using the Von kossa technique and notice formalin pigment had formed on the tissue during microscopic analysis of the section. What could explain this?

A

You used unbuffered formalin

72
Q

You stain a section of tissue using the Von kossa technique and notice the final products of the section are very brown. What could explain this?

A

The section was exposed to artificial lighting

73
Q

What is the purpose of the Alizarin Red S method?

A

Demonstrate calcium in tissue sections

74
Q

What is the principle of the Alizarin Red S method?

A

Calcium forms alizarin red S calcium complex through a chelation process.

75
Q

What are ideal fixatives to use for the Alizarin Red S method?

A
  • NBF
  • Alcoholic formalin
76
Q

What reagents are used for the Alizarin Red S method?

A
  • Alizarin red S
  • Fast green
77
Q

What will calcium look like following the Alizarin Red S method?

A

orange - red

78
Q

What will the background look like following the Alizarin Red S method?

79
Q

What is the benefit of using the Alizarin Red S method in comparison to the Von kossa technique?

A

ARS - is more sensitive to small amounts of calcium, simply adjusting the pH of the alizarin red increases its specificity.

Histology Exam (8 decks)

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