Pigments, Minerals, & Cytoplasmic Granules Staining

Question 1

Define: Endogenous pigment.

Answer 1

Pigments or minerals that arise or are produced from within the body

Question 2

Define: Exogenous pigment

Answer 2

Pigments and minerals that originate from outside the body

Question 3

Define: Hematogenous pigment

Answer 3

Pigment derived from blood

Question 4

Define: Anthracotic pigment

Answer 4

An accumulation of carbon from inhaled industrial pollution, cigarette smoke, or coal dust. The pigment is seen most frequently in the lungs or in lymph nodes from that area

Question 5

Define: Endogenous, non-hematogenous pigment

Question 6

Define: Mineral

Answer 6

Any naturally occurring, homogeneous inorganic substance having a definite chemical composition and characteristic crystalline structure. Can be studied by microincineration an addition to the frequent demonstration by special stains

Question 7

Compare and contrast the argentaffin and argyrophil reactions.

Question 8

Prussian blue:
1. Substance demonstrated:
2. Preferred fixative:
3. Primary reagents & their purpose:
4. Results of the stain:
5. Control material:

Answer 8

1. Detection of ferric (Fe3+) iron in tissues. Aka hemosiderin normally found in liver, spleen, and bone marrow. Can be found idiopathically in idiopathic hemochromatosis (excess iron build up in atypical locations)
2. Alcohol or 10% NBF
3. Potassium ferrocyanide 2%:
   Hydrochloric acid solution 2%:
   Nuclear-fast red (Kernechtrot) solution:
4. Nuclei and hemofuchsin - bright red
   Hemosiderin - blue
   Background - pink
5. QC with section containing ferric iron

Question 9

Turnbull blue:
1. Substance demonstrated:
2. Preferred fixative:
3. Primary reagents & their purpose:
4. Results of the stain:
5. Control material:

Answer 9

1. Detection of ferrous (Fe2+) iron in tissues. Not normally stored in tissue in this form as it is toxic. Hemochromatosis can cause storage in this form in spleen and liver. Turnbull is also used during Schmorl technique.
2. Alcohol or 10% NBF
3. HCL 0.06N:
   Potassium ferricyanide:
   Acetic acid 1%:
   Nuclear-fast red:
4. Ferrous iron (Fe2+) - Blue
   Background - Pink-red
5. A section containing ferrous iron (obviously)

Question 10

Schmorl ferric-ferricyanide reduction test:
1. Substance demonstrated:
2. Preferred fixative:
3. Primary reagents & their purpose:
4. Results of the stain:
5. Control material:

Answer 10

1. Detection of reducing substances present in tissues, such as melanin, argentaffin granules, and formalin pigment.
2. 10% NBF
3. Ferric chloride stock 1%:
   Potassium ferricyanide stock 0.1%:
   Ferric chloride-potassium/ferricyanide working solution:
   Mayer mucicarmine solution:
   Mentanil yellow 0.25%:
4. Reducing substances - Blue/green
   Goblet cells, mucin - Rose
   Background - Yellow/green
5. A section containing melanin (skin) or argentaffin granules (appendix)

Question 11

Fontana-Masson:
1. Substance demonstrated:
2. Preferred fixative:
3. Primary reagents & their purpose:
4. Results of the stain:
5. Control material:

Answer 11

1. Demonstrates argentaffin substances such as melanin, argentaffin granules of carcinoid tumors and some neurosecretory granules.
2. 10% NBF; Alcohol is a no no because it dissolves argentaffin granules.
3. Silver nitrate 10% solution
   Fontana silver solution
   Gold chloride 0.2% solution
   Sodium thiosulfate 5% solution
   Nuclear-fast red
4. Melanin - Black
   Argentaffin granules - Black
   Nuclei - Pink
5. A section containing melanin (skin) or argentaffin granules (intestine/appendix)

Question 12

Grimelius:
1. Substance demonstrated:
2. Preferred fixative:
3. Primary reagents & their purpose:
4. Results of the stain:
5. Control material:

Answer 12

1. Demonstrates argyrophil granules in the neurosecretory tumors.
2. 10% NBF
3. Silver nitrate 1% solution
   Acetic acid 0.2M solution
   Sodium acetate 0.2M solution
   Acetic acid-sodium acetate buffer 0.2M solution
   Working silver solution
   Reducing solution
   Nuclear-fast red solution
4. Argentaffin granules - Dark brown to black
   Agyrophil granules - Dark brown to black
   Nuclei - Red
   Background - Pale yellow/brown
5. An argyrophil-positive carcinoid tumors is preferred, but section of small intestine can be used.

Question 13

Churukian-Schenk:
1. Substance demonstrated:
2. Preferred fixative:
3. Primary reagents & their purpose:
4. Results of the stain:
5. Control material:

Answer 13

1. Demonstration of argyrophil granules in neurosecretory tumors.
2. 10% NBF
3. Citric acid 0.3 solution:
   Acidified water:
   Silver nitrate 0.5% solution:
   Reducing solution:
   Nuclear-fast red solution:
4. Argyrophil granules - Black
   Argentaffin substances - Black
   Nuclei - Red
   Background - Yellow/brown
5. An argyrophil-positive carcinoid tumors is preferred, or a section of small intestine.

Question 14

Gomori methenamine-silver:
1. Substance demonstrated:
2. Preferred fixative:
3. Primary reagents & their purpose:
4. Results of the stain:
5. Control material:

Answer 14

1. Demonstration of urates in tissues. Caused by disturbances in the uric-acid metabolism (aka gout). Collections of urate crystals are known as gouty tophi.
2. Absolute alcohol is required.
3. Silver nitrate, 5% solution
   Methenamine 3% solution
   Stock methenamine-silver nitrate solution
   Sodium borate (borax) 5% solution
   Working methenamine-silver nitrate solution
   Sodium thiosulfate 3% solution
   Light green stock solution
   Working light green solution
4. Urates - Black
   Background - Green
5. A section containing urates (-_-)

Question 15

Hall (Fouchet):
1. Substance demonstrated:
2. Preferred fixative:
3. Primary reagents & their purpose:
4. Results of the stain:
5. Control material:

Answer 15

1. Demonstrates the presence of bilirubin in tissue, distinguishing bile pigments from other pigments in tissue.
2. 10% NBF
3. Ferric chloride 10% solution
   Fouchet reagent
   Van Gieson solution
4. Bile or bilirubin - Emerald green to olive drab
   Background - Yellow
5. Tissue containing bile must be used (gallbladder).

Question 16

Von Kossa:
1. Substance demonstrated:
2. Preferred fixative:
3. Primary reagents & their purpose:
4. Results of the stain:
5. Control material:

Question 17

Rhodanine:
1. Substance demonstrated:
2. Preferred fixative:
3. Primary reagents & their purpose:
4. Results of the stain:
5. Control material:

Question 18

Describe Prussian blue reaction and state how it differs from the Turnbull blue reaction.

Question 19

Identify 3 body sites where melanin is found.

Question 20

List at least 4 other granules or minerals not demonstrated by the techniques in (3) and give at least 1 method for demonstrating each.

Question 21

How can anthracotic pigment be differentiated from other dark brown to black pigments (eg, formalin, melanin)?