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Special Stains Flashcards

1
Q

What is the purpose of PAS?

A

The demonstration of polysaccharides, neutral mucosubtances, and basement membranes.

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2
Q

What is the principle of PAS?

A

Based on the oxidation of certain tissue elements to aldehydes by periodic acid
Periodic acid oxidizes glycols to aldehydes which are made visible with Schiff’s reagent

Carbohydrates: Magenta

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3
Q

What is the quality (positive) control for PAS?

A

Kidney is the most sensitive control

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4
Q

How do you test the quality of Schiff’s Reagent?

A

Add Schiff reagent to formaldehyde
If the solution quickly turns red/purple it is good
if the reaction is delayed and the color is a deep blue-purple it is bad

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5
Q

What is the purpose of PAS w/ diastase digestion?

A

The demonstration of glycogen in tissue sections

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6
Q

What is the principle of PAS w/ diastase digestion?

A

Diastase and alpha-amylase act on glycogen to depolymerize it into smaller sugar units that are washed out of the section

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7
Q

What is the quality (positive) control of PAS w/ diastase digestion?

A

2 control sections of liver containing glycogen

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8
Q

What is the purpose of Mayer Mucicarmine?

A

Staining of “epithelial” mucin in tissue sections

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9
Q

What is the principle of Mayer Mucicarmine?

A

Stains carboxylated and sulfonated mucins, but not neutral mucins
Aluminum forms a chelation complex with carmine which attaches to the acid groups of mucins

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10
Q

How do you ID adenocarcinomas?

A

Using Mayer Mucicarmine

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11
Q

What is the quality (positive) control of Mayer Mucicarmine?

A

A section of unautolyzed colon, small intestine, or appendix

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12
Q

What is the purpose of Alcian Blue, pH 2.5?

A

The demonstration of acid mucopolysaccharides

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13
Q

What is the principle of Alcian Blue, pH 2.5?

A

When used at pH 2.5, alcian blue stains both sulfated and carboxylated acid mycopolysaccharides and sulfated and carboxylated sialomucins (glycoproteins).

forms salt linkages with the acid groups of acid mucopolysaccharides

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14
Q

What is the quality (positive) control of Alcian Blue, pH 2.5?

A

A section of unautolyzed small intestine, appendix, or colon

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15
Q

What is the purpose of Alcian Blue, pH 1.0?

A

The demonstration of sulfated mucosubstances

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16
Q

What is the principle of Alcian Blue, pH 1.0?

A

When used at pH 1.0, alcian blue only stains sulfated acid mucopolysaccharides and sialomucins (glycoproteins).

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17
Q

What is the quality (positive) control of Alcian Blue, pH 1.0?

A

A section of unautolyzed small intestine, appendix, or colon

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18
Q

What is the purpose of Alcian Blue with Hyaluronidase?

A

To differentiate epithelial from connective tissue mucins

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19
Q

What is the principle of Alcian Blue with Hyaluronidase?

A

Staining will disappear or be dramatically reduced when tissue sections containing hyaluronic acid, chondroitin sulfate A, or chondroitin sulfate C are digested with testicular hyaluronidase

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20
Q

What is the quality (positive) control of Alcian Blue with Hyaluronidase?

A

2 sections of umbilical cord should be used

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21
Q

What is the purpose of Alkaline Congo Red?

A

The demonstration of amyloid in tissue

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22
Q

What is the principle of Alkaline Congo Red?

A

Hydrogen binding to carbohydrates
Elongated hydrophobic structure will bind amyloid fibrils under specific conditions
Pretreatment with alkali aids in the release of native internal hydrogen bonds between adjacent protein chains, making more potential sites available for dye binding.
Linear configuration of amyloid allows azo and amine groups of the dye to form hydrogen bonds with similarity space hydroxyl radicals of the amyloid

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23
Q

What is the quality (positive) control of Alkaline Congo Red?

A

Sections containing amyloid must be used

large long standing deposits give less intense reactions than small, newly formed deposits.

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24
Q

What is a unique feature of amyloid binding in Congo Red staining?

A

Dichroism
reddish staining in bright field
bright apple green birefringence under polarized light

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25
What is the purpose of Crystal Violet?
Good rapid screening method for amyloid but not as specific as Congo Red
26
What is the principle of Crystal Violet?
Metachromatic staining of amyloid is because of mucopolysaccharide content addition of the acid to the staining solution will prevent overstaining of cytoplasmic components Lacks specificity
27
What is the quality (positive) control of Crystal Violet?
A section containing amyloid must be used
28
How is ground substance of connective tissue proper demonstrated?
with carbohydrate staining techniques
29
What differentiates type 1 collagen from smooth muscle?
Masson and Gomori trichrome and van Gieson stain differentiate collagen from smooth muscle
30
What staining allows you to see elastic fibers?
They require Verhoeff iron hematoxylin, Weigert resorcin fuchsin, orcein, or Gomori aldehyde fuchsin stains
31
What staining techniques are used to visualize the basement membrane?
Techniques that demonstrate carbohydrate component
32
What is the glomerular basement membrane stained with?
PAS and Silver technique
33
What is the purpose of Masson's Trichrome?
To differentiate collagen from smooth muscle in tumors | to ID increases in collagenous tissue in diseases
34
What is the principle of Masson's Trichrome?
3 dyes are used Sections are first stained with an acid stain (like Biebrich Scarlet) (stains acidophilic tissue elements like cytoplasm, muscle, and collagen) Sections are then treated with phosphotungstic and/or phosphomolybdic acid (biebrich scarlet diffuses out of the collagen but not the cytoplasm)
35
What is the quality (positive) control of Masson's Trichrome?
Practically every tissue has an internal control, so no other control sections are needed If control is desired; uterus, small intestine, appendix, or fallopian tube will provide good material
36
What is the purpose of van Gieson Picric Acid - Acid Fuchsin Stain?
Excellent counterstain for methods like Verhoeff-van Gieson (VVG) can be used as a primary connective tissue stain
37
What is the principle of van Gieson Picric Acid - Acid Fuchsin Stain?
in strongly acidic solution, collagen is selectively stained by acid fuchsin (an acid aniline dye). Picric acid provides the acidic pH necessary and acts as a stain for muscle and cytoplasm. Low pH important, selective staining will not occur at higher pH levels. Addition of 0.25mL of HCL to 100 mL of van Gieson will sharpen differentiation between collagen and muscle If picric acid solution is not saturated, collagen may stain pale pink to pale orange, and collagen, cytoplasm, and muscle may stain the same color
38
What is the quality (positive) control of van Gieson Picric Acid - Acid Fuchsin Stain?
Practically every tissue has an internal control, so no other control sections are needed If control is desired; uterus, small intestine, appendix, or fallopian tube will provide good material
39
What is the purpose of Verhoeff Elastic Stain?
To demonstrate pathological changes in elastic fibers or normal elastic tissue to determine whether or not the blood vessels have been invaded by tumor
40
What is the principle of Verhoeff Elastic Stain?
tissue is overstained with a soluble lake of hematoxylin-ferric chloride-iodine. Ferric chloride and iodine serve as mordants and have an oxidizing function that converts hematoxylin to hematein. Regressive method. Differentiation is accomplished by using excess mordant to break the tissue-mordant-dye complex. Sodium thiosulfate is used to remove excess iodine. Van Gieson is the most common counterstain
41
What is the quality (positive) control of Verhoeff Elastic Stain?
A section of aorta embedded on edge or cross-section of muscular artery
42
What staining technique would be used to view cirrhosis of the Liver?
Masson's Trichrome
43
What technique is used to demonstrate nerve fibers and neurofibrils?
Silver Methods
44
What is used to demonstrate Myelin?
Luxol Fast Blue and Iron Hematoxylin are commonly used for demonstration of the myelin sheath
45
What is the purpose of Cresyl Violet Method I?
Identification of neurons in tissue sections | Demonstration of the loss of nissl substance (chromatolysis)
46
What is the principle of Cresyl Violet Method I?
Nissl Substance is very basophilic and will very sharply stain with basic aniline dyes Can be demonstrated by varying pH and degree of differentiation
47
What is the quality (positive) control of Cresyl Violet Method I?
A section of spinal cord
48
What is the purpose of Cresyl Violet Method II?
ID of neurons in tissue sections | Demonstration of loss of nissl sbstance
49
What is the principle of Cresyl Violet Method II?
Uses cresyl echt violet at an acid pH Staining is restricted to nuclei and to DNA- and RNA- containing structures contrast of nissl substance and nuclei with background is enhanced
50
What is the quality (positive) control of Cresyl Violet Method II?
A section of spinal cord
51
What stain is primarily used to demonstrate spirochete bacteria?
Silver Methods
52
What stain can be used to classify bacteria?
Gram stains can also be used to classify bacteria. Gram+ bacteria stain deep blue Gram- bacteria remain unstained or are stained by counterstain.
53
What is the purpose of Kinyoun Acid-Fast Stain?
To detect the presence of acid-fast mycobacteria in tissue sections
54
What is the principle of Kinyoun Acid-Fast Stain?
lipoid capsule of the acid-fast organism takes up carbol-fuchsin and resists decolorization with dilute mineral acid. Carbol-fuchsin is more soluble in the lipids of the cell wall than in acid-alcohol, but is readily removed from bacteria that lack the waxy capsule. Staining is enhanced by phenol and alcohol. Stains red for mycobacteria uses heat to permeabilize cell walls
55
What is the quality (positive) control of Kinyoun Acid-Fast Stain?
Tissue containing acid-fast organisms must be used for a control Millipore-filtered water must be used in the waterbath
56
What is the purpose of PAS for Fungi?
Demonstration for fungi in tissue
57
What is the principle of PAS for Fungi?
Polysaccharides in the fungal cell walls are oxidized by the periodic acid to aldehydes aldehydes react with schiff reagent to yield rose-colored fungi
58
What is the quality (positive) control of PAS for Fungi?
A section containing fungi
59
What is the purpose of Prussian Blue?
Detection of ferric iron in tissues
60
What is the principle of Prussian Blue?
Detects the ferric ion in loosely bound protein complexes (like hemosiderin). Strongly bound iron (like that in hemoglobin) will not react. Sections are treated with acidic solution of potassium ferrocyanide and any ferric iron present reacts to form an insoluble blue pigment.
61
What is the quality (positive) control of Prussian Blue?
A section containing ferric iron
62
What is the purpose of H&E?
Stain negatively charged items with hematoxylin | Stain positively charged items with eosin
63
What is the purpose of Reticulin?
Stains reticular fibers and BM
64
What stain is used to diagnose diseases of the GI tract?
Alcian Blue
65
How does Gram Stain work?
Stain with crystal violet Precipitate dye with iodine-potassium iodine solution Extract soluble dye with organic solvent (EtOH, Acetone, aniline) some bacteria will lose dye - Gram- Some bacteria will not - Gram+ Stain with safranin - will stain Gram- Fungal hyphae and spores are also Gram+
66
What is the purpose of Mucicarmine?
Stains epithelial mucins deep red | will also stain cryptococcus neofromans (fungus)
67
What is the principle of mucicarmine?
Carmine stains nuclei - is made from ground insects, has synthetic alternatives Mucicarmine adds aluminum, which changes chemistry
68
What is the purpose of Gomori Methamine Silver Nitrate?
Detects bacteria known as spirochetes - carbohydrates in fungal walls and gomori was looking for glycogens and mucus Uses: chromium trioxide, sodium metabisulfite, methenamine-silver stock (with borax for working solution) Stains fungi, cellulose, starch, and chitin black - also melanin granules, glycogen or mucus
69
What does PAS stain?
Carbohydrates
70
What does Cresyl Violet stain?
Nerves
71
What stain would you use to view liver fibrosis?
Masson's trichrome
72
What does Congo Red stain?
Amyloid
73
What does Crystal Violet Stain?
nerves, amyloid

Histo 2 (4 decks)

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