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2G SUMMER > HISTO ENZYME HIST > Flashcards

HISTO ENZYME HIST Flashcards

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1
Q

are biological catalysts that accelerates a chemical reaction

A

ENZYMES

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2
Q

The process of utilizing histochemical techniques to demonstrate enzymes in tissues

A

Enzyme Histochemistry

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3
Q

serves to detect early metabolic changes in biopsy and autopsy tissue before manifestation on H&E staining or IHC

A

Enzyme Histochemistry

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4
Q

identifies the presence and localization of specific proteins using antibodies, regardless of whether the proteins are enzymatically active

A

Immunohistochemistry

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5
Q

detects the activity of specific enzymes in tissues by using substrates that produce a visible reaction product

A

Enzyme Histochemistry

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6
Q

Detects activity of specific enzymes

A

Enzyme Histochemistry

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7
Q

Detects specific proteins using antibodies

A

IHC

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8
Q

Tissue requirement for Enzyme Histochemistry

A

Fresh or frozen tissue

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9
Q

Tissue requirement for Immunohistochemistry

A

Works well with formalin fixed, paraffin-embedded tissues

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10
Q

EHC has ______________ sensitivity and detects only ____________________

A

Moderate, active enzymes

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11
Q

IHC has ______________ sensitivity and detects ____________________

A

High, proteins regardless of enzymatic activity

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12
Q

Enzymatic Reaction produces ___________ reaction for it to be detected

A

Visible Reaction (color or precipitate)

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13
Q

IHC Reaction produces ___________ for it to be detected

A

Chromogen/ Fluorophore

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14
Q

IHC has ______________ specificity, while EHC has ________________ specificity

A

High, Limited

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15
Q

are tissue components that serve as catalyst for most biological reactions

A

Enzymes

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16
Q

Enzymes may be _________ to specific cell components, or may be ______ and soluble in the cytoplasm and body fluids

A

Bound, Free

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17
Q

Tissues in EHC must be frozen at ______ to maintain enzyme structure, prevent breakdown, and preserve enzyme activity

A

-70 deg C

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18
Q

Disadvantage of unfixed frozen sections in EHC

A
  • Mechanical Disruption
  • Uneven section thickness
  • Co-factors leading to loss of reproducibility
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19
Q

EHC study ideal for the usage of unfixed frozen sections

A

Dehydrogenase study

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20
Q

best fixative of choice for preserving enzyme activity and ACP

A

Chilled Acetone

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21
Q

fixative that causes the least inactivation of enzymes compared to other fixatives

A

Chilled Acetone

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22
Q

Preferred fixative for many enzymes besides those requiring freezing

A

Cold 90-100% Ethyl Alcohol

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23
Q

Fixative that provides better cell structure preservation (cytological fixation)

A

Cold 90-100% Ethyl Alcohol

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24
Q

The most common method to detect enzymes in tissues

A

Metal Precipitation

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25
Metal precipitation uses a process called _________ or ________________
simultaneous capture or coupling
26
Process where the invisible primary reaction product is dyed with diazonium salts for a colored final reaction
Simultaneous Coupling
27
Simultaneous Coupling example mixture
Azo dye method for Phosphatase
28
Substrate + Enzyme + invisible reaction + diazonium salts + Insoluble azo dye at the site of enzyme activity
Azo Dye Simultaneous Coupling
29
Azo Dye Simultaneous Coupling Incubation Mixture
Tris Buffer 9.1pH Distilled water Substrate Soln. Fast Garnet GBC salt
30
Two groups of enzymes
Oxidative Enzymes Hydrolytic Enzymes
31
Enzymes that help chemical reactions where the substrate reacts with oxygen from air
Oxidative Enzymes
32
Examples of Oxidative Enzymes
Tyrosinase & Thyroxidase
33
2 Tetrazolium Salts as Hydrogen Acceptors in Oxidative Enzymes
Monotetrazolium (MTT) Ditetrazolium (NBT)
34
Tetrazolium Salt that forms a lipid soluble, finely granular formazan
Monotetrazolium (MTT)
35
Tetrazolium Salt that forms a lipid-insoluble, highly colored formazan deposit at the site of enzymes activity
Ditetrazolium (NBT)
36
NBT meaning
Nitro Blue Tetrazolium
37
Three groups of Oxidative Enzymes
Dehydrogenases Oxidases Peroxidases
38
MECHANISM OF ACTION: -Remove hydrogen from Substrate -Transfer hydrogen along an oxidative (hydrogen-acceptor) Pathway -Coenzyme accepts H
Dehydrogenases
39
3 Main reducing compounds used to visualize dehydrogenase activity
Methylene Blue Tetrazolium Method Tellurite Method
40
Methylene blue produces _______ in Dehydrogenase detection
Colorless
41
Tetrazolium Method produces _______ in Dehydrogenase detection
Bright red, purplish, or blue formazans
42
Tellurite Method produces _______ in Dehydrogenase detection
Insoluble black elemental tellurium
43
A group of enzymes that catalyze oxidation reactions
Oxidases
44
Act on various substrates, primarily phenols and amines
Oxidases
45
Oxidases uses ___________ as the electron acceptor
Molecular Oxygen
46
Refers to copper-containing oxidoreductases acting on phenolics
Polyphenol Oxidases (PPO)
47
Examples of Oxidases
- Indophenol Oxidase (Cytochrome Oxidase) - Tyrosinases - Dopa Oxidase
48
Indophenol Oxidase (Cytochrome Oxidase) Color
Blue
49
Tyrosinases uses ____ to localize enzyme activity
DOPA
50
Dopa Oxidase Color
Dark brown-gray to black
51
are heme-containing enzymes, use H2O2 as an electron acceptor
Peroxidases
52
Involved in various oxidative reactions, and crucial for immune defense and biological processes
Peroxidases
53
Peroxidase, uses the following examples of substrates ________________________
Benzidine: Blue/brown dye Napthol: purple black dye Leuoco-dyes: Regain original eye color
54
Breaks down neurotransmitters, and prefers the use of unfixed tissues
Monoamine oxidase
55
Enzymes that use water to break down complex molecules into simpler units. Demonstrated via precipitation reactions
Hydrolytic Enzymes
56
Common Metal Enzymes
Calcium, Lead, Cobalt, Iron, Copper
57
Two groups of Hydrolytic Enzymes
- Alkaline phosphatases (Gomori Calcium) - Acid phosphatase (Gomori Lead)
58
Alkaline phosphatase is active at __pH, uses calcium phosphatase method
9
59
Acid phosphatase is active at __pH, uses calcium phosphatase method
5
60
This histochemical technique is used to detect alkaline phosphatase in tissues. The cells and regions contained the enzyme appear black under the microscope
Gomori Calcium Method
61
this technique is used to identify lysosomes by detecting their acid phosphatase content
Gomori Lead Method
62
5 nucleotidase + Adenosine-5-phosphate = phosphate ions + lead ions
Lead Method
63
Insoluble lead phosphate to make it visible brown lead sulfide precipitate
Lead Method
64
ATPase
used to differentiate skeletal muscle fiber
65
Phosphate reacts with lead to form lead phosphate, treated with treated with ammonia sulfide -> converted to brown/black lead sulfide precipitate
Lead Method
66
Phosphate reacts with calcium to form calcium phosphate
Calcium-Cobalt Method
67
INCUBATION OF ATPase: (glycine buffer) pH _____, highlights _____ fibers
9.4pH, Highlights type 2 fibers
68
INCUBATION OF ATPase: (veronal-acetate buffer) ___________ pH, highlights ________ fibers
4.2 & 4.6, Type 1 fibers and distinguishes 2A, 2B, and 2C subtypes
69
useful for detecting monocytes, macrophages, and histiocytes
Non-specific esterases
70
helps differentiate monocytes+ from granulocytes-
Non-specific esterases
71
Two methods to detect Non specific Esterases
- Indoxyl Acetate Method - Alpha-Napththyl Acetate Method
72
Indoxyl Acetate Method uses _________________ substrates
Bromo-indoxyl acetate
73
Alpha-Napththyl Acetate Method uses _________________ substrates
Alpha-napththyl Acetate
74
Aka Naphtol AS-D chloroacetate esterase stain
Chloroacetate Esterase
75
Used to identify neutrophil polymorphs in frozen and paraffin-embedded tissue sections
Chloroacetate Esterase
76
Enzyme that hydrolyzes acetylcholine into choline and acetic acid
Acetylcholinesterase
77
Enzyme catalyzing glycogen breakdown into glucose 1 phosphate in the presence of inorganic phosphate
Phosphorylases
78
enzyme key in liver and muscle glycogen metabolism
Phosphorylases
79
Phosphorylase active and less active form
Phosphorylases- A (active) Phosphorylase- B (Less Active)
80
enzymes that splits hexose diphosphate into two molecules of triose phosphate
Aldolases
81
Enzymes that hydrolyze sulfates of phenols and thioglucosides
Sulfatases
82
Sulfatases ppt are visualized using _________________
Beta-naphthoquinone sulfonate

2G SUMMER (10 decks)

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