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Histopath Midterms > Special Histopath Technique > Flashcards

Special Histopath Technique Flashcards

1
Q

Histopath Technique for

Routine

Special

A

Routine
-Manual/Automated Processing

Special

Frozen Section
Ag-Ab based
-Immunohistochemistry
-Immunofluorescence
Molecular Technique
-FISH

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2
Q

A type of tissue process in which tissue is rapidly freeze and sectioned in a cryostat machine and then stains for microscopic examination. The process takes 10-15 minutes.

A

Frozen section

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3
Q

For rapid diagnosis during intraoperative surgery. Provides a real-time information.

A

Frozen section

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4
Q

Preparation for immunohistochemistry (IHC) and immunofluorescence.

A

Frozen section

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5
Q

Quick-freeze makes the tissue hard enough to section with microtome.

A

Frozen section

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6
Q

Frozen Section process takes?

A

10-15 mins

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7
Q

a chamber which houses a microtome. Inside temperature ranges from -18 to -70˚C.

A

Cryostat

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8
Q

Temperature of Cryostat

A

-18 to -70C

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9
Q

Methods of freezing

A

Liquid Nitrogen
Isopentane
Carbon Dioxide gas
Aerosol sprays

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10
Q

Most common method of freezing

A

Liquid Nitrogen

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11
Q

Liquid Nitrogen Temp

A

-190C

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12
Q

Isopentane Temp

A

-150C

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13
Q

Carbon Dioxide Gas Temp

A

-70C

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14
Q

Aerosol spray temp

A

-50C

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15
Q

Method of freezing that is used in cold knife procedure

A

Carbon Dioxide Gas

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16
Q

Method of freezing for quick freezing spray

A

Aerosol Sprays

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17
Q

acts as a embedding medium. in cryostat

A

Water

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18
Q

Cold knife procedure, commonly used as freezing agent

A

CO2 gas

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19
Q

Cold knife procedure temperature of

Knife
Tissue
Environment

A

-40 to -60C
-5 to -10C
-10C

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20
Q

Different temperature is employed between tissues and the knife.

A

Cold knife procedure

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21
Q

Refrigerated apparatus used fresh tissue microtomy.

A

Cryostat procedure

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22
Q

Cryostat cold chamber is maintained at temperature of

A

-15 to -30 C.

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23
Q

Optimum working temperature for cryostat procedure

A

-18 to -20

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24
Q

Size of the tissue section in frozen section

A

5-10 micra

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25
Q

Stain Used in Frozen Section

A

Hematoxylin and Eosin Stain

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26
Q

It is also requested if lipids, fats, or some nervous system elements are to be demonstrated.

A

Frozen section

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27
Q

provides initial diagnosis thereby should not be a replacement for paraffin-embedded section as it has also disadvantages.

A

Frozen section

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28
Q

Special way of preserving tissues by rapid freezing of fresh tissue

A

Quenching -160C to -180C

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29
Q

Ice water molecules are the removed through desiccation or transferring the frozen tissue block in vacuum a higher temperature for 24-48 hours

A

Sublimation -30 to -40C

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30
Q

Special way of preserving tissues by rapid freezing of fresh tissue (Quenching -160C to -180C).
Ice water molecules are the removed through desiccation or transferring the frozen tissue block in vacuum a higher temperature for 24-48 hours (Sublimation -30 to -40C).

A

Freeze Drying

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31
Q

Quenching Temp

A

-160 to -180

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32
Q

Sublimation Temp

A

-30 to -40

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33
Q

Machine used in Freeze Drying

A

Laboratory Bench-Top Vacuum Freeze Dryer

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34
Q

Similar with freeze drying but with addition of dehydration step.

A

Freeze substitution

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35
Q

Frozen tissue is dehydrated in expensive vacuum drying apparatus fixed in _____ or in _______ dehydrated in ______

A

Rossman’s formula
1% acetone
absolute alcohol.

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36
Q

Machine used in Freeze Substitution

A

Leica Automated Freeze Substitution System

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37
Q

It is a microscopic-based technique used to demonstrate specific cellular components particularly the protein ( epitope or antigen) in tissue samples.

A

Immunohistochemistry

38
Q

Immunohistochemistry technique is based on

A

antigen and antibody principle for detection

39
Q

An enzyme labeled antibody (IgG) binds to antigenic biomarkers on the surface of target tissue then a change in color is observed under light microscope.

A

Immunohistochemistry

40
Q

Specimen for Immunohistochemistry

A

Formalin fixed paraffin embeddedd
Frozen section

41
Q

Components of Immunohistochemistry

A

Primary antibody
Secondary antibody
Chromogen

42
Q

Antibody-IgG type, binds specifically to the target antigen.

A

Primary antibody

43
Q

binds to only one epitope of the antigen (specific).

A

Monoclonal

44
Q

binds to multiple epitope of an antigen (sensitive).

A

Polyclonal

45
Q

binds to the primary antibody, conjugated with enzymes or fluorescence.

A

Secondary antibody

46
Q

a substrate when added produces insoluble colored precipitate.

47
Q

exposing the epitope of Ag to allow the binding of antibody. Done in FFPE tissues prior to Ab labelling.

A

Antigen Retrieval

48
Q

breaks down formalin cross-linking to allow better exposure of antigenic sites. Commonly used enzyme are Trypsin and Protease.

A

Proteolytic Enzyme Digestion-

49
Q

tissue section is subjected to microwave oven heating to retrieve antigen lost during tissue processing. Boiling usually for 20 minutes.

A

Microwave Antigen Retrieval-

50
Q

the same with microwave technique with less time required and better retrieval of antigen. Autoclave and water bath can also be used.

A

Pressure cooker Antigen Retrieval-

51
Q

Commonly used enzyme for Proteolytic Enzyme Digestion

A

Trypsin
Protease

52
Q

Immunohistochemistry work flow (Simple)

A

Ag retrieval
Addition of Primary Ab
Application of Secondary Ab
Addition of Chromogen

53
Q

Immunohistochemistry work flow (Complex)

A

FFPE tissue
Deparaffinization
Rehydration
Antigen Retrieval
Blocking
Primary Ab addition
Secondary Ab addition
Counterstain
Dehydration
Coverslip

54
Q

Immunohistochemistry techniques

A

Direct technique
Indirect technique
Peroxidase anti peroxidase (PAP) technique
Avidin-Biotin complex (ABC) technique
Labeled Streptavidin Avidin Biotin (LSAB) technique

55
Q

One step method
Uses primary labelled antibody directly attach to the target antigen on the surface of the tissue.

A

Direct technique (traditional)

56
Q

Label used for Direct Technique (traditional)

A

Fluorochrome or Horseradish Peroxidase (HRP)

57
Q

More sensitive than direct technique
It uses
1. primary unlabeled antibody
2. secondary labeled antibody directed to primary antibody.

A

Indirect Technique

58
Q

commonly used enzymes in Indirect Technique

A

Horseradish peroxidase

59
Q

Peroxidase Anti Peroxidase (PAP) technique components

A

Antigen molecule
Primary Ab
Secondary Ab
Peroxidase anti peroxidase complex

60
Q

A type of indirect method which uses
1.1. Peroxidase anti-peroxidase complex (Ab against HRP)
2. Secondary antibody
3. Primary unlabeled antibody (from rabbit)

A

Peroxidase anti peroxidase (PAP) technique

61
Q

bridge in PAP complex by secondary antibody.

A

primary unlabeled antibody

62
Q

This reaction visualized by the use of hydrogen peroxide as an enzyme substrate, and a chromogen Diaminobenzidine (DAB) resulting to insoluble dark brown reaction.

A

Peroxidase anti peroxidase (PAP) technique

63
Q

100-1000 X sensitive than indirect method since it uses more enzyme in the which give a higher degree of reaction.

A

Peroxidase anti peroxidase (PAP) technique

64
Q

Uses avidin (from egg white) that has affinity to biotin a low molecular weight vitamin that easily conjugate to Ab and enzymes.

A

Avidin-Biotin complex (ABC) technique

65
Q

Uses
1. Primary antibody
2. Biotinylated secondary antibody
3. Avidin-biotin enzyme complex or labeled streptavidin

A

Avidin-Biotin complex (ABC) technique

66
Q

When the chromogenic enzyme substrate is applied, it yields a colored precipitate at the site of the reaction.

A

Avidin-Biotin complex (ABC) technique

67
Q

Avidin-Biotin complex (ABC) technique composition

A

Antigen
Primary antibody
Biotinylated secondary antibody
Biotin
Avidin
Streptavidin

68
Q

Uses
1.Primary antibody
2. Boitinylated secondary antibody
3.Streptavidin-enzyme conjugate

A

Labeled Streptavidin Avidin Biotin (LSAB) technique

69
Q

gives a better amplification and detection than ABC technique.

A

Labeled Streptavidin Avidin Biotin (LSAB) technique

70
Q

Immunohistochemistry immunostains

A

High molecular weight Cytokeratin (HMWCK)-
CD30
CD31
CD56
CK20

71
Q

Immunohistochemistry immunostains that detects epithelial neoplasm.

A

High molecular weight Cytokeratin (HMWCK)-

72
Q

Immunohistochemistry immunostains that is used to diagnose Hodgkin lymphoma.

73
Q

Immunohistochemistry immunostains that is use to identify endothelial tumor.

74
Q

Immunohistochemistry immunostains for the diagnosis of leukemia and small cell carcinoma.

75
Q

Immunohistochemistry immunostains for merkel carcinoma and gastrointestinal carcinoma.

76
Q

Based also on antigen-antibody principle for detection.

A

Immunofluorescence

77
Q

Fluorochrome is used instead for enzyme.

A

Immunofluorescence

78
Q

Immunofluorescence Color for

  1. Fluorescein
  2. Rhodamine
A

Apple-Green
Rhodamine

79
Q

Reaction is observed as a fluorescence under fluorescence microscope.

A

Immunofluorescence

80
Q

Provides better image quality
Frozen section or culture cells can be used as a specimen.

A

Immunofluorescence

81
Q

A fluorochrome used to demonstrate mucin
Stain last only for 2 hours

A

Fluorescent Acridine Orange Technique

82
Q

Fluorescent Acridine Orange Technique stain result for

  1. Acid mucopolysaccharides
  2. Fungi
  3. Background
A

Black
Greenish Red
Reddish Orange

83
Q

Used for demonstration of certain type of carcinoma.

A

Cytokeratin (CK) 8

84
Q

Flourescent staining for DNA and RNA color result for

  1. Rhodamine
  2. Acridine Orange (DNA);(RNA)
A

Orange Red Emmision

DNA-Yellow-green
RNA- Orange Red

85
Q

Uses fluorescent-labelled probe (short single stranded nucleic acid) which is complimentary to the target DNA or mRNA in a tissue section

A

Fluorescence In-situ hybridization (FISH)

86
Q

Fluorescence In-situ hybridization (FISH) uses?

A

fluorescent-labelled probe

87
Q

It is based on the principle of hybridization- complementary binding between a probe and target nucleic acids.

A

Fluorescence In-situ hybridization (FISH)

88
Q

It targets the nucleic acids instead to the antigen express on the surface of the cell.

A

Fluorescence In-situ hybridization (FISH)

89
Q

Frozen section, paraffin section and smears can be used.
Detection is through radioactivity or fluorescence microscope.

A

Fluorescence In-situ hybridization (FISH)

90
Q

Fluorescence In-situ hybridization (FISH) uses what type of section?

A

Frozen section
Paraffin Section
Smears

Histopath Midterms (4 decks)

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