IHC Flashcards
(110 cards)
1
Q
It is carried out in disease diagnosis
A
IMMUNOHISTOCHEMISTRY
2
Q
this involved antigen - antibody reaction
A
Immunohistochemistry
3
Q
Purpose of IHC
A
- To identify tissue/cellular antigens or phenotypic markers
- Detect antigen in cells/tissue
- Detection of organism in cytologic preparation
4
Q
Found in cells; the one detected
A
Antigen
5
Q
Belonging to class of serum protein that immunoglobulins
A
Antibodies
6
Q
Detector
A
Antibody
7
Q
Most common form of antibody used jn IHC
A
IgG
8
Q
Types of antibodies used in IHC
A
Polyclonal
Monoclonal
9
Q
Produced by different cells
A
Polyclonal
10
Q
It reacts with various epitopes
A
Polyclonal
11
Q
Structure of antigen react/bind with antibody
A
Epitope
12
Q
Main source of polyclonal
A
Rabbit
13
Q
Other sources of polyclonal
A
Goat, Pigs, sheep
14
Q
Produced from individual clone of plasma cell/one cell
A
Monoclonal
15
Q
Reacts with only one specific type of epitope
A
Monoclonal
16
Q
Main source of monoclonal
A
Mice
17
Q
Labelling of Antibodies
A
Use of enzyme
Use of fluorochrome label
Use of colloid metal
Radio label
18
Q
Enzymatic method in IHC
A
Enzyme label
19
Q
Enzyme used in labelling antibodies
A
HRP: Horse Radish peroxidase
AP: Alkaline Phosphatase (alternative)
20
Q
Purpose of chromogen
A
Color developer
21
Q
Enzyme requires chromogens?
A
Yes
22
Q
Chromogens used in enzyme labelling
A
3,3’ DAB - Diaminobenzidine
EAC - 3- amino-9- ethylcarbazole
23
Q
Color produce by DAB
A
Brown
24
Q
Color produce by EAC
A
Brick red
25
Chromogens require the use of ___
Substrate
26
Substrate used in HRP
Hydrogen peroxide
27
Substrate used in EAC
Phosphate
28
Antibodies are label with enzyme after the reaction? T/F
False. It is label BEFORE the reaction
29
Advantage of using enzyme
1. Standard sample can be stored for a long time
2. Hematoxylin can be used as counterstain
3. Can identify using LM/EM; Not requiring FM
30
Compatible counterstain of DAB
Hematoxylin
Methyl green
Methyl blue
31
Compatible counterstain of EAC
Hematoxylin
Methyl blue
32
Chromogens used in Alkaline phosphatase
BCIP - 5-bromo-4-chloro-3-indolyl-phosphate
NBT - Nitro blue tetrazolium
FR salt - Fast Red
33
Produce Bluish violet/ black violet
BCIP/NBT
34
Suitable counterstain of BCIP/NBT
nuclear fast red & brilliant green
35
Produced rose color
FR SALT
36
Suitable counterstain of FR salt
None
37
Traditional counterstain/ nuclear stain
Hematoxylin
38
Stain produce reddish violet
Mayers
39
Stain where tissue morphology is well defined
Harris
40
Stain produce red color
Harris
41
Incubation and temperature to link antibody with PEROXIDASE
30-60 minutes at room temperature
42
Immunofluorescence technique
Fluorochrome label
43
This requires fluorescence microscope
Fluorochrome label
44
Derivative of fluorochrome label that attach to protein
FITC - fluorescence isothiocyanate
45
Example used in colloidal metal
Gold (pink product)
46
It is used to detect antigen; binds to tissue carbohydrates
Lectins
47
Specimen for IHC
Frozen section
Processed specimen like
48
Preferred fixative for IHC
Formaldehyde
49
Prepared on cryostat
Frozen section
50
Frozen section must be fixed with ___
Methanol / Acetone
51
Specimen requested for rapid diagnosis
Frozen section
52
It is carried out intraoperatively
Frozen section
53
Purpose of Frozen section
1. Preserve positive of antigen
2. To prevent destruction of labile antigen site
54
Processed specimen like
1. Formalin fixed
2. Paraffin embedded
55
Increase accessibility of antibody to antigen
Antigen retrieval
56
This process must be done in processed specimen like
Antigen retrieval
57
Methods of antigen retrieval
1. Heat induced epitope retrieval
2. Protease induced epitope retrieval
58
Heat induced epitope retrieval
Microwave antigen
Pressure cooker antigen retrieval
Autoclave heating, water bath heating, steamer heating
Decloaker heating, combination of microwave and enzyme digestion
(MPAWS, CD)
59
Protease induced epitope retrieval
Proteolytic enzyme retrieval
60
Enzyme used in PIER
tyrosine
Protease
61
This enzyme can break formalin crosslink
Protease
62
Pre treatment must be done on what type of specimen
Processed specimen like
1. Formalin fixed
2. Paraffin embedded
63
Block endogenous peroxidase; 3% hydrogen peroxide
Peroxidase block
64
Blocks all nonspecific sites & reduces background; 10% normal serum
Protein block
65
Purpose of control
1. Test for specificity of antibodies involved
2. Prevent misinterpretation due to false + and false - results
66
Tissue section containing antigen being detected
Positive control
67
How to prepare negative control
Omit the primary antibody from staining
Replace primary antibody by an Ig that is directed against unrelated antigen
68
Internal tissue control is also known as ___
Built in control
69
Contains target antigen not only on tissue element but also in adjacent normal tissue element
Internal tissue control
70
Uses only 1 antibody that is labeled
Direct IHC
71
It is the improvement of traditional method
EPOS - Enhanced Polymer Ones Step Staining Method
72
EPOS requires addition of ___
Dextran polymer
73
Method of detection that is lore sensitive
EPOS
74
2 step method; use 2 antibody
Indirect IHC
75
Function of secondary antibody
Attach to Fe portion of primary antibody increasing sensitivity
76
It has greater consistency of staining; faster and accurate
Automation
77
Less man power; decrease use of reagent
automation
78
Found in tissue; tumor of malignancy
Tumor markers
79
Epithelial tumor markers
Keratin
EMA
CEA
TTF-1
PSA
ER/PR
80
Marker of ET cells; diagnosis of carcino
Keratin
81
T/F: all types of carcinoma will be positive for CK
False. Not all type
82
(+) lungs, uterus, kidney
CK 7 (Cytokeratin)
83
(+) colon cancer, stomach cancer
CK 20
84
This determine the site of tumor
EMA
85
Useful in detecting adenocarcinoma
EMA
86
(+) breast, lung, kidney
EMA
87
Oncofetal antigen
CEA
88
Differentiate adenocarcinoma from mesothelioma
CEA
89
Elevated in GIT cancer; monitoring colon, pancreas, cervix, breast cancer
CEA
90
Found in thyroid neoplasm
TTF-1
91
Differentiate lung adenocarcinoma from mesothelioma
TTF-1
92
Induced prostate infection
PSA
93
Elevated in prostate, pancreas & malignancy
PSA
94
Normally found in breast ET cell
ER/PR
95
ER/PR found in myoepithelial cells?
No
96
Determining prognosis breast cancer and identification of metastatic breast carcinoma
ER/PR
97
Indication of positive results in ER/PR
Good prognosis
98
Intermediate filament markers
Actin
Vimentin
Desmin
Glial fibrillary acidic protein - GFAP
99
Tumors derived from smooth, cardiac, & skeletal muscle
Actin
100
Present in normal mesenchymal cells
Vimentin
101
Vimentin: neoplastic counterpart
Sarcoma, melanoma, leukemia, seminoma schwannomas
102
Cancer involving schwann cells
Seminoma schwannomas
103
Highly specific for myogenic tumor
Desmin
104
Confirms diagnosis of astrocytoma
GFAP
105
Marker for choriocarcinoma
Human chorionic gonadotropin (HCG)
106
HCG is synthesized by
Placental syncitiotrophoblast
107
Marker for embryonal carcinoma & teratomas; hepatocellular carcinoma
AFP - Alpha feto protein
108
Best screening marker for lymphoma
Lymphomas
109
Lymphomas also known as
CD45
110
Most common marker to assess proliferation of tumor cells
KI & MIB-1
