Immunohistochemistry

Question 1

Used for identification of specific and highly selective cellular EPITOPES or ANTIGENS in tissues.

Answer 1

Immunohistochemistry

Question 2

Can also be used for detection of organisms in CYTOLOGIC PREPARATION.

Answer 2

Immunohistochemistry

Question 3

Uses ANTIGEN-ANTIBODY INTERACTIONS.

Answer 3

Immunohistochemistry

Question 4

Found in CELLS.

Answer 4

Antigens

Question 5

Utilizes IMMUNOGLOBULIN: IgG

Answer 5

Antibody

Question 6

[2] Types of antibody

Answer 6

1. Polyclonal
2. Monoclonal

Question 7

Immunizing an animal with a purified specific molecule (IMMUNOGEN)

[antibody]

Answer 7

Polyclonal antibody

Question 8

Involves collection of immunoglobulin rich serum after production of HUMORAL ANTIBODY against antigen.

[antibody]

Answer 8

Polyclonal antibody

Question 9

Immunochemically NOT IDENTICAL to each other = CROSS REACT

[antibody]

Answer 9

Polyclonal antibody

Question 10

Products of an individual clone of plasma cells.

[antibody]

Answer 10

Monoclonal antibody

Question 11

Produced by HYBRIDOMA and CLONING TECHNIQUES.

[antibody]

Answer 11

Monoclonal antibody

Question 12

Antibody produce are immunochemically identical that DO NOT CROSS REACT.

[antibody]

Answer 12

Monoclonal antibody

Question 13

Propagation: culture medium or transplantation of hybridoma in MICE.

[antibody]

Answer 13

Monoclonal antibody

Question 14

[2] Labels

Answer 14

1. Enzyme
2. Fluorochrome labels

Question 15

Use of HORSE RADISH PEROXIDASE + DIAMINOBENZIDINE

[labels]

Answer 15

Enzyme labels

Question 16

Use of FLUORESCEIN ISOTHIOCYANTE

[labels]

Answer 16

Fluorochrome labels

Question 17

CRYOSTAT FROZEN SECTION and fixed in a few seconds in ABSOLUTE METHANOL or ACETONE.

Answer 17

Specimen prepatration

Question 18

[3] Specimen preparation

[PMP]

Answer 18

1. Proteolytic Enzyme Digestion
2. Microwave Antigen Retrieval
3. Pressure Cooking Antigen Retrieval

Question 19

FUNCTION: to breakdown formalin cross linking, unmask and allow certain antigenic sites to be exposed.

[specimen prep]

Answer 19

Proteolytic Enzyme Digestion

Question 20

Useful for demonstrating HEAVY CHAIN IMMUNOGLOBULINS, COMPLEMENT, SPECIFIC ANTIGENS (CYTOKERATIN)

[specimen prep]

Answer 20

Proteolytic Enzyme Digestion

Question 21

0.1% trypsin in 0.1% calcium chloride (pH 7.8)

[proteolytic enzyme digestion]

Answer 21

Trypsin

Question 22

0.05 – 0.1 in distilled water (pH 7.8)

[proteolytic enzyme digestion]

Answer 22

Protease

Question 23

OPTIMAL LENGTH OF EXPOSURE: 10 – 60 minutes

[specimen prep]

Answer 23

Microwave Antigen Retrieval

Question 24

Involves BOILING OF FORMALIN fixed deparaffinized sections.

[specimen prep]

Answer 24

Microwave Antigen Retrieval

Question 25

Optimal length of exposure: [Microwave Antigen Retrieval]

Answer 25

10 - 60 mins

Question 26

Most satisfactory time: [Microwave Antigen Retrieval]

Answer 26

20 mins

Question 27

Demonstrates proliferation markers (Ki 67 and MIB-1), hormone receptors (ER and PR), growth factor receptors (HER-2/neu). [specimen prep]

Answer 27

Microwave Antigen Retrieval

Question 28

Not subjected to “HOT SPOTS” and “COLD SPOTS” [specimen prep]

Answer 28

Pressure Cooking Antigen Retrieval

Question 29

More CONSISTENT in recovery of many antigens. [specimen prep]

Answer 29

Pressure Cooking Antigen Retrieval

Question 30

Less time consuming. [specimen prep]

Answer 30

Pressure Cooking Antigen Retrieval

Question 31

[3] Controls

Answer 31

1. Positive 2. Negative 3. Internal tissue

Question 32

Section known and proven to have the ANTIGEN IN QUESTION. [control]

Answer 32

Positive control

Question 33

OMIT the PRIMARY BODY from the staining schedule. [control]

Answer 33

Negative control

Question 34

REPLACE the SPECIFIC PRIMARY ANTIBODY by an Ig that is directed against unrelated antigen. [control]

Answer 34

Negative control

Question 35

"BUILT IN" control [control]

Answer 35

Internal tissue

Question 36

Eliminates the variable of tissue fixation between specimens and controls. [control]

Answer 36

Internal tissue control

Question 37

Contains the TARGET ANTIGEN but not in the tissue elements under investigation. [control]

Answer 37

Internal tissue control

Question 38

[5] Epithelial Tumor Markers [KEC, TP]

Answer 38

1. Keratin 2. Epithelial Membrane Antigen (EMA) 3. Carcinoembryonic Antigen 4. Thyroid Transcription Factor -1 (TTF-1) 5. Prostate Specific Antigen (PSA)

Question 39

Highly sensitive marker for epithelial cells epithelial tumors vs. non-epithelial tumors. [epithelial tumor markers]

Answer 39

Keratin

Question 40

High molecular weight protein. [epithelial tumor markers]

Answer 40

Epithelial Membrane Tumors