IMMUNOHISTOCHEMISTRY (IHC)

Question 1

A technique for identifying ANTIGENS by means of ANTIGEN-ANTIBODY INTERACTIONS, the site of antibody binding being identified either by direct labeling of the antibody, or by use of a secondary labeling method

Answer 1

Immunohistochemistry

Question 2

Primary antibody binds directly to:

Answer 2

UNKNOWN ANTIGEN

Question 3

Secondary antibody binds to the:

Answer 3

PRIMARY ANTIBODY, often conjugated with a detection enzyme or fluorochrome

Question 4

Most commonly used antibody for immunohistochemistry

Answer 4

IgG

Question 5

Most frequently used animal to produce POLYCLONAL ANTIBODIES

Answer 5

RABBIT

Rabbit>Goat>Pig>Sheep>Horse>GUINEA PIG

Question 6

Most frequently used animal to produce MONOCLONAL ANTIBODIES:

Answer 6

MICE

Question 7

Fixative of choice for IHC

Answer 7

FORMALIN

Question 8

Adhesive for IHC

Answer 8

POLY-L-LYSINE

Question 9

Most commonly used antibody for IHC

Answer 9

IgG

Question 10

Most commonly used animal for retrieving monoclonal antibodies

Answer 10

MICE

Question 11

Most commonly used animal for retrieving polyclonal antibodies:

Answer 11

RABBIT

Question 12

Most commonly used enzyme label for IHC

Answer 12

HORSE RADISH PEROXIDASE

Question 13

A common fluorochrome used in IHC

Answer 13

FLUORESCENCE ISOTHIOCYANATE

Question 14

Color of chromogen Diaminobenzidine (DAB)

Answer 14

BROWN

Question 15

Color of chromogen Aminoethylcarbazole (AEC)

Answer 15

RED

Question 16

Secondary/nuclear counterstain used in IHC

Answer 16

HEMATOXYLIN

Question 17

In IHC, _____________ is raised against specific cellular _________and then conjugated with a __________

Answer 17

In IHC, ANTIBODY is raised against specific cellular ANTIGEN and than conjugated with a VISUAL MARKER

Question 18

Best positive control for IHC

Answer 18

IDENTICAL TO PATIENT TISSUE

Question 19

Size of paraffin sections for IHC

Answer 19

3-5 micra

Question 20

Microscopes used in IHC

Answer 20

LIGHT OR FLUORESCENT

Question 21

IMMUNOHISTOCHEMISTRY TECHNIQUES:

Answer 21

1. Direct technique
2. Indirect technique
3. Peroxidase-antiperoxidase (PAP) technique
4. Avidin-Biotin Complex (ABC) technique
5. Labeled Streptavidin Avidin Biotin (LSAB) technique
6. Immunofluorescence Methods

Question 22

Traditional technique; Conjugate the PROIMARY ANTIBODY directly to the label such as fluorochrome or horseradish peroxidase

Answer 22

DIRECT TECHNIQUE

Question 23

More sensitive than the traditional direct technique; TWO or THREE STEP PROCEDURE that involves application of unconjugated primary antibody, followed by a labeled antibody directed against the first antibody

Answer 23

INDIRECT TECHNIQUE

Question 24

The most commonly used enzyme for indirect antibody enzyme-complex techniques

Answer 24

HORSERADISH PEROXIDASE (HRP)

Question 25

An INDIRECT ANTIBODY ENZYME-COMPLEX TECHNIQUE where the soluble peroxidase-antiperoxidase complex is bound to unconjugated primary antibody by a second layer of "bridging" antibody, usually a swine anti-rabbit antibody, that then binds to both the primary antibody and the rabbit PAP complex

Answer 25

PEROXIDASE-ANTIPEROXIDASE (PAP) TECHNIQUE

Question 26

In PAP, combining horseradish peroxidase with the most common chromogen ________________ results in a stable insoluble ____________________ end product when antigen is present in the tissue

Answer 26

In PAP, combining horseradish peroxidase with the most common chromogen DIAMINOBENZIDINE (DAB) results in a stable insoluble DARK BROWN REACTION end product when antigen is present in the tissue

Question 27

REMEMBER!!!! PAP is the peroxidase staining procedure in which reagents are link exclusively by antigen-antibody reactions WITHOUT ANY CONJUGATION STEPS

Question 28

In Immunoperoxidase staining, efficiency of the chromogen-substrate reaction step can be checked by:

Answer 28

OMITTING THE BLOCKAGE OF ENDOGENOUS PEROXIDASE

Question 29

Uses AVIDIN derived from egg white because of its marked affinity for BIOTIN, a low molecular weight vitamin that can be easily conjugated and enzyme markers

Answer 29

AVIDIN-BIOTIN COMPLEX (ABC) TECHNIEUQ

Question 30

A method that is found to be 4 to 8 times MORE SENSITIVE than the old ABC method; an immunohistochemistry labelling method in which a molecule of avidin is directly labeled with an enzyme

Answer 30

LABELED STREPTAVIDIN AVIDIN BIOTIN (LSAB) Technique

Question 31

The most common tissue processing method for fluorescence microscopy techniques is frozen section fixed in ice cold acetone

Answer 31

IMMUNOFLUORESCENCE METHODS

Question 32

Tissues most often test for diagnostics using fluorescence-labeled-antibodies:

Answer 32

KIDNEY and SKIN

Question 33

For solid tissue biopsies; tissue is reacted with FLUORESCEIN-CONJUGATED ANTIBODY specific for the material being sought within the tissue

Answer 33

DIRECT IMMUNOFLUORESCENCE

Question 34

Used for the detection of AUTOANTIBODIES in the patient's serum including anti-nuclear antibody (ANA), anti-mitochondrial antibody (AMA), and liver-kidney microsomal antibody

Answer 34

INDIRECT IMMUNOFLUORESCENCE

Question 35

Based on the specificity of the interaction of a probe with the target NUCLEIC ACIDS; The detection procedure for an in-situ hybridization probe is SIMILAR to IHC. However it is based on the specificity of the interaction of a probe with the target NUCLEIC ACID, rather than the target protein or immunogen

Answer 35

IN-SITU HYBRIDIZATION