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HISTOPATHOLOGY (TopnotcherQT) > IMMUNOHISTOCHEMISTRY (IHC) > Flashcards

IMMUNOHISTOCHEMISTRY (IHC) Flashcards

1
Q

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

A

Immunohistochemistry

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2
Q

Primary antibody binds directly to:

A

UNKNOWN ANTIGEN

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3
Q

Secondary antibody binds to the:

A

PRIMARY ANTIBODY, often conjugated with a detection enzyme or fluorochrome

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4
Q

Most commonly used antibody for immunohistochemistry

A

IgG

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5
Q

Most frequently used animal to produce POLYCLONAL ANTIBODIES

A

RABBIT

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

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6
Q

Most frequently used animal to produce MONOCLONAL ANTIBODIES:

A

MICE

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7
Q

Fixative of choice for IHC

A

FORMALIN

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8
Q

Adhesive for IHC

A

POLY-L-LYSINE

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9
Q

Most commonly used antibody for IHC

A

IgG

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10
Q

Most commonly used animal for retrieving monoclonal antibodies

A

MICE

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11
Q

Most commonly used animal for retrieving polyclonal antibodies:

A

RABBIT

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12
Q

Most commonly used enzyme label for IHC

A

HORSE RADISH PEROXIDASE

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13
Q

A common fluorochrome used in IHC

A

FLUORESCENCE ISOTHIOCYANATE

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14
Q

Color of chromogen Diaminobenzidine (DAB)

A

BROWN

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15
Q

Color of chromogen Aminoethylcarbazole (AEC)

A

RED

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16
Q

Secondary/nuclear counterstain used in IHC

A

HEMATOXYLIN

17
Q

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

A

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

18
Q

Best positive control for IHC

A

IDENTICAL TO PATIENT TISSUE

19
Q

Size of paraffin sections for IHC

A

3-5 micra

20
Q

Microscopes used in IHC

A

LIGHT OR FLUORESCENT

21
Q

IMMUNOHISTOCHEMISTRY TECHNIQUES:

A
  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
22
Q

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

A

DIRECT TECHNIQUE

23
Q

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

A

INDIRECT TECHNIQUE

24
Q

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

A

HORSERADISH PEROXIDASE (HRP)

25
Q

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

A

PEROXIDASE-ANTIPEROXIDASE (PAP) TECHNIQUE

26
Q

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

A

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

27
Q

REMEMBER!!!!

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

A
28
Q

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

A

OMITTING THE BLOCKAGE OF ENDOGENOUS PEROXIDASE

29
Q

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

A

AVIDIN-BIOTIN COMPLEX (ABC) TECHNIEUQ

30
Q

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

A

LABELED STREPTAVIDIN AVIDIN BIOTIN (LSAB) Technique

31
Q

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

A

IMMUNOFLUORESCENCE METHODS

32
Q

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

A

KIDNEY and SKIN

33
Q

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

A

DIRECT IMMUNOFLUORESCENCE

34
Q

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

A

INDIRECT IMMUNOFLUORESCENCE

35
Q

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

A

IN-SITU HYBRIDIZATION

36
Q
A

HISTOPATHOLOGY (TopnotcherQT) (26 decks)

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