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HTL > Immunohistochemistry > Flashcards

Immunohistochemistry Flashcards

1
Q

An antigen is a substance that triggers the production of:

A

immunoglobins

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2
Q

A light chain present in some antibodies is:

A

kappa

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3
Q

Different molecular sites on antigens are known as:

A

epitopes

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4
Q

When an antibody labeled with a chromogen is reacted with tissue from a patient, the IHC technique is called:

A

direct

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5
Q

Which of the following is a fluorochrome:

a. alkaline phosphatase
b. rhodamine
c. fast red
d. diamineobenzidine

A

rhodamine

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6
Q

In the indirect IHC method the…

A

patients serum is added to tissue sections containing known antigens

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7
Q

In the avidin-biotin methods, the primary antibody is followed by:

A

biotinylated secondary antibody

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8
Q

A dye that absorbs light and then emits its own light at a longer wavelength is known as:

A

a fluorochrome

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9
Q

In some IHC tehcniques, alkaline phosphatase functions as the:

A

enzyme

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10
Q

Horseradish peroxidase is used in some avidin-biotin methods as the:

A

enzyme

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11
Q

A substrate for alkaline phosphatase-labeled antibodies is commonly:

A

napthol-AS-phosphate

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12
Q

Which of the following chromogens is INSOLUBLE in alcohol:

a. fast red TR
b. 3,3’-diaminobenzidine tetrahydrochloride (DAB)
c. 3-amino-9-ethylcarbazole (AEC)
d. fast blue BBN

A

b. 3,3’-diaminobenzidine tetrahydrochloride (DAB)

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13
Q

IF 3-amino-9-ethylcarbazole is used as a chromagen, the hematoxylin used for counterstaining should be:

A

alcohol free

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14
Q

Tissue for immunoflourescence must be:

A

frozen and unfixed

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15
Q

A neoplasm is defined as a/an:

A

new growth of uncontrolled cell multiplication

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16
Q

What is NOT a proper approach to validation of a new antibody?

A

using only the manufacturers recommendations

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17
Q

When the vimentin stain is completely negative on formalin-fixed tissues, this indicates that most likely the tissue has been:

A

overfixed

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18
Q

Besides the heat employed, another important factor in head-induced epitope retrieval (HIER) us the:

A

composition of the solution used

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19
Q

Ficin is used in IHC for:

A

enzyme-induced epitope enhancement

20
Q

What has been eliminated with polymeric IHC staining methods?

A

serum and avidin-biotin blocking steps

21
Q

In the peroxidase-antiperoxidase (PAP) method of antigen detection, the PAP complex is made in the same species as:

A

the primary antibody

22
Q

Imidazole may be used in peroxidase techniques to:

A

intensify the DAB reaction

23
Q

What link antibodies would most likely follow as a monoclonal primary antibody?

A

rabbit anti-mouse

24
Q

T/F: Prediluted antibodies should always be used as provided by the mansufacturer.

A

False.

25
Q

T/F: Negative tissue antigen controls may be run by substituting for the primary antibody the diluent in which the antibody is prepared.

A

True

26
Q

T/F: One standard staining protocol may be written to cover all specimens.

A

False.

27
Q

T/F: Zinc formalin preserves immunoreactivity well.

A

True.

28
Q

T/F: Multilink antibodies can be used only with monoclonal primary antibodies.

A

False.

29
Q

T/F: Blocking reactions are used to block endogenous activity of the same enzyme as that used for the enzyme immune complex.

A

True

30
Q

T/F: The enzyme label for immunoperoxidase methods contain horseradish peroxidase.

A

True

31
Q

T/F: Monoclonal antibodies are often preferred over polyclonal antibodies because there is no batch-to-batch variability.

A

True.

32
Q

T/F: DAB is an alcohol-soluble chromagen.

A

False.

33
Q

T/F: Harris hematoxylin should not be used with 3-amino-9-ethylcarbazole (AEC) because of the alcohol present.

A

True.

34
Q

T/F: Regulation regarding predictive marker staining of the tissue places responsibility for documenting time in the fixative on the laboratory.

A

True.

35
Q

T/F: The only method of heat-induced epitope retrieval involves using a microwave oven.

A

False.

36
Q

T/F: Optimal dilutions for each antibody must be determined by your lab.

A

true

37
Q

T/F: Metal salts involving nickel, copper, and osmium may be used to intensify 3-amino-9-ethylcarbazole reaction.

A

False. Metal salts intensift DAB.

38
Q

T/F: Alkaline phosphatase label can be substituted for peroxidase label in most IHC methods.

A

True

39
Q

T/F: Precut control slides may be stored at room temp indefinitly.

A

False.

40
Q

In the bloody areas of a tissue section stain with the immunoperoxidase technique, there is a marked reaction of the red blood cells. This is most likely the result of:

A

failure to use hydrogen peroxide. It is essential to reduce non-specific staining of RBCs.

41
Q

Paraffin sections stained with the immunoperoxidase technique show excess background staining. What could explain this?

A

Forgetting to apply the non-immune serum. If the first protein solution applied to the tissue is the primary antibody, nonspecific binding can occur.

42
Q

The skin control for S-100 was stained with the immunoalkaline phosphatase technique using fast red TR as the chromogen. It shows negative staining. One possible explanation is:

A

sections were dehydrated and cleared. With prolonged dehydration, the alkaline phosphatase chromogen can break down.

43
Q

At the end of the immunoperoxidase staining procedure, it was realized that an anti-goat linking antibody was used with monoclonal HMB-45 antibody. Microscopic results should show a:

A

negativity stained positive control and a negative stained specimen since monoclonal antibodies are most often prepared by mice or rabbits. An anti-mouse, anti-rabbit or multilink linking secondary must be used.

44
Q

Both the known positive control and the specimen are negative following immunoperoxidase staining with LCA primary antibody. DAB was used as a chromogen. What could be the cause?

A

wrong secondary antibody was used. The secondary antibody must be targeted to the primary for linking to occur.

45
Q

Excessive background is noted on both the control and specimen stained with immunoperoxidase. This might be due to:

A

insufficient washing with buffer

46
Q

Excessive background is noted on the specimen but not the control stained with immunoperoxidase. This is most likely due to:

A

free antigen in the tissue

47
Q

Both the positive control and the specimen stained with immunoalkaline phosphatase using CEA antibody show very weak staining. Why?

A

incorrectly diluted antibody.

HTL (18 decks)

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