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B. Final exam 405 > Exam 4 > Flashcards

Exam 4 Flashcards

1
Q

cause of background staining on a negative reagent control

A

Reagent contamination

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2
Q

A preferred method for the storage of control slides to prevent loss of antigenicity would be

A

Dipped in paraffin

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3
Q

This method of amplification involves the repeating or reapplying of amplification reagents

A

Cycled TSA

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4
Q

Loss of antigenicity due to slide oxidation more readily affects which type of antigen?

A

Nuclear

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5
Q

There is no staining on both the specimen and control on the same slide. The protocol was performed on an automated IHC stainer. The slide was recut and the same protocol repeated on the stainer using the same control. The results were optimal. What could be a possible explanation for the first slide

A

Instrument malfunction

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6
Q

In tyramide signal amplification, the tyramide conjugate is applied

A

After the HRP detection enzyme

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7
Q

Which of the following could be a cause of background staining on the patient specimen, control (both on the same slide) and on the slide surrounding the tissue

A

Waterbath adhesive or additive was used

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8
Q

Macrophages have ingested the target antigen and are staining positive. This would be referred to as

A

Antigen diffusion

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9
Q

Which would be an appropriate counterstain for HRP-AEC detection

A

Mayer hematoxylin

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10
Q

The purpose of this type of control is to determine if the IHC reagents in general are contributing to non-specific staining

A

Negative reagent control

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11
Q

The introduction of polymer-based detection systems has had what effect in negative reagent controls

A

Reduced the need

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12
Q

Disadvantage of TMA is

A

Assembly time

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13
Q

Amplification is used to

A

Increase the staining affinity

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14
Q

A specimen that is underfixed will show what type of staining pattern?

A

Optimal staining on the edge, no staining to weak staining in the center

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15
Q

Of the following, which would be the best choice for a single tissue type control

A

A single tissue type known to contain both positive and negative components

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16
Q

Once activated the tyramide conjugate is covalently attracted to

A

Proteins

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17
Q

Which may be considered an advantage of TMAs

A

Allows for sampling of many different tissue types

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18
Q

For a positive tissue control, it is acceptable, but not optimal, to run

A

A batch control

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19
Q

Slide oxidation can be prevented by

A

Storing in an airtight container

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20
Q

Which of the following is not a type of control used in immunohistochemistry

A

Positive reagent control

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21
Q

An IHC stained slide shows optimal intensity in the specimen, but the control is weak (both on the same slide). What could be the cause

A

The control tissue is oxidized

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22
Q

The control tissue fell off the slide during IHC staining, but the patient specimen remains (they were both placed on the same slide). How might the slide still be usable for diagnosis

A

If the specimen contains areas that do and do not contain the antigen

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23
Q

After IHC staining, the control stains adequately, but the specimen shows unexpected weak intensity, especially in the center. What is the most likely cause

A

The specimen was not properly fixed

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24
Q

There is non-specific staining in only the red blood cells of the negative reagent control, tissue control and the specimen. The protocol uses HRP-DAB detection. What is the most likely explanation

A

Hydrogen peroxide blocking was omitted from the protocol

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25
Q

The degradation of protein, DNA and RNA in paraffin sections is called

A

Slide oxidation

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26
Q

There is weak staining intensity with a concentrated antibody. How could the protocol be modified to correct this

A

Decrease the primary antibody dilution

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27
Q

The expected localization pattern of an antigen

A

Can be found in the Antibody Specification Sheet

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28
Q

Which would be the best choice for the negative reagent control

A

Non-immune serum of the same isotype

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29
Q

Which of the following counterstains would be used with fluorescent detection

A

DAPI

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30
Q

In tyramide signal amplification, the tyramide conjugate is activated by

A

Horseradish peroxidase

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31
Q

The optimal method for utilizing a negative reagent control would be to

A

Run a separate patient tissue section for each block

32
Q

Which of the following would be a disadvantage of TMAs

A

Human error

33
Q

Negative reagent controls should be used for avidin-biotin based detection systems to detect

A

Endogenous reactivity

34
Q

Upon microscopic review of an IHC stained slide using LSAB detection, there are areas of inconsistent background staining in the specimen and control tissue (both are on the same slide). What is a possible cause

A

Biotin blocking was omitted

35
Q

A negative tissue control is used to

A

Detect primary antibody cross reactivity

36
Q

Which of the following is used for Ig amplification

A

Intermediary antibodies targeting the primary antibody

37
Q

Which of the following would be an advantage to TMA

A

Donor tissue is conserved

38
Q

When using Ig amplification with a mouse primary antibody, you would use

A

Rabbit anti-mouse IgG

39
Q

Which of the following factors would not influence IHC results

A

Antigen localization

40
Q

An optimal positive tissue control is one that is

A

Fixed and processed identically as the patient

41
Q

Which antibody is typically used as a tissue processing control to determine adequate fixation

A

Vimemtin

42
Q

This type of control would confirm that the IHC protocol has been performed correctly

A

Positive tissue control

43
Q

There is usually a high chance of false positive or negative results with cytology specimens because

A

Controls treated identically to cytology specimens are not readily available

44
Q

Which of the following is considered the best practice for positive and negative tissue controls

A

Multi-tissue block

45
Q

Non-specific or background staining on the negative reagent control may indicate

A

Reagent contamination

46
Q

On an IHC stained slide, the patient specimen has weak positivity. The control stains adequately. Both are on the same slide. What could cause this

A

A fixative other than 10% NBF was used to fix the patient tissue

47
Q

Both the control and patient tissue unexpectedly show non-specific or background staining (both are on the same slide). The most likely reason is

A

Endogenous activity was not blocked

48
Q

There is optimal intensity with non-specific staining. What protocol parameter would you suggest changing

A

Decrease the antigen retrieval aggressiveness

49
Q

Which type of tissue control is commercially prepared to contain negative, weak and strongly positive cells to improve standardization

A

Cell line control

50
Q

This type of tissue control can improve standardization across many different laboratories

A

Cell line control

51
Q

A pathologist orders two IHC tests on the same specimen block. The antigen retrieval for each protocol is different. If you were asked to run only one negative reagent control, which would be true

A

Choose the most aggressive antigen retrieval of the two

52
Q

There is incomplete cell membrane staining in cells known to contain the antigen meaning all of the antigen is not being detected. This would be referred to as

A

Weak specificity/sensitivity

53
Q

A spectrum of staining in the positive tissue control is important to

A

Detect subtle changes in antibody sensitivity

54
Q

When selecting positive tissue control, it is optimal to select tissue with

A

Low to high intensity staining

55
Q

The purpose of amplification is to

A

Increase intensity

56
Q

Specific staining in an unexpected area of localization on the positive tissue control may indicate

A

Antigen diffusion

57
Q

A pathologist returns an IHC slide to the histology lab stating that there is non-specific staining in the specimen, however the control is optimal (both are on the same slide). The patient tissue is liver. What could be an explanation for this

A

Reactivity with hepatitis antigen

58
Q

When creating a TMA, the tissue core is removed from the

A

Donor block

59
Q

An IHC result shows weak staining in the correct area of localization in both the patient and control tissue on the same slide. This is an example of

A

Optimal specificity, low intensity

60
Q

Autopsy tissue is a poor choice for positive tissue controls because

A

The tissue is often subject to autolysis and putrefaction

61
Q

Positive tissue controls are necessary to

A

Assess the performance of the primary antibody

62
Q

Upon microscopic review the control tissue demonstrates optimal staining, and the patient specimen is weak (both are on the same slide). What could be a cause

A

The IHC reagents did not adequately cover the entire slide

63
Q

An isotype control is also referred to as a/n

A

Negative reagent control

64
Q

An IHC stained slide shows weak intensity, but good specificity on the control and specimen (both on the same slide). A possible cause is

A

The primary antibody dilution is too high

65
Q

The most common type of hematoxylin used as a counterstain for IHC is

A

Mayer

66
Q

Non-specific binding of serum (Ig) proteins will not be detected with this type of negative reagent control

A

Diluent

67
Q

A positive tissue control is also used to

A

Verify that the protocol was performed correctly

68
Q

Intensity that is too dark in the patient specimen, but optimal in the control (both on the same slide) can be indicative of

A

Over incubation of DAB chromogen

69
Q

Control tissues with low to high levels of antigen expression are necessary to

A

Reduce the risk of false positive or negative results

70
Q

After completing an IHC protocol, positive staining is absent in the control and specimen (both on the same slide). What could be a possible cause

A

Inadequate deparaffinization

71
Q

Which of the following would be omitted for the negative reagent control

A

Primary antibody

72
Q

The staining intensity is too dark and the specificity is too weak. What are two potential protocol modifications that could improve results

A

Increase the antigen retrieval time, decrease the antibody incubation time

73
Q

Which of the following is not a purpose for controls in immunohistochemistry

A

Determine the appropriate section thickness

74
Q

There is excessive background on an IHC stained patient specimen, however the control is adequate (both on the same slide). What could be a cause

A

The specimen is necrotic

75
Q

When the patient tissue contains tissue elements that express both positive and negative staining, it is called a(n)

A

Intrinsic control

B. Final exam 405 (5 decks)

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