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Histology QIHC Exam > Chapter 12: Immunohistochemistry > Flashcards

Chapter 12: Immunohistochemistry Flashcards

1
Q

An antigen is a substance that triggers the production of:

A

Antibody (immunoglobulins).

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2
Q

A light chain present in some antibodies is:

A

Kappa and Lambda.

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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

What are two examples of a fluorochrome?

A

Rhodamine and Fluorescein Isothiocyanate (FITC).

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6
Q

Please describe the indirect IHC method:

A

Patient serum is added to tissue sections containing known antigens.

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7
Q

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

A

A biotinylated secondary Ab.

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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

A fluorochrome.

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9
Q

In some IHC techniques, alkaline phosphate functions as the:

A

The enzyme.

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10
Q

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

A

The enzyme.

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11
Q

The substrate for alkaline phosphatase labeled Ab is usually:

A

Napthol-AS-phosphate.

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12
Q

What is a common chromogen that is insoluble in alcohol?

A

DAB (3,3’ diaminobenzidine tetrahydrochloride)

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13
Q

If 3-amino-9-ethylcarbazole is used as the chromogen, the hemaxotylin used for the counterstaining must be:

A

Alcohol-free such as Mayer hematoxylin.

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14
Q

Tissue for immunofluorescence MUST be:

A

Frozen and unfixed.

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15
Q

A neoplasm is defined as:

A

A neoplasm is defined as a new growth of tissue in which cell multiplication is uncontrolled and progressive, it can either be malignant or benign.

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16
Q

What is NOT the proper approach to validating a new Ab?

A

Validating it ONLY with the manufacturers instructions, the lab should never assume that the Ab is immediately ready for use.

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17
Q

When the Vimentin stain is completely negative on formalin fixed tissues, this indicates that the tissue has been:

A

Unfixed.

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18
Q

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

A

The composition of the retrieval solution.

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19
Q

How is Ficin used in IHC?

A

Ficin is used as the enzyme in induced epitope enhancement.

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20
Q

0.5mL of a 1:50 dilution of Ab is needed. How many microliters of the primary would be needed?

A

10.0 microliters would be needed.

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21
Q

What has been eliminated with polymeric IHC staining methods?

A

The serum and avidin-biotin blocking steps have been eliminated, which improves turnaround times.

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22
Q

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

A

The primary Ab, or the secondary Ab won’t link them together.

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23
Q

How is Imidazole used in the PAP method of Ag detection?

A

As an intensifier to the Dab kit.

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24
Q

What link Ab would follow a monoclonal kappa primary Ab?

A

A rabbit anti-mouse because kappa is usually prepared in either mice or rabbits.

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25
Q

What is CK20 an Ab for?

A

Colon cancer.

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26
Q

Negative control slides that are run with each stain omit:

A

The primary Ab.

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27
Q

T/F: Prediluted Ab should always be provided by the manufacturer.

A

False, they should always be validated before use.

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28
Q

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

A

True, negative controls are those that emit the Ab during the primary incubation step and substitute for it.

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29
Q

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

A

False, a detailed procedure must be written for each tissue type.

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30
Q

T/F: Zinc formalin preserves immunoreactivity very well.

A

True.

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31
Q

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

A

False, they can be used with both monoclonal and polyclonal Ab.

32
Q

T/F: Blocking reactions are used to block endogenous activity of the same enzyme

A

True.

33
Q

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

A

True.

34
Q

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

A

True.

35
Q

T/F: DAB is an alcohol soluble chromogen.

A

False, DAB is not soluble in alcohol.

36
Q

T/F: Harris hematoxylin should not be used with AEC because of the alcohol present.

A

True, AEC is soluble in alcohol.

37
Q

T/F: Regulations regarding predictive marker staining of tissue places responsibility for documenting fixation times on the lab.

A

True.

38
Q

T/F: The only method of heat induced epitope retrieval involves the use of the microwave oven.

A

False. Other acceptable methods are heating are: pressure cookers, vegetable steamers and a circulating water bath.

39
Q

T/F: Optimal dilutions for each Ab must be determined in your lab.

A

True.

40
Q

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

A

AEC is not intensified, but the DAB is.

41
Q

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

A

True.

42
Q

T/F: Precut control slides may be stored at room temperature indefinitely.

A

False, storing them at -20C is ideal.

43
Q

T/F: The procedure for staining cytology smears is the same as the one for staining parrafin sections.

A

False, they need different protocols.

44
Q

What does CD3 test for?

A

T cell lymphomas.

45
Q

What does CD20 test for?

A

B cell lymphomas.

46
Q

What does HMB45 test for?

A

Melanoma.

47
Q

What does HER2 test for?

A

Breast carcinoma.

48
Q

What does GFAP test for?

A

Glioblastoma.

49
Q

What does AE1/AE3 test for?

A

Carcinomas.

50
Q

In the bloody areas of a tissue section stained with the immunoperoxidase technique, there is a marked reaction of the RBCs. What happened?

A

Failure to use hydrogen peroxide.

51
Q

Paraffin sections stained with the immunoperoxidase technique show excess background staining. What happened?

A

The nonimmune serum was not applied.

52
Q

The skin control for S100 was stained with the immunoalkaline phosphatase using fast red TR as the chromogen. It shows negative staining. What happened?

A

The sections were dehydrated and cleared with the rest of the IHC slides.

53
Q

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

A

Negatively stained positive control and a negatively stained specimen because mice or rabbit would have to be used instead.

54
Q

Both the known positive control and the specimen are negative following immunoperoxidase staining with LCA primary Ab. Dab was used as the chromagen. What happened?

A

The wrong secondary Ab was used, it should have been targeted to the primary Ab.

55
Q

Excessive background staining is noted in both the control and the specimen stained with immunoperoxidase. What happened?

A

The slides weren’t washed sufficiently with reaction buffer.

56
Q

Excessive background staining is noted on the specimen but not on the positive control stained with immunoperoxidase. What happened?

A

Free antigen in the patient tissue due to autolysis, necrosis or degeneration will cause this. Pam calls it mucin.`

57
Q

Both the positive control and the specimen stained with immunoalkaline phosphatase using CEA Ab shows very weak staining. What happened?

A

The primary Ab was incorrectly diluted.

58
Q

Why are acid-alcohol decolorizers generally recommended over aqueous decolorizers for use with AFB?

A

Acid-alcohol decolorizers allow for more uniform decolorization.

59
Q

What hematoxylin solution contains ethylene glycol and aluminum sulfate?

A

Gill hematoxylin.

60
Q

The bacterium Legionella pneumophilia can be demonstrated using which stain?

A

The Steiner technique.

61
Q

In the Jones and Gomori methenamine technique for basement membranes, sections are oxidized with what?

A

Periodic acid.

62
Q

Nissl substances can be demonstrated with what stain?

A

Thionin and cresyl echt violet stain.

63
Q

In the Wilder technique for demonstrating reticulin, uranyl nitrate acts as what?

A

The sensitizer.

64
Q

Microscopic evaluation of an H&E stained section reveals dark deposits of material dispersed irregularly over the tissue. What happened?

A

The hematoxylin was not filtered before use.

65
Q

Microscopic evaluation of a GMS stained section shows connective tissue fibers and RBCs stained black. What happened?

A

Overexposure to the hot methenamine silver.

66
Q

During microscopic evaluation of a fungus stain, extraneous fungal elements are seen above rather than in the section. What happened?

A

The staining solutions were cotaminated.

67
Q

Sections of lung stained with both GMS and PAS show positive staining of what appear to be fungal organisms. However, the same oval shaped bodies are seen in sections of the tissue where they shouldn’t be. Polarization of the bodies reveal a Maltese cross configuration. What is the “positively” stained material?

A

Talcum powder from either the tech or the pathologist.

68
Q

What are spirochetes?

A

Filamentous bacteria that are known to be argyrophilic.

69
Q

Following H&E staining, slides are dehydrated through ascending strengths of alcohol and cleared in xylene, but the first xylene is milky white. What happened?

A

The xylene has been contaminated with water, all dehydrating and clearing solutions should be changed.

70
Q

If hematoxylin is unavailable, what dye would make an appropriate substitute in a routine H&E?

A

Celestine blue.

71
Q

The phosphotungstic acid-hematoxylin (PTAH) stain is useful for demonstrating what?

A

Muscle striations.

72
Q

Hemosiderin is thought to be composed of:

A

Protein.

73
Q

Whether collagen fibers are found in dense regular arrangement or in an irregular arrangements varies with the:

A

They vary due to the location and function of the organ or tissue.

74
Q

Microscopic review of sections stained with H&E reveals very pink cytoplasm and pale reddish-brown nuclei. What happened?

A

The hematoxylin has become oxidized and needs to be replaced.

75
Q

In silver impregnation techniques, the use of improperly cleaned glassware results in:

A

Nonselective silver precipitation, chemically cleaned glassware should be used with silver stains ONLY.

Histology QIHC Exam (9 decks)

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