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Histopath > Steps for antigen detection > Flashcards

Steps for antigen detection Flashcards

1
Q

What are the possible methods in labelling antibodies? (4)

A
  1. Use of Enzymes
  2. Use of Fluorochrome Dyes
  3. Use of Colloidal Gold
  4. Use of Radioactive Isotopes
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2
Q

This is commonly used to label antibodies

A

Enzymatic method

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3
Q

Commonly used enzymes in labelling antibodies: (2)

A
  1. Horseradish Peroxidase (HRP)
  2. Alkaline Phosphatase
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4
Q

Match:

  1. Horseradish Peroxidase (HRP)
  2. Alkaline Phosphatase

a. Alternative
b. Widely used

A
  1. HRP - b. Widely used
  2. ALP - a. Only an alternative
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5
Q

This is required when using enzymes for labelling.

A

Chromogens

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6
Q

This is a color developer to produce a colored end product

A

Chromogens

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7
Q

The end product of enzymatic method can be observed using which type of microscope?

A

Light microscope

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8
Q

The use of chromogens will require the use of which substance?

A

Substrate

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9
Q

This is the one consumed by the enzyme

A

Substrate

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10
Q

Which acts as substrate in these enzymes?

  1. Horseradish Peroxidase (HRP)
  2. Alkaline Phosphatase
A
  1. HRP - Hydrogen Peroxide
  2. ALP - Phosphate
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11
Q

Technique used in Fluorochrome dyes

A

Immunofluoresence

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12
Q

Identify the following:

Horse Radish Peroxidase

Substrate:

Chromogen: 3,3’ diaminobenzidine (DAB)

Color of Chromogen:

Counterstain: (3)

A

Substrate: Hydrogen Peroxide

Chromogen: 3,3’ diaminobenzidine (DAB)

Color of Chromogen: Brown

Counterstain: Hematoxylin,
Methyl Green, &
Methyl Blue

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13
Q

Identify the following:

Horse Radish Peroxidase

Substrate:

Chromogen: 3-amino-9-ethylcarbazole (AEC)

Color of Chromogen:

Counterstain: (2)

A

Enumerate the following:

Horse Radish Peroxidase

Substrate: Hydrogen Peroxide

Chromogen: 3-amino-9-ethylcarbazole (AEC)

Color of Chromogen: Brick-red

Counterstain: Hematoxylin &
Methyl Blue

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14
Q

In HRP, hematoxylin serves as _________ counterstain, and methyl green and methyl blue as __________ counterstain.

A

Traditional, alternative.

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15
Q

Identify the following:

Alkaline Phosphatase

Substrate:

Chromogen: 5-Bromo-4-Chloro-3-Indolyl-Phosphate (BCIP)

Color of Chromogen:

Counterstain: (2)

A

Identify the following:

Alkaline Phosphatase

Substrate: Phosphate

Chromogen: 5-Bromo-4-Chloro-3-Indolyl-Phosphate (BCIP)

Color of Chromogen: Bluish Violet

Counterstain: Nuclear Fast Red & Brilliant Green

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16
Q

Identify the following:

Alkaline Phosphatase

Substrate:
Chromogen: Nitro Blue Tetrazolium (NBT)

Color of Chromogen:

Counterstain:

A

Identify the following:

Alkaline Phosphatase

Substrate: Phosphate

Chromogen: Nitro Blue Tetrazolium (NBT)

Color of Chromogen: Black Violet

Counterstain: Nuclear Fast Red & Brilliant Green

17
Q

Identify the following:

Alkaline Phosphatase

Substrate:

Chromogen: Fast Red TR Salt

Color of Chromogen: Black Violet

Counterstain:

A

Identify the following:

Alkaline Phosphatase

Substrate: Phosphate

Chromogen: Fast Red TR Salt

Color of Chromogen: Rose-colored

Counterstain: None

18
Q

Forms of Hematoxylin used for IHC: (2)

A
  1. Mayer’s Hematoxylin
  2. Harris Hematoxylin
19
Q

What is the advantage of using Harris Hematoxylin?

A

Issue morphology are
well-defined

20
Q

Form of hematoxylin with reddish-violet resulting color.

A

Mayer’s Hematoxylin

21
Q

Form of hematoxylin with red nucleus and commonly used.

A

Harris Hematoxylin

22
Q

Incubation time to link antibody with peroxidase:

A

30-60 mins

23
Q

Incubation temperature to link antibody with peroxidase:

A

Room temperature

24
Q

This label is detected using a Fluorescence Microscope

A

Fluorochrome Label

25
Q

This label is detected using a Light Microscope

A

Colloidal metals

26
Q

This is also used to detect antigens aside from antibodies

A

Lectins

27
Q

Sources of lectins: (2)

A

Plant
Animal

28
Q

Lectins can be labeled just like antibodies. These can bind only with antigens.

a. First statement is true, second statement is true.
b. First statement is true, second statement is false.
c. First statement is false, second statement is true.
d. First statement is false, second statement is false.

A

b. First statement is true, second statement is false.

Lectins can also bind with tissue carbohydrates

29
Q

This is used in Fluorochrome label

A

FITC – fluorescein isothiocyanate

Histopath (12 decks)

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