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Histopath LAB > IHC > Flashcards

IHC Flashcards

1
Q

is used in histology to detect the presence of specific protein marker that can assist with accurate tumor classification and diagnosis

A

Immunshstochemistry (IHC)

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2
Q

It is used in the pathology laboratory as an aid in the differential diagnosis and classification of cancer, and for certain other diseases, including infections.

A

IHC

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3
Q

The factors that influence the immunohistochemical staining result start in the _____and end at the____ by the pathologist, which ultimately leads to treatment decision by the oncologist

A

surgery operating room

interpretation of the stain

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4
Q

Pre analytical steps

BAG TES

A

Biopsy
Acessioning
Grossing
Tissue processing and Embedding
Sectioning

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5
Q

Immunshstochemistry (IHC) is used in histology to detect the presence of specific______ that can assist with accurate tumor classification and diagnosis,

A

protein marker (antigen)

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6
Q

(surgically removed tissue specimen or needle biopsy) from the surgery room arrives in fixative at the pathology laboratory.

A

BIOPSY

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7
Q

the sample details are entered into the_______. A barcoded label can ensure track and trace capabilities

A

ACCESSIONING

laboratory information system (LIS)

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8
Q

the specimen is visually examined for suspicious areas that require further examination.

Samples from the specimen that require further microscopic testing are excised as tissus blocks and placed barcoded cassetes

A

Grossing

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9
Q

The steps where the tissue block is processed into a form and condition suitable for making ultrathin microscopic sections.

Typically, the tissue is fixed in formalin then dehydrated before it is embedded in paraffin

A

TISSUE PROCESSING AND
EMBEDDING

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10
Q

The fine art of cutting the parafin-embedded tissue blocks into ultrathin (-4 um) sections and placing them onto glass sides.

A_____ on the slide can ensure traceability and may also contain protocol information for the requested test for that particular section.

A

SECTIONING

barcode

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11
Q

ANALYTICAL STEPS

A

Antigen retrieval
Blocking
Primary antibody
Enzyme HRP

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12
Q

It is performed to recover the antigens
that may have been altered by fixation,

A

Antigen Retrieval

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13
Q

Endogenous enzymes are blocked (this step can also be performed after
primary antibody incubation)

A

Blocking

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14
Q

A_______ is applied that specifically binds to the antigen of interest

A

primary antibody

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15
Q

The______ antibody carries the label (enzyme) upon application it binds to the primary antibody

A

Enzyme-HRP

secondary

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16
Q

ARS

A

Antigen Retrieval Solution

17
Q

Antigen retrieval is not necessary in

A

Frozen sections

18
Q

Performed to visualize nuclei and overall tissue architecture

A

Counterstaining

19
Q

DAB

A

diaminobenzidine

20
Q

Post-Analytical Steps
• In the post-analytical process, the pathologist interprets the stains in context with________ controls, using ______microscopy.
• The results are reported to the_____ for treatment decision.

A

positive and negative tissue

bright field

oncologist

21
Q

SECTIONING
Use High Quality Sections

Take particular care to use thin, flat sections that have been thoroughly dried onto the slide. Preferably use __________ or _______slides for IHC.

Uneven, poorly-adhering sections stain unevenly with variable background staining

A

charged slides or APES coated

22
Q

Again, these controls must be processed identically to the specimen but contain the target protein.

In some cases, it will be advantageous to have this control tissue stain only marginally positive as to monitor not only for the presence of the antigen, but also for any possible loss of sensitivity.

This loss might not be apparent if only intensely staining controls are used.

Controls for loss of sensitivity would be particularly important, for example, when staining tumors.

A

INTERNAL TISSUE CONTROLS

23
Q

Chromogen

A

DAB

Brown

24
Q

IHC STAINING PROCEDURE
MATERIALS

A

• DRYING OVEN
• CONTROL TISSUE (+)
• HUMID CHAMBER
• STAINING JAR
• TIMERS
• COVERSLIPS
• DISTILLED WATER
• CHARGED SLIDES
• REAGENTS FOR DEPARAFFINIZATION
• WATER BATH 95-99 DEGREES CELCIUS
• PIPETTES AND TIPS
• DAKO FLEX Ready-to-use Primary Antibodies or
• Dako Concentrated Primary Rabbit or mouse
Antibodies
• Dako Antibody Diluent
• Envision FLEX Target Retrieval Solution (high pH)
• Envision FLEX Target Retrieval Solution (low pH)
• Wash buffer
• Hematoxylin

25
Q

• Envision FLEX Target Retrieval Solution (high pH)

• Envision FLEX Target Retrieval Solution (low pH)

A

Blue

Pink/ Magenta

26
Q

Counterstaining

A

Hematoxylin

27
Q

For ________the sensitivity and specificity are the core elements.

A

primary antibodies

28
Q

Ideally, the primary antibody must provide both a______ and______ to produce an accurate and robust IHC

A

high sensitivity and a high specificity

29
Q

have become widely used because of their high specificity, consistency, purity and commercial availability.

A

Monocional antibodies

30
Q

One binding site

A

Monoclonal antibody

31
Q

Monoclonal antibodies, produced in______according to the ______method developed by_____.

A

mice

In vitro hybridoma

Kohler

32
Q

, typically produced in_____ by ________techniques, with booster immunizations to maximize the reactivity against the target antigen, frequently give a higher______ (avidity) compared to monoclonal antibodios, as the many antibody “species’ present react with more antigen sites.

A

Polycional antibodies

Rabbits

traditional immunization techniques

sensitivity

33
Q

The major drawback of Formalin-Fixed Paraffin-Embedded (FFPE) tissue is that formalin-induced molecular modification of proteins (antigens) may result in loss of the ability of the antibody to react with the antigen, a loss that can only be corrected by the restoration (retrieval) of the
“formalin-modified’ antigen molecular structure

A

TARGET RETRIEVAL SOLUTION

34
Q

These improve antigen expression of your samples by breaking down formalin induced______, re-exposing epitopes on the antigen to antibody binding.

A

TRS

antigen cross-linking

35
Q

Heat and enzyme retrieval are both employed, with______ now being the most commonly used.

In simple terms HIER involves heating your slides in buffer at___ or ____(depending on your antibody) using a microwave or a pressure cooker.

A

heat induced epitope retrieval (HIER)

pH6 or pH9

36
Q

TRS

A

Target Retrieval Solution

37
Q

HIER PROCEDURE

1.Place a staining jar containing diluted____ in awater bath. Cover the staining jar with lid to stabilize the temperature and prevent evaporation then cover the water bath.

2.Heat the water bath and the staining jar filled with TRS to_____

3.Measure temperature inside the staining jar with a calibrated thermometer to ensure correct temperature. Immerse the rack slides in the Preheat bed PRS in the staining jar. Cover the staining Jar and the
water bath

4.Bring the temperature of the water bath and the TRS back to 95-99°C. Incubate for____ minutes at 95-99°C.

5.Remove the entire staining jar with slides from the water bath. Allow the slides to cool in the solution for___ minutes at room temperature.

  1. Immediately soak slides in a staining jar with diluted_____
A

TRS

90-95°C.

25

20

Wash Buffer (WB).

38
Q
  1. Tap and wipe off excess buffer and draw a circle around the Uissue section with_____.
  2. Apply two drops (100 ul) or enough __________ Incubate for 5 mlnutes.
  3. Put sildes in the_______.
  4. Rinse gently with wash buffer solution and place in a fresh wash buffer bath for five minutes.
    5.Tap off excess buffer and apply two drops or enough______, Incubate for 30 minutes.
  5. Rinse gently as in Step 4.
  6. Tap off excess buffer and apply two drops or enough______, Incubate for 30 minutes.
  7. Rinse gently as in Step 4.
  8. Tap off excess buffer and apply two drops or enough _______Incubate for 10 mlnutes.
  9. Rinse gently with distilled water and place in a fresh distilled water bath for 5 minutes.
    11.Tap off excess water and apply two drops or enough______. Incubate then wash with distilled water and soak in wash buffer for 1 minute.
    uh distilled-water-twise.
    STAINING FROCEDURE - one rhanel and Xylene twice each for 1 min.
    nountina medlum
A

Dako pen

Dual Endogenous Enzyme Block

humidity chamber

Dako Primary Antibody

Labeled-polymer HRP

DAB+ Substrate Working Solution

hematoxylin

Histopath LAB (19 decks)

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