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POM (Principles of Medicine) > Histopathology > Flashcards

Histopathology Flashcards

1
Q

What is histopathology?

A

Deals with tissues, examining the architecture of the tissue; observations used in diagnosis, as well as towards informing the efficacy of a particular treatment

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2
Q

What are tissue samples?

A

Biopsies, resection specimens, frozen sections, and post-mortems

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3
Q

What are biopsies?

A

Concerns small sections of tissues removed from the patient; immersed in a formalin solution that preserves the tissues by cross-linking proteins.

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4
Q

What is formalin?

A

Preserves tissues by cross-linking proteins, samples are embedded in paraffin wax, enabling thin sections to be cut by a microtome.
Sample mounted on a microscope slide for preparation prior to analysis

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5
Q

What is the main function of a biopsy?

A

As a diagnostic tool

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6
Q

Which chemical stains are used for cells in the biopsy section?

A

Haematoxylin and Eosin staining used to identify nuclei and cytoplasmic granules of leukocytes

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7
Q

Which type of stain will stain acid-fast bacteria red, aiding in diagnosis for TB infection?

A

Ziehl-Neelsen stain

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8
Q

What are resection specimens?

A

Sourced from tissue removed from surgical procedure, processed as for a biopsy

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9
Q

What are resections used for?

A

To identify the stage and progression of the disease (cancers, diagnostic too for further treatment)

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10
Q

Why are tissues donated to biobanks?

A

Used to inform genomic studies of the disease process

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11
Q

Which sample is taken during surgical procedures and examined by pathologists in real time?

A

Frozen sections

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12
Q

How are sections frozen?

A

By cryostat, cut then mounted on glass slide and stained for biopsies

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13
Q

Why are frozen sections used?

A

Rapid diagnosis, relayed back to the surgeon to inform the surgery. Establishes presence & quantity of cancerous tissue, as well as identifying other pathological processes
Fresh tissue sample w/o formalin

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14
Q

How long are frozen sections stored and used for?

A

30 minutes

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15
Q

What is the timescale for biopsies?

A

2-3 days

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16
Q

What is the timescale for resection specimens?

A

5-7 days

17
Q

What is cytopathology?

A

The study of disease at a cellular level.
Cytopathologists identify cells, collected and smeared onto a microscope slide
The slide is stained and examined.

18
Q

What is used to analyse for a smear when inserted into a lesion & aspriate?

A

Fine needle aspirates

19
Q

Why are fine needle aspirates used?

A

The needle penetrates inaccessible tissue (thyroid nodule), and assess the suspect mass without the surgical requirement

20
Q

What is the disadvantage with fine needle aspirates?

A

Cytology concerns individual cells, and thereby unable to determine the architecture of the tissue and cellular influences on adjacent cells.

21
Q

What does brown coloration indicate during a skin biopsy?

A

Antibody recognizing the endothelial cell marker CD31 certifies the biopsy as an endothelial cell tumor.
Combining biopsies with analysis of fine needle aspirate taken from an enlarged lymph node, a diagnosis of reactive lymphadenopathy can be established due to mixed cell population observed

22
Q

What is used in immunohistochemistry?

A

Antibodies can be used to specifically detect molecules in the process of immunochemistry.

23
Q

Which region of the antibody is used to attach with markers?

A

Fc region.

24
Q

What are the four main attachments to antibody Fc regions?

A

Enzymes
Fluorescent probes
Magnetic beads
Drugs

25
Q

Which enzymes are attached to Fc regions of antibodies?

A

Peroxidases, and alkaline phosphatase used in conjunction with a colorless substrate to produce a colored product- CD31 staining; in addition to the detection of estrogen receptors on breast cancer tissue biopsies.

26
Q

Why are fluorescent probes used in antibody diagnostics?

A

Rapid measurement of the levels of molecules within a sample.
Multiplexing allows a measure of several molecules ina single clinical sample

27
Q

What is multiplexing?

A

Several antibodies with different fluorescence

28
Q

Why are magnetic beads used in antibody diagnostics?

A

Cells can be depleted of containment, Anti-CD3 antibodies used to deplete bone marrow of T cells prior to use in bone marrow grafts

29
Q

When are antibody diagnostics used?

A

Blood group serology, immunoassays (detection of hormones) circulating antibodies/antigens
Immunodiagnosis - presence of circulating anti-HIV antibodies suggests infection with HIV
High circulating levels of antibody suggest myeloma
IgE classes suggestive of the allergic phenotype

30
Q

What does ELISA stand for?

A

Enzyme-linked immunosorbent assay

31
Q

What is ELISA?

A

Serum samples allowed to adhere to a plastic plate; probed with a specific antibody raised against the molecule of interest
Antibody conjugated with an enzyme catalyses colourless substrate into a colour product
Reference to a standard curve, relatively absorbance of the solution can be used to determine precise levels of the molecule being studied

32
Q

What is the first stage of ELISA?

A

Antibody coating: Specific capture antibody is immobilized on high-protein binding plates, incubated
Plates are blocked with irrelevant protein (alubmin)

33
Q

What is protein capture during ELISA?

A

Samples and standard dilutions are added to the wells, and will be captured by the bound antibodies.

34
Q

What is a detection antibody during ELISA?

A

Specific biotinylated detection antibody inserted to wells, enables detection of the captured protein, Wash unbound detection antibodies.

35
Q

Which enzyme is conjugated with the wells?

A

Strepavidin-enzyme conjugate with alkaline phosphates is added to the wells and will bind to the biotinylated antibody

36
Q

What happens upon addition of substrate to ELISA?

A

Colorimetric substrate is added to the wells, and will form a coloured solution when catalysed by the enzyme

37
Q

How is the absorbance measured during ELISA?

A

By ELISA reader = amount of protein established.

38
Q

What is flow cytometry?

A

Allows detection of specific cells using fluorescently conjugated specific antibodies that associate to CD specific receptor sites on the lymphocyte cell surface membrane

39
Q

How does flow cytometry work?

A

Cells labeled with differently conjugated antibodies, run as a stream of single cells through a laser beam. Colour of emitted light and forward/side scatter denotes the identity of the cell surface molecules expressed, the size and granularity of cells.

POM (Principles of Medicine) (34 decks)

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