Histopathology Flashcards

1
Q

What does a histopathologist do?

A

Deals with tissues, examines them and notes the architecture of the tissue and identify what it tells us about a particular condition

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2
Q

What does a cytopathologist do?

A

Deals with cells, takes them from the patient and prepares them for examination

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3
Q

What are the four tissue samples a histopathologist will work with?

A
  1. Biopsies
  2. Resection specimens
  3. Frozen Sections
  4. Post-Mortems
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4
Q

What solution are biopsy tissues placed in for preservation?

A

Formalin

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5
Q

How are biopsy samples preserved?

A

The formalin they are preserved in cross links the proteins

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6
Q

What are biopsy tissues embedded in to allow very thin sections to be cut by a microtome?

A

Paraffin wax

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7
Q

What cuts the biopsy tissue into very small sections?

A

Microtome

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8
Q

What stain is used to identify the nuclei and cytoplasmic granules of leukocytes within tissues?

A

Haemotoxylin and Eosin (H&E)

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9
Q

What is used to stain acid fast bacteria?

A

Ziehl-Neelsen stain

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10
Q

What stain is used to aid the diagnosis of tuberculosis infection?

A

Ziehl-Neelsen

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11
Q

How long does the results of a biopsy take to reach the clinician?

A

2-3 Days

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12
Q

How long does it take for the results of a frozen section to reach the clinician?

A

30 Minutes

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13
Q

How long does it take for results of a resection specimen to reach the clinician?

A

5-7 days

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14
Q

What are resection specimens?

A

specimens are taken from tissue that has been removed as part of a surgical procedure and can be processed as for a biopsy.

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15
Q

What are resection specimens primarily used to do?

A

Look at the stage of disease

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16
Q

Why may tissue samples from resection specimens be donated to biobanks?

A

They may be used to inform genomic studies of the disease process

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17
Q

What is a frozen section?

A

A sample taken during a surgical procedure and examined by pathologists in real time

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18
Q

What is used to freeze the tissue?

A

Cryostat

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19
Q

What are the requirements for tissue samples to be frozen?

A

Fresh and free of preservatives like formalin

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20
Q

What is a fine needle aspirate?

A

When a fine needle is inserted into a lesion and used to aspirate the cells out the lesion for anaylsis

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21
Q

Why is fine needle aspiration useful?

A

The needle can easily penetrate relatively inaccessible tissues without the need for surgery

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22
Q

What is the downside of fine needle aspiration?

A

the cytopathologist is only looking at cells and is unable to comment upon the likely architecture of the tissue.

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23
Q

Give an example of when both histopathology and cytopathology are used?

A

In the diagnosis of reactive lymphadenopathy

fine needle aspirate of enlarged lymph nodes and skin biopsy

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24
Q

What does Oil Red O stain?

A

Lipids

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25
Q

What does congo red stain?

A

ECM components like elastin and collagen

26
Q

Which molecules are conjugated to the Fc region of an antibody for their use in flow cytometry?

A

Fluorophores

27
Q

When requesting a test from the pathology lab to aid with a diagnosis, it is important to appreciate just how long the various procedures take. Put the following pathology lab tests in order of the time taken to get the result back to the clinician (quickest to slowest).



  1. Resection tissue
  2. Biopsy
  3. Frozen section
A

3 - 2 - 1

28
Q

What is Kaposi’s sarcoma?

A

Rare cancer and is often indicative of immunodeficiency (AIDS)

Spindly resembling cells evidently penetrates the collagen fibres

29
Q

What four forms can antibody conjugations come in?

A
  1. Enzymes
  2. Fluorophores
  3. Magnetic beads
  4. Drugs
30
Q

What 2 types of detection can antibodies be used for?

A

Direct - antibody with marker binds to antigen
Indirect - Indirect - antibody with marker binds to another antibody which binds to antigen

31
Q

What are conjugations?

A

Attachments to the Fc region of the antibody

32
Q

What are two examples of enzyme attachments?

A

Peroxidase and alkaline phosphatase

33
Q

When may enzyme attachments be used clinically?

A

The detection of oestrogen receptors on breast cancer tissue receptors

34
Q

What is multiplexing and what does it allow?

A

When several antibodies with different fluorescence are used, allows the measurements of several molecules in a single clinical sample

35
Q

What part of the antibody are conjugates attached to?

A

The Fc region

36
Q

How does enzyme conjugation work?

A

Antibody binds to antigen, and colourless substrate is added, the colourful product can be detected wherever the immune complex is

37
Q

Why do fluorescent probes allow for the rapid measurement of the levels of molecules within a sample?

A

because the colour is already there and unlinke enzyme conjugation does not need to be formed

38
Q

Which is a good choice for an antibody conjugate when you have a small sample?

A

Fluorescent probes are you can perform multiplexing

39
Q

What can magnetic beads be used for?

A

The purification of cell types

40
Q

Give an example of how magnetic beads are used as a clinical conjugate?

A

deplete the bone marrow of T cells before bone marrow graft

41
Q

What can drugs as an antibody conjugate be used to do?

A

Directly deliver the drugs to the cells that are malfunctioning

42
Q

Describe an example of the clinical use of drugs as an antibody conjugate?

A

Kadcyla, anti-HER antibody - used in the treatment of breast cancer as HER2 is overexpressed in over 30% of breast cancers

43
Q

What makes antibodies useful as a diagnostic tool?

A

The unique specificity of the antibodies

44
Q

What other molecules can antibodies detect?

A

Proteins, carbohydrates and lipids

45
Q

Describe how indirect detection of antibodies works?

A

The antibody with the antigen bound to it is detected by a secondary antibody which is conjugated with an enzyme of fluorescent probe

46
Q

What can manufactured antibodies be used for?

A

Blood group serology
Immunoassays
Immunodiagnosis

47
Q

What can immunoassays be used for?

A

Detection of hormones or circulating antibodies/antigens

48
Q

What can be detected via immunodiagnosis using manufactured antibodies?

A

IgE antibodies suggestive of allergic phenotype
Infectious diseases due to presence of anti-disease antibodies like HIV

49
Q

What is immune complex glomerulonephritis?

A

Disease caused by the presence of IgG-antigen immune complexes and Complement C3 from kidney biopsy

50
Q

What does flow cytometry notably allow the detection of?

A

Lymphocyte subpopulations

51
Q

How does flow cytometry work?

A
  1. Cells labelled with differently conjugated antibodies
  2. These cells then run as a stream of single cells through laser beams
  3. The colour of the light emitted and the scatterpatterns of the laser beam can denote the identify of the cell surface molecules expressed and therefore the type and size of cell
52
Q

What is flow cytometry highly informative to?

A

The nature of a particular inflammatory reaction, as the tagged antibodies allow the detection of specific white blood cells like CD4 helper cells etc

53
Q

Anti CD3+ antibodies are used to detect what cell?

A

T cell - pan T cell marker

54
Q

Anti CD4+ antibodies are used to detect what cell?

A

T helper cells

55
Q

Anti CD8+ antibodies are used to detect what cell?

A

Cytotoxic T cells

56
Q

Anti CD19+ antibodies are used to detect what cell?

A

B cells

57
Q

Anti CD56+ antibodies are used to detect what cell?

A

NK cells

58
Q

What does ELISA mean?

A

Enzyme Linked ImmunoSorbent Assay

59
Q

What is the purpose of ELISA?

A

Used to determine the precise concentrations of molecules in a sample

60
Q

How does ELISA work?

A
  1. Serum sample taken and allowed to adhere to a plastic plate -
  2. Antibody with enzyme as a conjugate is added, which can turn a colourless substrate a particular colour
  3. Wash away excess, unbound antibody
  4. Measure absorbance of light by coloured product - relative absorbance of the solution can be used to determine the precise levels of the molecule which is being studies