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1.1 POM Cells > Histopathology > Flashcards

Histopathology Flashcards

1
Q

What is histopathology?

A

Examination of tissues

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2
Q

What is cytopathology?

A

Examination of individual cells

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3
Q

What are the 3 types of tissue samples?

A
  1. Biopsies
  2. Resection specimens
  3. Frozen specimens
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4
Q

What do histopathologists do?

A

Examine tissues
- Analyse biopsies, resection specimens, frozen specimens and post-mortems

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5
Q

What do cytologists do?

A

Examine individual cells

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6
Q

What are the 2 purposes of tissue sections?

A
  1. Diagnoses
  2. Determine efficacy of treatment
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7
Q

What is a biopsy

A

Small section of tissue removed from patient

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8
Q

How are biopsies taken? (5)

A
  1. Small section of tissue surgically removed
  2. Preserved in formalin
  3. Embedded in paraffin wax
  4. Microtome slices to thin sections (2-3 micrometres)
  5. Mounted on a slide —> further prep eg. staining —> analysed under microscope
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9
Q

What solution is used to fix tissues during biopsy and why?

A

Formalin

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10
Q

Why are biopsies embedded in paraffin wax?

A

Allow microtome to slice very thing sections

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11
Q

What is used to obtain think slices from a biopsy?

A

Microtome

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12
Q

What is a microtome?

A

Instrument used to cut extremely thin slices (biopsy)

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13
Q

How can a biopsy be used in cancer diagnosis?

A

Diagnose type of cancer

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14
Q

What is a resection specimen?

A

Tissue sample removed during a surgical procedure

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15
Q

What are resections primarily used for?

A

See stage of disease

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16
Q

What can resection specimens be donated to?

A

BioBanks —> research pathology

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17
Q

What are frozen sections?

A

Fresh tissue frozen and analysed during a surgical procedure

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18
Q

When are frozen sections analysed?

A

Whilst a surgical procedure is occurring

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19
Q

How long do frozen sections take?

A

30 mins

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20
Q

How are frozen sections taken? (3)

A
  1. Fresh tissue taken
  2. Cryostat freezes tissue
  3. Tissue cut, mounted and stained —> analysed under microscope
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21
Q

What machine is used to obtain frozen sections?

A

Cryostat

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22
Q

What are the 2 conditions of the tissue needed for a frozen section?

A
  1. Fresh
  2. No preservatives (no formalin used)
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23
Q

How long do biopsies take?

A

2-3 days

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24
Q

How long do resection sections take?

A

5-7 days

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25
Q

What is a tissue smear?

A

Thin layer of tissue/cells spread onto a microscope slide

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26
Q

What are the 2 modes of cytology analysis?

A
  1. Smears
  2. Fine needle aspirate
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27
Q

What is an aspirate?

A

Sample of tissue draw out of body using a needle

28
Q

How are fine needle aspirates taken?

A

Thin, fine needle used to withdraw sample of cells/fluid a mass or lesion

29
Q

What is the advantage of fine needle aspirates?

A

Can reach usually inaccessible tissues without surgery
- eg. Thyroid nodule

30
Q

What is the disadvantage of fine needle aspirates?

A

Looks at isolated cells only —> can’t see tissue architecture

31
Q

What proportion of pathological diagnoses test use antibody conjugation?

A

Over half (53/86)

32
Q

Which method is most used for pathological diagnosis?

A

Conjugation of antibodies

33
Q

Which 3 dsDNA producing diseases are diagnosed using antibodies and why?

A
  1. SLE (lupus)
  2. Sjogren’s syndrome
  3. Rheumatoid arthritis
34
Q

What is immunohistochemistry?

A

Technique used to visualise and identify specific proteins using tagged antibodies (markers)

35
Q

Where are antibody conjugates binded to?

A

Fc region (constant)

36
Q

What are the 4 antibody conjugates?

A
  1. Enzymes
  2. Fluorescent probes
  3. Magnetic beads
  4. Drugs
37
Q

What are 2 examples of enzyme conjugated antibodies?

A
  1. Peroxidase
  2. Alkaline phosphatase
38
Q

Why do fluorescent probe conjugated antibodies allow for rapid analysis?

A

Immediately detected —> don’t need extra step

39
Q

What is multiplexing?

A

Simultaneous analysis of multiple proteins using multiple different antibodies

40
Q

When is multiplexing used?

A

When tissue sample is valuable/difficult to obtain —> get out as much information from single sample as possible

41
Q

When are magnetic beads used as antibody conjugates?

A

Cell type isolation (purification)

42
Q

How do magnetic beads antibody conjugates allow for cell type isolation?

A
  1. Magnetic bead surrounded by antibody
  2. Specific cell type binds to antibodies
  3. Magnet used to attract magnetic beads —> takes antibodies and bound cells with it —> only specific cells towards magnet
43
Q

What is an example of a magnetic bead conjugated antibody?

A

Isolation of T cells from bone marrow using anti-CD3

44
Q

Why may drugs be conjugated to antibodies?

A

Target specific tissues/sites of diease

45
Q

What is an example of a drug conjugated antibody?

A

Kadcyla = anti-HER2 + emtansine
- Anti-HER2 —> binds to HER2 receptors
- Emtansine —> cytotoxic chemical (drugs)
- Used to treat HER2-positive breast cancer

46
Q

What are the 2 types of antibody detection?

A
  1. Direct
  2. Indirect
47
Q

What is direct antibody detection?

A

Detection of antibodies directly in a sample
- Primary antibody binds to antigen —> detected

48
Q

What is a primary antibody?

A

Antibody that directly binds to a specific antigen

49
Q

What is indirect antibody detection?

A

Use of a secondary marked antibody to detect presence of primary antibody
- Primary antibody binds to antigen —> secondary antibody added to bind to primary antibody —> detected

50
Q

What is a secondary antibody?

A

Antibody that binds to primary antibody constant region

51
Q

What are 3 uses of manufactured antibodies

A
  1. Blood group serology
  2. Immunoassays
  3. Immunodiagnosis
52
Q

What are 3 examples of when antibodies are used for immunodiagnosis?

A
  1. See presence of pathogen
  2. See body’s antibody levels —> see if infected
  3. Test for IgE presence —> see if hypersensitivity reaction
53
Q

What is the ELISA test?

A

Enzyme Linked ImmunoSorbent Assay

54
Q

What are the 4 steps of the ELISA test?

A
  1. Sample adhered to plastic plate
  2. Probe with antibody specific to molecule of interest
  3. If antibodies bind —> enzyme conjugate colours solution
  4. Standard curve used to determine concentration of molecule (colorimetry)
55
Q

What is flow cytometry?

A

Cells with differently conjugated antibodies analysed via laser beam

56
Q

What is the use of flow cytometry?

A

Immunophenotyping (analyse number of each immune cell present depending on differing surface proteins)

57
Q

What are the … steps of flow cytometry?

A
  1. Antibodies attached to cells
  2. Fluid containing antibody-labelled cells —> single-file stream through laser
  3. Light through cell —> colour and scatter pattern analysed
58
Q

What is identified via flow cytometry? (2)

A
  1. Identity of cell surface molecules (that added antibody is bound to)
  2. Cell size and granularity
    —> Identity of cell
59
Q

Which properties of the laser beam are used in flow cytometry analysis?

A
  1. Colour
  2. Scatter pattern
60
Q

What are the 5 examples of antibodies used in flow cytometry and what do they detect?

A
  1. anti-CD3+ —> all T cells
  2. anti-CD4+ —> Th cells
  3. anti-CD8+ —> Tc cells
  4. anti-CD19+ —> B cells
  5. anti-CD56+ —> NK cells
61
Q

How can T cells be detected in flow cytometry?

A

anti-CD3+

62
Q

How are Th cells detected in flow cytometry?

A

anti-CD4+

63
Q

How are cytotoxic T cells detected in flow cytometry?

A

anti-CD8+

64
Q

How are B cells detected in flow cytometry?

A

anti-CD19+

65
Q

How are NK cells detected in flow cytometry?

A

anti-CD56+

66
Q

Which type of reaction is flow cytometry usually used to analyse?

A

Immune

1.1 POM Cells (15 decks)

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