Histopathology Flashcards

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1
Q

Define histopathology.

A

The study of changes in tissue associated with disease.

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2
Q

Define Cytopathology

A

The study of changes in cells associated with disease.

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3
Q

What are the purpose of histopathological samples being taken?

A
  • Screening programmes highlight some abnormalities.
  • To aid diagnostic processes.
  • To aid therapeutic processes.
  • To determine prognosis and direct treatment.
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4
Q

When is bowl cancer screening needed?

A

If there is blood in the faces.

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5
Q

When is cervical screening carried out?

A

If the HPV virus is detected.

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6
Q

What are the guidelines for screening programmes?

A

Benefits should outweigh negatives, target population should be large.

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7
Q

What is the aim of histology?

A

To generate a microscope slide for a pathologist to make a diagnosis from.

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8
Q

What are the steps to produce a histological sample?

A
  • Receipt/ booking in
  • Fixation
  • Dissection
  • Processing
  • Embedding
  • Microtomy
  • Staining
  • Special Stains
  • Immunohistochemistry
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9
Q

Where do histopathological samples come from?

A

GPs surgeries, outpatient clinics or from surgery.

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10
Q

Describe the labelling on histopathological samples.

A

At least 3 forms of labelling are critical to ensure the sample and its identity correspond.

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11
Q

What substance are most histological samples fixed in and what is this?

A

Formalin - a pungent gas soluble in water.

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12
Q

Why may formalin not be used in the fixation process of pathology?

A

Formalin impedes some studies of proteins by cross-linking with them. So it is often not used if specific protein structure and function needs to be maintained.

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13
Q

What is the aim of the dissection process of histopathology?

A

To transfer pieces of tissue onto cassettes.

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14
Q

What does the technique used in dissection depend on?

A

Sample size

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15
Q

Describe dissection of small samples

A

Does not require any dissection or sampling.

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16
Q

Describe dissection of intermediate specimens

A

Require dissection and sampling, some may have representative pieces taken.

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17
Q

Describe dissection of large specimens.

A

Dissection by consultant pathologists.

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18
Q

What is the aim of the processing part of the histological process ?

A

To remove water from the tissue and infiltrate with molten paraffin wax.

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19
Q

What are the reagents for processing in histopathology?

A

Alcohol
xylene
Molten wax

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20
Q

What is the aim of embedding in histopathology.

A

To transfer the tissue into a wax block.

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21
Q

What factor is important in the embedding process in histopathology?

A

Sample orientation

22
Q

What is the aim of microtomy in the histological process?

A

To shave a thin section of tissue from the wax block and mount onto a microscope slide so it can be stained.

23
Q

What piece of equipment is used in microtomy?

A

A microtome

24
Q

How thick should samples be after microtomy?

A

3-4 microns thick typically

25
Q

What dyes are typically used for staining in histopathology?

A

H&E

26
Q

Describe how H&E staining works.

A

Haematoxylin is a basic due which stains acidic structure such as DNA in the cell nucleus (stains blue)

Eosin is an acidic dye which stains basic structures pink

27
Q

What does is immunohistochemistry used to stain?

A

Antigens in tissue

28
Q

What is immunohistochemistry useful for ?

A

Determining what specific type of cancer a person may have.

29
Q

Outline what happens in immunohistochemistry.

A

Histological, immunological and biochemical techniques are combined for the identification of specific tissue components by means of specific antibody/antigen reaction tagged with. a visible label. It involves the process of selectively identifying antigens in cells of a biological specimen by exploiting the principle of antibodies binding specifically to antigens in biological tissues.

30
Q

Why is formalin used for fixation?

A
  • It preserves tissue in a ‘life like state’.
  • Prevents cell autolysis.
  • Kills bacteria and viruses
  • It forms cross links with proteins and preserves them.
31
Q

What virus does formalin not kill?

A

CJD

32
Q

What issues can occur during fixation?

A

Fixation can introduce artefacts (any structure or feature in the histological section, which is not normally present in the normal tissue, and it may come from outside sources).

33
Q

What are the possible mechanical transmission methods of prion disease?

A

Acquired, sporadic and familial

34
Q

What symptoms can occur due to prion diseases?

A

o Myoclonus, ataxia
o Often presents with personality change
o Gait rapidly affected
o Aphasic, no eye contact, no memory

35
Q

What methods are used to diagnose prion diseases?

A

MRI
EEG
CSF analysis

36
Q

What is good about immunohistochemistry?

A
  • Antibodies bind to antigen in specific manner
  • Can be used to locate particular cells and proteins where protein expression varies in space
  • Can be used to identify cellular events such as apoptosis where protein expression varies over time
  • It is possible to visualise multiple targets simultaneously using immunohistochemistry.
37
Q

In histochemistry, immunoglobulins will ‘stick’ specifically to the antigens against which they are raised in the fixed tissues/ cells. But how do we see them?

A

Add something that permits visualisation – enzyme, fluorophore etc. But why modify every antibody you make when you can create a generic ‘secondary antibody’ directed at the unchanging part of the ‘primary’ antibody.

38
Q

Describe direct immunohistochemsitry.

A
  • Primary antibody only
    o A cell with an antigen attached to it. The antibody binds to this which is conjugated to a fluorescent marker and allows detection.
39
Q

Describe indirect immunohistochemistry.

A
  • Uses primary and secondary antibodies
    o Primary antibody binds to antigen on cell surface. Fluorescently labelled secondary antibody then binds to primary antibody.
  • This allows more specific detection
  • Allows amplification of signal
40
Q

What are the benefits of indirect immunohistochemsitry ?

A

Allows more specific detection and signal amplification.

41
Q

Describe how colposcopy works.

A
  • Uses Acetic acid:
    o Abnormal areas appear white due to higher DNA content
    o Normal areas do not stain
  • Uses Lugol’s Iodine:
    o Normal cells appear brown due to high glycogen content
    o Abnormal cells wont stain
42
Q

What is colposcopy ?

A

An examination of the cervix that is carried out if there are any concerns of HPV or cervical cancer.

43
Q

Describe the treatment of HPV.

A
  • Remove the area of concern with a clear margin around it. This will hopefully be curative and prevent the development of cervical cancer.
  • This is done outpatient in colposcopy
44
Q

Describe the structure of the lobes and ducts of a healthy breast.

A
  • Lobes and ducts form a tree like structure where the ducts are like the branches of trees and the lobules are the leaves.
  • A lobule has multiple acini.
    Healthy histology
  • 2 layers of epithelial tissue
  • Myoepithelial – contractile ( help excrete milk)
  • Luminal – secretory
45
Q

What are the different possible causes of breast lumps?

A
  • Lipoma
  • Fibroadenoma
  • Carcinoma
  • Phyllodes tumours
  • Cyst
  • Fat necrosis
46
Q

What is a lipoma?

A

Bening proliferation of adipose tissue.

47
Q

What is a fibroadenoma?

A

Benign tumour with epithelial and stroll involvement.

48
Q

In what people are fibroadenomas common?

A

Women in their 20s and 30s.

49
Q

In what people are carcinomas common?

A

Patients 60+ or with a genetic mutation

50
Q

Describe the histopathology of carcinoma.

A

Loss of myoepitheial layer

51
Q

Describe the immunohistochemsitry of carcinoma.

A

 The antibody p63 positively stains the myoepithelial layer
 There is no p63 staining in breast carcinoma because the myoepithelial layer has been lost