Histochemistry in Disease Diagnosis Flashcards

1
Q

Write about the liver biopsy
(5)

A

Its a small core/s of tissue
Used for the investigation of liver disease

Used to confirm a suspected clinical diagnosis

Used to establish extend of liver disease/monitor disease progression

Used to investigate malignancy (primary or metastatic)

Might be investigating a hepatocellular carcinoma -> usually investigating metastasis

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2
Q

Explain how a liver biopsy would be taken
(5)

A

Biopsy taken with a small gauge needle

Biopsy put in small pot of formalin

Need to be careful not to snap the tissue

Might see fatty change, inflammation etc

Are there any nests of malignancy?

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3
Q

What stains are often carried out on liver biopsies

A

H&E
Masson Trichrome -> looking for fibrosis
Gordon and Sweet’s silver stain

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4
Q

What is fibrosis

A

Fatty change intracellular in hepatocytes

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5
Q

Why might a silver stain be used on liver biopsies

A

Staining for reticulin

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6
Q

What might cause tissue alteration in liver biopsy

A

Infections e.g. Hep C or Hep B or parasites
Alcohol
Drugs
Metabolic issues
Autoimmune conditions
Haemachromatosis
Wilson’s disease
Malignancy

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7
Q

Why would we stain for alpha-1-antitrypsin
(3)

A

A deficiency in an enzyme causes deposition of AAT in the liver
We use Diastase and PAS to stain for AA1

The Diastase digests the glycogen so we can see the PAS positive material left

This is often seen in paediatric hospitals as it is a metabolic disease that affects neonates

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8
Q

How do we stain for Wilson;s disease

A

Rubeanic acid stain

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9
Q

How do we stain for Wilson;s disease

A

Rubeanic acid stain

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10
Q

How do we investigate malignancy in liver
(5)

A

H+E for morphology

Use IHC to confirm if malignancy is primary or metastatic

Use IHC to confirm if hepatocellular or cholangiocarcinoma

90% of liver cancer is metastatic

If metastatic need to identify cell or origin -> need IHC panel

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11
Q

Why might a renal biopsy be taken
(3)

A

Only done in specialised hospitals with a renal clinic

Only taken on clinical suspicion of renal disease -> usually diagnostic imaging is done first

suspicion of glomerulonephritis, nephropathy, chronic kidney disease

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12
Q

How do we process renal biopsies?
(3)

A

Need to determine if we have the structure we need for staining in the biopsy e.g. we need glomeruli if looking for glomerulonephritis

Biopsies usually come in fresh on moistened gauze in a petri dish

There are three different processes:
- Light microscopy - FFPE
- Fluorescent microscopy - frozen section
- Electron microscopy - resin

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13
Q

How do we examine glomeruli
(3)

A

Stain for basement membrane

Can use silver -> sensitise basement membrane then apply the silver

Look for alterations e.g. thickening or projections -> tramlines in the basement membrane

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14
Q

How do we examine nephronopathy?
(4)

A

Membranous staining

Look for projections

Fluorescence -> granular appearance

Normally linear stain in basement membrane in fluorescence

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15
Q

How would you be able to diagnose membranoproliferative glomerulonephritis

A

Tram tracking
Double layer of basement membrane

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16
Q

How do we investigate amyloidosis

A

Congo red
Fluorescence -> apple-green biorefringence

17
Q

What are the main components of the GIT?

A

Salivary glands
Oesophageal biopsy
Gastric biopsy
Small intestine
Large intestine

18
Q

What do we stain for in the GIT

A

Mucins
Infections

19
Q

Where are salivary glands usually investigated and how is this done

A

Head and Neck clinic

Mucin stains applied -> glandular structures with mucins

20
Q

What stains do we use to investigate H. pylori?
(2)

A

Warthin Starry

Giemsa

21
Q

Describe how H. pylori is investigated

A

Bacteria seen on surface of epithelium or upper layer of glandular cells, might be seen in crypts

22
Q

When is histochemistry used in cytopathology
(3)

A

Infections such as PCP, TB or fungi

Mucins such as mucin secreting adenocarcinoma

Amyloid

23
Q

How is PCP investigated?

A

Pneumocystis jirovecii

Bronchiole washes or brushings

Grocott’s silver

24
Q

How is TB investigated

A

ZN stain

25
Q

Why would we see adenocarcinomas in cytology

A

If you don’t have enough cells for immunohistochemistry

Could get a resection but this would take time

26
Q

What are signet cells

A

Adenocarcinoma of the stomach
Characteristic vaculation of cells
Nucleus gets pushed to one end of the cell
Can stain for mucin to confirm as mucin secreting carcinoma

27
Q

Write about histochemistry in malignancy
(7)

A

Alterations to cell/tissue components
- mucins
- connective tissue alteration
- amyloid

Assessment of invasion
- basement membrane

Immunohistochemistry utilised more for specific protein and biomarker of prognosis and therapeutics

28
Q

Quality Control and Trouble-shooting
(6)

A

Good control material
- know positive tissues for specific entities

Check each slide and control
- Quality of tissue section
- Quality of stain

Trouble-shoot
- slide/tissue sections/control
- protocols
- reagents

29
Q

Write about control in the lab

A

Need control material that is positive for the component your staining for

Need to check all of each test slides and the controls

Some labs will cut a control onto the same slide as the test -> usually pathologists do this and prefer this -> they have something to compare against

Need to be able to trouble shoot issues -> need to be able to identify issues and why they happened e.g. is there an issue with microtomy

30
Q

Give some examples of protocols

A

Have you done everything in order
If using automated stainer -> are the times correct
Are the protocols still valid
Check your reagent
Check stock control
If making up reagent have you use the right components and concentrations