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Cellular Pathology > Stains and diagnosis > Flashcards

Stains and diagnosis Flashcards

1
Q

Congo red

A

Stains amyloid red

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2
Q

Alcian blue

A

Stains acidic mucins blue

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3
Q

Perl’s Prussian blue

A

Histochemical stain for iron/haemosiderin

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4
Q

Grocott

A

Fungi

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5
Q

PAS

A

Fungi

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6
Q

Giemsa

A

Differentially stains human and bacterial cells purple and pink respectively, platelets/erythrocytes pink and lymphocytes sky blue

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7
Q

Ziehl-Neelson

A

Bacteria, especially tubercules

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8
Q

Martius Scarlett Blue

A

Tinctorial stain for connective tissues - Red fibrin, Blue collagen, Yellow RBCs

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9
Q

Bacteria diagnosis stains

A

H&E, Gram, Giemsa, Ziehl-Neelson

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10
Q

Fungi stains

A

H&E, PAS, Grocott

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11
Q

Argyrophil and argentaffin

A

Argyrophil cells are capable of being impregnated with silver, but need a reducing agent to reduce it to a visible metallic silver; argentaffin cells can be impregnated with silver AND reduce the silver

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12
Q

Toluidine blue

A

Metachromatic (changes colour when bound to tissue), used for carcinoids, acidophilic

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13
Q

H&E

A

Haematoxylin is basic and purplish/blue (basophilic - nucleus/ribosomes/nucleoprotein)
Eosin is reddish/pink and acidic/cationic (acidophilic structures - cytoplasm, cell walls, ECM)
Haematoxylin oxidised to haematein, combined with mordant to stain cells

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14
Q

Liver stains - PAS/D

A

Glycogen - For glycogen storage diseases - Diastase (alpha-amylase) digests glycogen and starches to glucose. Periodic acid oxidizes glycols (and glucose??) to aldehydes, detected by Schiff reagent

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15
Q

Liver stains - Perl’s

A

Haemosiderin - For haemochromatosis or haemosiderosis

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16
Q

Liver stains - Orcein

A

HepB surface antigens, Elastin, Copper-bound proteins (Wilson’s)

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17
Q

Liver stains - Rhodamine

A

Copper - For Wilson’s disease

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18
Q

Liver stains - Masson Trichrome

A

Collagen - For fibrosis/cirrhosis

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19
Q

Fontana Masson

A

Argenaffinic substances - Often melanin

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20
Q

LFT panel

A

Bilirubin - jaundice, obstructions…
ALP - Bile duct system
AST, ALT, GGT - Damage or disease of hepatocytes
Albumin - made by liver

21
Q

Types of liver biopsy

A

Percutaneous - Through skin, usually w/ imaging/USG/MRI
Transvenous - Catheter via jugular vein, navigate to hepatic veins (clotting issues, ascites)
Laparoscopic/open wedge - Invasive, one large or multiple small incisions, greater visualisation

22
Q

Alcoholic liver disease (ALD) - stages

A

AFattyLD (steatosis) -> Alcoholic hepatitis (inflammation) -> Cirrhosis (scarring and fibrosis)

23
Q

Alcoholic liver disease (ALD) - ethanol mechanism

A

Ethanol -> ADH -> Acetaldehyde -> ROS -> OxStress -> Steatosis
Also - Inflammation -> Kupffer cells -> NADPH oxidase -> ROS -> OxStress -> Steatosis

24
Q

Alcoholic liver disease (ALD) - treatment options

A

Alcohol cessation, antioxidants, GFs, anti-caspase, anti-fibrotics, anti-inflammatories, TLR agonists

25
Q

Wilson’s disease

A

Autosomal recessive ATP7B mutation increases circulating copper -> Chronic hepatitis and cirrhosis + RBC haemolysis
Penicillamine and trientine are two chelating agents used to treat Wilson disease

26
Q

IHC markers - pulmonary, adenocarcinoma, squamous cell carcinoma

A

TTF1 shows pulmonary origin, adenocarcinoma is P63 -ve, squamous cell carcinoma is P63 +ve

27
Q

Druggable gene mutations associated with lung cancer, with assays

A

EGFR (FISH gold standard, can use IHC), ALK1 (IHC), PDL1 (IHC), ROS1 (IHC & FISH)

28
Q

IHC marker - breast

A

CK14

29
Q

Normal andrology values - semen volume, pH, sperm per ml/ejaculation, total motility, vitality, morphology

A
  • semen volume min 1.5 ml
  • pH min 7.2
  • sperm min 15 mil per ml
  • therefore total sperm min 39 mil per ejaculation (~ 15 x 1.5??)
  • total motility min 40%
  • vitality min 58% live spermatozoa
  • morphology min 4% normal forms of spermatozoa
30
Q

Breast cancer - Nottingham Prognostic Index

A

Nottingham Prognostic Index (NPI) = = [0.2 x S] + N + G

Where:
S is the size of the index lesion in centimetres
N is the node status (0 nodes = 1, 1-3 nodes = 2, >3 nodes = 3)
G is the grade of tumour (I to III - 1 to 3)

31
Q

Breast cancer - NPI - Grading criteria

A

Differentiation, variation in size/shape of tumour cells, mitotic counts

32
Q

CRC - Screening tests

A

Foecal Occult Blood (FOB) test for non-visible blood in stool - All men and women 60-74

Bowel scope test (colonoscopy) - All men and women over 55

FOB being replaced by FIT (Foecal Immunochemical Test)

Younger individuals in high-risk groups also invited - Hereditary Non-Polyposis Colorectal Cancer (HNPCC), family history, IBD

33
Q

Masson Trichrome

A

Red keratin and muscle fibres, blue or green collagen and bone, light red or pink cytoplasm, and dark brown to black cell nuclei

34
Q

Gömöri trichrome

A

Histological stain used on muscle tissue, can be used to test for certain forms of mitochondrial myopathy - Stains nuclei black, muscle fibre and cytoplasm red, collagen blue

35
Q

Demonstrating nucleic acids

A

Nucleic acids can be demonstrated in histological sections using the special stain methyl green-thionine (or methyl-green pyronin) - RNA stains red; DNA stains blue-green

For DNA - Feulgen reaction can also be used - Warm HCl, then Schiff’s reagent - DNA turns red as its hydrolysis releases aldehydes which make red complexes(?) with Schiff

36
Q

IHC markers - SCLC vs NSCLC

A

TTF1 for lung origin
Small cell lung cancer is CK7-, Non-small cell is CK7+
Both are CK20-

37
Q

IHC markers - CRC vs others

A

TTF- CK7-
CK20+ CDX2+: Colorectal carcinoma
CK20-:

38
Q

Ishak grading

A

For hepatitis/liver injury:

Interface hepatitis 0-4 + Confluent necrosis 0-6 + Necrosis/apoptosis/lobular inflammation 0-4 + Portal inflammation 0-4

1-3 is minimal, 4-8 mild, 9-12 moderate, 13-18 severe

39
Q

EWSR1 - FISH vs RT-PCR

A

FISH analysis using a break-apart probe is highly sensitive in detecting EWSR1 rearrangements but does not identify its translocation partner, whereas RT-PCR analysis is a more specific test in identifying the EWSR1-FLI1 fusion gene but has suboptimal sensitivity (54%) in formalin-fixed, paraffin-embedded tissue

40
Q

SRBCTs - Diagnosis and fusions

A

Diagnostic IHC used for subtyping, and alongside RT-PCR for diagnosis
IHC/RT-PCR - Expression of CD99 + FLI1 (EWS/pPNETs), myogenin (myogenic transcriptional regulator - RMS), keratin/EMA (SS), PAX5 (EWS/pPNET and RMS)
FISH - Fusions - PAX3/7-FKHR (FKHR == FOXO1) for RMS, SYT-SSX for SS, EWS-FLI1 for EWS/pPNETs
NOTE – FLI1 is also expressed in many lymphomas, including lymphoblastic lymphoma, which is also CD99 positive, so further analysis and consideration is needed

41
Q

IHC for breast cancer - Scoring methods

A

H-score = SUM[ (1 x % cells weakly stained) + (2 x % cells moderately stained) + (3 x % cells strongly stained) ]
Allred QuickScore = Proportion score + Intensity score
Proportion of staining (0 - no staining, 1 - 1 nucleus, 2 - 1 to 10 nuclei, 3 - 11 to 33 nuclei, 4 - 34 to 66 nuclei, 5 - 66 to 100 nuclei)
Intensity score (0 - negative, 1 - weak, 2 - intermediate, 3 - strong)

42
Q

Breast cancer NPI+

A

FFPE and IHC - Molecular classification via ER, PR, CK5/6/7/8, EGFR (AKA HER1), HER2/3/4, p53, Mucin 1
Then different NPI-like algorithms (size, shape, LN stage) for each class
More personalized and better prognostic ability, but more time-consuming and requires more tests (cost)

43
Q

Breast cancer - SLNB

A

Sentinel lymph node biopsy - Radioactive tracer or blue dye injected near tumour site allows ID of the first lymph node draining the breast, and so the first lymph node the cancer is likely to spread to
This is then biopsied, dissected, and assessed for presence of tumour cells to determine if the cancer has become invasive or not yet -> STAGING and prognosis
BC, melanoma, penile cancer
SNLs sectioned at levels throughout the node - less samples but still representative
IHC can be used at the same time to help locate small deposits of metastatic tumour cells -> Broad-spectrum CKs - AE1/AE3 and CAM5.2

44
Q

Breast cancer - TDLU IHC

A

The outer basal layer (of myoepithelial cells) stains brown with CK14 IHC in normal breast, and not the inside. CD44 can also stain rare but vital breast stem cells within the double-walled structure of the TDLU.

45
Q

Breast cancer - Diagnostic techniques

A

X-rays - Densities, (micro)calcifications, age-related atrophy of glandular tissue (and replacement by adipose tissue)
FNA - Cannot distinguish in situ vs invasive, not much prognostic info or functional measures (e.g. ER/PR/HER2 status)
Needle core biopsy may be preferred > FNA
Sentinel lymph node biopsy

46
Q

GI diagnostic techniques

A

Radiology, manometry (pressure), endoscopy (can take biopsies!), EUS (endoscopic ultrasound), scintigraphy (radiotracers)
Endoscopy - white light, narrow band (filter to block long-wavelength, deep-penetrating red light, keep blue/green light -> better visualisation of superficial morphology), chromoendoscopy (stains and dyes used during endo)

47
Q

Colorectal adenocarcinoma IHC markers

A

TTF1 & CK7 - ; CK20 & CDX2 +

48
Q

MSI tests in CRC

A

PCR for length/number of a panel of 5 microsatellite regions (BAT25/26, etc..), IHC for MMR gene protein expression.

Cellular Pathology (4 decks)

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