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Advanced Haematology > Week 13 > Flashcards

Week 13 Flashcards

1
Q

Was is Cytochemistry?

A

In vitro staining of cells

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2
Q

What are some elements that may be stained in cytochemistry?

A

Leucocytes, erythrocytes

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3
Q

In Cytochemisty it is difficult to determine blast cell origin from morphology what is it helpful with?

A

Acute leukaemias

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4
Q

Even though cytochemisty is now largely replaced by Immunophenotyping, high resolution cytogenetic studies, molecular genetics. It is still used to some extent in differentiating what?

A

Differentiate M0 from M1 leukaemia (Myeloperoxidase, Sudan Black B)

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5
Q

List some specimen types that can be used – smears and imprints:

A
  • Bone marrow (most common)
  • Lymph node
  • Spleen
  • Peripheral blood
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6
Q

List 2 things that can affect enzymatic techniques.

A
  • Some fixatives

* Age of specimen

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7
Q

What is the main use for the Kleihauer – Betke test?

A

• Evaluate foeto-maternal haemorrhage
Guide dosage for anti-D administration
• Rh(D) negative mother with Rh(D) Positive baby

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8
Q

Do maternal cells stain more strongly or do foetal cells?

A

Foetal stain more strongly

Maternal stain like ghosts

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9
Q

List the steps of the Kleihauer – Betke test for preparing maternal blood film

A
  • Fix in ethanol
  • Immerse in acid-citrate buffer (pH 3.3)
  • Stain
  • Determine % foetal cells
  • Calculate volume of bleed
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10
Q

What cells is myeloperoxidase enzyme is found in?

A

Neutrophil and eosinophil primary granules

Monocytes - to some extent

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11
Q

What cells is myeloperoxidase enzyme not found in?

A

Lymphocytes

Erythrocytes

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12
Q

Myeloperoxidase is useful for differentiating blasts from which leukaemias?

A

AML blasts from ALL blasts

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13
Q

Sudan Black B is useful for differentiating which leukaemias?

A

AML from ALL

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14
Q

Sudan Black B has similar results to myeloperoxidase however it’s a little more sensitive for what?

A

early myeloid cells

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15
Q

What does sudan black B stain? give examples

A

lipids eg: • Sterol fats, neutral fats, phospholipids

Note: SBB is lipid soluble

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16
Q

Where are the lipids found that Sudan Black B stain?

A
  • Primary and secondary granules of neutrophils

* Lysosomal granules of monocytes

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17
Q

How would the following cell types stain?
Granulocytes
Monocytic cells
Lymphoid cells

A

Granulocytes positive from myeloblast onwards
Monocytic cells negative to weakly positive
Lymphoid cells Negative - note burkitts cell vacuoles may stain positive

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18
Q

What would the myeloperoxidase and sudan black b results be for M0 Acute myeloid leukaemia minimally differentiated?

A
  • Myeloperoxidase negative

* Sudan Black B negative

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19
Q

What would the myeloperoxidase and sudan black b results be for M1 Acute myeloid leukaemia without maturation?

A
  • Myeloperoxidase positive

* Sudan Black B positive

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20
Q

What are esterases used for?

A

Used to differentiate granulocyte vs monocyte cell lines.

21
Q

How many esterase isoenzymes are present in leucocytes?

A

Nine

22
Q

Esters used as substrates may be “Specific”, meaning

A

Only granulocyte series stain positive

• Napthol AS-D chloroacetate (specific esterase)

23
Q

Esters used as substrates may be “Non-specific”, meaning

A
  • Monocyte series and some other cell lines stain positive
  • a-napthyl acetate (non-specific esterase)
  • a-napthyl butyrate (non-specific esterase)
24
Q

Leucocyte alkaline phosphatase is used to differentiate what?

A

• Chronic myeloid leukaemia and Leukaemoid reaction in severe infection

25
Q

Would an untreated CML have an elevated or decreased LAP?

A

Decreased LAP

26
Q

Would a Leukaemoid reaction have an elevated or decreased LAP?

A

High normal to increased LAP

27
Q

Would the following conditions have a High, Normal. or low LAP score?
• 3rd trimester of pregnancy
• Polycythaemia vera

A

High scores

28
Q

Would Secondary polycythaemia have a High, Normal. or low LAP score?

A

Normal score

29
Q

Would the following conditions have a High, Normal. or low LAP score?
• Paroxysmal nocturnal haemoglobinuria
• Sideroblastic anaemia
• Myelodysplastic disorders

A

Low score

30
Q

Acid phosphatase enzyme present in what cell lines?

A

Myeloid, monocytic, lymphoid, plasma cell, platelets

31
Q

Acid phosphatase is tartrate __________

A

resistant

32
Q

Hairy cells produce isoenzyme ____

A

5

33
Q

tartaric acid inhibits all isoenzymes except

A

5

34
Q

In ACP the addition of L-(L+)-tartaric acid make
• Haemopoietic cells positive or negative
• Hairy cells remain positive or negative

A
  • Haemopoietic cells negative

* Hairy cells remain positive

35
Q

Masson’s trichrome stain Used to study

A

connective tissues, muscle, collagen fibres

36
Q

Masson’s trichrome stain is used to demonstrate increased collagen in ________

A

myelofibrosis

37
Q

Silver stain is used to stain _______ ______ that replace normal marrow stroma in _________

A

reticulin fibres, myelofibrosis

38
Q

Reticulin fibres comprised of Type ___ collagen

A

III

39
Q

What 3 things would you check if control cells do not exhibit expected staining pattern ie:
• No colour when they should be positive
• Incorrect colour

A

Reagents - correct or expired
Specimen - age, storage
Method - correct steps

40
Q

Smears for which of the following could be stable for several months at room temperature

a) Myeloperoxidase
b) Sudan Black B
c) Leucocyte Alkaline Phosphatase
d) a-Napthyl butyrate esterase

A

b) Sudan Black B

41
Q

Which cytochemical stain can be used to differentiate AML from ALL?

a) TRAP
b) Leucocyte Alkaline Phosphatase
c) Myeloperoxidase
d) Alpha-napthyl acetate

A

c) Myeloperoxidase

42
Q

Sudan Black B stains which cellular components?

a) Glycogen
b) Lipids
c) Structural proteins
d) Enzymes

A

b) Lipids

43
Q 
Hairy cells produce an abundance of which isoenzyme of acid phosphatase?
How can you demonstrate this isoenzyme?
a) 1
b) 3b
c) 4
d) 5
A

d) 5

• Demonstrate tartrate resistance

44
Q

An LAP score of 250 would be consistent with a diagnosis of:

a) Normal cells
b) CML
c) Hairy cell leukaemia
d) Leukaemoid reaction

A

d) Leukaemoid reaction

45
Q

The cytochemical stain a-napthyl butyrate is a non-specific esterase stain that reacts with cells of which lineage?

a) Erythroid
b) Monocytic
c) Granulocytic
d) Lymphoid

A

b) Monocytic

46
Q

Which of the following stains is used to evaluate mature neutrophils and bands when discriminating reactive changes from CML?

a) Acid phosphatase
b) Leucocyte Alkaline Phosphatase
c) Romanowsky
d) Sudan Black B

A

b) Leucocyte Alkaline Phosphatase

47
Q

Cytochemical staining on a spleen imprint stained positive for acid phosphatase. The stain was repeated after treating with tartrate, and remained positive. This suggests which condition?

a) Erythrocytic leukaemia
b) AML
c) Hairy cell leukaemia
d) Acute megakaryocytic leukaemia

A

c) Hairy cell leukaemia

48
Q

A normal blood film can act as a suitable positive control for testing all of the following staining methods, except:

a) Myeloperoxidase
b) Alpha-napthyl acetate esterase
c) Acid phosphatase with tartrate pre-treatment
d) Leucocyte alkaline phosphatase

A

c) Acid phosphatase with tartrate pre-treatment

49
Q

Which of the following is not a suitable fixative for slides that are to be cytochemically stained?

a) Phosphate buffered saline
b) Methanol
c) Acetone
d) Formaldehyde

A

a) Phosphate buffered saline

Advanced Haematology (13 decks)

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