PRELIM LABORATORY: CYTOCHEM REACTION AND INTERPRETATION

Question 1

Enzyme that catalyzes the oxidation of substances by
hydrogen peroxide

Answer 1

Peroxidase

Question 2

What does the peroxidase enzyme catalyze?

Answer 2

the oxidation
of substances by hydrogen peroxide

Question 3

Peroxidase catalyzes the oxidation
of substances by hydrogen peroxide, to form what precipitate? (describe the precipitate)

Answer 3

an insoluble
reddish-brown precipitate.

Question 4

Recommended method for the demonstration of Auer rods

Answer 4

myeloperoxidase (mpo)

Question 5

T/F: Myeloperoxidase helps differentiates acute myelogenous/monocytic leukemia from ALL

Answer 5

true

Question 6

Fixative reagent of myeloperoxidase

Answer 6

buffered formalin-acetone

Question 7

Counterstain reagent of myeloperoxidase

Answer 7

giemsa

Question 8

How many seconds for the buffered formalin-acetone of myeloperoxidase?

Answer 8

30 seconds

Question 9

How many minutes for the incubation mixture of myeloperoxidase?

Answer 9

15 minutes

Question 10

How many minutes for the counterstain of myeloperoxidase?

Answer 10

40 minutes

Question 11

Positive control of myeloperoxidase activity

Answer 11

neutrophil

Question 12

Negative control of myeloperoxidase activity

Answer 12

lymphocytes

Question 13

Myeloperoxidase is primarily expressed in and associated with these cells (cell specificity)

Answer 13

granulocytes & monocytes

Question 14

Color if positive (+) for MPO

Answer 14

reddish brown/dark brown

Question 15

Stains various lipids such as sterols, phospholipids and neutral fats

Answer 15

sudan black b

Question 16

T/F: Myeloperoxidase is more sensitive than sudan black b

Answer 16

false (sbb is believed to be more sensitive)

Question 17

Reagents of sudan black b

Answer 17

• Formalin-acetone
• Sudan Black B solution
• 70% ethanol
• Nuclear Fast red

Question 18

Reagents of MPO

Answer 18

• buffered formalin-acetone
• incubation mixture
• giemsa

Question 19

Fixative of sudan black b

Answer 19

Formalin-acetone

Question 20

Counterstain of sudan black b

Answer 20

nuclear fast red

Question 21

Sudan black b is found in which granules?

Answer 21

• primary/secondary granules of the neutrophils
• lysosomal
  granules of Monocyte

Question 22

Most specific for identification of AML

Answer 22

myeloperoxidase

Question 23

Most sensitive cytochemical stain in identifying AML

Answer 23

sudan black b (non-enzymatic test for lipids)

Question 24

Used as transport by enzymes

Answer 24

substrate

Question 25

Marker for mature and immature neutrophil/mast cells

Answer 25

chloroacetate esterase

Question 26

T/F: Chloroacetate esterase is used to differentiate granulocytes from monocytes

Answer 26

true

Question 27

Chloroacetate esterase is (less or more?) sensitive than peroxidase stain in terms of primitive myeloid

Answer 27

less

Question 28

Positive control for chloroacetate esterase

Answer 28

neutrophils

Question 29

Color of granules for chloroacetate esterase positive cells

Answer 29

bright red granules

Question 30

Useful in identifying monocytes, promonocytes, and monoblasts

Answer 30

alpha naphthyl butyrate esterase

Question 31

T/F: Alpha naphthyl butyrate esterase can identify acute/chronic myelomonocytic, acute monocytic, from other nonlymphocytic leukemia

Answer 31

true

Question 32

Cells that alpha naphthyl butyrate esterase do not stain

Answer 32

- lymphoblast - plasma cell - megakaryocytes

Question 33

Indication for alpha naphthyl butyrate esterase positive

Answer 33

dark red precipitate

Question 34

LYMPHOCYTE indication for alpha naphthyl acetate esterase positive

Answer 34

show focal dotlike staining

Question 35

MONOCYTE positive indication for alpha naphthyl acetate esterase

Answer 35

red-brown

Question 36

Present in all HSCs and is found in lysosomes. This also provides a simple test for identification of T-acute lymphocytic leukemia.

Answer 36

acid phosphatase

Question 37

Acid phosphatase provides a simple test for identification of what?

Answer 37

T-acute lymphocytic leukemia

Question 38

Acid phosphatase identification of T-acute lymphocytic leukemia should be evaluated with what 2 enzymes?

Answer 38

peroxidase & esterase

Question 39

Color of granules positive for acid phosphatase

Answer 39

discrete purplish to dark red granules

Question 40

In this non-enzymatic staining, carbohydrates present in blood cells are oxidized to glycogen by?

Answer 40

periodic acid

Question 41

Schiff’s reagent then reacts with the aldehydes to form what precipitate?

Answer 41

an insoluble red colored precipitate

Question 42

Periodic acid schiff pattern of reaction

Answer 42

- diffuse - granular - or a mixture of the two

Question 43

Common stain used for carbohydrates

Answer 43

Periodic acid schiff

Question 44

Color if positive for periodic acid schiff

Answer 44

bright fuchsia pink

Question 45

Help in diagnosing ALL and erythroid type of AML(M6)

Answer 45

periodic acid schiff

Question 46

When differentiating ALL and erythroid type of AML(M6), periodic acid schiff positive (+) indicates for?

Answer 46

AML(M6)

Question 47

When differentiating ALL and erythroid type of AML(M5),

Answer 47

acute leukocytic

Question 48

Dye that can bind with acid mucopolysaccharide in blood cells to form metachromatic complexes

Answer 48

toluidine blue

Question 49

Most useful in recognition of mast cell disease and acute or chronic basophilic leukemia

Answer 49

toluidine blue

Question 50

Reagents for toluidine blue

Answer 50

- Mota’s fixative - 0.1% Toluidine Blue (2 mins)

Question 51

How many minutes for toluidine blue mota’s fixative

Answer 51

1 minute

Question 52

How many minutes for 0.1% toluidine blue

Answer 52

2 minutes

Question 53

Positive control for toluidine blue

Answer 53

buffy coat precipitation of normal peripheral blood

Question 54

Interpreted as positive for dorm

Answer 54

Nuclei will stain light blue metachromatic granules will appear reddish violet

Question 55

Leukocyte alkaline phosphatase (LAP) is also known as

Answer 55

neutrophil alkaline phosphatase

Question 56

Why is it called neutrophil alkaline phosphatase

Answer 56

because only Neutrophils are the only leukocyte that normally contains various alkaline phosphatase

Question 57

LAP helps in differentiating what disorders?

Answer 57

Chronic myelocytic leukemia from leukemoid reactions or other myeloproliferative disorders

Question 58

Explain the principle of alkaline phosphatase

Answer 58

Alkaline phosphatase activity is present in varying degrees in the neutrophil and band form. The amount of dye precipitated is proportional to the amount of enzyme present. Generally, a high WBC with increased neutrophils is associated with increased enzyme.

Question 59

Reagents of LAP

Answer 59

-formalin and methanol - napthol - fast blue BBN - nuclear fast red

Question 60

Fixative of LAP reagent

Answer 60

formalin and methanol

Question 61

Substrate solution of LAP reagent

Answer 61

naphthol

Question 62

Working solution of LAP reagent

Answer 62

fast blue bbn

Question 63

Counterstain of LAP

Answer 63

nuclear fast red

Question 64

When performing LAP test, what must be the temperature when immersing blood films in cold? How long?

Answer 64

4-10 degrees C & 30 seconds

Question 65

Provide definition of grading: 0+

Answer 65

No granules

Question 66

Provide definition of grading: 1+

Answer 66

very few granules (faint, diffuse staining)

Question 67

Provide definition of grading: 2+

Answer 67

moderate granules scattered

Question 68

Provide definition of grading: 3+

Answer 68

numerous granules

Question 69

Provide definition of grading: 4+

Answer 69

cytoplasm is packed

Question 70

Normal LAP values according to steininger

Answer 70

40-100

Question 71

Identify if increased/decreased in LAP: - leukemoid reaction - pregnancy - polycythemia vera - aplastic leukemia

Answer 71

increased

Question 72

Identify if increased/decreased in LAP: - CML - Paroxysmal Nocturnal Hemoglobinuria - Sickle cell anemia - Hypophosphatasia

Answer 72

decreased

Question 73

Test to screen CGD (Chronic Granulomatous disease)

Answer 73

nitroblue tetrazolium test (NBT)

Question 74

A yellow, water-soluble dye, NBT on reduction is converted into?

Answer 74

an insoluble blue formation formazan

Question 75

In NBT, (+) Blue color indicates?

Answer 75

granulomatous absorb the formazan blue

Question 76

Primary color of NBT

Answer 76

(-) Red

Question 77

At least 2 px slide are obtained with 2 observers & should count how many neutrophils scoring from 0 to 4+ on the basis of the quantity & intensity of the precipitated red dye in their cytoplasm?

Answer 77

50

Question 78

Procedure for LAP test

Answer 78

1. Immerse blood films in cold (4-10 degrees Celsius fixative for 30 seconds) 2. Wash well in either tap or distilled water 3. Immerse fixed slides in working solution for 20 minutes at room temperature 4. Wash well in tap water

Question 79

Control for LAP test is blood drawn from a woman in her ___ trimester, or within ___ post-partum

Answer 79

- third - 2 days

Question 80

Principle for NBT

Answer 80

- 0.1 mL of blood is mixed with 0.1 mL of 0.2 % nitro blue tetrazolium in saline and 0.1 mL phosphate buffered saline - This mixture is incubated at 37% for 15 min - Then held at room temperature for 15 min.