q3: cytochemistry / cytochemical stains

AML in MPO

a. +

b. -

a. +

AML in SBB

a. +

b. -

a. +

AML in TDT

a. +

b. -

b. -

ALL in MPO

a. +

b. -

b. -

ALL in SBB

a. +

b. -

b. -

ALL in TDT

a. +

b. -

b. -

MPO reactions and Sudan black B reactions frequently __ to each other

direct / parallel

Stains ALP, present in the neutrophil and to a small degree, in certain B cells.

Leukocyte alkaline phosphatase

Helpful in differentiating chronic myelogenous from a leukemoid reaction or polycythemia vera

Leukocyte alkaline phosphatase

Stains peroxidases present in the granulocytes and monocytes

Peroxidase

Used to differentiate acute myelogenous and monocytic leukemia from acute lymphocytic leukemia

Peroxidase

Stains lipids present in the monocytes and granulocytes

Sudan Black B

Used to differentiate acute myelogenous and myelomonocytic leukemias from acute lymphocytic leukemia

Sudan Black B

Stains esterases present in the granulocytes

Chloroacetate esterase

Used to differentiate granulocytic cells from monocytic cells

Chloroacetate esterase

Stains esterases present in the monocytic cells, macrophages, megakaryocytes and platelets.

Non-specific esterase

Used to differentiate monocytic leukemias from granulocytic leukemias

Non-specific esterase

Stain mucoproteins, glycoproteins, and high molecular weight carbohydrates normally present in almost all blood types except pronormoblast

Periodic Acid Schiff

Used to help in the diagnosis of DiGugleilmo's syndrome and may be an aid, when used in conjunction with other stains, to classify some acute leukemias

Periodic Acid Schiff

Stains ACP present in the myelogenous cell, lymphocytes, plasma cells, monocytes and platelets

Acid phosphatase

Using L (+) tartaric acid, the stain is helpful in diagnosing hairy cell leukemia

Acid phosphatase

Fresh capillary blood

Alternatively, blood may be collected using heparin as anticoagulant.

a. LAP

b. Peroxidase

c. SBB

d. Chloroacetate esterase

e. Non-specific esterase

f. Periodic Acid Schiff

g. Acid phosphatase

a. LAP

Fresh blood smears made from capillary blood are recommended, or use of fresh whole blood anticoagulated with EDTA or heparin

a. LAP

b. Peroxidase

c. SBB

d. Chloroacetate esterase

e. Non-specific esterase

f. Periodic Acid Schiff

g. Acid phosphatase

b. Peroxidase

Air dried bone marrow smears

a. LAP

b. Peroxidase

c. SBB

d. Chloroacetate esterase

e. Non-specific esterase

f. Periodic Acid Schiff

g. Acid phosphatase

c. SBB

Air dried blood or bone marrow smears. Blood anticoagulated with EDTA or heparin may also be used

a. LAP

b. Peroxidase

c. SBB

d. Chloroacetate esterase

e. Non-specific esterase

f. Periodic Acid Schiff

g. Acid phosphatase

d. Chloroacetate esterase

Air-dried blood or bone marrow smears. Blood anticoagulated with EDTA or heparin may also be used

a. LAP

b. Peroxidase

c. SBB

d. Chloroacetate esterase

e. Non-specific esterase

f. Periodic Acid Schiff

g. Acid phosphatase

e. Non-specific esterase

Air dried blood or bone marrow smears

a. LAP

b. Peroxidase

c. SBB

d. Chloroacetate esterase

e. Non-specific esterase

f. Periodic Acid Schiff

g. Acid phosphatase

f. Periodic Acid Schiff

Air-dried blood or bone marrow smears, or use blood anticoagulated with heparin

a. LAP

b. Peroxidase

c. SBB

d. Chloroacetate esterase

e. Non-specific esterase

f. Periodic Acid Schiff

g. Acid phosphatase

Increased LAP, Decreased CML

leukocyte alkaline phosphatase

Stain for granulocyte and monocyte

Esterase

Added in non-specific esterase to inhibit monocyte

sodium fluoride

Non enzyme stains (3)

Sudan Black B

Periodic Acid Schiff

Toluidine Blue

Marker for primary granules of auer rods

Myeloperoxidase

Peroxidase activity produces dark brown granules in cytoplasm of granulocytes and monocytes

Myeloperoxidase

Stain should be done only on fresh specimens

Myeloperoxidase

Marker for phospholipids and lipids

Sudan Black B

Dark purple black granules in neutrophil precursors

Lymphoblasts are negative

Sudan Black B

Can be done on stored specimens Parallel

MPO For interpretation

Sudan Black B

DNA polymerase immunoperoxidase

Terminal deoxyribonucleotidyl transferase

Is present in 90% cases of ALL

Terminal deoxyribonucleotidyl transferase

Used to differentiate AML to ALL

Terminal deoxyribonucleotidyl transferase

Marker for glycogen, glycoproteins, mucoproteins and high molecular weight carbohydrates

Periodic Acid Schiff

Activity results in bright fuchsia pink or magenta red

Pattern of staining varies with each cell type

Periodic Acid Schiff

Lymphoblastic leukemia shows blocky or chunky pattern

L1 and L2 block pattern

Erythroblasts in M6 leukemia is positive

Periodic Acid Schiff

Marker for mature and immature neutrophils and mast cells

Naphthol AS-D chloroacetate Esterase

Enzyme activity results in bright red granules in cytoplasm of neutrophils, neutrophil precursors and mast cells

Naphthol AS-D chloroacetate Esterase

Known as specific esterase

Stable enzyme that lasts for months

Naphthol AS-D chloroacetate Esterase

Marker for monocytes, megakaryocytes, and plasma cells

Alpha- napthyl acetate esterase

Monocytic stain red-brown

Alpha- napthyl acetate esterase

Known as nonspecific esterase

Alpha- napthyl acetate esterase

Identifying monocytes, promonocytes, and monoblasts

Alpha- naphthyl Butyrate esterase

Enzyme activity results in dark red precipitates in cytoplasm

Alpha- naphthyl Butyrate esterase

Present in all hematopoietic cells and found on lysosomes

Acid phosphatase

Activity is indicated by purple to red granules

Acid phosphatase

Cannot be stored

Acid phosphatase

Marker for hairy cell leukemia

Tartrate resistant acid phosphatase

Activity is indicated by purple to dark red granules in cytoplasm

Tartrate resistant acid phosphatase

Excellent marker for hairy cell leukemia

Tartrate resistant acid phosphatase

Neutrophils is the only leukocyte that contain this activity

Leukocyte alkaline Phosphatase (LAP)

100 cell count is done. Neutrophils are scored from with no activity to 4 with a large amount of activity

Leukocyte alkaline Phosphatase (LAP)

Used to differentiate CML from a leukemoid reaction

Leukocyte alkaline Phosphatase (LAP)

Binds with acid mucopolysaccharides in blood cells

Toluidine Blue

Strongly metachromatic

Toluidine Blue

Useful for recognition of mast cells and tissue basophils

Toluidine Blue

Stain intensity: None

a. 0

b. 1+

c. 2+

d. 3+

e. 4+

a. 0

Faint to moderate

a. 0

b. 1+

c. 2+

d. 3+

e. 4+

b. 1+

Moderate to strong

a. 0

b. 1+

c. 2+

d. 3+

e. 4+

c. 2+

Strong

a. 0

b. 1+

c. 2+

d. 3+

e. 4+

d. 3+

Brilliant

a. 0

b. 1+

c. 2+

d. 3+

e. 4+

e. 4+

Amount: none

a. 0

b. 1+

c. 2+

d. 3+

e. 4+

a. 0

Amount: 50

a. 0

b. 1+

c. 2+

d. 3+

e. 4+

b. 1+

Amount: 50 t0 80

a. 0

b. 1+

c. 2+

d. 3+

e. 4+

c. 2+

Amount: 80 to 100

a. 0

b. 1+

c. 2+

d. 3+

e. 4+

d. 3+

Amount: 100

a. 0

b. 1+

c. 2+

d. 3+

e. 4+

e. 4+

an enzyme found in the membranes of secondary granules of neutrophils

LAP

Lymphoma, undifferentiated

a. Stem cell leukemia

b. Acute lymphoblastic

c. Chronic lymphocytic

d. Monocytic leukemia

e. Acute myelogenous

f. Plasma cell leukemia

a. Stem cell leukemia

Lymphoma, poorly leukemia differentiated; lymphocytic

a. Stem cell leukemia

b. Acute lymphoblastic

c. Chronic lymphocytic

d. Monocytic leukemia

e. Acute myelogenous

f. Plasma cell leukemia

b. Acute lymphoblastic

Lymphoma, well leukemia differentiated; lymphocytic

a. Stem cell leukemia

b. Acute lymphoblastic

c. Chronic lymphocytic

d. Monocytic leukemia

e. Acute myelogenous

f. Plasma cell leukemia

c. Chronic lymphocytic

Reticulum cell sarcoma

a. Stem cell leukemia

b. Acute lymphoblastic

c. Chronic lymphocytic

d. Monocytic leukemia

e. Acute myelogenous

f. Plasma cell leukemia

d. Monocytic leukemia

Chloroma granulocytic leukemia

a. Stem cell leukemia

b. Acute lymphoblastic

c. Chronic lymphocytic

d. Monocytic leukemia

e. Acute myelogenous

f. Plasma cell leukemia

e. Acute myelogenous

Myeloma

a. Stem cell leukemia

b. Acute lymphoblastic

c. Chronic lymphocytic

d. Monocytic leukemia

e. Acute myelogenous

f. Plasma cell leukemia

f. Plasma cell leukemia