WHITE BLOOD CELLS GRANULOPOEISIS

CFU-GEMM  CFU-GM (via CSF-GM)

BONE MARROW SPECIMENS CFU-GM  Myeloblast/Monoblast (CSF-G/M)
 Aspirate – Evaluation of HSCs *MITOTIC POOL (Proliferative Compartment) –
- Aspiration Needle development of Myeloblast to Myelocyte.
- University of Illinois Needle 1. MYELOBLAST
 Biopsy – Evaluation of non-HCSs of BM o 15-20 um
- Trephine Needle o Basophilic cytoplasm
- Jamshidi Needle o Round/Slightly oval nucleus
- Westerman-Jensen Needle o Fine Chromatin
SITE FOR SPECIMEN COLLECION o 2-5 nucleoli
1. Posterior Superior Illiac Crest o NO CYTOPLASMIC GRANULES
2. Anterior Superior Illiac Crest 2. PROMYELOCYTE
3. Sternum o 15-21 um
4. Anterior medial surface of the tibia o Basophilic cytoplasm
5. Spinous process of vertebrae o N:C = 3:1
BM CELLULARITY o 2-3 nucleoli
M: E RATIO – numeric expression comparing o Coarse chromatin
the relative number of granulocytic precursors o NON-SPECIFIC GRANULES
with the relative erythrocytic precursor in BM (2- a. Lysosomal Enzymes: Acid Hydrolase
4:1) Acid Phosphatase
-glucuronidase
NON-HEMATOPOEITIC CELLS OF BONE b. Elastase
MARROW c. Arysulfate
1. Macrophage 5. Adipocytes d. Cationic Bacterial Proteins
2. Osteoblast 6. Endothelial Cells e. Myeloperoxidase
3. Osteoclasts 7. Reticular Cells 3. MYELOCYTE
4. Mast Cells o 15-18um
o Coarse chromatin
COMMONLY USED CD MARKERS o No nucleolus
CD 2,3 Lymphoid, pan T cells o N:C = 1:1
CD 10 CALLA o Cytoplasm: Neutrophilic – rose pink
CD 4 T-helper Cells Eosinophilic – orange-red
CD 8 T-suppressor Cells Basophilic – Dark blue/Purple
CD 13 Pan Myeloid Cells o FORMATION OF SPECIFIC GRANULES
CD 11c, 14 Monocytes
CD 19, 20 Lymphoid B Cells A. NEUTROPHIL SPECIFIC GRANULES
CD 33 Pan Myeloid Cells a. Lysozymes f. Gelatinase
CD 34 Stem Cell Marker b. Lactoferrin g. Aminopeptidase
CD 16, 56 Natural Killer Cells c. Collagenase h. -microglobulin
d. Plasminogen Activator
LEUKOPOEISIS – takes 14 days e. ALKALINE PHOSPHATASE
- Differentiation of progenitor cells to
WBCs General Rule: B. EOSINOPHIL SPECIFIC GRANULES
1. As cells mature, size decrease a. Large Granules
2. As cell mature, chromatin coarsens and -Major Basic Protein
eventually clumps. -Eosinophil-Derived Neurotoxin
3. Nucleus DIVIDE or forms lobe/s. -Eosinophil-protein X
4. Cytoplasm transits from basophilic to -Eosinophil-cationic protein
less basophilic/acidic. Depends on cell -Acid Hydrolase
type. -Peroxidase
5. N:C ration decreases. -Phospholipase
-Cathepsin
-Histaminase
*Charcot-Leyden Crystals-formed in Eosinophil-PROMYELOCYTE
o Ground Glass Appearance
b. Small Granules
o Round/kidney shaped (horse shoe) nucleus
-Arylsulfate
o Medium Cells in Flow Cytometry
-ACP
o Scavenger Cells
C. BASOPHILIC SPECIFIC ENZYME
LYMPHOCYTES
a. Histamine d. Peroxidase
Pluripotential Cells Common Lymphoid Progenitor
b. Eosinophilic-chemotactic factor A
CLP  Lymphoblasts
c. Heparin
1. LYMPHOBLASTS
*POST MITOTIC POOL (NON-MITOTIC)
o 10-18 um
4. METAMYLOCYTE (JUVENILE)
o Moderate dark blue cytoplasm
o 10-15 um
o N:C = 4:1
o Indented nucleus/Kidney/Bean shaped
o 1-2 nucleoli
o Clumped chromatin
2. PROLYMPHOCYTE
5. BAND CELLS (STAB/STAFF CELLS)
o Similar/smaller than lymphoblasts
o Youngest WBC in circulation
o Moderate to dark blue cytoplasm
o 9-15um
o Round/oval nucleus
o Elongated, sausage shaped
o 1-2 nucleoli
6. SEGMENTED NEUTROPHIL
o For T-cells, this is the phase it migrates
o 9-15 um
to Thymus for maturation
o Pink-Rose violet specific granules 3. LYMPHOCYTES
o 2-5 nuclear lobes
o Small – 8 to 10 um
o Kamikazee cells
o Medium – 10 to 12 um
*FERRATA CELLS – tissue neutrophils
o Large – 12 to 16 um (rarest, Large
EOSINOPHILS Granular Lymphocytes AKA NK Cells)
o 9-15 um
o Reddish-orange granules T Lymphocytes (60-80%) – 4 to 10 years lifespan
o Usually 2 lobes B Lymphocytes (10-20%) – 3 to 4 days
o Less efficient phagocyte Null Lymphocytes (10%) – matures via antigenic
o Affected by ACTH stimulation
BASOPHILS
*T Cells differentiate in THYUS via
o 10-16 um
CYTOKINE stimulation.
o Dark-purple/black blue cytoplasm
Pro-T  Pre-T  Immature T
o Generally unsegmented (rarely 3-4 lobes)
CD 4 (+) = T-helper/inducer cells
o Obscured nucleus because of coarse granules CD 8 (8) = T-suppressor/cytotoxic cells
o EFFECTOR CELLS OF Hypersensitivity 1 Naïve T-cells  migrate to Secondary
Lymphoid Organs; wait for antigenic simulation for
MONOCYTES
maturation.
CFU-GM  Monoblast (CSF-M)
1. MONOBLAST
*B Cells differentiate in the BONE
o 12-20 um MARROW Pro-B  Pre-B 
o Basophilic cytoplasm Immature B
o Non-granular Immature B cell  migration to Secondary lymphoid
o 1-2 lobes organs; wait for antigenic stimulation ad matures to
o N:C = 4:1 to 5:1 PLASMA CELLS/MEMORY CELLS.
2. PROMONOCYTE
o 14-18 um
PLASMA CELLS
o Blue-gray cytoplasm 1. Plasmacytes/Plasmacytoid Cells
o N:C = 3-2:1
o 8-20 um
3. MONOCYTE
o Cell shape may be oval
o Largest cell in PBS
o Deeply basophilic cytoplasm
o 14-20um
o Large, well-defined perinuclear halo (hof)
o Fine azure granules (ACP, Peroxidase) *hof = Golgi apparatus
o Eccentric nucleus with cartwheel pattern
 o WBC ABNORMALITIES
I. Nuclear Abnormalities
A. HYPERSEGMENTATION
- Greater than 6 lobes
a. Hereditary
o Undritz Syndrome
- Autosomal recessive
o Myelokathexis
- Pyknotic hypersegmented nucleus
- Neutropenia due to
hypoplastic myeloid cells.
b. Acquired
o Megaloblastic Anemia
*Ineffective Erythropoesis (low Retic Ct.)
- Pancytopenia (low WBC, RBC and Plt ct)
- due to deficient DNA synthesis
B. HYPOSEGMENTATION
- Dumbbell shaped nucleus
- Spectacle Like
- Peanut Shaped
- Pince-Nez
a. Pelger Huet – autosomal dominant
o Homozygous – no
segmentation/oval nucleus
o Heterozygous – bilobed (pince nez)
b. Pseudo-Pelger Huet – associated with
Myeloproliferative disorders. Ie: CML
II. Cytoplasmic Abnormalities
A. Alder-Reily Anomaly
- Ader-Reily Inclusion/Reily Inclusion
- Mucopolysaccharide deposit on
granulocytic cytoplasm
- MAY RESEMBLE
TOXIC GRANULATION
- In severe forms, may also be present
in monocytes and lymphocytes
- Also associated with Hurler Syndrome
and Hunter Syndrome
B. Auer Rods
- Fused primary granules
- Present in monoblasts and myeloblasts
- (+) Peroxidase reaction
- Associated with AML 1,2 and M5
*Faggot Cells – cells with
aggregates(nest)of Auer Rods = M3 (APL)
C. Chediak-Higashi Anomaly
- Large Lysosomal Inclusion
- (+) Peroxidase Reaction [Turgeon]
- Impaired chemotaxis and delayed killing
of bacteria
- Associated with Albinism
- Storage Pool defect – deficient
dense granules.
D. May-Hegglin Anomaly
- Pale-blue inclusions (RNA Remnants)
- DOHLE-LIKE BODIES
- Associated with Thrombocytopenia
and Giant Platelets
GROUP OF ASSOCIATED DISEASE:
1. May-Hegglin Anomaly
2. Sebastian Syndrome
3. Fetchner Syndrome
4. Epstein Syndrome
E. Ehrlichia
- Due to Ehrlichia sp. Infection
- MORULAE – Ehrlichia-vacuole
bound colonies of dividing organisms
- Present in PHAGOCYTES
F. Dohle Bodies (Dohle-Amato Bodies)
- Light blue inclusion in neutrophils (may
be seen singly or multiply) peripherally.
- MAY be present in Monocytes
and Lymphocytes
- Aggregates of RER and/or RNA
- Due to INFECTION and
TOXIC CONDITIONS.
G. Toxic Granulation ad Vacuolation
- Due to infections or toxic conditions
- Prominent granulation (primary
granules) and precipitates of RNA
- Vacuolation is associated to
phagocytosis and degranulation.
- Seen in Neutrophils and may be present
in Monocytes.
III. Function Abnormalities
A. Job Syndrome/Hyperimmunoglobulinemia E
- Associated with elevated levels of IgE
- NORMAL random activity
- ABNORMAL directional/chemotactic
activity
B. Lazy Leukocyte Syndrome
- Extremely rare, assoc. with neutropenia
- ABNORMAL random, directional
and chemotactic activity.
- Abnormal cellular actomysin
component and surface receptors.
C. Chediak-Higashi Syndrome
- Inability to degranulate and kill bacteria
- Abnormal fusion of primary and secondary
granules
D. Chronic Granulomatous Disease
- Inability to kill ingested microbe due to:
 - Impaired NADPH oxidase 
impaired oxidative metabolism 
impaired RESPIRATORY BURST
(-) Nitroblue Tetrazolium Reduction Test
No reduction of Nitroblue Tetrazolium (yellow) to
Formazan (Blue)
E. Leukocyte Adhesion Disorder
- Neutrophils and monocytes are unable
to adhere to the blood vessel wall
- Poor prognosis because of inability
to transmigrate to the site of infection
- Presents with Glanzmann
Thrombasthenia like infection.
- Poor bacterial recognition.
F. Myeloperoxidase Disorder
- AKA Alius-Grignaschi Anomaly
- Abnormal or deficient myeloperoxidase 
H2O2
- impaired oxidative burst
- most commonly associated with DM
IV. Abnormal Phagocytes
A. LE Cells
- Neutrophils with homogenous round bodies
- Smooth and evenly stained
B. Tart Cells
- Monocyte with ingested lymphocytes
- Rough and unevenly stained
V. Abnormal Lymphocytes
A. Atypical/Reactive/Stimulated/Variant
AKA DOWNEY CELLS
a. Type 1 – plasmacytoid lymphocytes or Turk’s
irritation Cells
- With large block of
chromatin b.Type 2 – encountered
during I.M.
- Round mass of chromatin
- Ballerina skirt appearance
c. Type 3 – vacuolated
- Swiss cheese/moth eaten appearance
B. Basket Cells
o Pressure in smear preparation that can be
corrected by adding BOVINE
ALBUMIN.
o Chronic Lymphoblastic Leukemia
- Super sensitive lymphocytes
- Fragile cells
- Many smudge cells
a. Basket Cells – nuclear remnant that looks
like basket
b. Smudge cells – thumbprint appearance
C. Hairy Cells
- Originally B-cells but with hair-
like projections
- Characterized by (+) TRAP reaction.
D. Seizary Cells
- With brain like appearance
- Ceribriform nucleus
- Associated with Mycosis fungoides
- Seizary Syndrome
VI. Abnormal Plasma Cells
A. Flame Cells
- Red/Pink Cytoplasm
- Increased cytoplasmic
glycogen/amorphous matter
B. Russel Bodies
- Cytoplasmic aggregates of immunoglobulin
C. Grape/Berry/Morulla/Mutt Cells
- Aggregates of Russel Bodies
D. Dutcher’s Bodies
- Intranuclear protein inclusions
VII. Abnormal Macrophage
A. Gaucher’s Disease
- Accumulation of glucocerebroside
in monocytes
- Deficiency in glucocerebrosidase
- Characterized by crumpled/wrinkled
cytoplasmic appearance (Chicken
Scratch)
- With 1 to 3 nuclei
B. Neimann-Pick Disease
- Deficiency in sphingomyelinase
- Accumulation of sphingomyelin in
the cytoplasm of monocytes (Pick
Cells)
- “Pick Cells” – foamy cytoplasm
C. Tay-Sach’s Disease
- Deficiency in Hexoaminidase
- Accumulation of glycolipid and ganglioside
- Vacuolated cytoplasm
D. Sand-Off’s Disease
- Deficiency in Hexoaminidase A and B
- Accumulation of glycolipid and ganglioside
- Vacuolated cytoplasm
E. Sea-Blue Histiocytes
- Unknown deficient enzyme
- Blue-green cytoplasm
WBC COUNT AND PERIPHERAL BLOOD
SMEAR
WBC Ct = 4.5 to 11.5 x 109/L (adult)
= 10.0 to 30.0 x 109/L (Newborn)
**Drop Below Normal: Virus (measles), brucellosis,
typhoid fever, Infectious Hepatitis, Rheumatoid Arthritis,
Liver cirrhosis and SLE.
**Slightly higher in afternoon than in morning. Elevated
in exercise, stress and anxiety.
 DIFFERENTIAL COUNT
DILUTING FLUIDS (Weak Acids – for RBC lysis) o Segmenters o Monocytes
o 1-3% Glacial Acetic Acid Relative: 45 – 75% Relative: 0 to 10%
o 1% Hydrochloric Acid Absolute:1.8 to 7.8 x 109/L Absolute: 0 to 0.8 x 109/L
o Turk’s Fluid (Gentian Violet + AcetOH, H2O)
o Bands o Basophils
DILUTION FACTOR: 1:20 (25uL blood:475uL dil. sol) Relative: 0 to 7% Relative: 0 to 2%
WBC ct 9
Absolute: 0 to 0.7 x 10 /L Absolute: 0 to 0.2 x 109/L
= (cell count x dilution factor)/(area factor x depth
factor) o Eosinophils o Lymphocytes
Relative: 0 to 6% Relative: 20 to 40%
Correction for WBC ct. is required if: Absolute: 0 to 0.6 x 109/L Absolute: 1.0 to 4.8 x 109/L
- > 5 nRBC/100 Diff ct. cells in adult specimen
- >10 nRBC/Diff ct cells in child specimen

FORMULA: ABNORMAL COUNTS

Corrected WBC ct  NEUTROPHILIA
= (uncorrected WBC x 100) / (100+nRBC) o Left Shift – elevated ct of immature cells
o Rught Shift – increased nuclear segmentation
PREPARATION AND STAINING OF SMEARS o Shift Neutrophilia (Physiologic) – exercise,
1. Cover Glass Smear (Ehrlich) pain and pregnancy.
2. Cover Glass and Slide (Beacon) o Pathologic Neutrophilia – left shift with toxic
3. Wedge Smear/Push/Spreader Slide changes
Angle bet. 2 slides = 25 degrees (30 to 40) - Bacterial infection, tissue destruction,
Blood drop: 2-3 mm in diameter general infection, growth factors, drugs
Slide: 76 x 25 x 1 to 1.2 mm and toxins/
*Ideally, the distance bet the drop and the edge is o LEUKOMOID REACTION - >50x109/L
0.25 cm to 1 cm  NEUTROPENIA
o Chronic severe bacterial invasion
THIN Smear: “LSS” o Hypersplenism
Low angle o BM aplasia
Slow Spreading o Deficient DNA Synthesis
Small drop of blood o Viral infection
THICK Smear: “FIL”  EOSINOPHILIA
Fast Spreading o Infestation by tissue-invading parasite
Increased Angle o Allergic reactions
Large drop of blood o Pulmonary infiltrates with eosinophilia
STAINS: Romanowsky Stain o Tropical eosinophilia
- Wright’s o Hypersensitivities
- Giemsa o Malignancies of Myeloid cells
- Wright-Giemsa o Scarlet fever
- Leishmann o Periateritis nodosa
- Jenner  EOSINOPENIA
- Mary Gruwald o Decreased production (aplasia)
o Acute bacterial infection
o ACTH administration
o Hypoadrenalism
 MONOCYTOSIS
o Bacterial infection Inflammation
o Tuberculosis Surgical trauma
o Syphilis Collagen vascular dse
o Gastrointestinal disease
o Recovery from neutropenia
o Myeloproliferative disorders
EOSINOPHIL COUNT (50-350 x 109/L) FAB CLASSIFICATION OF LEUKEMIAS
DILUTING FLUIDS: Phloxine/Neutral Red/Iodide
Propylene Glycol A. ACUTE LYMPHOBLASTIC LEUKEMIA
NaCO3 a. L1 – most common in children
Heparin - Small homogenous blast cells
Pilot’s - phloxine - Mostly T-cells
Manner’s - phloxine b. L2 – most common in adults
Randolph’s - phloxine - Large heterogenous blast cells
Hinklemann’s - phloxine c. L3 – rarest. Large and homogenous vacuolated
Tannen’s – phloxine blasts
- Burkitt Type of Non-hodgik lymphoma
THORN’s TEST (assess adrenocortical function) - highly associated with EBV.
Eosinophil Ct #1: Fasting Specimen - Poor prognosis
Eosinophil Ct #2: 4hrs after ACTH administration - Most commonly B cells
Normal: 1 > 2 (50% lower)
Hypoadrenalism: 1 = 2 B. ACUTE NON-LYMPHOBLASTIC LEUKEMIA
a. M0 –Acute Undifferentiated Leukemia
>30% Blasts in BM
LEUKEMIA b. M1 – Acute Myeloblastic
- Abnormal uncontrolled proliferation Leukemia WITHOUT maturation
and accumulation of one or more HSCs. >30% Myeloblasts in BM
<10% Granulocytes in BM
CLASSIFICATIONS: c. M2 – Acute Myeloblastic Leukemia
1. Duration of untreated disease with maturation
a. Acute Leukemia - days to months >30% Myeloblasts in BM
b. Subacute Leukemia - 2mos to 6mos >10% Granulocytes in BM
c. Chronic Leukemia - minimum of 1 year d. M3 – Acute Promyelocytic Leukemia
>30% Promyelocytes in BM
2. Number of WBCs in PBS >10% Granulocytes in BM
a. Leukemic Leukemia - <15,000/uL *faggot cells
b. Subleukemic Leukemia - >15,000/uL (with *assoc with DIC
blasts/immature cells) e. M4 – Acute Myelomonocytic Leukemia
c. Aleukemic Leukemia - >15,000/uL 20 to 80% Myelo/Monocytes in BM
*Naegeli’s leukemia
3. Type of WBCs involved f. M5a – Acute Monoblastic
a. Acute Leukemia- Increased BLAST cells Leukemia WITHOUT maturation
b. Chronic Leukemia - Increased mature cells >80% Monocytes/Monoblasts in BM
*Schilling’s Leukemia
4. French-American-British Classification *Common in Young Adults
a. >/= 30% Blasts in BM g. M5b – Acute Monoblastic Leukemia
b. Cell morphology with maturation
c. Cytochemical stain reaction >80% Monocytes/Monoblasts in BM
*Common in Middle Age
5. World Health Organization Classification h. M6 – Acute Erythroleukemic Leukemia
>> STANDARD FOR DIAGNOSIS >30% Myeloid cells
a. >/= 20% Blasts in BM >50% Erythoid Cells
b. Cell morphology *Erythremic Myelosis
c. Cytochemical stain reaction *DiGugliemo’s Syndrome
d. Immunologic probes i. M7 – Acute Megkaryocytic Leukemia
e. Cytogenetics >30% Myeloid Cells
f. Abnormal/Clinical manifestations >30% Megakaryocytes
 MYELODYSPLASTIC SYNDROMES 4. POLYCYTHEMIA VERA
DYSMYELOPOETIC SYNDROMES o Increased RBC, WBC and platelet count.
o Clonal abnormality in HSCs o Panmyelosis – Increased HSCs in BM
o Preleukemic state o Pancytosis – Increased Blood cells in PBS
*Dyserythropoesis – binucleated erythroblastic cells o Increased or Normal LAP Score
*Dyemyelopoiesis – nuclear-cytoplasm asynchrony
*Dysmegakaryopoeisis – micromegakaryoblasts RELATIVE POLYCYTHEMIA
Blasts Ringed 1. Diminished Plasma Volume –
MDS Blasts in PBS in Sideroblas dehydration, shock and anxiety
BM ts 2. Spurious Polycythemia – stress or GAISBOCK
Refractroy SYNDROME
Anemia ABSOLUTE POLYCYTHEMIA
< 1% < 5% +/-
(Refractory o Secondary to increased EPO production
Cytopenia) (RA or - Depleted oxygen/tissue oxygen
RC) - Neoplasms (Wilm’s Tumor)
RA w/ Ringed - Post-renal transplant
>
Sideroblasts < 1% < 5% - Acute Hepatitis
15%
(RARS) o Genetic Polycythemia
RA w/ Excess 5 to - Mutated EPO receptor
< 5% +/-
Blasts (RAEB) 20% - CHUVASH polycythemia
RAEB w/ o Primary marrow disorder
20 to
transformation > 5% +/- - Polycythemia rubrovera
30%
(RAEB-t) - Erythemia
Chronic < 5% - Vasquez-Osle Disease
5 to - Panmyelosis
Myelomonocytic (Persistent -
20% -
Leukemia (CMML) Monocytosis)
MYELOPROLIFERATIVE LYMPHOPROLIFERATIVE
DISORDERS DISORDERS
o Group of acquired, malignant disorders that develop 1. T and B cell Leukemia (ALLs)
from the proliferation of an abnormal pluripotential 2. Lymphomas – proliferation of
stem cell. neoplastic lymphocytes
1. CHRONIC MYELOGENOUS LEUKEMIA A. NON-HODGKIN’S LYMPHOMA
o Stem cell disorder affecting the granulocytic, - B cell neoplasia (Burkitt’s)
monocytic, erythrocytic and megakaryocytic B. HODGKIN’S LYMPHOMA
cell lines. - Large cells with large nucleoli
o Common in adults - Owl’s eye appearance, usually
o Increased WBC ct. but LOW LAP SCORE binucleated “RH” – Reed-Sternberg
o PHILADELPHIA CHROMOSOME – - Hodgkin’s Lymphoma
translocation of Chromosome 22 to o RYE CLASSIFICATION
Chromosome 9. - Based on histologic appearance of lymph
o (-) Philadelphia Chromosome have poor node biopsy.
prognosis. o ANA ARBOR STAGING
2. MYELOFIBROSIS with - Based on extent of tissue involvement.
MYELOID METAPLASIA C. HAIRY CELL LEUKEMIA
o Fibrosis in BM with granulocytic hyperplasia o AKA Leukemic Reticuloendotheliosis
o Idiopathic o B Cells with Hair Like projections
o Anisocytosis, DACRYOCYTOSIS o TRAP (+) [Tartrate Resistant
and leukoerythroblasic anemia. Acid Phosphatase]
3. ESSENTIAL THROMBOCYTHEMIA D. MYCOSIS FUNGIODES/SEZARY SYN.
o Marked elevation of platelet count - Neoplastic T-cells (with
o >1000 x 109/L Ceribriform/Brainlike cytoplas)
o Giant dysfunctional platelets E. CHRONIC LYMPHOBLASTIC LEUKEMIA
- Fragile lymphocytes/smudge/basket cells
Key features of the Major Myeloid Leukemia (AML)
Cell Surface
Category Cell Morphology Cytogenetics Prognosis
Markers
Large blasts with abundant
basophilic cytoplasm, Auer More favorable than
CD 13, 33, 117, t (8;21) (q22;q22)
AML with t (8;21) Rods, dysplasia, abnormal AML without recurrent
19 and 34 AML1/ETO
granules, maturing fenetic abnormality
granulocytes
Blasts with monocytic and
neutrophilic differentiation, inv (16) (p13;
CD 13, 33, 14,
AML with inv (16) increased q22) or t (16;16) Superior to other AML
4, 64
eosinophils/immature (p13; q22)
eosinophils
t (15:17) (q22; q12)
Promyelocytes with CD 13, 33
Acute Promyelocytic PMLIRARa Good if responsive to
azurophilic granules, Auer
Leukemia Variants all ATRA
Rods CD 2+/-, DR-
involve 17q12
CD 13, 33, 34,
+/- 56, 57
Variable. Monocytic t (9;11) (p22; q23), Less favorable than
AML with 11q23 CD 14, 4, 36
differentiation seen in t(9;11) others any other AML
with monocytic
differentiation
11q23
abnormality seen
Multilinage dysplasia, RS, CD 13, 33, 34, with Median survival 3
AML, therapy related
increased basophils +/- 56, 57 topoisomerase II years
inhibitor-
associated AML
AML, not otherwise categorized
CD 13, 33, 117
AML, minimally M0 in FAB. Myeloblasts, <3% Nonspecific/not
in > 20% of Poor
differentiated positive SBB, MPO or ANA available
blasts
M1 in FAB. Myeloblasts >90%
Nonspecific/Not
AML without maturation of non-erythroids in BM, >3 CD 13, 33, 117 Poor
avalable
(+) with MPO, SBB
Variable. Less
M2 in FAB. Same as for AML Nonspecific/Not
AML with Maturation CD 13, 33, 117 favorable than AML
with t(8;21) available with inv (16)
M4 in FAB. Monocytic cells CD 13,33, 14, 4, Variable. Less
Acute Myelomonocytic Nonspecific/Not
20-79%, granulocytic 30-80% 11b, 11c, 64, favorable than AML
Leukemia available
of non-erythroids 36, 68 with inv (16)
M5 in FAB. >80% of t(8;16)(p1I;p13)
Acute CD 13, 33, 117,
erythroids are monoblasts seen in some acute
Monoblastic/Monocytic 14, 4, 11b, 11c, Poor
(M5a) or show monocytic diff monocytic
Leukemia 64, 36, 68
(M5b) leukemia cases
M6 in FAB >50% GLycophorin A,
Erythroblasts and >20% Hb A in erthroid
Erythroleukemia myeloblasts on non-erythroid. blasts; CD13, Nonspecific Poor
Pure erythroleukemia without 33, 117 in
myeloid blasts are rare myeloblasts
Polymorphic megakaryoblasts, Nonspecific in
Acute Megakaryoblastic may simulate lymphoblasts. PB CD41, 61, 36, adults. Infants Poor, especially with
Leukemia may show fragments of often CD13, 33 may show t (1;22)
megakaryocyte t (1:22) (p13; q13)
CYTOCHEMICAL STAINS
1. MYELOPEROXIDASE 8. TOLUIDINE BLUE
o MPO is present on the Primary Granules o Mtd: Corper and Cruickshank
of Myeloid Cells. o Staining of Basophil and Mast
(+) AML (-) ALL cell metachromatic granules.
o (+) purplish red. Increased in CGL
2. SUDAN BLACK B
o Parallels MPO reaction. Stains Lipids, 9. NITROBLUE TETRAZOLIUM TEST
Neutral fats, phospholipids o Reduction of Nitroblue tetrazolium (Yellow) to
(+) AML (-) ALL Formazan (Blue) by NADPH oxidase.
o Normal: 10% or less
3. ESTERASES o Infection: 70%
o Esterases are present in primary granules o CGD: Decreased or 0
of granulocytes and monocytes but not in *In CGD, there is an impairment of NADPH.
lymphocytes
a. SPECIFIC ESTERASE 10. FEULGEN STAIN
*-naphthol-SD-chloroacetate o Specific stain for DNA (+) Howel-Jolly Bodies
(+) AML except M5
( - ) ALL and M5 11. PRUSSIAN BLUE
b. NON-SPECIFIC ESTERASE o For hemosiderin (+) siderocytes, RARS
*-napthtyl-acetate/butyrate
(+) M5 ONLY 12. SUPRAVITAL STAINS
( - ) AMLs and ALL a. New Methylene Blue – for reticulum of Retics
b. Crystal Violet – Heinz Bodies
4. PERIODIC ACID SCHIFF c. Brilliant Cresyl Blue – Hemoglobin H
o Stains glycogen.
o Normally, ALL cells are (+) except Erythroblast
**ABNORMAL ERYTHROBLASTS are (+)
which is indicative of M6 (APL)
**In ALL there is a Block like (+) ity

5. Tdt (TERMINAL
DEOXYRIBONUCLEOTIDYL
TRANSFERASE)
o Marker for immature
lymphocytes (+) ALL(-) AML

6. ACID PHOSPHATASE
o All cells stain (+) with acid phosphatase [ACP]
o BUT ACP’s activity is inhibited by TATRATE
o Upon addition of tartrate, ACP will be ( -
) normally.
o Only HAIRY CELLS HAVE
TARTRATE RESISTANT ACP (TRAP
+)

7. LEUKOCYTE ALKALINE PHOSPHATASE

o ALP is a specific granule of Neutrophil
o LAP scoring is done using the KAPLOW
Count (NV= 30-185 LAP score)
o 100 segmenters are counted; and are
graded according to staining reaction (0 to
4+)
o Multiply the grading by the number of
cells counted on that grading, get the total.