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Hematology
EDTA
(Lavender top)

Modified Westergren ESR
(Black top tube)
Citrate
(Light blue top tube)

Polycythemic patients

Oxalate
Heparin

Order of Draw
(Henry 21st Edition)

Order of Draw
(Syringe method)
EDTA containing tubes

MUST TO KNOW IN HEMATOLOGY
Greek:
-Haima = Blood
-Logos = Study/science
Chelates calcium
Inversion: 8x
Anticoagulant of choice for hematology cell counts and cell morphology
Blood smear: prepare w/in 2 hrs
Preferred anticoagulant for platelet count:
= In some patients w/ EDTA anticoagulated blood – platelet satellitism
= Platelet satellitism: platelets adhere to neutrophils
♫ Effect to automated platelet count  Decreased
♫ Remedy: Repeat platelet count using citrate (Rodak: Platelet count x 1
...
5mL NSS/Citrate
Ratio = 1:4 (Anticoagulant-to-Blood)
For coagulation and platelet studies
= Preserves labile factors V and VIII
= Buffered 3
...
109M) citrate
Inversion: 3-4x
Ratio = 1:9 (Anticoagulant-to-Blood)
 Hct
 Excess Citrate =  PT,  APTT
Remedy: Reduce the volume of citrate
Amount of citrate = [(100-Hct)÷(595-Hct)] x mL WB
Double/balanced oxalate (Ratio = 2:3): Maintained cell structures
a
...
Ammonium oxalate (Wintrobe’s) = swell cells
Inactivation of thrombin
Anticoagulant for osmotic fragility test
Inversion: 3-4x
Not for blood film preparation:
= Distorts cells
= Produces bluish background on Romanowsky’s stain
Not for coagulation
= Inhibits thrombin and all stages of coagulation
Evacuated tube:
1
...
Citrate (blue)
3
...
Heparin (green)
5
...
Fluoride (gray)
1
...
Other anticoagulated tubes
3
...
Fingertips
2
...
Lateral portion of the plantar surface of the foot: <1 year old
Difference from venous specimen:
 WBCs
 Hgb, Hct, RBCs, platelet
Veins in the arms (antecubital region):
1
...
Cephalic (lateral)
3
...
5 inches
18 = pink
21 = green
22 = gray
23 = blue/light blue/turquoise
Venipuncture: 150
BB: 450  10-200 once in the skin
3-4 inches above the site (7
...
Gower 1 = Zeta2 + Epsilon2
b
...
Gower 2 = Alpha2 + Epsilon2
3rd month of gestation
Fetal liver = Granulopoiesis, Erythropoiesis, Megakaryopoiesis
Spleen, thymus, lymph nodes
Hemoglobin production:
a
...
HbA1 = Alpha2 + Beta2
c
...
5-3%
HbF = <2%
Page | 178

Neonates
Marrow specimens

Posterior iliac creast
M:E ratio

BM Cellularity

Yellow BM
Red BM
Marrow differential
Metamyelocyte/Juvenile
granulocyte
Stem cells
Osteoblasts
Osteoclasts
CD2, CD3
CD19, CD20
CD34
CD16, CD56
CD10
Erythropoietin
Thrombopoietin
Erythropoiesis
IL-3
1
...
Basophilic normoblast

HbF = 60-80%
HbA = 20-40%
1
...
Aspiration
= Aspiration needle (University of Illinois sterna needle)
Safest site for BM aspirate/biopsy
Numeric expression comparing the relative number of granulocytic precursors
w/ the relative erythroid precursors in the BM
NV = 2:1 to 4:1 (Ave
...
50%)
Fats
Hematopoietic cells
Recommended that at least 500, preferably 1000 cells be counted for a marrow
differential
Predominant cell (WBC) in adult BM (up to 32%)
<1% cells in BM
Bone forming cells
Confused w/ plasma cells
Waterbug or comet appearance
Bone destroying cells
Confused w/ megakaryocytes
T cells
B cells
Stem cell marker (lymphoid and myeloid precursor)
NK cells
CALLA (Common ALL Antigen)
Produced by the kidney
Primary regulator of erythropoiesis
Produced by kidney and liver
Regulator of thrombopoiesis
1’ stimulus = Hypoxia
Multi-CSF (Colony Stimulating Factor)
Stimulates hematopoietic cells
Proerythroblast/rubriblast
N/C ratio = 8:1
Nucleoli = 1-2
Can produce up to 16 RBCs per 1 pronormoblast/rubriblast
Prorubricyte
Intensely basophilic cytoplasm
N/C ratio = 6:1
Nucleoli usually not visible
Page | 179

3
...
Orthochromic
Metarubricyte
normoblast
Small pyknotic nucleus (dark, small, nonfunctional)
N/C ratio = 1:2
5
...
Mature RBC
Discocyte
6-8 μm in diameter
Life span: 120 days
3-5 days
BM: Pronormoblast  Reticulocyte
1-2 days
PB: Reticulocyte  RBC
General Cell Maturation Characteristics for Leukocytes
Immature Cells
Mature Cells
Larger
Smaller
(+) Nucleoli
(-) Nucleoli
Chromatin: fine and delicate (most reliable)
Chromatin: coarse and clumped (most reliable)
Nucleus: large and round
Nucleus: round
...
Myeloblast
Earliest recognizable stage in granulocytic series
N/C ratio = 4:1
Nucleoli = 2-5
2
...
Myelocyte
Youngest cell in the series wherein a granulocyte can be identified
a
...
Eosinophil myelocyte = orange-red granules
c
...
Metamyelocyte
Juvenile granulocyte
Not capable of mitosis (post-mitotic pool)
Indented/kidney-shaped nucleus
Predominant WBC in BM
5
...
Segmented neutrophil

6b
...
Basophil

Monopoiesis
Monocyte

Lymphopoiesis
Lymphocyte

T lymphocytes
B lymphocytes

Resembles Pelger-Huet cells
= PH cells: coarser chromatin than stab cells
Rose-pink granules
Nucleus: 2-5 lobes
Diurnal variation (PM)
Specific granules:
a
...
Lactoferrin
c
...
Plasminogen activator
e
...
Major basic protein
b
...
Cathepsin
d
...
Eosinophil-derived neurotoxin
e
...
Phospholipase
[Smaller]
a
...
Acid phosphatase
Dark purple to blue-black granules (water-soluble)
Nucleus: generally unsegmented or bilobed (rare: 3 or 4)
Specific granules:
a
...
Heparin
c
...
Monoblast
2
...
Monocyte
Largest cell in PB
14-20 μm in diameter
Blue-gray cytoplasm
Many azurophilic granules (ground glass appearance)
Nucleus: kidney/horse-shoe shaped, may be folded (brainlike)
1
...
Prolymphocyte
3
...
Megakaryoblast
2
...
Granular megakaryocyte
4
...
Metamegakaryocyte
6
...
Plasmablast
2
...
Plasmacyte/Plasma cell
Large well-defined hof/perinuclear halo (light staining area in the cytoplasm
near the nucleus)
Eccentric nucleus
Deeply basophilic cytoplasm (Red/pink cytoplasm: Flame cell [Abnormal])
Chromatin: “Cart wheel pattern”
5 days (Megakaryoblast  Platelets)
N/C ratio = 10:1
Nucleus: may show slight lobulation (Endomitosis)
N/C ratio = 4:1 to 7:1
Largest cell in BM
Cytoplasm contains coarse clumps of granules aggregating into little bundles,
which bud off from the periphery to become platelets
Multiple nuclei
N/C ratio = <1:1
Disintegrated cell surrounded by platelet
1-4μm in diameter
Light blue to purple
Very granular
a
...
Hyalomere: surrounds the chromomere, nongranular and clear to light blue
Life span: 8-11 days
Circulating platelets
Platelets stored in the spleen
Nuclear division w/o cytoplasmic division
# of platelets a megakaryocyte can produce
1 mol O2
4 mol O2
Early and late heme synthesis
Alpha
Beta, Gamma, Delta, Epsilon, Zeta
Alpha, Zeta
Beta, Gamma, Delta, Epsilon
Normal = Sigmoid in shape
X-axis = Hgb concentration in g/dL | Y-axis = OD
 CO2
 Temperature
 2,3-DPG
 pH
 Affinity of Hgb for O2
 HbF
CO2
 Temperature
 2,3-DPG
 pH
 Affinity of Hgb for O2
Page | 182

Heme synthesis

Arterial blood
Venous blood
P50
Bohr effect
Oxyhemoglobin (HbO2)
Deoxyhemoglobin (HbCO2)
Carboxyhemoglobin
(HbCO)

Methemoglobin/
Hemiglobin (Hi)

Succinyl coenzyme A + Glycine + Pyridoxal PO4
↓
(ALA synthase)
↓
D-Aminolevulinic acid
↓
(ALA dehydrase)
↓
Porphobilinogen
↓
(Uroporphyrinogen synthase)
(Uroporphyrinogen cosynthase)
↓
Uroporphyrinogen
↓
(Uroporphyrinogen decarboxylase)
↓
Coproporphyrinogen
↓
(Coproporphyrinogen oxidase)
↓
Protoporphyrinogen
↓
(Protoporphyrinogen oxidase)
↓
Protoporphyrin IX + Fe2+
↓
(Ferrocheletase)
↓
HEME
O2 saturation = 95%
pO2 = 95 mmHg
O2 saturation = 70%
pO2 = 40 mmHg
pO2 = 26
...

Blood: Chocolate brown color
Reversible
Fe2+  Fe3+
Methemoglobin reductase deficiency
Exposure to chemicals/drugs
Exposure to nitrite, chlorine and nitrate
Page | 183

Sulfhemoglobin

Erythrocyte membrane

Embden-Meyerhoff
pathway
Hexose monophosphate
shunt
(Pentose PO4 pathway)
Rapoport-Luebering
pathway
Methemoglobin reductase
pathway
Pitting
Culling
RBC lysis
Extravascular hemolysis

Intravascular hemolysis

Anisocytosis
RDW

HbM (Abnormal hemoglobin)
Blood: Mauve lavender
Irreversible
Cannot transport O2
Can combine w/ CO forming carboxysulfhemoglobin
Mixture of oxidized, partially denatured forms of hemoglobin that form during
oxidative hemolysis  green hemochrome
Sulfonamides, aromatic amine drugs
Bacteremia: Clostridium perfringens
Enterogenous cyanosis
50% protein:
1
...
Peripheral protein: Spectrin and Actin
= Responsible for biconcave shape of red cells
= Abn
...
Phospholipids
2
...
5-14
...
B9)
deficiency
Polychromasia

Spherocytosis
Microcytic, hypochromic

Variation in Hgb content (Ex
...
Acute blood loss
2
...
Aplastic anemia
1
...
Thalassemia = N-RDW
3
...
Hookworm infections) |  RDW
4
...
Chronic disease
1
...
Nonmegaloblastic anemia = Liver disease, alcoholism
Congenital = Fanconi’s anemia
Acquired = radiation, chemical (benzene), drugs (chloramphenicol)
Pancytopenia = WBCs, RBCs, Retics Plts
Differentiates IDA (TIBC) from other microcytic, hypochromic anemia
Normal = Area of palor 1/3 of the cell diameter
1+ = Area of palor 1/2 of the cell diameter
2+ = Area of palor 2/3 of the cell diameter
3+ = Area of palor 3/4 of the cell diameter
4+ = Thin rim of hemoglobin
Oval macrocytes
Howell-Jolly bodies
Hypersegmented neutrophils
Ineffective erythropoiesis (pancytopenia)
♫ w/ CNS problems
1
...
D
...
Vegetarian diet
4
...
Pregnancy
2
...
Steatorrhea, sprue
Reticulocytosis
Visible on Wright’s stain
Blue-gray coloration, pink cytoplasm
Indicates young RBCs
 Erythropoietic activity
a
...
Hemolysis
EOFT
EOFT
Page | 185

Poikilocytosis
Macrocytes
Oval macrocytes
Macroovalocytes
Acanthocyte
Spur cell
Thorn cell
Echinocyte
Burr cell
Sea urchin cell
Crenated RBCs

Codocyte
Target cell
Mexican hat cell

Leptocyte
Spherocyte
Ball
Bronze cell

AIHA vs
...
B12 or folic acid deficiency
Asynchronous development: mature cytoplasm but immature nucleus
Irregular spikes/spicules
Abnormal L:S ratio
♫ Abetalipoproteinemia
Liver diseases
Echinocyte: artifactual
Burr cell: pathologic
RBCs w/ projections of equal length and distribution
 ATP
♫ Exposure to hypertonic solution
♫ Artifact in air drying
♫ Anemia associated w/ renal insufficiency
Greek: Kodon = bell
Central hemoglobinized area (bull’s eye)
Scanning EM: Bell/tall hat shaped
 Surface membrane to volume ratio
 Cholesterol & phospholipid
♫ Hemoglobinopathies
♫ Thalassemia
♫ Liver disease, postsplenectomy, IDA
Thin variant of a codocyte
Spheroid
Biconvex
 Surface area to volume ratio
Defect of loss of membrane:
♫ Hereditary spherocytosis = Post-splenectomy:  Spherocytes
♫ Hemolytic anemia
♫ Burns
♫ Banked blood stored for a long time
AIHA = OFT, MCHC, (+) DAT
HS = OFT, MCHC, (-) DAT
Greek: Stoma = mouth
Mouth, slit-like pallor area
Bowl-shaped
 Permeability to Na+ (Normal:  permeability to K+)
♫ Hereditary stomatocytosis
♫ Rhnull disease
Rod/cigar shaped
Narrower than ovalocytes
Defect in cytoskeleton
 Membrane protein band 4
...
Fine stippling =  polychromatophilia ( production of RBCs)
2
...
vivax = Worldwide
P
...
falciparum rings
Cold agglutinin disease (anti-I)
Primary atypical pneumonia (anti-I)
RBCs in stack of coins arrangement
 plasma globulin
♫ Multiple myeloma: proliferation of Ig-producing plasma cells (ESR)
♫ Macroglobulinemia
Disperses rouleaux formation
True agglutination remains intact
Hemoglobinopathies
Qualitative defect in Hgb
Ex
...
Adult = β4 = HbH (deletion of 3/4 alpha genes)
b
...
6
Migration: (Cathode > Anode)
C > S > F > A1 > Barts > I > H
E D
O G
A2 Lepore
Normal: HbA1 is the fastest (most anodal)
Abnormal: HbH is the fastest (most anodal)
Acid pH: 6
...
3
Migration: (Cathode > Anode)
F > A |Origin| O > S > C
E
D
G
1
...
Solubility test
= Sodium thiosulfite
= (+) Turbidity
 in β-thalassemia
Quantitation: Anion exchange microchromatography
Alkali resistant
(+) HiCN
Tests:
1
...
Betke (NaOH)
b
...
Acid elution test
= HbF resists acid-elution
= Cells w/ HbF = deep pink color
= Cells w/ N-HbF = ghost cells
1
...
Isopropanol precipitation test: 17% solution
Sample Criteria for Erythrocyte Morphology Evaluation
w/in Normal
1+
2+
3+
Limits (OIO)
(per OIO)
(per OIO)
(per OIO)
0-5
5-10
10-20
20-50
0-5
5-10
10-20
20-50
0-2
3-10
10-50
50-75
0-2
3-10
10-20
20-50
-1-5
6-10
>10
Numerous
-Agg
...
of 5-10 RBCs
aggregates

4+
(per OIO)
>50
>50
>75
>50
--Page | 189

Pelger-Huet

Hypersegmentation

Alder-Reilly granules

Toxic granules
Toxic vacuoles
Auer rods
Faggot cells
Chediak-Higashi granules

May-Hegglin inclusion

Dohle bodies
Dohle-Amato bodies

IT: Infections, Toxic states

Job’s syndrome
Lazy leukocyte syndrome
Chronic Granulomatous
Disease (CGD)

Nuclear Abnormalities
Hyposegmentation (neutrophil)
Bilobed nucleus: Dumb-bell shaped/spectacle/peanut-shaped/”Pince-nez”
Resembles Stab cell (To differentiate: PH cell has more clumped chromatin)
♫ Pelger-Huet anomaly = Autosomal Dominant
♫ Pseudo-Pelger-Huet = Acquired in myeloproliferative disorders
≥ 6 lobes (neutrophil)
Abnormal DNA synthesis
♫ Undritz anomaly = hereditary hypersegmentation
♫ Megaloblastic anemia
Cytoplasmic Abnormalities
Large purple-black coarse cytoplasmic granules
Accumulation of degraded mucopolysaccharides (all leukocytes)
♫ Alder-Reilly anomaly = Autosomal Recessive
♫ Mucopolysaccharidoses: Hurler, Hunter, Sanfilippo syndrome
Resemble toxic granules (IT)
Large purple to black granules resembling ALR granules
♫ Infections
♫ Toxic states
Infections
Toxic states
Pink or red rod shaped structures
Fused primary granules (peroxidase positive)
Myeloid and monocytic series only
w/ mass of Auer rods
M3 (APL) = associated w/ DIC
Giant red, blue to grayish round inclusions (large lysosomal granules)
Seen in lymphocyte, neutrophil and monocyte
Lysosomal defects
Platelets lack dense granules
♫ Chediak-Higashi syndrome = Autosomal Recessive (Albinism)
Pale blue inclusions derived from RNA
♫ May-Hegglin anomaly
= Autosomal Recessive
= Giant platelets
= Thrombocytopenia
Resemble Dohle bodies (IT)
Single or multiple blue inclusions
Aggregates of free ribosomes of rough ER
Resembles
♫ Infections
♫ Toxic states
Dohle bodies
Toxic granules
Toxic vacuoles
Abnormalities in Function
Normal random activity
Abnormal chemotactic activity
Abnormal random and chemotactic activity
Inability of phagocytes to kill ingested microorganisms
Impaired NADPH oxidase
Impaired oxidative metabolism/respiratory burst
Page | 190

LE cell

Tart cell

Test: NBT dye test
Cells Exhibiting Phagocytosis
Neutrophil w/ large purple homogeneous round inclusion
Believe to be a neutrophil that ingested another neutrophil
Buffy coat
Smooth and evenly stained
♫ SLE
Monocyte w/ ingested lymphocyte
Rough and unevenly stained

Abnormalities Involving Lymphocytes
a
...
Type II
= Infectious mononucleosis: caused by EBV (target: B cells [CD21])
= Atypical lymphocyte in IM: T cells reacting to B cells infected w/ EBV
c
...
MPO: Myeloperoxidase
= (+) AML
= (-) ALL
2
...
TdT: Terminal Deoxyribonucleotidyltransferase
= Marker for immature lymphocyte
= (+) ALL
= (-) ANLL
Acute Lymphoblastic Leukemias (ALL)
Lymphoblasts are small and homogeneous (vary little in size)
Childhood ALL
Lymphoblasts are large and heterogeneous (vary in size)
Adult ALL
Page | 192

L3

Burkitt-type
Rare
Lymphoblasts are large but homogeneous, and vacuolated
Acute Nonlymphocytic Leukemias (ANLL)
M1
Acute myeloblastic leukemia w/o maturation (AML w/o mat)
BM:
>30% blasts
<10% granulocytic cells
M2
Acute myeloblastic leukemia w/ maturation (AML w/ mat)
BM:
>30% blasts
>10% granulocytic cells
M3
Acute promyelocytic leukemia (APL)
>30% blasts
>10% granulocytic cells
>30% or >50% promyelocytes
(+) Faggot cells = Associated w/ DIC
M4
Acute myelomonocytic leukemia (AMML)
Naegeli’s leukemia
20% to <80% monocytic cells
M5a
Acute monoblastic leukemia w/o maturation
Schilling’s leukemia
>80% monocytic cells (>80% monoblasts)
M5b
Acute monoblastic leukemia w/ maturation
>80% monocytic cells (<80% monoblasts)
M6
Erythroleukemia
Erythremic myelosis
Di Guglielmo’s syndrome
>30% blasts
>50% erythrocytic precursors
M7
Acute megakaryocytic leukemia
>30% blasts
>30% megakaryocytic cells
Chronic Myeloproliferative Disorders
MPD
Proliferation of abnormal pluripotential stem cell
Stem cell differentiates into the granulocytic (myeloid stem cell),
megakaryocytic and erythroid cell lines
1
...
Chromosome studies
b
...
Myelofibrosis w/ myeloid Fibrosis and granulocytic hyperplasia of BM, w/ granulocytic and
metaplasia (MMM)
megakaryocytic proliferation in the liver and spleen (extramedullary)
(+) Dacryocytes
 LAP
BM aspirate = impossible (dry tap)
BM biopsy = appropriate
3
...
Polycythemia Vera (PV)
BM: Panmyelosis
PB: Pancytosis/Pancythemia
Page | 193

RBCs, WBCs, Plts
 LAP (Other polycythemia: N-LAP)
Polycythemia
1’ Absolute polycythemia
Other names: Polycythemia Vera, Polycythemia Rubravera, Vaquez Osler
disease, Panmyelosis
 RBC mass ( Hct)
 RBCs,  WBCs,  Platelets
 Erythropoietin (EPO)
2’ Absolute polycythemia
In response to hypoxia
w/ appropriate production In patients w/ pulmonary/cardiac disease
of EPO
 RBCs,  WBCs,  Platelets
 EPO
2’ Absolute polycythemia
In patients w/ tumors of kidney, liver, brain, adrenal and pituitary gland
w/ inappropriate
 RBCs, N-WBCs, N-Platelets
production of EPO
 EPO
Relative polycythemia
Spurious/Gaisböck polycythemia
Associated w/ stress and anxiety
N-RBC mass
 Hct because of decreased plasma volume
RBC mass
Differentiate absolute from relative polycythemia
 RBC mass = Absolute polycythemia
N-RBC mass = Relative polycythemia
Myelodysplastic Syndrome/Dysmyelopoietic Syndrome
MDS
Clonal abnormalities in hematopoietic cells
“Pre-leukemia”: can progress to ANLL if not treated
<30% blast
Differentiation
% Blasts in PB
% Blasts in BM
Comments
Refractory anemia
Mildest type
<1%
<5%
(RA)
RA w/ ringed sideroblast
<5%
RS: Ringed sideroblast
<1%
(RARS)
>15% RS
RA w/ excess blast
<5%
5-20%
(RAEB)
RAEB in transformation
>5%
20-30%
(RAEBt)
Chronic Myelomonocytic
Persistent monocytosis
<5%
5-20%
Leukemia (CMML)
Lymphoproliferative disorders
LPD
Proliferation of cells derived from lymphoid stem cell  T/B cells
1
...
Lymphoma
Malignancy involving lymphoid tissue
a
...
Hodgkin’s lymphoma
= proliferation of cells reacting to neoplasm
= (+) Reed-Sternberg cell: large cell w/ large nucleoli (Owl’s eye)
♫ Diagnosis: Lymph node biopsy
= Rye classification: based on histologic appearance of lymph node biopsy
= Ann-Arbor: staging based on the extent of tissue involved
3
...
Mycosis fungoides and
Sezary syndrome
Myeloperoxidase (MPO)

Sudan Black B (SBB)

Terminal
deoxyribonucleotidyl
Transferase (TdT)
Periodic Acid-Schiff (PAS)

Naphthol AS-D
Chloroacetate esterase

α-Naphthyl Acetate
esterase

α-Naphthyl Butyrate
esterase
Acid phosphatase

Tartrate resistant acid
phosphatase (TRAP)
Leukocyte alkaline
phosphatase (LAP)/
Neutrophil ALP (NAP)

Neoplastic T cells
Sezary cells: w/ cerebriform nucleus (Brainlike)
Special Stains
Marker for primary granules and Auer rods
(+) AML
(-) ALL
Stain should only be done on fresh specimens
Marker for phospholipids and lipids
(+) AML
(-) ALL
Can be done on stored specimens
Parallels MPO for interpretation
DNA polymerase immunoperoxidase
Marker for immature lymphocytes
(+) ALL
(-) AML
Marker for glycogen, glycoproteins, mucoproteins and HMW CHO
a
...
Abnormal: erythroblasts are PAS (+) = M6
(+) L1 and L2 = blocklike positivity
(-) L3
(+) M6
Specific esterase
Marker for mature and immature neutrophils and mast cells
(+) AML
(-) AMoL [M5]
(-) ALL
Lasts for months
Nonspecific esterase
Marker for monocytes, megakaryocytes and plasma cells
(+) AMoL [M5] = inhibited by fluoride
(+) AMegL [M7]
(-) AML
Nonspecific esterase
Identifies monocytes, promonocytes and monoblasts
(+) AMoL [M5] = inhibited by fluoride
(-) AML
Present in all hematopoietic cells and found in lysosomes
(+) Hairy cell leukemia = TRAP (+)
(+) T cell leukemia
(-) Non-T cell leukemia
Cannot be stored
Excellent marker for hairy cell leukemia
Neutrophil is the only leukocyte that contains this activity
Found in 3’ granules (Metamyelocytes)
 Leukomoid reaction, MMM, PV, 3rd trimester of pregnancy
 CML, CGL, PNH
Should be done on fresh specimen
♫ Kaplow count:
Page | 195

Toluidine blue

Cooper & Cruickshank
Nitroblue tetrazolium test
(Original)

Modified NBT

Feulgen reaction
Prussian blue stain
Supravital stain (RHH)

BCB

= Count 100 neutrophils
= Grade 0 to 4+
NV = 30-185
LAP Score:
0 = No stain
1+ = Faint stain
2+ = Pale stain
3+ = Strong stain
4+ = Deep (Very intense) stain
Binds w/ acid mucopolysaccharides in blood cells
Strongly metachromatic
Useful for recognition of mast cells and tissue basophils (metachromatic
granules)
 CGL: Chronic granulocytic leukemia
Stain for basophils
Reagent: Toluidine blue
Test for CGD
NBT: Colorless to pale yellow ---(Toxic O2 molecules)---> (+) Blue formazan
Test:
♫ Heparinized blood  Centrifuge  Buffy coat (WBC) + NBT  (+) Formazan
NV = ≤10% formazan (+)
CGD = 0 to negligible
Infection = 70%
w/ stimulating agent
Test:
♫ Buffy coat (WBC) + Bacterial suspension ---(NBT)---> (+) Formazan
NV = 80-100% formazan
CGD = <50% formazan
Specific reaction for DNA
Howell-Jolly bodies = Feulgen (+)
Stain for hemosiderin
(+) Sideroblast
(+) Siderocytes
RHH = NCB
Reticulum = New methylene blue
Heinz bodies = Crystal violet
Hemoglobin H = Brilliant cresyl blue
Stain for automated cell counter

Page | 196

Primary hemostasis
Platelets

Secondary hemostasis
Arteries
Veins
Capillaries
Maturation Stage
Megakaryoblast
Promegakaryocytes
Megakaryocytes
Metamegakaryocytes
Platelet structure

Platelet Adhesion

Bernard-Soulier syndrome
(Giant platelet syndrome)
Von Willebrand disease
Platelet Activation

HEMOSTASIS
Involves blood vessels and platelets
Formation of platelet plug
Test: Bleeding time
Functions:
Adhesion
Activation
Release
Aggregation
Involves the coagulation factors
Formation of stable fibrin clot
Test: Clotting time
Carry blood from the heart to the capillaries
Primary Hemostasis
Return blood from the capillaries to the heart
Injured vessel: vasoconstriction
= initiated by serotonin and thromboxane A2 derived from platelets and
endothelial cells
Cytoplasmic
Cytoplasmic
Nuclear
Thrombocytes
Granules
Tags
Features
Visible
(-)
(+)
Single nucleus
(-)
Fine chromatin
(+) Nucleoli
Few
(+)
2 nucleus
(-)
Numerous
Usually (-)
2 or more nuclei
(-)
Aggregated
(-)
4 or more nuclei
(+)
60% proteins
30% lipids
8% carbohydrate
Various minerals, water, nucleotides
1
...
Glycocalyx = outer surface
b
...
Submembranous area
2
...
Microfilaments: actin & myosin (actomyosin/thrombostenin)
= responsible for clot retraction
b
...
Organelle zone
= alpha & dense granules
= mitochondria, lysosomal granules
4
...
Dense tubular system = site of arachidonic acid metabolism
b
...
Alpha granules
= Platelet factor
= Platelet derived growth factor
= Fibrinogen
= Factor V
= vWF
= β-thromboglobulin
= Thrombospondin
= Fibronectin
= Albumin
b
...
Alpha-granule deficiency
♫ Gray platelet syndrome
♫ Quebec platelet disorder = (-) Factor V binding protein (multimerin)
b
...
PL)
Promote vascular repair
Platelet-derived growth factor (α) = promotes smooth muscle growth
β-thromboglobulin (α) = chemotactic for fibroblasts
Other systems affected
Plasminogen (α)
α2-antiplasmin (α)
C1 esterase inhibitor (α)
Platelet aggregation
Platelet attachment to each other
Requires fibrinogen and Ca2+
Glanzmann’s
(-) gpIIb/IIIa complex: receptor for fibrinogen
thrombasthenia
Petechiae
Pinpoint hemorrhagic spots
Page | 198

Purpura
Ecchymosis
Epistaxis
Hemarthrosis
Hematemesis
Hematoma
Hematuria
Hemoglobinuria
Melena
Menorrhagia
Hereditary hemorrhagic
telangiectasia
(Oslwer-Weber-Rendu
disease)
Congenital hemangiomata
(Kasabach-Merritt
syndrome)
Ehler-Danlos syndrome
Marfan’s syndrome
Pseudoxanthoma elasticum
Senile purpura
Scurvy
Henoch-Schonlein purpura
Thrombocytopenia

Thrombocytosis

Platelet adhesion

Acquired

Hemorrhage of blood into small areas of skin
Blood escapes into large areas of skin
Nosebleed
Leakage of blood into joint cavities
Vomiting of blood
Swelling or tumor in the tissues or a body cavity that contains clotted blood
RBC in urine
Hgb in urine
Stool containing dark red or black blood
Excessive menstrual bleeding
Vascular Disorders
Most common inherited vascular disorder
Blood vessels are thin & lack smooth muscle
Tumor composed of blood vessels
 vascular fragility
Elastic fibers are calcified & structurally abnormal
Degradation of collagen & elastin
(-) Vitamin C = for collagen synthesis
Defective synthesis of collagen
Immunologic damage to endothelial cells
Quantitative Platelet Disorders
 Platelet production of BM = aplastic anemia
 Survival time =  platelet destruction (DIC, ITP)
 Platelet sequestration by the spleen = splenomegaly
Dilution of platelet count = Thrombocytopenia α # of units transfused
 Units transfused =  Thrombocytopenia
Multiple transfusion: stored blood contains nonviable platelets
Reactive = moderate increase, asymptomatic (after hemorrhage, splenectomy)
Autonomous = marked increase, associated w/ thrombotic/hemorrhagic
complications (Ex
...
vWD = (-) vWF
♫ Platelet aggregation test:
= Normal: Epinephrine, Collagen, ADP
= Abnormal: Ristocetin
2
...
Storage pool defects
= Gray platelet (α), HP, WAS, CHS (d)
Uremia:  toxic metabolites
Paraproteinemias: coating of platelet membrane w/ abnormal protein
AML: abnormal megakaryocytes
MPD
Drugs: Aspirin = inhibits COX
Page | 199

Platelet count

Unopette

Platelet estimate
(Wedge smear)
WBC estimate (HPF)
Platelet Estimate of
0-49,000/μL
50,000-100,000/μL
100,000-150,000/μL
150,000-199,000/μL
200,000-400,000/μL
401,000-599,000/μL
600,000-799,000/μL
>800,000μL
N-Plt count,  BT
 Plt count, N-BT
 Plt count,  BT
Platelet aggregation test

Laboratory Tests for Primary Hemostasis
1
...
count/mm3 = # plt x AF x DCF x DF = # plt x 1 x 10 x 200 = # plt x 2,000
(RBC pipette = 1:200 dilution)
♫ 1 platelet uncounted = -2,000 plt/mm3
a
...
Guy and Leake
- Sodium oxalate
- Formaldehyde
- Crystal violet
c
...
Unopette
2
...
Dameshek
b
...
Olef’s
Diluent: 1% Ammonium oxalate
Stand moist chamber for 15-20mins to allow platelets to settle
1
...
02mL blood
TV = 2mL
Dilution = 1:100
Plt
...
Epinephrine
b
...
ADP
d
...
D
...
1 = routine collection method
Plt ct
...
Castor oil/Hirschboeck
(+) Dimpling/droplet like serum on the surface of blood drop
NV = 15-45mins
b
...
MacFarlane
CRT = (vol
...
systolic & diastolic pressure for 5 mins)
+1 = 0-10 petechiae (few)
+2 = 10-20 petechiae (many)
+3 = 20-50 petechiae (multiple)
+4 = >50 petechiae (confluent)
NV = 0 to occasional
Not repeated on the same arm for 7 days
In vivo measure of primary hemostasis
a
...
Ivy method = uses blood pressure cuff (40mmHg)  puncture forearm
Secondary Hemostasis
All are produced in the liver except Factor VIII
In liver disease, all coagulation factors are  except factor VIII complex
Factor VIII complex:
a
...
vWF: produced by megakaryocytes and endothelial cells
Activated at cold temp
...
K independent
Involved in the contact phase
XII ---(Collagen)---> XIIa (small amount)
PK ---(XIIa)--------> Kallikrein
XII ---(Kallikrein+HMWK)---> XIIa (large amount)
XI ---(XIIa)---------> XIa
I, V, VIII, XIII
Ca2+ dependent
Vit
...
K dependent
First: VII  IX  X  II: Last
Adsorbable factors: removed by adsorbing agents [BaSO4, Al(OH)3]
(+) in plasma
(-) in serum
Page | 202

Diseases
BT

Disease of 1’ hemostasis
Fibrinogen deficiency
N*
Prothrombin deficiency
N
Parahemophilia
N
Factor VII deficiency
N
Hemophilia A
N

von Willebrand disease
Hemophilia B
N
Factor X deficiency
N
Hemophilia C
N
Factor XII deficiency
N
Factor XIII deficiency
N

DIC
*BT may be prolonged in afibrinogenemia
Fresh Plasma

Aged Plasma

PT
N




N
N
N

N
N
N


APTT
N



N






N

Adsorbed
Plasma
+
+
+
+
+
+

Stypven
N



N
N
N
N

N
N
N

Fresh Serum

TT
N

N
N
N
N
N
N
N
N
N
N


Duckert’s
N
N
N
N
N
N
N
N
N
N
N
Abn
Abn
Aged Serum

I
+
+
II
+
+
+ (<20%)
V
+
VII
+
+
+
+
VIII
+
IX
+
+
+
+
X
+
+
+
+
XI
+
+
+
+
XII
+
+
+
+
XIII
+
+
Prothrombin:
80% is consumed during coagulation
<20% residual prothrombin
Disorders of Coagulation Causing Clotting Factor Deficiencies
Inherited Coagulopathies
Acquired Coagulopathy
Factor
Inheritance Pattern
Coagulopathy
I
Autosomal recessive
Afibrinogenemia
Liver disease
Autosomal dominant
Dysfibrinogenemia
DIC
Fibrinolysis
II
Autosomal recessive
Prothrombin deficiency
Liver disease
Vit
...
K deficiency
Anticoagulant therapy
VIII
X-linked recessive
Hemophilia A
DIC
Autosomal dominant
vWD
Fibrinolysis
IX
Autosomal recessive
Hemophilia B
Liver disease
Christmas disease
Vit
...
K deficiency
Anticoagulant therapy
Unclear

Unclear

Liver disease
DIC
Fibrinolysis
PK
Autosomal recessive
Fletcher trait
Unclear
HMWK
Autosomal recessive
Fitzgerald trait
Unclear
Factor VIII deficiency
Common inherited coagulation factor deficiency
a
...
vWD = most frequently inherited coagulation disorder
Major sites of coagulation
Endothelium
inhibition
Platelet
Protein C
Degrades factor Va and VIIIa
Protein S
Vit
...
Capillary blood method
= Drop or slide
= Capillary tube/Dale and Laidlaw
= Blue capillary tube
NV = 2-4mins
b
...
Activators:
a
...
Ellagic acid
c
...
Kaolin
2
...
CaCl2
Begin timing for clot formation on the addition of CaCl2 rgt
NV = 25-35 secs
Stypven time/Russel viper
East Indian viper venom Vipera russelli: directly activates factor X
venom time
Detect coagulation factor deficiencies involving common pathway
PPP + Stypven rgt: platelin-CaCl2 rgt
NV = 6-10 secs
Page | 204

Thrombin time

Reptilase time

Duckert’s test
5M urea solubility test

Circulating anticoagulants
Lupus inhibitor
Tilt tube method
Fibrometer
Photo-optical detection of
clot formation

Fibrinolysis
Plasminogen activators

Prolonged in:
a
...
Presence of FDP or FSP
c
...
streptokinase)
d
...
Fibrinogen deficiency
b
...
Presence of thrombolytic agent
Not affected by heparin
PPP + Atroxin
Begin timing for clot formation upon the addition of atroxin
NV = 10-15 secs
For factor XIII deficiency
Rgt: 5M Urea
Substitutes for urea:
- 1% monochloroacetic acid
- 2% acetic acid
Normal = Clot is insoluble to urea (24 hrs)
F
...
= Clot is soluble to urea (24 hrs)
Prolonged APTT and PT not corrected
Against the phospholipid portion of prothrombinase complex (Va-Xa-Ca2+-PL)
Instrumentation for Tests of Hemostasis
Visual detection of fibrin clot formation
Manual technique
End point = clot formation
Electromechanical detection of fibrin clot formation
Fibrin strand formation is detected using a wire loop or hook w/c has been
incorporated into a semi-automated mechanical instrument
Detection of fibrin clot formation depends on  in light scattering associated
w/ conversion of fibrinogen  fibrin
♫ Semi-automated instruments:
- Electra 750 and 750A
- Fibrintimer series
- FP 910 coagulation analyzer
♫ Automated instruments:
- Ortho Koagulab 16S and 40A
- Coag-A-Mate X2 and XC
- MLA Electra 700 and 800
Digestion of fibrin clot
Occurs when plasminogen  plasmin
1
...
Tissue type
- Urokinase-like PA
3
...
meningitidis
♫ Neoplasm
♫ Snake bite
♫ HTR
Laboratory Evaluation of Fibrinolysis
Clot should remain intact for approximately 48 hrs at 37’C
Clot lysis prior to 48 hrs indicates excessive systemic fibrinolysis
Euglobulins: proteins that ppt
...
euglobulin
Euglobulin + thrombin  Clot euglobulin
Euglobulin  Dissolved in buffer
Normal = No clot lysis (2 hrs)
 Fibrinolysis = Clot lysis <2 hrs
Page | 206

Protamine sulfate test
Ethanol gelation test
Latex D-dimer assay
Test
WBCLT
Euglobulin clot lysis
Ethanol gelation
Protamine sulfate
D-dimer
Heparin

Warfarin
Coumarin
Coumadin
INR

ISI

Detects the presence of fibrin monomers (2’-DIC) in the plasma
Plasma + Protamine sulfate + ETOH  (+) gel-like clot (paracoagulation)
Detects the presence of fibrin monomers (2’-DIC) in the plasma
Plasma + NaOH (pH) + ETOH  (+) pptn/gel
Most specific test for DIC
Primary Fibrinolysis
Secondary Fibrinolysis
< 48 hrs
< 48 hrs
< 2 hrs
< 2 hrs
+
+
+
Injected
Action: inhibits thrombin
Monitoring: APTT = sensitive, method of choice (CAP)
Neutralize w/ protamine sulfate
Oral
WARF: Wisconsin Alumni Research Foundation
Action: Vit
...
5-3
...
1N HCl
Comparing the brownish yellow color of solution to standard comparator block
Rgt: 0
...
34mL O2
1g Hgb = 3
...
02mL blood: 5mL rgt (1:251)
Reagents:
Drabkin’s reagent (Brown container)
a
...
Potassium cyanide = Hi  HiCN
c
...
Sodium bicarbonate (Orig
...
Dihydrogen potassium phosphate (Mod
...
High WBC count: to correct, centrifuge  read supernatant
b
...
Lipemic blood: prepare patient’s blank (pt
...
Wintrobe & Landsberg = Double oxalate
2
...
Haden = Sodium oxalate
4
...
Bray = Heparin
Centrifuge: 2000-2300g for 30mins
Layers (Spun Hct):
1st: Fatty layer = barely visible unless the patient is lipemic
2nd: Plasma
3rd: Buffy coat (1mm = 10,000 WBCs/mm3)
Bottom: Packed cells
Length = 11
...
0mm
Calibration = 0-100
a
...
Right: 100-0 (Hct)
Capillary tube:
Length = 7
...
5cm (70-75mm)
Bore = 1mm
Anticoagulant: Red (Heparin)
No anticoagulant: Blue
Centrifuge: 10,000-15,000g for 5mins
Trapped plasma: plasma that remains in RBC portion after spinning
=  in poikilocytosis
♫ Advantages:
a
...
Less blood
c
...
) NSS
2
...
8% Sodium citrate
3
...
) Hayem’s
5
...
) Bethell’s
7
...
5:100 (Blood: Diluent) = 1:200
RBC/mm3 = # RBC x AF x DCF x DF = # RBC x 5 x 10 x 200 = # RBC x 10,000
WBC Count
AM, PM
Hypotonic solutions: lyse non-nucleated RBCs
1
...
) 1% HCl
3
...
5:10 (Blood: Diluent) = 1:20
Leukocytosis = Use RBC pipette (1:100 or 1:200)
WBC/mm3 = # WBC x AF x DCF x DF
Counting Chamber
2 counting areas
Each CA w/ 16 1mm2 squares
Depth = 0
...
2mm x 2 = 6
...
2mm3
4 counting areas
Each CA w/ 10 1mm2 squares
Depth = 0
...
2mm x 4 = 8mm3
Volume/counting chamber = 2mm3
2 primary squares
Each 1’ square w/ 9 1mm2 2’ squares
Depth = 0
...
1mm x 2 = 1
...
9mm3
Center square:
w/ 25 3’ square
Each 3’ square w/ 16 small squares
25 x 16 = 400
5 (counted) x 16 = 80 small squares
4 corners:
Each 2’ square w/ 16 3’ squares
4 x 16 = 64 3’ squares
Not lysed by WBC diluents
Falsely counted as WBCs
Page | 210

NV:
Adult = ≥5 nRBC/100 WBC differential
Newborn = ≥10 nRBC/100 WBC differential
Formula for WBC
Corrected WBC = uncorrected WBCs x 100
correction
100 + NRBCs
Preparation and Staining Procedures for the Blood Smear
Techniques
1
...
Cover glass and slide (Beacom)
3
...
Spun smear/Spun/Spinner’s technique = Automated: Hemaspinner
0 (30-400)
25
Angle between 2 slides
22 x 22mm
Square coverslip
2-3mm
Drop of blood
0
...
5 inch
Smear terminates near the end of the slide (automated spreader)
Buffy coat smear
For patients w/ WBC count of <1 x 109/L
Demonstration of LE cell
Thick blood smear
For blood parasites
Methanol
Fixative for blood and BM smears
Toxic, causes permanent blindness
Romanowsky’s stains
Wright’s
Giemsa = preferred stain for blood parasites
Modified Wright’s-Giemsa
Leishman
Jenner
May-Grunwald
-----------------------------------------------------------------------------------------------------Contains:
= Methylene blue (or Azure B - oxidized): basic
= Eosin: acidic
w/in 2-3 hours of specimen Time blood smears should be stained
collection
pH 6
...
2
Malarial parasite staining
Excessively blue stain
Thick films
Prolonged staining time
Inadequate washing
Too high alkalinity of stain
Diluents tends to cause excessive basophilia
Excessively pink stain
Insufficient staining
Prolonged washing time
Mounting coverslips before they are dry
Too high acidity of the stain
Buffer may cause excessive acidophilia
Cross-sectional or
Blood film is moved from side to side
crenellation method
Longitudinal method
WBCs are counted from the tail toward the head of the smear
Battlement method
Near the tail on a horizontal edge: count 3 consecutive horizontal edge fields,
count 2 fields towards the center of the smear, count 2 fields horizontally, count
2 fields vertically to the edge
WBC Counting
100 cells = routine
50 cells = if patient WBC count <1
...
8-7
...
0-4
...
01-0
...
6 x 109/L
Relative = 0-1%
Absolute = 0-0
...
spiralis)
Eosinophilic leukemia
Viral infections
Whooping cough
IM
Lymphocytic leukemia
Lymphoma
Brucellosis
Tuberculosis
Monocytic leukemia
SBE
Typhoid
Rickettsial infections
Collagen disease
Hodgkin’s disease
Gaucher’s disease
(+) immature granulocytic cells
Leukemia
Bacterial infections
Hypersegmented neutrophils (≥6 lobes)
NV:
Adult = 0
...
5% (Ave: 1
...
45L/L])
NV = 1
General indicator of the rate of erythrocyte production increase above normal
in anemias
Indicates BM response to anemia
RPI = CRC ÷ Maturation time of retics in the blood
NV = 1 (Hct: 45%)
1
...
5 days = Hct: 35 ± 5%
2
...
5 days = Hct: 15 ± 5%
Adequate response of BM to anemia
- Chronic hemolysis
- Recent hemorrhage
- Response to therapy
Inadequate response of BM to anemia
- Aplastic anemia
- Ineffective erythropoiesis (megaloblastic anemia)
% Retics = Retics (A) ÷ [RBC (B) x 9] x 100
NV = 50-350 x 106/L
Allergic reactions
Parasitic infections
Brucellosis
Leukemias
Hyperadrenalism (Cushing’s disease)
Shock
Administration of ACTH
Composition:
a
...
Propylene glycol = lyses RBCs
c
...
Heparin = prevents clumping
Diluting fluids:
1
...
Manner’s = phloxine
3
...
Hinkleman = eosin
5
...
count)
Normal: Eo
...
count: 1 = 2 (hypoadrenalism)
Erythrocyte Indices (Wintrobe Indices)
Classify anemia according to RBC morphology
MCV = (Hct ÷ RBC) x 10
NV = 80-100 fL (old: μm3)
MCH = (Hgb ÷ RBC) x 10
NV = 27-32 pg (old: μμg)
Page | 213

MCHC
Defective centrifuge

MCHC >38% does not occur
MCHC will not fall <22%
ESR
 ESR

 ESR

Stages of ESR

Wintrobe & Landsberg

Standard/Original
Westergren

Rarely used
MCHC = (Hgb ÷ Hct) x 100
NV = 31-36% (31-36 g/dL)
Values affected:
= Hct
= MCV
= MCHC
Incorrect calculation
(+) cold agglutinins
Lipemia
(+) HbS & HbC
Erythrocyte Sedimentation Rate (ESR)
Nonspecific measurement used to detect & monitor an inflammatory response
to tissue injury
Erythrocytes:
*Macrocytes
*Anemia
Plasma composition: most important determinant
*Fibrinogen
*α1-globulin
*α2-globulin
*β-globulin
*γ-globulin
*Cholesterol
Technical factor:
*Tilting = 30 angle = 30% error
*Temp
...
Initial rouleaux
40 mins = 2
...
Final sedimentation of RBCs
60 mins = Total
Requires smaller amount of blood
Involves no dilution
Length: 11
...
0mm
Anticoagulant: Double oxalate
Most sensitive
Requires more blood
Length: 300mm
Internal bore: 2
...
15mm
Anticoagulant: Citrate (black)
Anticoagulant-to-Blood ratio = 1:4
Page | 214

Modified Westergren
Zeta Sedimentation Ratio
(ZSR)
Erythrocyte Osmotic
Fragility test
(Griffin and Sanford
method)

Ascorbate cyanide
screening

G6PD fluorescent screening
Paroxysmal nocturnal
hemoglobinuria (PNH)
Sucrose hemolysis test

Ham’s acidified serum test

Patient has received normal
RBCs
PCH
Donath-Landsteiner test

Autohemolysis test

Anticoagulant: 2mL EDTA + 0
...
02
Add RBCs, stand for 2hrs at room temp
Check for hemolysis (pink/red supernatant)
NV:
- Initial hemolysis = tube 21 or 22 (0
...
44%)
- Complete hemolysis = tube 16 or 17 (0
...
34%)
Detects deficiencies in the pentose phosphate pathway:
- G6PD
- glutathione peroxidase
- glutathione reductase
Rgts:
- Na ascorbate
- Na cyanide
Normal = red
(-) Enzyme = brown
G6P + NADP ---(RBC: G6PD)---> 6-phosphogluconate + NADPH (fluorescence)
Normal: Max fluorescence at 10mins
G6PD def: Little or no fluorescence
Acquired disorder in w/c red cells are abnormally sensitive to complement
(-) DAF
Screening test for PNH
Patient RBCs + ABO compatible serum + sucrose solution
Normal = (-) Hemolysis
PNH = (+) Hemolysis
Confirmatory test for PNH
Tube 1: Patient RBCs + normal serum + weak acid (0
...
2N HCl)
Tube 3: Patient RBCs + normal inactivated serum + weak acid (0
...
2 to 2%
Blood + glucose ---> Hemolysis: 0-0
...
9%
-----------------------------------------------------------------------------------------------------G6PD deficiency (PPP) = corrects w/ glucose only
PK deficiency (EMP) = corrects w/ ADP only
H
...
Technicon autoanalyzer
Blood cells are nonconductors of electricity
...
Sysmex counter, Coulter counter
Triplicate count (3x)
a
...
Blood is diluted 1:251 (hypotonic)
♫ Lymphocytes = 35-90fL
♫ Monocytes = 90-160fL
♫ Granulocytes = 160-450fL
RBCs, WBCs, plts
X-axis
- Horizontal/abscissa
- Size of cells
Y-axis
- Vertical/ordinate
- Number of cells
V=IxR
Where:
V = voltage
I = current
R = resistance
 Count: “BEA”
♫ Bubbles
♫ Extraneous electrical pulses
♫ Aperture plug
 Count
♫ Improper setting of aperture error
% of RBCs that are polychromatophilic
Slight = 1%
1+ = 3%
2+ = 5%
3+ = 10%
4+ = >11%
Page | 217

Normocytic, Normochromic
RBCs

Hemolytic anemias

1
...
Abnormal hemoglobin, increased destruction of RBCs
*Certain acquired hemolytic anemia
*PNH
*Sickle cell anemia
*HDN
*Anemia of chronic renal insufficiency
1
...
Hereditary – membrane defects
**Spherocytosis
**Elliptocytosis
**Acanthocytosis
**Stomatocytosis
**Rh null disease
b
...
Hereditary – hemoglobinopathies
**Sickle cell disease
**Hemoglobin C disease
d
...
Hereditary – defective globin synthesis
**Thalassemia
f
...
Extracorpuscular causes: nonimmune acquired hemolytic anemias
*Chemicals, toxins, venoms
*Physical trauma: disorders causing fragmentation (burns, cardiac replacement
valves, MAHA, HUS)
3
...
Miscellaneous
*Anemia of liver disease
*Sulfhemoglobinemia
*Porphyrias
*Methemoglobinemias

Page | 218



---