Hematology Review

By

Felicia Magee Tardy, M.S.,

MT (ASCP)

Department of Clinical Pathology

University of Mississippi Medical Center

Jackson, Mississippi

Introduction

• Hematology: the study of blood and blood forming tissues.

• Blood consists of 55% plasma and 45% formed elements.

• Formed elements include erythrocytes, leukocytes, and thrombocytes.

Erythrocytes• Normal range 4.2-

5.5 million per mm3 in adults.

• Biconcave shape.• Diameter 7

microns.• Cells for transport

of O2 and CO2.

• Life span 120 days.

Leukocytes• Normal range 4 -

11 thousand per mm3 in adults.

• Five types.• Size 8-20 microns.• Involved in fighting

infection, combatting allergic reactions, and immune responses.

Thrombocytes

• Smallest cells in the blood.

• Normal range 130,000-400,000.

• Active role in coagulation and hemostasis.

Routine Hematology

Anticoagulant of choice: EDTA• Complete Blood Counts (CBCs)

• Manual WBC Differentials

• Erythrocyte Sedimentation Rates (ESRs)

• Sickle Screens

• Reticulocyte Counts

Automated Counting

Coulter Principle

Electrical impedance: resistance or change in current when cell passes between two electrodes in NaCl solution.

Automated Counting

Flow Cytometry

Uses lasers to measure both forward and side scatter.

Forward scatter measures size.

Side scatter measures granularity.

Sources of Error• Inadequate mixing of specimen.• Hemolyzed specimens.

• Lipemic specimens.

• Cold agglutinins.

• Clotted specimens.

• Platelet clumps or platelet satellitosis.

• Diluted specimens.

Know Normal Ranges!!!

WBC

RBC

HGB

HCT

MCV

MCH

MCHC

PLT

MPV

SEGS

LYMPHS

MONOCYTES

EOSINOPHILS

BASOPHILS

Erythrocytic Maturation Series

Rubriblast

Prorubricyte

Rubricyte

Metarubricyte

Reticulocyte

Erythrocyte

RBC Morphology

Elliptocytes Target Cells

Tear Drops Stomatocytes

Sickle Cells Schistocytes

RBC Inclusions

Howell-Jolly Bodies

• Round, purple inclusions in RBCs.

• Composed of DNA.• Commonly seen in

in patients with hypofunctioning spleens.

• Splenectomy.

Basophilic Stippling

• Numerous, small purple inclusions in RBCs.

• Aggregates of ribosomal RNA.

• Most commonly seen in lead poisoning.

Pappenheimer Bodies

• Clusters of dark blue granules, irregular in size and shape.

• Composed of iron and ribosomal RNA.

• Seen in sideroblastic and hemolytic anemias.

Classifications of Anemias

Microcytic, Hypochromic– Iron deficiency– Sideroblastic– Chronic disease, Inflammation– Lead poisoning– Thalassemia trait

Microcytic, Hypochromic

• Many RBCs smaller than

nucleus of normal

lymphocytes, increased central pallor.

• Iron deficiency, thalassemias, anemia of chronic disease.

Classifications of Anemias

Normochromic– Hereditary Spherocytosis– Hereditary Elliptocytosis– PNH– G6PD deficiency– Aplastic anemia– Acute blood loss

Classifications of Anemias

Macrocytic– Vitamin B12 deficiency– Folate deficiency– Liver disease

Macrocytic RBCs

• Most RBCs larger than nucleus of normal

lymphocytes, increased MCV.

• Folate or Vitamin B12 deficiencies, alcoholism, and liver disease.

Reticulocytes• Immature RBCs.• Contain residual

ribosomal RNA.• Reticulum stains

blue using a supravital stain (new methylene blue).

• Counted and expressed as % of total red cells.

Reticulocyte Count

Uses supravital stain which stains cells in the living state.

Retic % = # retics per 1000 RBCs

10

Corrected retic= % retics x pt. HCT

45

Hemoglobinopathies

Beta Chain Substitutions

Hgb S: Valine for glutamic acid

(6th position, beta chain)

Hgb C: Lysine for glutamic acid

(6th position, beta chain)

Hemoglobinopathies

Alkaline Electrophoresis

- C S F A +

A2 D

E GO

WBCEvaluation

MyelocyticMaturation Series

Myeloblast

Promyelocyte

Myelocyte

Metamyelocyte

Band Neutrophil

Segmented Neutrophil

Toxic Granulation

• Increased basophilic granules

in neutrophils.• Seen in severe

infections, burns, malignancies, and pregnancy.

• Distinguish from basophils.

Dohle Bodies

• Sky blue inclusions in cytoplasm of neutrophils.

• Seen in infections, burns, myleproliferative disorders, and pregnancy.

• Composed of RER and glycogen granules.

Pelger-Huet Anomaly

Special Stains

PAS: ALL, Erytrholeukemia

LAP: CML v. Leukomoid Reaction

Myeloperoxidase: AML, AMMoL (weak+)

Sudan Black: AML, AMMoL (weak+)

Specific Esterase: AML, AMMoL

Non-specific Esterase: AMML, AMoL

TRAP: Hairy Cell Leukemia

TDT: ALL

Prussian Blue: Sideroblastic Anemia

CML versusLeukomoid Reaction

Characteristic CML Leukomoid

LAP score Decreased Increased

Toxic gran. Decreased Increased

Dohle bodies Absent Present

Philadelphia May be present Absent

chromosome

WBC Disorders

Alder-Reilly: Large azurophilic granules, increased mucopolysaccharides

Chediak-Higashi: Membrane defect of lysosomes, large primary granules in segs

May-Hegglin: Familial disorder with large platelets and Dohle bodies

Pelger-Huet: Hyposegmented neutrophils

FAB Classifications

M1: Myeloblastic without maturation

M2: Myeloblastic with maturation

M3: Promyelocytic

M4: Myelomonocytic

M5a: Monocytic, poorly differentiated

M5b: Monocytic, well differentiated

M6: Erythroleukemia, DiGuglielmo’s

M7: Megakaryoblastic

Acute Myeblastic Leukemia (M1)

Chronic MyelogenousLeukemia

FAB Classifications

L1: Small, uniform lymphoblasts

L2: Large, pleomorhphic lymphoblasts

L3: Burkitt’s type (vacuolated and

deeply basophilic cytoplasm)

Acute Lymphoblastic Leukemia

Chronic Lymphocytic Leukemia

Body Fluids

Types of Body Fluids

• Cerebrospinal Fluid (CSF)

• Pleural Fluid

• Pericardial Fluid

• Peritoneal Fluid

• Synovial Fluid

• Amniotic Fluid

• Miscellaneous Fluids

Body Fluids Testing

• Physical examination

• RBC and WBC counts

• WBC differential

• Crystal examination*

• Rope’s test*

*Refers to synovial fluids only.

Cerebrospinal Fluid (CSF)

• Examined to determine the presence of meningitis or intracranial hemorrhage.

• Multiple tubes drawn (4).• Differentiate between traumatic tap

and intracranial hemorrhage.• Xanthochromia is indicative of

intracranial hemorrhage.

Quick Review

• Differentiate between traumatic tap and intracranial hemorrhage.

• Differentiate between transudate and exudate.

• Crystal identification and associated conditions.

Traumatic Tap v. Intracranial Hemorrhage

Traumatic Tap• Decreasing amounts

of blood with each tube drawn

• May contain clots• No xanthochromia• Supernatant clear

Intracranial Hemorrhage• Blood evenly distributed

throughout collection tubes

• No clots• Xanthochromia• Hemosiderin, hematoidin

crystals

Transudates v. Exudates

Characteristic Transudate Exudate

Appearance Clear, colorless Yellow, turbid, purulent, bloody

Specific gravity <1.015 >1.015

Protein <3 g/dL >3 g/dL

LD <200 IU >200 IU

Cell count <1000/uL >1000/uL

Conditions Congestive Infections,

Heart failure Malignancies

Crystal Identification

• Monosodium urate (uric acid)Yellow when parallel to the compensator,

and blue when perpendicular.

Seen in gout.

• Calcium pyrophosphateBlue when parallel to the compensator,

and yellow when perpendicular.

Seen in pseudogout.

Crystal Identification

Good Luck!!!!