Hematology Review

1
Hematology Review

• By
• Felicia Magee Tardy, M.S.,
• MT (ASCP)
• Department of Clinical Pathology
• University of Mississippi Medical Center
• Jackson, Mississippi

2
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.

3
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.

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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.

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Thrombocytes

• Smallest cells in the blood.
• Normal range 130,000-400,000.
• Active role in coagulation and hemostasis.

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Routine Hematology

• Anticoagulant of choice EDTA
• Complete Blood Counts (CBCs)
• Manual WBC Differentials
• Erythrocyte Sedimentation Rates (ESRs)
• Sickle Screens
• Reticulocyte Counts

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Automated Counting

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

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Automated Counting

• Flow Cytometry
• Uses lasers to measure both forward and side
  scatter.
• Forward scatter measures size.
• Side scatter measures granularity.

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Sources of Error

• Inadequate mixing of specimen.
• Hemolyzed specimens.
• Lipemic specimens.
• Cold agglutinins.
• Clotted specimens.
• Platelet clumps or platelet satellitosis.
• Diluted specimens.

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Know Normal Ranges!!!

• WBC
• RBC
• HGB
• HCT
• MCV
• MCH
• MCHC

• PLT
• MPV
• SEGS
• LYMPHS
• MONOCYTES
• EOSINOPHILS
• BASOPHILS

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Erythrocytic Maturation Series

• Rubriblast
• Prorubricyte
• Rubricyte
• Metarubricyte
• Reticulocyte
• Erythrocyte

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RBC Morphology
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Elliptocytes Target Cells
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Tear Drops Stomatocytes
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Sickle Cells Schistocytes
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RBC Inclusions
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Howell-Jolly Bodies

• Round, purple inclusions in RBCs.
• Composed of DNA.
• Commonly seen in in patients with hypofunctioning
  spleens.
• Splenectomy.

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Basophilic Stippling

• Numerous, small purple inclusions in RBCs.
• Aggregates of ribosomal RNA.
• Most commonly seen in lead poisoning.

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Pappenheimer Bodies

• Clusters of dark blue granules, irregular in size
  and shape.
• Composed of iron and ribosomal RNA.
• Seen in sideroblastic and hemolytic anemias.

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Classifications of Anemias

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

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Microcytic, Hypochromic

• Many RBCs smaller than
• nucleus of normal
• lymphocytes, increased central pallor.
• Iron deficiency, thalassemias, anemia of chronic
  disease.

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Classifications of Anemias

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

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Classifications of Anemias

• Macrocytic
• Vitamin B12 deficiency
• Folate deficiency
• Liver disease

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Macrocytic RBCs

• Most RBCs larger than nucleus of normal
• lymphocytes, increased MCV.
• Folate or Vitamin B12 deficiencies, alcoholism,
  and liver disease.

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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.

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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

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Hemoglobinopathies

• Beta Chain Substitutions
• Hgb S Valine for glutamic acid
• (6th position, beta chain)
• Hgb C Lysine for glutamic acid
• (6th position, beta chain)

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Hemoglobinopathies

• Alkaline Electrophoresis
• - C S F A
• A2 D
• E G O

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WBCEvaluation
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MyelocyticMaturation Series

• Myeloblast
• Promyelocyte
• Myelocyte
• Metamyelocyte
• Band Neutrophil
• Segmented Neutrophil

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Toxic Granulation

• Increased basophilic granules
• in neutrophils.
• Seen in severe infections, burns, malignancies,
  and pregnancy.
• Distinguish from basophils.

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Dohle Bodies

• Sky blue inclusions in cytoplasm of neutrophils.
• Seen in infections, burns, myleproliferative
  disorders, and pregnancy.
• Composed of RER and glycogen granules.

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Pelger-Huet Anomaly
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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

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CML versusLeukomoid Reaction

• Characteristic CML Leukomoid
• LAP score Decreased Increased
• Toxic gran. Decreased
  Increased
• Dohle bodies Absent Present
• Philadelphia May be present
  Absent
• chromosome

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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

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FAB Classifications

• M1 Myeloblastic without maturation
• M2 Myeloblastic with maturation
• M3 Promyelocytic
• M4 Myelomonocytic
• M5a Monocytic, poorly differentiated
• M5b Monocytic, well differentiated
• M6 Erythroleukemia, DiGuglielmos
• M7 Megakaryoblastic

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Acute Myeblastic Leukemia (M1)
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Chronic MyelogenousLeukemia
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FAB Classifications

• L1 Small, uniform lymphoblasts
• L2 Large, pleomorhphic lymphoblasts
• L3 Burkitts type (vacuolated and
• deeply basophilic cytoplasm)

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Acute Lymphoblastic Leukemia
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Chronic Lymphocytic Leukemia
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Body Fluids
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Types of Body Fluids

• Cerebrospinal Fluid (CSF)
• Pleural Fluid
• Pericardial Fluid
• Peritoneal Fluid
• Synovial Fluid
• Amniotic Fluid
• Miscellaneous Fluids

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Body Fluids Testing

• Physical examination
• RBC and WBC counts
• WBC differential
• Crystal examination
• Ropes test
• Refers to synovial fluids only.

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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.

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Quick Review

• Differentiate between traumatic tap and
  intracranial hemorrhage.
• Differentiate between transudate and exudate.
• Crystal identification and associated conditions.

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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

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Transudates v. Exudates

• Characteristic Transudate
  Exudate
• Appearance Clear, colorless Yellow,
  turbid, purulent, bloody
• Specific gravity lt1.015
  gt1.015
• Protein lt3 g/dL gt3 g/dL
• LD lt200 IU gt200 IU
• Cell count lt1000/uL gt1000/uL
• Conditions Congestive Infections,
• Heart failure Malignancies

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Crystal Identification

• Monosodium urate (uric acid)
• Yellow when parallel to the compensator, and blue
  when perpendicular.
• Seen in gout.
• Calcium pyrophosphate
• Blue when parallel to the compensator, and yellow
  when perpendicular.
• Seen in pseudogout.

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Crystal Identification
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Good Luck!!!!