Introduction to Hematology

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 Introduction to Hematology
 Hematology is about relationships
• Bone marrow to the systemic circulation
• Plasma environment
to the red blood
cell life span
• Hemoglobin to
the red blood cell

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 Introduction to Hematology
 Blood
• sticky, opaque, metallic taste
• bright red to dark red (oxygenation level)
• pH 7.35-7.45
• 38oC
• 4-6 L of blood
 Study of blood and related
disorders
 Staining, counting, analysis of components

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 History
 Blood has been viewed for centuries
 Anton van Leeuwenhook described RBCs
 Giulio Bizzozero described platelets
 Wright Stain* – J. Homer Wright
 Analyzers now categorize and enumerate RBCs,
WBCs, and Platelets
 Technologist still responsible for morphology and
overall description of hematology smears

*Romanowsky style stain = polychromatic mixture of acid and

basic dyes
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 Red Blood Cells
 Anucleate
 Biconcave
 Reddish protein (Hgb)
 Transports oxygen and carbon dioxide

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 Red Blood Cells
 For years, cells were counted to detect
anemia and polycythemia
 whole blood would be
diluted* with saline (0.85%)
 1:200 dilution
 Performed in a hemocytometer
 50s = Electronic counters

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 White Blood Cells
 Leukocytes
 Protect from infection
 Heal injuries
 Counted in similar fashion to RBCs
 1:20 dilution with acid
 lyses RBCs
 Seen in peripheral blood, bone marrow,
body fluids, cerebral spinal fluid

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 White Blood Cells
 Divided into 5 general groups

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 Platelets
 Thrombocytes
 Repair damaged vessels – thrombosis
 “Cell fragments”
 Small element but responsible for major
conditions

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 Complete Blood Count
 CBC and CBCD
 automated analyzers should give differential
 Blood film examination by technologist not
always required
 Manual differential not
always required either

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 Coagulation
 Platelet function and plasma coagulation
 Coagulation is a complex cascade of a series
of proteins, compounds and elements
 Responsible for clot formation after platelet
plug initiation
 Tests include: Prothrombin Time (INR),
Activated Partial Thromboplastin Time, D-
Dimer, Fibrinogen

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 Coagulation
 Helps monitor anticoagulant therapy of
patients
 Specimen integrity*
 Liquid anticoagulant requiring specific
dilution with WB
 Special attention to hemolyzed, lipemic,
icteric samples depending on method of
detection (optical vs mechanical), clots

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 Coagulation
 Prothrombin Time (PT)
• monitor oral anticoagulation
• Warfarin/Coumadin
• PT in seconds converted to INR
 Activated Partial Thromboplastin Time
• APTT, aPTT, PTT
• monitors IV anticoagulation (usually admitted)
• Heparin
Can also evaluate factor deficiencies and
screen for inhibitors
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 Hematology Department
 Other parts of the department will vary
depending on level of lab complexity and
location… but can include:
• Flow cytometry (Immunophenotyping cells)
• Bone marrow (specimen prep, special stains, etc.)
• Coagulation (thrombin time, clotting factors, etc.)
• Platelet studies (functional testing)
• Erythrocyte Sedimentation Rate (ESRs)
• Reticulocyte count

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 Hematology Department
 Other parts of the department will vary
depending on lab and location… but can
include:
• Manual testing (malarial parasites, infectious
mononucleosis, sickle cell disease, osmotic
fragility
• Body fluid analysis: CSF, synovial fluid, pericardial
fluid, pleural fluid, peritoneal fluid
 Many of these tests are found in higher level
facilities and require specialized/advanced
training and technologists
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 Microscope
 Eye piece/ocular  Iris diaphragm
 Objectives • Increases or decreases
• 10x, 40x, 60x, 100x light from the microscope
light source
• Magnification number
• Numerical Aperture  Stage
number (NA)  Adjustment knobs
‒ Resolution power • Coarse
• Tube length • Fine
‒ Distance from eye piece to
• Bring the image into focus
objective

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 Microscope (continued)

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 Microscope Problem-Solving
 Use the correct side of the slide
 Open both diaphragms for maximum light
 Wipe off 40x lens with lint-free paper
 Clean eye piece with lens cleaner to remove
dust particles
 Remove dust from light source

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 Microscope
 Brightfield microscopy for smear
examination
 Tungsten lamps with blue filter
 Khoeler illumination
 Especially important on oil-immersion lenses

*Please be careful when adjusting between

lenses that you don’t get oil on the non-oil
lenses
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 Standard Precautions
 Treat all specimens as a potential source of
infection
 Personal Protective Equipment (PPE)
• Gloves
• Gowns/laboratory coats
• Splash shields
 Hand washing

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 Standard Precautions
 Great risk for exposure to blood and body
fluids
 Many aspects involve manipulating open
blood specimens/manual manipulation with
the cap removed from sample
 Wide variety of sample types: blood, all sorts
of body fluids, CSF

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 Standard Precautions
 Hand Washing: Soap and Water vs. Alcohol
based
 Needles, capillary tubes, cap piercers

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 Personal Protective Equipment

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 Safety
 Use puncture-resistant containers for sharps
 No mouth pipetting, eating, smoking,
drinking, or gum chewing
 Keep loose papers and notebooks out of lab
 No dangling jewelry
 Use closed-toe shoes
 Personal hygiene
• Keep nails short
• Keep facial hair trimmed
• Tie long hair back
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 Hazards
 Be aware and adhere to chemical spill action
plans
 Fire hazard (large quantities of methanol,
other flammable chemicals)
 Electrical hazards
 Radioactive hazards (not found in all
hematology departments)
 Physical hazards (heavy reagent packs*),
always use trolleys and lift properly

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 Quality Assurance
 Ensures laboratory testing reliability
 Examples of Q A indicators
• Number of labeling errors
• Errors in data entry
• Testing turnaround times
• Proficiency testing performance
• Standardized Competency testing

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 Quality Control
 Monitors accurate analysis of laboratory
results
 Quality control: important components
• Standards/calibrators
• Control materials (normal and abnormal) –

Pos/Neg -

Low/Norm/High -

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 Quality Control
 Quality control: important components
• Statistics
‒ Mean, mode, and median control values
‒ Standard Deviation – precision measurement
‒ Coefficient of Variation – SD expressed as %
‒ Accuracy
‒ Precision

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 Quality Control
 Complex in comparison to other
departments
 Cell measurements are not conducive to
chemical standardization
 Difficult matrix effects, reference intervals
 Many systems employ a moving average
 Morphological grading and technologist
specific descriptions and comments are
usually site specific

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 Important Values
 Reference values
• Normal values determined by patient population
 Delta checks
• Historical check of lab results
• Key to identifying preanalytical errors
 Reflex testing
• Additional testing to verify abnormal results
 Critical values
• Results that are markedly decreased or increased
from the reference range
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 Sample Collection
 Phlebotomy safety must always be followed
 Physical variables can potentially affect some
(time of collection must always be accurately
recorded on sample)
 Ethylenediaminetetraacetic acid
 Sodium Citrate (2 different volumes)
 Order of Draw is crucial. Why?

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 Sample Collection
Venipuncture Heel prick
Blood culture Blood gas
Coagulation EDTA microtainer
Serum (without/with gel) Other anticoagulant microtainers
Heparin (without/with gel) Serum microtainers
EDTA
Sodium fluoride

• Must follow rejection criteria for all collections…

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 Sample Collection
 Most hematology analyzers use whole blood
 Rare chemistry test uses whole blood
 EDTA prevents clotting giving plasma.
How?
 Sample will separate upon standing
therefore mixing required before analysis
 Blood smear should be made ASAP when
required (morphology can change)

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 Sample Collection
 Plasma descriptions
• normal plasma is light yellow – dark yellow
• icteric often dark yellow – brown

• Lipemic appears as cloudy

• Hemolyzed is pink- red

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 Preanalytical Factors
 Factors that affect the sample before testing
• Specimen identification
• Sample labeling
• Proper collection methods
• Time of collection
• Specimen integrity

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 Postanalytical Factors
 Factors that occur after testing
 Result documentation
 Reporting critical values
• Determined by each lab
• Reported immediately to caregiver to ensure
proper patient medical attention

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 Definitions and Terms
 Anemia
 Polycythemia
 Leukocytopenia
 Leukocytosis
 Thrombosis
 Thrombocytopenia
 Thrombocytosis
 Microscope Terms and Troubleshooting
 Quality Control Terms
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 Homework
 Read Chapter 1
 Review definition/terms list
 Hematology Problem Set 1
 Check out the links on moodle

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