September 9, 2003

SalesBasic Hematology

TechnicalTraining

September 9, 2003

Our goal . . .

To heighten your awareness of…

Basic hematology•blood cell types•associated tests•manual methods

September 9, 2003

Basic Hematology Objectives:

By the end of this session, you will be able to… Cite the definition of hematology and why its

study is important. List the major components found in blood. Describe the function of an erythrocyte,

leukocyte and thrombocyte. Recognize the terminology used to describe

abnormalities associated with erythrocytes, leukocytes and thrombocytes.

Cite the “RBC Indices” and explain their purpose. Cite the purpose of an anticoagulant and define

the anticoagulant used in hematology studies. Define the different types of samples

used for hematology studies.

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WHYstudyHematology?

September 9, 2003

- To communicate more effectively with the end user of our instrumentation

- To gain an understanding of the material being tested on our equipment

- To work safely and efficiently with blood

- To understand troubleshooting on a system from your customer’s point of view

- To establish relevance for this type of testing and why it is necessary for good patient care

Why study Hematology….

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What is Blood?

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What is BLOOD? The fluid that circulates

through the heart, lungs, arteries, veins and capillaries.

Carries oxygen and nutrients to the tissues

Carries away carbon dioxide and wasted products produced by the tissues

Veins shown in blue Arteries shown in red

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What is BLOOD? A diagnostic tool for the clinician

to assess patient status Since blood is exposed to virtually

all tissues in the body, it may be considered the “barometer” of the condition of the body.

In normal, healthy individuals the blood contains a normal number of blood cells.

If, however, there is an abnormal condition / disease process, the blood typically reflects a change from normal by either raising or lowering the number of blood cells and their relative proportions.

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Composition of Blood Blood is comprised of

approximately 55% plasma and 45% cellular component.PLASMA 55%

Water 90%NutrientsClotting ProteinsAntibodiesHormonesSaltsWastes

CELLS 45%Red Blood CellsWhite Blood CellsPlatelets

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Who Studies What About Blood?

CHEMISTRY department

COAGULATION department

FLOW CYTOMETRY department

HEMATOLOGY department

Red Blood Cells White Blood CellsPlatelets

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Normally for every white blood cell, there are 1000 red blood cells and 20 platelets

The Blood

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Basic Cell Structure

Cell Membrane

CytoplasmNucleus

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Blood Cells Seen Through Microscope

wb:white blood cells

rb:red blood cells

p:platelets

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Development of Blood Cells

The Johns Hopkins Human Anatomy

Series

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Where Blood Cells Are Produced

Embryo / Infants up to 4 months:

Blood Islands (Yolk Sac)

Liver Spleen

Children (>4 months and Adults)

Bone Marrow

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Development of Blood CellsGeneral Observations

•Nucleus of immature cell is round or oval•As the cell matures, the nucleus decreases in relative size and takes on various shapes•Some cells loose their nucleus entirely•The nucleus is the initiator of cellular division•Cells are produced, mature and take on the function and appearance of a mature cell in the bone marrow•As cells mature, they usually become smaller in size•As cells mature, they are released into the “peripheral blood”•An immature cell is NOT capable of carrying out the specific functions of a mature cell, therefore, the presence of immature cells in the peripheral blood typically indicates a problem

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Red Blood Cells

September 9, 2003

Red Blood Cells Are also referred to as erythrocytes or RBCs Their biconcave disk shape allows for more

surface area (efficient oxygen transfer) Their flexibility (ability to deform) allows

their passage through small capillaries The quantity of red blood cells is expressed

as the quantity of rbc’s or as a ratio of red blood cells to whole blood

The quantity of red blood cells is expressed as the red blood cell count (cells/microliter)

The ratio of red blood cells to the volume of whole blood is defined as the hematocrit

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Red Blood Cells (RBCs)

Erythrocyte Biconcave Disc

Shape Ability to deform Size: 7.5 - 9.5

micron Volume: 90 fl

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Red Cell Count (RBC)

DefinitionNumber of red cells in one cubic millimeter of whole blood

Adult Males: 5.4±0.7 million/cu mm

Adult Females: 4.8±0.6 million/cu mm

Significance of Abnormal ValuesBelow normal: Anemia

blood loss hemolysis deficiency of iron,

vitamins B6, B12 Expanded plasma

volume (e.g. pregnancy)

Above normal: Polycythemia

Severe dehydration (e.g. burns)

Polycythemia vera

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Manual RBC

Calculation of RBC

Cells actually counted: 500

Dilution: 1:200

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Manual RBC

Tedious procedure

Technique dependent

Accuracy?

Precision?

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Hematocrit (Hct)

DefinitionRelative volume of packed red cells in a given volume of whole blood.

Normal ValuesAdult males: 47±5%Adult females: 42±5%

Significance of Abnormal ValuesDecreased: AnemiaIncreased: Severe dehydration , Polycythemia vera

PLASMA

WHITE CELLS AND PLATELETS

(BUFFY COAT)

PACKED RED CELLS

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Hematocrit

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Hematocrit

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Cell & Plasma separation Left: Healthy

person Right: Person with

Iron Deficiency Anemia

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Hematocrit

Major consideration:

Trapped plasma

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What within the Red Blood Cell carries the oxygen?

HEMOGLOBIN

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HEMOGLOBIN = “BLOOD PROTEIN”

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Hemoglobin (Hgb) Concentration Function:

Transport of oxygen Structure

Main component of mature red cell (=1/3)

DefinitionGrams of Hemoglobin in 100 ml of whole blood

Normal ValuesAdult males: 16 ± 2 grams/100ml

Significance of Abnormal ValuesBelow Normal: AnemiaAbove Normal: Severe dehydration Polycythemia vera

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AnemiaIs caused by

• too few RBCs

• too little hemoglobin

• or any impairment in the ability of the hemoglobin or RBC to effectively provide oxygen to the tissues

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Hemoglobin Concentration

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Hemoglobin Concentration

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Red Cell IndicesMEAN CORPUSCULAR VOLUME

(MCV)

Normal values: Male 87 +/- 7cu micron (3)* Female 90 +/- 9 cu microns (3)

Microcytic = Smaller than normal

Normocytic = Normal

Macrocytic = Larger than normal

(fL)s femtolitermillions) (inRBC

10*HctMCV

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Normocytic Red Blood Cells wb:

white blood cells

rb:red blood cells

p:platelets

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Microcytic Red Blood Cells

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Macrocytic Red Blood Cells

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Red Cell Indices Continued

MEAN CORPUSCULAR HEMOGLOBIN (MCH)

Normal value: 29±2 micromicrograms (g): Normochromic

Less than Normal: Hypochromic

g)(grams micromicromillions) RBC(in

10*HgbMCH

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Normochromic Red Blood Cells

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Hypochromic Red Blood Cells

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Red Cell Indices Cont’d

MEAN CORPUSCULAR HEMOGLOBIN CONCENTRATION (MCHC)

Normal value: 34 ± 2%Hypochromic = Less Hgb than normal Normochromic = Normal

(percent) Hct100*HgbMCHC

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Normohromic, Normocytic Red Cells

wb:white blood cells

rb:red blood cells

p:platelets

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Macrocytic Hypochromic Red Cells

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Microcytic Red Cells

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Reticulocytes

Non -nucleated immature red blood cells

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Measuring Reticulocytes

Manual method: Stain cells with a “supravital” dye such as New Methylene Blue, make a slide, count 1000 red blood cells while enumerating the number of reticulocytes…..

Automated method: VCS technology!

September 9, 2003

Reticulocytes

Measuring reticulocytes is an effective means of assessing bone marrow function. If the tissues need more oxygen, the signal is given to the bone marrow to increase the production of red blood cells...

September 9, 2003

Reticulocytes

In a state of hypoxia, the bone marrow will release an increased number of reticulocytes.

Measuring reticulocytes assesses effective or ineffective hematopoeisis.

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The Blood

Platelets

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Platelets (PLT)

Thrombocytes Cellular

Fragments Function in

Hemostasis Primary

Hemostatic Plug

Size:2-4 microns

Volume:2-20 fL

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Platelets and Hemostasis

September 9, 2003

PlateletsSignificance of Abnormal Values Below normal:

(thrombocytopenia)some anemias, acute leukemia certain drugs and radiation drug idiosyncrasy“idiopathic” purpura

Above normal: (thrombocythemia)acute blood loss or surgerysome anemias iron deficiency) chronic leukemia metastatic carcinoma polycythemia vera

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Normal Platelets

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Example of Abnormal Platelets

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The Blood

White Blood Cells

September 9, 2003

White Blood Cells (WBCs) Leukocytes Function

Defend the body from infection

Differentiated into 5 types

Mononuclear-lymphocytes and monocytes

Polymorphonuclear- neutrophils, eosinpohils and basophils

Each plays individual roles

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Mononuclear WBCs: Monocytes Lymphocytes

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Polymorphonuclear WBCs: Neutrophils Eosinophils

Basophils

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White Cell Count (WBC)Significance of Abnormal Values Lower than normal:

viral infectionsendocrine disorderstreatment with certain drugsradiation

Higher than normal:bacterial infectionsacute inflammatory conditionsneoplastic disease (e.g. leukemia)

Differential White Cell Count Definition: Relative proportion of different types of leukocytes

September 9, 2003

Manual WBC

Calculation of WBC

Cells actually counted: 100

Dilution: 1:20

Volume Counted: 0.4mm3

WBC=100 X 20 = 5000/mm3(5.0 x 103) 0.4

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WBC Differential Smear of Whole Blood

Stained with Wright’s Stain

100 Cells Counted

Classified into Cell Types Morphologically

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WBC Differential continued

WBC Differential

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Relative Numbers Expected

Neutrophils 50% to 75% Lymphocytes 20% to 45% Monocytes 3% to 11% Eosinophils 1% to 3% Basophils 0% to 1%

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WBC: Absolute Versus Relative For example, if the laboratory’s WBC normal range lies

between 5,000 and 10,000 and the lymphocyte range is between 20% and 40% of the total count, then the normal absolute lymphocyte count should be between 1,000 and 4,000/L.

Patient A Patient BLymphocyte % 90% 90%Lymphocyte # 13,500 3,600WBC / L 15,000 4,000Lymph% x total WBC = absolute number of lymphs

Patient A and B both show a relative lymphocytosis of 90%. However, only patient A has an absolute lymphocytosis with a lymphocyte count of 13,500/L. Patient B, on the other hand, has a normal absolute lymphocyte count of 3,600/L.

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Laboratory Measurement of Blood

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Specimen:

EDTA

run within 24 hour from collection

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Tube / Anticoagulant Summary

September 9, 2003

In summary, Hematology is the study of blood. We study hematology to communicate and work with our

instrumentation more effectively. Because blood is exposed to virtually all of the tissues in the

body, it becomes a “barometer” of the condition of the body. Under normal conditions, there are a normal number and relative proportions of circulating blood cells. In abnormal conditions, the numbers and relative proportions are altered.

Our instruments provide the clinician with information on the patient’s status based on the numbers and relative proportions of circulating cells.

Our systems are an integral part ofproviding good patient care.

September 9, 2003

Any questions?

September 9, 2003

Acknowledgements:

Washington State University

Johns Hopkins Printing

Beckman Coulter Miami Education Center staff

September 9, 2003

The End!