Basic Hematology Technical

Sales

Training

September 9, 2003

Our goal . . . To heighten your awareness of

Basic hematology
blood cell types associated tests manual methods

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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 September 9, 2003 used for hematology studies.

Hematology?
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WHY study

Why study Hematology.

- 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 customers point of view

- To establish relevance for this type of testing and why it is necessary for good patient care
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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% Nutrients Clotting Proteins Antibodies Hormones Salts Wastes CELLS 45% Red Blood Cells White Blood Cells Platelets

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Who Studies What About Blood?

CHEMISTRY department COAGULATION department

FLOW CYTOMETRY department

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HEMATOLOGY department Red Blood Cells White Blood Cells Platelets

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

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Basic Cell Structure

Cytoplasm Nucleus

Cell Membrane

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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 Cells General 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

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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 rbcs 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)

Significance of Abnormal Values Below normal: Anemia

blood loss hemolysis deficiency of iron, vitamins B6, B12 Expanded plasma volume (e.g. pregnancy)

Definition Number of red cells in one cubic millimeter of whole blood Adult Males: 5.40.7 million/cu mm Adult Females: September 9, 2003 4.80.6 million/cu mm

Above normal: Polycythemia

Severe dehydration (e.g. burns) Polycythemia vera

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)

Definition
Relative volume of packed red cells in a given volume of whole blood.

PLASMA

Normal Values
Adult males: 475% Adult females: 425%

WHITE CELLS AND PLATELETS (BUFFY COAT)

Significance of Abnormal Values

Decreased: Anemia Increased: Severe dehydration , Polycythemia vera

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) Definition Grams of Hemoglobin in 100 ml of whole blood Normal Values Adult males: 16 2 grams/100ml Significance of Abnormal Values Below Normal: Anemia Above Normal: Severe dehydration Polycythemia vera

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Anemia
Is 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 Indices

MEAN CORPUSCULAR VOLUME (MCV)
Hct * 10 MCV femtoliter s (fL) RBC (in millions)
Normal values: Male 87 +/- 7cu micron (m3)* Female 90 +/- 9 cu microns (m3)
Microcytic = Smaller than normal Normocytic = Normal
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Macrocytic = Larger than normal

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)
MCH Hgb * 10 micromicro grams ( mmg) RBC(in millions)

Normal value: 292 micromicrograms (mmg): Normochromic

Less than Normal: Hypochromic

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Normochromic Red Blood Cells

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Hypochromic Red Blood Cells

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Red Cell Indices Contd

MEAN CORPUSCULAR HEMOGLOBIN CONCENTRATION (MCHC)
Hgb * 100 MCHC Hct (percent)

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

Normochromic = Normal

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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!
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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...

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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

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Platelets
Significance 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 surgery some 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

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White Blood Cells (WBCs)

Leukocytes Function Defend the body from infection Differentiated into 5 types Mononuclearlymphocytes and monocytes Polymorphonuclearneutrophils, 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 infections endocrine disorders treatment with certain drugs radiation Higher than normal: bacterial infections acute inflammatory conditions neoplastic disease (e.g. leukemia)

Differential White Cell Count Definition:

Relative proportion of different types of leukocytes

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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 Wrights Stain 100 Cells Counted

Classified into Cell Types Morphologically
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WBC Differential continued

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WBC Differential

Relative Numbers Expected

Neutrophils Monocytes Eosinophils Basophils

50% to 75% 3% to 11% 1% to 3% 0% to 1%

Lymphocytes 20% to 45%

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WBC: Absolute Versus Relative

For example, if the laboratorys 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/mL. Patient A 90% 13,500 15,000 Patient B 90% 3,600 4,000

Lymphocyte % Lymphocyte # WBC / mL

Lymph% 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/mL. Patient B, on the other hand, has a normal absolute lymphocyte count of 3,600/mL.

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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

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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 patients status based on the numbers and relative proportions of circulating cells. Our systems are an integral part of September 9, 2003 providing good patient care.

Any questions?

September 9, 2003

Acknowledgements:

Washington State University

Johns Hopkins Printing

Beckman Coulter Miami Education Center staff

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The End!

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