Conceptos de Hematologia Basica

HEMATOLOGY FUNDAMENTALS SERVICE TRAINING COURSE
BASIC HEMATOLOGY
BASIC HEMATOLOGY
OBJECTIVE
Given the Basic Hematology course materials:
Training Guide
The books titled Hematology: Principles and Procedures by
Barbara A. Brown and BLOOD by Dennis W. Ross
The Johns Hopkins Development of Blood Cells cell maturation
chart.
Read, study, review, and be able to answer the Hematology Basic

Knowledge Check questions. The knowledge check will be
considered completed when the student can achieve a minimum
overall score of 90%.
KNOWLEDGE CHECK
You will be given a Knowledge Check and asked to answer the

questions using all the course materials provided.
MODULE RESOURCES
To Complete this module you will need:
Books:
Hematology Fundamentals Service Training Course Training
Guide
HEMATOLOGY: Principles and Procedures by Barbara A.
Brown
BLOOD by Dennis W. Ross
Johns Hopkins Development of Blood Cells cell maturation chart
Hematology Fundamentals Service Training Course 1

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What to Do
Read and study all the course materials provided. When you feel
confident that you know the material, ask your Instructor for the
Knowledge Check. Complete the Knowledge Check and have your
Instructor check it and review any mistakes with you.
Although it is recommended that you read and study all the course
materials, if at any time you feel confident you know the material
well enough to complete the Knowledge Check, feel free to do so.
2 Hematology Fundamentals Service Training Course

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HEMATOLOGY FUNDAMENTALS SERVICE TRAINING COURSE
BASIC HEMATOLOGY
BASIC HEMATOLOGY
Introduction
This session is intended to provide you with a basic introduction to

Hematology. Cell structure, cell maturation, methods for measuring
components of blood samples and common terms used in
hematology will be presented. How Beckman Coulter
instrumentation is used in the hematology setting will also be
addressed.
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 customer's
point of view
• To establish the relevance for this type of testing and why it is
necessary for good patient care
What is Blood?
• The fluid that circulates through the heart, lungs, arteries, veins
and capillaries carrying oxygen and nourishment to the tissues
and carrying away carbon dioxide and waste products produced
by the tissues.
• A diagnostic tool for the clinician to assess patient status.
Since blood is exposed to virtually all tissues in the body, it
becomes 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 or disease process, the
blood typically reflects a change from normal by either

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raising or lowering the number of blood cells and their

relative proportions.
• It important to study blood to assist in screening or identifying a
normal versus an abnormal condition of the body.
Composition of Blood
• Blood is comprised of approximately 55% plasma and 45%
cellular components.
Water 90%
Nutrients
PLASMA 55% Clotting Protiens
Antibodies
Hormones
Salts
Wastes
Red Blood Cells

CELLS 45% White Blood Cells
Platelets
Figure 1 Composition of Blood
• Plasma is comprised of 90% water and 10% salts, nutrients,

hormones, antibodies, clotting proteins and waste products.
• The cellular components are red blood cells, white blood cells
and platelets.
Who Studies What About Blood?

• Chemistry Department:
• Coagulation Department:
• Flow Cytometry Department:
• Hematology Department:

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The three cellular components are red blood cells, white

blood cells and platelets.
Plasma and the three cellular components comprise the
four major components of blood.
Normally for every white blood cell, there are 1000 red
blood cells and 20 platelets.
Figure 2 Formed Elements of Blood

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Basic Cell Structure

• Any cell consists of 3 basic parts:
Figure 3 Basic Cell Structure
Cell Membrane
• A semipermeable separation between the various internal cellular

components, the organelles, and the surrounding environment.
• Antigens are bound to the surface of cell membranes.
• Maintains the cellular integrity of the interior of the cell by
controlling the passage of materials in and out of the cell.
Cytoplasm
• Contained within the cell membrane.

• Composed of a variety of small cellular structures called
organelles which are the functional units of the cell.
Smaller organelles can only be seen with an electron
microscope.
Larger organelles can be seen on stained blood preparations
using a light microscope. These larger organelles give the

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cytoplasm a characteristic "look" that can help identify the

cell.
Nucleus
• Functions as the control center of the cell and is essential for

long-term survival.
• Controls the cell division process to produce an exact replicate of
the cell.
Many cells retain the ability to divide and replicate
themselves throughout the life of the cell; for example, skin,
gut, lining of the mouth, etc.
As a blood cell matures, the nucleus either decreases in
relative size or is extruded (pushed out of the cell) and, as a
result, the blood cell loses its ability to divide and replicate.
Lymphocytes are the only blood cells that continue to
replicate themselves even after maturity.
• In hematology, identification of a cell and its maturity is typically
based on:
Cell Size
Nuclear appearance
Granularity
Blood Cell Maturation Characteristics
Overall Cell Size
• Overall cell size is usually compared with the size of a mature

red cell.
• As a cell matures, it usually becomes smaller in size.
Nuclear to Cytoplasmic Ratio
• N:C or the amount of space occupied by the nucleus in

relationship to the space occupied by the cytoplasm.
• Nucleus of an immature cell tends to be round or oval and is very
large in proportion to the rest of the cell.
• As the cell matures, the nucleus decreases in relative size and
may take on various shapes.

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Some cells loose their nucleus entirely. Mature red blood

cells and platelets are the two cell types in the circulating
blood that do not have a nucleus.
Nuclear Characteristics
• Chromatin pattern, nuclear shape and the presence or absence of

nucleoli are important nuclear features that can aid in the
identification of cell type.
Chromatin
Physical basis of heredity.

Loose and fine in most immature cells.
Clumped with a dark appearance in more mature cells.
Nucleoli
Found only in immature cells.
Shape
Either round or oval in young cells.

In cells that retain their nucleus as they mature, nuclear
shapes become very distinctive for particular cell types.
Where Blood Cells are Produced
Refer to the Development of Blood Cells chart
• In an adult, blood cells are formed in the bone marrow.

• The nucleus of the immature cell is typically round or oval and is
very large in proportion to the rest of the cell.
• As the cell matures, the nucleus decreases in relative size and
may take on various shapes.
• Some cells loose their nucleus entirely.
• The nucleus is the initiator of cell division.
Notice that the nucleus of a mature blood cell is very coarse
and condensed.
This is because the nucleus of most mature blood cells is no
longer functioning in cell division.

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• Cells are produced, mature and take on the appearance and

function of a mature cell in the bone marrow.
• As cells mature, they usually become smaller in size.
• As cells mature in structure and function, they are released into
the blood circulating through the veins and arteries.
This circulating blood is commonly referred to as the
peripheral blood.
• An immature cell is not capable of carrying out the specific
functions of a mature cell.
Because mature cells have a specific role and function when
circulating in the peripheral blood, only mature cells should
be circulating in the peripheral blood.
The presence of immature cells in the peripheral blood
typically indicates a problem.
Red Blood Cells

• Red blood cells are also referred to as erythrocytes or RBCs.
• The nucleus is completely extruded from the cell by maturity.
Mature RBCs have no nucleus or remnants of one.
• Their biconcave disk shape allows for more surface area,
therefore more oxygen carrying capacity.
Figure 4 Red Blood Cell
• Their flexibility (or ability to deform) also allows the RBCs to

squeeze through small capillaries to the tissues.

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• The quantity of RBCs may be expressed as a red blood cell count

or as a ratio of the volume of red cells to the volume of whole
blood.
The quantity of red cells is expressed as an RBC count.
The ratio of the volume of red cells to volume of whole blood
is defined as the hematocrit (Hct).
Figure 5 Hematocrit
• The Reference Range (also referred to as "Normal Range")

for an RBC count for males is 4.7 to 6.1 million cells per µL
(microliter) of whole blood.
• The Reference Range for an RBC count for females is 4.2 to
5.4 million cells per µL of whole blood.
• The Reference Range for hematocrit (Hct) for males is 42%
to 52%.
• The Reference Range for hematocrit (Hct) for females is 37%
to 47%.
Hemoglobin
• The portion of the RBC that transports oxygen from the lungs to
the tissues and carbon dioxide from the tissues to the lungs, is a
protein called hemoglobin.
(hemo = blood ; globin = protein)

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• The red cell membrane serves as a retaining barrier for the

hemoglobin molecule but is also permeable to permit oxygen and
carbon dioxide to pass freely.
• Normally, the Hemoglobin (Hgb) value is approximately
three times the RBC count ( RBC x 3 = Hgb ).
• The Reference Range for Hgb for a male is 14-18 g/dL (grams
per deciliter).
• The Reference Range for Hgb for a female is 12-16 g/dL.
• Normally, the Hemoglobin (Hgb) value times three equals the
Hematocrit (Hct) approximately ( Hgb x 3 = Hct ).
RBC Indices
• In Hematology, indices (pronounced: n˜di ss) refers to calculated
values used for describing red cell properties.
• There are three RBC Indices MCV, MCH, MCHC
• The indices remain constant in a stable patient population,
therefore, the laboratory can use the indices to monitor
instrument performance. This forms the basis of XB Analysis.
MCV
• Mean Corpuscular Volume (MCV), which describes the

volume of the average red cell in a given sample of blood, is one
of the RBC indices.
Hct
• The calculation for MCV is ---------- x 10
RBC
• The Reference Range for MCV for males is 87 + 7 fL
(femtoliters); for females 90 + 9 fL.
• Microcytic (micro = small ; cytic = cell) red cells decrease the
MCV result while macrocytic (macro = large ; cytic = cell) red
cells increase the MCV result.
If an MCV for a male or female is 120 fL, the red cell
population is considered macrocytic; however, an MCV of 98
fL is considered macrocytic only for a male.
• A variation in the sizes of red cells is called anisocytosis
(an = without ; iso = same ; cyto = cell ; osis = condition of).
In other words, a condition of "unlike" cells.

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ACTIVITY
Consider the effect of the diluent on the size of the RBC.
1. If the RBC size is responsive to its supporting media, what

would be the outcome if red cells are suspended in water?
2. What would be the outcome if red cells are suspended in a

highly concentrated salt solution?
3. What do you think the saline concentration in Coulter's diluting

fluid (ISOTON(r) III) is?
ACTIVITY ANSWER KEY

1. In effort to equilibrate the internal and external environment, the
RBCs tend to take in water from the suspending media, swell
and burst.
2. The opposite effect would be observed. The "water" from inside

the cell will pass through the membrane into the suspending
media thereby shrinking the RBC. In this case, the RBC volume
(MCV) would be decreased.
3. COULTER ISOTON III diluent has been formulated to best

mimic plasma to keep cells in their near-native state.
MCH
• Mean Corpuscular Hemoglobin (MCH) describes the average

weight of hemoglobin in the red blood cell.
Hgb
• The calculation for MCH is ----------- x 10
RBC

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• MCH is directly proportional to the size of the red blood cell and
the concentration of hemoglobin in the cell.
• The Reference Range for MCH is 27 to 31 pg
(picograms or µµg / micro-micrograms).
MCHC
• Mean Corpuscular Hemoglobin Concentration (MCHC)

describes the average concentration of hemoglobin in the red
blood cells.
• MCHC gives the ratio of the weight of the hemoglobin to the
volume of the red blood cell expressed in a percentage.
• MCHC is expressed as an average concentration by dividing the
hemoglobin value by the hematocrit and multiplying by 100.
Hgb
• The calculation for MCHC is---------- x 100
Hct
• The Reference Range for MCHC for males and females is 33
to 37%.
• One of the best parameters for indicating either instrument
malfunction or an improper or unusual specimen.
Calculation is dependent on all measured RBC parameters
[Hgb and Hct (which is based on the RBC and MCV)].
Any problem with the Hgb, RBC or MCV is often reflected
by an abnormal MCHC.
Specimens with MCHCs outside the normal range should
therefore be suspected.
An MCHC above 38% should never occur.
An MCHC should never fall below 22% even when
hypochromia is present.
If the frequency of either high or low MCHCs increases, an
instrument problem is most likely present.
RBC Evaluation
• The hematology technologist evaluates red cells by their form,
shape and appearance as observed through a microscope.

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• Morpho is a prefix that means form, shape and appearance; logy

is a suffix meaning the study of. Therefore, the study of red cell
size, shape and appearance is red cell morphology.
• The presence of a variety of shapes of red cells is called
poikilocytosis.
ACTIVITY
1. View Kodachromes of the normal and abnormal RBCs and
classify as Normochromic, Hypochromic, Normocytic,
Macrocytic, Microcytic, Anisocytosis or Poikilocytosis.
2. Describe what the following terms indicate about the RBC:
Normochromic ______________________________________
Hypochromic _______________________________________
Normocytic ________________________________________
Macrocytic _________________________________________
Microcytic _________________________________________
Anisocytosis _______________________________________
Poikilocytosis_______________________________________
ACTIVITY ANSWER KEY
Normochromic RBCs having normal color / MCH normal

Hypochromic RBCs having less than normal color / MCH
decreased
Normocytic RBCs having a normal size / MCV normal
Macrocytic RBCs larger than normal / MCV increased

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Microcytic RBCs smaller than normal / MCV decreased

Anisocytosis RBCs varying in size
Poikilocytosis RBCs varying in shape
LECTURE / DISCUSSION
Reticulocytes
• A reticulocyte is the last stage before a red cell is considered
mature.
• A reticulocyte, when stained with Wright Stain, has a bluish cast
to it and is slightly larger than a normal mature RBC.
Reticulocytes have residual RNA, which when stained with a
supravital stain (a stain that stains the cells while they are still
living) becomes visually evident when viewed
microscopically.
New Methylene Blue is an example of a commonly used
supravital stain.
• A normal reticulocyte count is considered to be
approximately 0.7 - 2.8% of a normal RBC count.
Platelets
• Platelets may also be referred to as Thrombocytes or Plts

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Figure 6 Formation of Platelets
• The normal life span of a platelet is 10 days + 1 day
Refer to the Development of Blood Cells chart. Locate the

Megakaryoblast on the far right and top of the page. Notice as the
cell matures, small pieces of the cytoplasm "bud" off (platelets).
• The clotting process is an extremely complex system.

• Coagulation instruments can test for various components in this
system. It is important to be aware that platelets play a
significant role in the clotting process.
• The Reference Range for a Platelet count is about 150,000 -
400,000 cells/µL.
• Since platelets are consumed in the clotting process, clotted
blood specimens or poorly collected fingerstick samples have a
low platelet count. It is unacceptable to use clotted blood samples
for hematology studies.
• Platelets appear in the peripheral blood as small, disc-shaped
cellular fragments about 2 to 4 microns in diameter. RBCs are
larger than platelets.

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White Blood Cells

• White Blood Cells are also referred to as Leukocytes or WBCs
• The normal life span of a WBC is 11 days in the bone marrow to
mature then hours to years in the tissues.
• The Reference Range for a WBC count is 4,000 to 10,000
cells/µL.
• Leukocytes can be classified or differentiated in a number of
ways. The most common is morphologically (form, shape and
appearance when looked at through a microscope).

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Types of WBCs
• Mononuclear
Lymphocytes
Monocytes
• Polymorphonuclear
Granulocytes
- Neutrophils
- Eosinophils
- Basophils
Figure 7 White Blood Cells

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ACTIVITY
• Refer to the Development of Blood Cell chart.

Observe the different coloration of Neutrophils compared to
Basophils compared to Eosinophils.
Observe the granulocytes from the immature stem cell to the
more mature cell.
Notice the nucleus change from a large round nucleus in the
immature cells to a "multi-lobed" nucleus in the mature cell.
Mononuclear Leukocytes
Lymphocytes
• Lymphocytes are produced in the bone marrow and migrate to

the lymph tissues (lymph glands) where they become involved
with the body's immune function.
• These cells can be subclassified by use of "cell surface markers"
as T cells, B cells or Natural Killer (NK) cells.
• Classifying lymphocytes is very important in the diagnosis and
management of AIDS patients.
• Determining cell surface markers can be described as
"fingerprinting" the lymphocytes. This type of testing uses
antibodies manufactured against certain antigens on the
lymphocyte cell membrane (see Figure 3).
The antigens, in this case, are proteins attached to the
lymphocyte cell membrane.
Antibodies are special proteins that attach to only one
specific type of antigen . . . like a lock and key).
Certain kinds of lymphocytes have specific kinds of antigens,
so if we use a known type of antibody and it attaches to the
antigen on the cell, we can identify the type of lymphocyte it
is!
A flow cytometry instrument can perform this special type
of lymphocyte subclassification.

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Monocytes
• Monocytes are produced in bone marrow but they migrate to

body tissues to serve as scavenger cells that "eat" foreign
particles and digest them.
Polymorphonuclear Leukocytes
Granulocytes
• Granulocytes are polymorphonuclear cells meaning the nucleus

has more than one lobe.
• Described by the granules in their cytoplasm and their staining
characteristics when stained with Wrights/Giemsa stain.
Neutrophils - granules stain "neutrally" and appears as a
slight pink color in the cytoplasm
Eosinophils - granules stain a reddish / orange color
Basophils - granules stain dark bluish-black
• Granulocyte functions differ in relation to their granule content.
Neutrophils - granules contain digestive enzymes to destroy
bacteria
Eosinophils - granules contain histamine and enzymes and
are involved in the latter stages of inflammation, allergic
reactions and parasitic reactions
Basophils - granules contain histamine, heparin and heparin
like substances; although the function of basophils has been
debated, they do seem to be associated with allergic reactions
WBC Differential Analysis

• WBC differential analysis, which is one of the fundamental
analyses of hematology, is a diagnostic tool that can act as a
pointer to aid the physician in the diagnosis and/or monitoring of
a multitude of disease states.
• In the differential blood count procedure, leukocytes are
identified and manually enumerated from their morphological
appearance on stained blood smears or by automated instruments
that use a combination of cellular characteristics and
histochemical reactions.

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Relative Numbers (%)
• Differential results are generally reported in percentages which

means the enumeration of a particular type of white blood cell is
actually relative to all the other cell types that are present.
• In other words, a lymphocyte percentage of 40% means that in
the total number of white blood cells counted (100%), 40% of
those cells were lymphocytes while the other 60% of the cells
were some other type of white cell.
• Relative numbers expected in normal circulating blood:
Neutrophils 50% to 75%
Lymphocytes 20% to 45%
Monocytes 3% to 11%
Eosinophils 1% to 3%
Basophils 0% to 1%
Absolute Numbers (#)
• Another way to report differential results.

• Individual absolute numbers are computed based on the total
WBC count and the individual differential percentage (%).
• The availability of absolute numbers helps resolve the difficulties
sometimes posed by the use of percentages alone.
• 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/µL.
Patient A Patient B
Lymphocyte % 90% 90%
Lymphocyte # 13,500 3,600
WBC / µL 15,000 4,000
Lymph% x total WBC = absolute number of lymphs

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• 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/µL. Patient B, on the other hand, has
a normal absolute lymphocyte count of 3,600/µL.
• In other words, the patient may have a relative increase in
lymphocytes not because they are producing more but because
they lack other cells types such as neutrophils. On the other hand,
an absolute increase in lymphocytes means there is an actual
increase in the production of lymphocytes.
Terminology
• The suffix penia means a severe decrease; therefore, leukopenia
indicates a decrease in the number of WBCs.
• The suffix cytosis means an increase of; therefore, leukocytosis
indicates an increase in the number of WBCs.
• Granulocytosis indicates there is an increase in granulocytes;
lymphocytosis, an increase in lymphocytes.
• Lymphopenia indicates a decrease in lymphocytes;
granulopenia, a decrease in granulocytes.
• Because of the relative relationship among the white cells,
granulopenia is generally accompanied by lymphocytosis while
granulocytosis is generally accompanied by a lymphopenia.
Laboratory Measurement of Blood
Specimen Collection
• A Technologist or Technician uses a peripheral blood specimen

to study the blood cells in vitro (outside the body).
Specimens are typically collected by a phlebotomist who is
specifically trained in blood collection.
The phlebotomist may obtain the specimen from a vein
(venipuncture) or from capillaries (skin puncture of the
finger, heel or ear lobe).
A venipuncture collected in an evacuated collection tube is
the preferred specimen.
• The normal inclination of blood is to clot once released from the
natural environment of the body . . . this is nature's way of
protecting us from bleeding.

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• If a patient has blood collected into an empty tube, the platelets

and special clotting proteins cause the sample to clot.
All cell types become enmeshed in the clot.
The straw colored fluid that is expressed from the clot is
called serum.
Serum is typically used in chemistry studies in the laboratory.
• If blood is drawn into a tube containing a special chemical
additive to prevent clotting, the blood remains in a fluid state.
This chemical additive is called anticoagulant (to prevent
coagulation or clotting).
If anticoagulated is allowed to sit without mixing for several
hours , the clear fluid that appears in the upper portion of the
tube is the plasma.
Anticoagulants
• Anticoagulants may be dry-powder form or liquid.

• There are a variety of anticoagulants used and can be identified
by the color of the tube stopper.
• In hematology, the anticoagulant of choice is K3EDTA
(ethylenediamine tetra acetic acid) and is found in a purple or
lavender stoppered tube.
• Calcium is a required component to the clotting process. EDTA
has the ability to bind up calcium in the blood. This process of
binding up calcium is commonly referred to as "chelating
calcium." The bottom line . . . no calcium, no clotting!
• Most anticoagulants (Citrate, Oxalates) work by the same
mechanism as EDTA (binding calcium to prevent clotting.)
• Heparin acts as an antagonist to the normal clotting reactions of
the clotting cascade.
• The choice of anticoagulant in Hematology is determined by
what mixes quickly and efficiently with the blood and also what
maintains the cells in their near native state.
• Beckman Coulter recommends K3EDTA as the optimal
anticoagulant because all testing and studies done on our
instrumentation was performed using K3EDTA samples.
• K3EDTA is cited in our instrument manuals as the preferred
anticoagulant; however, the major manufacturers of blood
specimen tubes are in the process of converting from glass to

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plastic tubes containing the anticoagulant K2EDTA. Therefore, in

the near future, testing and claims made by Beckman Coulter
Inc. will cite K2EDTA as the preferred anticoagulant.
ACTIVITY
• Observe the variety of tubes available. Notice the
presence/absence/volume of anticoagulant associated with the
color stoppered tube (VACUTAINER and HEMOGARD by
Becton-Dickinson as well as VENOJECT by Terumo).
• Observe the corresponding blood filled tubes (2 of each; mix
only one). Associate the effects of the presence/absence of
anticoagulant and observe the difference between serum and
plasma.
TUBE / ANTICOAGULANT SUMMARY
Department Where
Tube Color Stopper Anticoagulant
Commonly Using
Chemistry
Red None
Blood Bank
Hematology
Lavender EDTA (Na2 , K2 , K3)
Flow Cytometry
Blue Sodium Citrate Coagulation
Green Heparin Chemistry
ACD (Acid Citrate Flow Cytometry

Yellow
Dextrose) Blood Bank
Potassium Oxalate-
Gray Chemistry
Sodium Fluoride
Specimen Collection Devices

• To increase safety for laboratory personnel, manufacturers are
producing stoppers different from the traditional rubber stopper
in an effort to minimize aerosolization and splattering of blood
when removing the stopper. (HEMOGARD and Terumo tubes
are examples.)

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• For children, babies and difficult patients, microcollection

devices allow the finger, heel or earlobe to be pricked and blood
collected manually.
Be aware of the potential for clotting in this type of specimen.
Specimen Collection Precautions

• Blood tubes must be properly filled to assure the appropriate
blood to anticoagulant ratio is maintained.
• When blood is drawn with a syringe, it must be transferred to a
blood specimen tube. When this transfer occurs, it is very easy to
either overfill or underfill the specimen tube.
Effects of Overfilling a Specimen Tube
• Overfilling overcomes the function of the additive /

anticoagulant.
• Overfilled tubes are not properly anticoagulated and, as a result,
microclots can form.
Any clotting in a specimen tube makes it an unacceptable

sample for hematology and analysis on any Beckman Coulter
Hematology Instrument. The specimen must be redrawn.
Effects of Underfilling a Specimen Tube
• Underfilled specimen tubes have an excess of anticoagulant.

• Excess anticoagulant may cause cells to crenate and shrink.
Sample Handling
• Varies depending on the type of tube and the testing plans for the
sample.
• Department requirements / storage requirements:
Chemistry
Coagulation

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Flow Cytometry
Hematology
- Mixing Requirements
• Equilibration cells go through in response to the anticoagulated
environment. ( shrink / swell )
• Key Points
Laboratory results from any instrument are only as good
as the specimen they come from.
Never overlook the integrity of the sample(s) being run
through the system.
CBC, the Complete Blood Count

• One of the most frequently requested laboratory tests.
• Patient results are considered against a "Normal Range."
Normal Range is established in the laboratory by taking a
sampling of population to determine the average number and
quantity of each CBC parameter.
Once the average is established, a range is mathematically
determined by calculating a 2SD (Standard Deviation) from
the mean of the collected data.
On printed tickets from Coulter (and in any Hematology
text), normal ranges are given but are generic.
• CBC typically includes:
• A WBC differential includes:
• Clinical utility of a CBC:

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Methods of Performing a CBC

• Manual, semi-automated and automated methods are available
for performing a CBC.
Manual Methods
• Locate the procedures for manual blood cell counts starting on

page 89 of your textbook, HEMATOLOGY: Principles and
Procedures by Barbara Brown.
Hemoglobin Measurement
• When a specified volume of red blood cells is exposed to a

chemical (lytic agent) that destroys the red cell membrane,
hemoglobin is released.
• The released hemoglobin reacts with the cyanide in the reagent
and is converted to a stable pigment.
• The color of the "lysed" solution can be measured
spectrophotometrically.
• By comparison with standard solutions, a value for hemoglobin
is calculated.
• Procedures for a manual hemoglobin determination starts on
Hematocrit Measurement
• When an anticoagulated whole blood sample is placed in a straw

like glass tube and centrifuged, the ratio of volume of RBCs to
volume of whole blood can be determined. This is defined as the
hematocrit.
• The procedure for a manual hematocrit determination begins on
Calculation of the RBC Indices
• RBC Indices can be calculated manually by inserting the RBC,

Hgb and/or Hct results into the following equations:

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BASIC HEMATOLOGY
MCV = ( Hct / RBC ) x 10
MCH = ( Hgb / RBC ) x 10
MCHC = ( Hgb / Hct ) x 100
WBC Differential
Manual Microscopic Method
• The procedure for a manual differential count starts on page 102

of your textbook, HEMATOLOGY: Principles and Procedures
by Barbara Brown.
• Indicate what the following terms refer to in terms of the
associated parameter and whether high or low:
Hypochromic _________ parameter would be _________.
Macrocytic _________ parameter would be _________.
Leukocytosis _________ parameter would be _________.
Anemia _________ parameter would be _________.
Thrombocytosis _________ parameter would be _________.
ACTIVITY ANSWER KEY
Hypochromic MCH parameter would be LOW.
Macrocytic MCV parameter would be HIGH.
Leukocytosis WBC parameter would be HIGH.
Anemia RBC parameter would be LOW.
Thrombocytosis PLATELET parameter would be HIGH.

04/11/2001
BASIC HEMATOLOGY
Most laboratories no longer perform their daily CBCs using the

manual methods to measure WBCs, RBCs, Hgb, Hct, platelets and
calculate the RBC indices.
ACTIVITY
1. List potential sources of error when performing a manual CBC:
__________________________________________________
__________________________________________________
__________________________________________________
__________________________________________________
2. Based on your knowledge of COULTER automated hematology

systems, list the advantages of an automated CBC:
__________________________________________________
__________________________________________________
__________________________________________________
__________________________________________________

04/11/2001
BASIC HEMATOLOGY
ACTIVITY ANSWER KEY

1. Answers may vary:
Dilutions may be made improperly or with incorrect reagents

Dilutions are incorrectly charged on the hemacytometer
Inconsistent methods of counting cells on the hemacytometer
from tech to tech
Fatigue in counting
Hematocrit sample may be drawn up into the
microhematocrit tube incorrectly yielding bubbles
Improper centrifugation of the sample
Inconsistent reading of the results from tech to tech
Hemoglobin calibration is incorrect
Reagent expired
Sample dilution error
WBC Differential inconsistently read from tech to tech (very
subjective); poor quality of wedge smear and/or staining
2. No subjectivity, better precision, accuracy, consistent pipetting
and dilution preparation, consistent counting / measuring
method.
The Laboratory
Departments
• A typical hospital laboratory consists of many departments.

Hematology, Chemistry, Blood Bank, Urinalysis,
Coagulation, Flow Cytometry, etc.
Licensing Agencies
• ASCP (American Society of Clinical Pathologists)

04/11/2001
BASIC HEMATOLOGY
• ASCLS (American Society of Clinical Laboratory Science)

formerly ASMT (American Society of Medical Technologists)
• NCA, National Certification Agency for Medical Laboratory
Personnel
• ISCLT, International Society for Clinical Laboratory Technology
• HEW (Health, Education and Welfare)
Personnel
• Titles associated with different laboratorians:

MT Medical Technologist 4 year degree
including 12 months of clinical training /
licensed
MLT Medical Laboratory Technician 2 years of
college credit including 12 months training
in an approved hospital-related school /
licensed
CLA Certified Laboratory Assistant 12 months
training in an approved hospital related
school / licensed
Phlebotomist Individual responsible for collecting blood
samples for the laboratory
Key Operator
• As a Coulter representative, it is imperative that you

communicate with the appropriate person about the instrument
you are working on.
• The person who generally knows the most about the operation of
the instrument and its performance is referred to as the Key
Operator. Most of the time, this person has attended a Customer
training course at the Education Center.
• Although we recognize that the Supervisor or the Laboratory
Director and/or Pathologist have an interest in the COULTER
instrument and its operation, you should always direct your
communication initially with the "key operator".

04/11/2001
BASIC HEMATOLOGY

04/11/2001

Conceptos de Hematologia Basica

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Conceptos de Hematologia Basica

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HEMATOLOGY FUNDAMENTALS SERVICE TRAINING COURSE

Given the Basic Hematology course materials:

Read, study, review, and be able to answer the Hematology Basic

You will be given a Knowledge Check and asked to answer the

To Complete this module you will need:

Johns Hopkins Development of Blood Cells cell maturation chart

Hematology Fundamentals Service Training Course 1

2 Hematology Fundamentals Service Training Course

This session is intended to provide you with a basic introduction to

Why Study Hematology?

Hematology Fundamentals Service Training Course 3

raising or lowering the number of blood cells and their

Red Blood Cells

Figure 1 Composition of Blood

• Plasma is comprised of 90% water and 10% salts, nutrients,

Who Studies What About Blood?

4 Hematology Fundamentals Service Training Course

The three cellular components are red blood cells, white

Figure 2 Formed Elements of Blood

Hematology Fundamentals Service Training Course 5

Basic Cell Structure

Figure 3 Basic Cell Structure

• A semipermeable separation between the various internal cellular

• Contained within the cell membrane.

6 Hematology Fundamentals Service Training Course

cytoplasm a characteristic "look" that can help identify the

• Functions as the control center of the cell and is essential for

Blood Cell Maturation Characteristics

Overall Cell Size

• Overall cell size is usually compared with the size of a mature

Nuclear to Cytoplasmic Ratio

• N:C or the amount of space occupied by the nucleus in

Hematology Fundamentals Service Training Course 7

Some cells loose their nucleus entirely. Mature red blood

• Chromatin pattern, nuclear shape and the presence or absence of

Physical basis of heredity.

Found only in immature cells.

Either round or oval in young cells.

Where Blood Cells are Produced

Refer to the Development of Blood Cells chart

• In an adult, blood cells are formed in the bone marrow.

8 Hematology Fundamentals Service Training Course

• Cells are produced, mature and take on the appearance and

Red Blood Cells

Figure 4 Red Blood Cell

• Their flexibility (or ability to deform) also allows the RBCs to

Hematology Fundamentals Service Training Course 9

• The quantity of RBCs may be expressed as a red blood cell count

• The Reference Range (also referred to as "Normal Range")

(hemo = blood ; globin = protein)

10 Hematology Fundamentals Service Training Course

• The red cell membrane serves as a retaining barrier for the

• Mean Corpuscular Volume (MCV), which describes the

Hematology Fundamentals Service Training Course 11

Consider the effect of the diluent on the size of the RBC.

1. If the RBC size is responsive to its supporting media, what

2. What would be the outcome if red cells are suspended in a

3. What do you think the saline concentration in Coulter's diluting

ACTIVITY ANSWER KEY

2. The opposite effect would be observed. The "water" from inside

3. COULTER ISOTON III diluent has been formulated to best

• Mean Corpuscular Hemoglobin (MCH) describes the average

12 Hematology Fundamentals Service Training Course

• Mean Corpuscular Hemoglobin Concentration (MCHC)