BLOOD

Properties of Blood
• Only Fluid Tissue of the Body
• Color: Scarlet (oxygen-rich) – dark red (oxygen-
poor)
• More dense than water and five times more
viscous
• pH: 7.35-7.45
• Temperature: 38 o C (100oF)
• 8% of body weight
• 5-6 L in males, 4-5 L in females
 Functions of Blood
• Transportation
– Of Gases, Nutrients and Hormones
• Regulation
– Maintain body temperature
– Maintain normal pH (buffering)
– Maintain adequate fluid volume (Proteins, Salts)
• Protection
– Preventing Blood Loss (Clotting)
– Preventing Infection (WBC and Antibody)
Components of Blood
• Plasma – liquid component of the
blood
• Formed elements – solid component
of the blood
– Buffy Coat – Leukocytes and Platelets
– Erythrocytes - Red Blood Cells

• Hematocrit – volume percentage of

red blood cells in blood
– Normal Values:
• Adult male: 40.7 %– 50.3%
• Adult female: 36.1 % - 44.3%
• Newborn: 45 % - 61%
• Infant: 32 % - 42 %
– High hematocrit (>53% in men
and >51% in women) is termed
POLYCYTHEMIA
Whole Blood vs Serum vs Plasma
Whole Blood
(used in CBC)

Undergoes Does not undergo

Clotting and clotting,
Centrifugation Undergoes
Centrifugation
Formed Elements

Serum (used in Serology Plasma (used in

and Blood Chemistry) Coagulation Studies)

PLASMA CAN CLOT WHILE SERUM CANNOT

 Components of Plasma
Constituent Description and Importance
Water 91% of plasma volume; solvent; absorbs heat
Solutes
Plasma Proteins 7% by weight of plasma volume; albumin (60%) contributes to
osmotic pressure) , globulins (36%) are either for transport
(alpha and beta) or for immune response (gamma). Fibrinogen
(4%) is for clotting
Nitrogenous Non-protein by products of cellular metabolism such as urea,
substances uric acid, creatinine and ammonium salts
Organic Materials absorbed from digestive tract including glucose,
Nutrients carbohydrates, amino acids, fats
Electrolytes Na+, K+, Cl-, PO4-, SO4- HCO3-
Respiratory Oxygen and Carbon Dioxide
Gases
Hormones Steroid and Thyroid Hormones
 Cellular Elements
Constituent Duration of Development Description and Importance
and Life Span

Erythrocytes 15 days; 100-120 days Salmon colored biconcave anucleate disks ; Transport
oxygen and carbon dioxide

Leukocytes

Neutrophils 14 days; 6 hrs – few days Granulocyte; Multilobed nucleus with inconspircuous
cytoplasmic granules; Phagocytize bacteria

Eosinophils 14 days; 5 days Granulocyte; Bilobed nucleus with red cytoplasmic

granules; Antiparasitic and downregulates
inflammation
Basophils 1-7 days; few hrs- few days Granulocyte; Lobed nucleus with large purplish black
granulues; Mediates inflammation, has heparin

Lymphocyte Days – weeks; hours – Agranulocyte; spherical/indented nucleus ; humoral

years and cell-mediated immune response

Monocyte 2-3 days; months Agranulocyte; U- or kidney-shaped nucleus;

phagocytosis

Platelets 4-5 days; 5-10 days Discoid cytoplasmic fragments; for clotting
 Cellular Elements

Erythrocytes Neutrophils Eosinophils Basophils

Lymphocytes Monocyte Platelets

 Hematopoiesis
• Blood Cell Production
• Encompasses the Overall Interactions of Celuular
Proliferation, Differentiation Morphogenesis,
Maturation and Death
• Modulated by poietins, colony stimulating factors
and interleukins
• Three Developmental Periods
– Mesoblastic Period
– Hepatic Period
– Myeloid Period
 Marrow Hematopoiesis
• Totipotent/Omnipotent – stem cells that can
be any cell
• Pluripotent – stem cell that can be any type of
blood cell
• Reticulocyte – very young RBC’s with nucleus
• Blood Dust/ Hemoconia – debris or remnants
of cells; small refractive colorless particles in
the blood that are granules from cells in blood
or minute fat globules.
 Erythropoiesis
• Erythroblastic island – unit of erythropoiesis in
the marrow
• Hemoglobin synthesis begins at early normoblast
stage and is finished by reticulocyte stage.
• Red cell development is characterized by three
main features:
– Decreasing cell size and nucler extrusion
– Progressive loss of organelles (ribosomes) and
decrease in basophilia
– Increase in hemoglobin content and subsequent
eosinophilia
 Reticulocyte Count
Why? To determine if RBC’s are being created in the bone marrow at
an appropriate rate
Principle Reticulocytes are young RBC’s that have not fully extruded
their organelles yet. Staining these young RBC’s with methylene blue
will reveal the organelles and would provide a good way to measure
the production of new red blood cells.
Methodology

Blood Smear
Counting Calculations
Preparation

Normal Values: 0.5%-1.5% (adults), 3%-6% (newborns)

High Values: Hemolytic Anemia, Bleeding, Erythroblastosis Fetalis,
High EPO, Pregnancy, Treatments for anemia, Recent move to high
altitudes
Low Values: Bone marrow failure, Cirrhosis, Iron Deficiency Anemia,
Chronic Kidney Disease, Anemia by low levels of Vit B12 or folate
Erythrocyte Sedimentation Rate
Why? To determine presence and extent of Inflammation in Body
Principle RBC’s settle more rapidly during some diseased states such
as increase in fibrinogen, immunoglobulins and acute-phase
reaction proteins. Fell into disuse as more specific methods for
following inflammation have been discovered.
Methodology
– Westergren Method (more precise)
– Wintrobe Method
Normal Values: Healthy men (<15 mm/hr) Healthy women (<20
mm/hr); higher in old age in both genders
High Values: Increased level of fibrinogen, gamma globulins, tilted
tubes, high room temp, Rouleaux formation
Low Values: abnormal RBC’s, short ESR tubes, low room temp, delay
in test performance, clotted blood, excess anticoagulant, bubbles
 Granulopoiesis
Myeloblast – earliest recognizable stage

Promyelocyte – development of
azurophilic granules

Myelocyte - development of specific

granules

Metamyelocyte – nuclear segmentation

Stab / Band Forms – ring /horseshoe

nucleus
 Lymphopoiesis
• Two recognizable stages: Lymphoblast and
prolymphocytes
• Marked by progressive diminution in cell size
• Lymphocytes proliferate outside bone marrow,
specifically in thymus, spleen and other
immune organs
• T-lymphocytes and B-lymphocytes
 Monopoiesis
• Monocytes and granulocytes may have a
common progenitor and colony-stimulating
factors may be the determining factor which
matures to which.
• Two morphological precursors are recognized:
monoblasts and promonocytes.
• Characterized by decrease in cell size and
progressive indentation of nucleus.
• Monocytes spend 3-4 days in circulation
before they enter tissues where they mature
into macrophages.
Other Notes on Hematopoiesis
• Platelets – fragmentation from
megakaryocyte; not true cells.
• Natural Killer Cells – found to have memory
cells and thus can be part of innate or
adaptive immunity
• Some animals (frogs and chicken) retain the
nucleus of their Red Blood Cells even in
maturity.
 References
Marieb, E. N. & Hoehn, K. (2011). Human Anatomy and Physiology.
Pearson Benjamin Cummings.
Young, B., Lowe, J., Stephens, A., Heath, J. (2006). Wheater’s
Functional Histology. Elsevier Churchill Livingstone.
Hubbard, R. (2010). A Concise Review of Clinical Laboratory Science.
Lippincott Williams & Wilkins.
Manual Reticulocyte Count Procedure. Retrieved at
http://webmedia.unmc.edu/alliedhealth/CLS/CLS416%2011/Hemo
SL%20Lab%20Procedures%202011%20Manual%20Retic.pdf
Reticulocyte Count. Retrieved at
http://www.nlm.nih.gov/medlineplus/ency/article/003637.htm
Erythrocyte Sedimentation Rate. Retrieved at
http://www.medicine.mcgill.ca/physio/vlab/bloodlab/esr.htm
• one of the most commonly ordered blood
tests by doctors
• measures the quantity of all the different
types of cells in the blood (e.g. RBCs, WBCs,
platelets)
• may be done as part of a regular physical
examination
• Find the cause of symptoms such as fatigue,
weakness, fever, bruising, or weight loss
• Check for certain conditions (e.g. anemia,
polycythemia, leukemia, allergy, asthma)
• Check how the blood cells and their counts are
affected if there is abnormal bleeding
• Check how the body is dealing with some types
of drug or radiation treatment
• Screen for high and low values before a surgery
• White blood cell count (WBC or Leukocyte count)
• WBC differential count
• Red blood cell count (RBC or erythrocyte count)
• Hematocrit (Hct)
• Hemoglobin (Hbg)
• Mean corpuscular volume (MCV)
• Mean corpuscular hemoglobin (MCH)
• Mean corpuscular hemoglobin concentration (MCHC)
• Red cell distribution width (RDW)
• Platelet count
• Mean platelet volume (MPV)
• WBC count
– More than normal:
• Leukocytosis – due to infection or inflammation
• Malignancy – unlimited reproduction (e.g. leukemia,
lymphoma)
• use of corticosteroids, underactive adrenal glands, thyroid
gland problems, certain medicines, or removal of the spleen
– Less than normal:
• Leukopenia – due to chemotherapy and reactions to other
medicines, aplastic anemia, viral infections, malaria,
alcoholism, AIDS, lupus, or Cushing's syndrome
• Large spleen
• WBC differential count
– Eosinophilia: allergic reactions, autoimmune
diseases, parasitic infections
– Neutrophilia: bacterial infections
• segmented neutrophils: “shift to the right”
• band form of neutrophils: “shift to the left”
– Lymphocytosis: viral infections
– Monocytosis: chronic infections
– Basophilia: allergic reactions
• RBC count, Hbg, Hct
– More than normal:
• Polycythemia vera
• Smoking, CO exposure, alcoholisms, some diseases of the
lungs, kidney, heart and liver
• Dehydration, diarrhea or vomiting, excessive sweating,
severe burns, and the use of diuretics,
– Less than normal:
• Anemia, bleeding, malnutrition
• Removal of spleen
• RBC Indices
– MCV
• Less than normal: Iron deficiency
• More than normal: Folic acid or Vit B12 deficiency
• Platelet count
– More than normal:
• Thrombocytosis – result from bleeding, iron deficiency,
inflammation or blood malignancy, or problems with
the bone marrow
– Less than normal:
• Thrombocytopenia – pregnancy
• Viral hemorrhagic fevers (Dengue)
• computerized, highly specialized machines
• include programmable automated alarm
systems for indicating results outside the
reference range
Schematic design of microfluidic device for chip-based, complete blood count
test. Cells are first sorted by size and then counted with an impedance
measurement. Due to the variable and elastic nature of red blood cells, optimal
filter trapping design was explored empirically rather than through modeling.
Image courtesy Bradley Layton, Drexel University, and SB Microsystems.
• Hemocytometer
– precision-made slide for performing manual cell
counts with the aid of a microscope
– counting chamber that contains two
microscopically ruled area
marked off by lines
– Depth = 0.1 mm
• Hemocytometers are used when:
– automated cell counters and hematology
analyzers are unavailable
– blood cell counts are extremely low
– to get a cell count for body fluids (spinal fluid,
joint fluid, semen counts, and other bodily fluids)
• Hemocytometer
– total lined area on
each side is made of
a large square (3 X 3
mm)
– large square is
divided into nine
equal squares
each 1 mm2
– total area of all the
squares is 9 mm2
• Dilution Pipettes – for dilution of samples
– WBC pipette
• with white bead inside a small bulb; smaller than the RBC
pipette
– RBC pipette
• with red bead inside a big bulb
• Cover slip
– Much thicker than typical cover slip
• Manual differential counter – used for
counting different blood cells
• First, determine if the liquid needs to be diluted.
• Clean the chamber and cover slip with a lens
paper.
• Cover slip is placed over the counting surface
prior to putting on the cell suspension.
• The suspension is introduced into one of the V-
shaped wells with a pipette.
• Leave the hemocytometer for a few minutes to
allow the cells to settle.
• The charged counting chamber is then placed on
the microscope stage and the counting grid is
brought into focus at low power.
• Counting chamber
– 9 squares
• WBC count
– 4 large corner squares (each
has 16 smaller squares)
• RBC count
– large center square (has 25
smaller squares)
• four corner squares and center
square
• Platelet count
– entire large center square
• WBC count
– Diluent: Acetic acid (to dilute the amount of WBC
for counting and to lyse the RBC and platelets)
– Dilution Factor (DF)  0.5 : 10 = 1 : 20
(e.g. 50 µL of blood + 950 µL diluent)
– No. of squares counted: 4
– Volume of one square: 0.1 mm

No. of cells counted x Dilution factor

WBC/  L =
No . of squares counted x Volume of one square
• RBC count
– Diluent: Hayem’s fluid or normal saline solution
(to reduce the number of RBC; to maintain the
normal disk shape of the RBC; to prevent
autoagglutination)
– Dilution Factor (DF)  0.5 : 100 = 1 : 200
– No. of squares counted: 5
– Volume of one square: 1/25 x 0.1 mm3
No. of cells counted x Dilution factor
RBC /  L =
No . of squares counted x Volume of one square
• PLT count
– Diluent: 1% ammonium oxalate solution (to lyse
RBC)
– Dilution Factor (DF)  0.5 : 50 = 1 : 100
– No. of squares counted: 1
– Volume of one square: 0.1 mm3

No. of cells counted x Dilution factor

PLT /  L =
No . of squares counted x Volume of one square
• Normal reference values:
– Total WBC count
• Men and nonpregnant women: 5,000 – 10,000/mm3
– Total RBC count
• Women: 4,000,000 – 5,000,000/mm3
• Men: 4,500,000 – 5,500,000/mm3
• Children: 3,800,000 – 6,000,000/mm3
• Newborn: 4,100,000 – 6,100,000/mm3
– Total PLT count
• Adults: 140,000 – 400,000/mm3
• Children: 150,000 – 450,000/mm3
• Diluent and blood should be properly mixed
before filling the hemocytometer.
• Suspensions should be dilute enough so that the
cells or other particles do not overlap each other
on the grid, and should be uniformly distributed.
• The hemocytometer must be properly filled to
avoid erroneous results in manual cell counting.
• RBCs are much smaller than WBCs. Platelets are
the smallest.
• Each side of the chamber should be counted and
an average of the two should be taken.
• Experimental Biosciences. 10 August 2012. Using a
Counting Chamber. [online]. Available from:
http://www.ruf.rice.edu/~bioslabs/methods/microscopy/ce
llcounting.html [Accessed 25 August 2014].
• Nabili, S. T. 28 May 2014. Complete Blood Count. [online].
Available from:
http://www.emedicinehealth.com/complete_blood_count_
cbc/article_em.htm [Accessed 25 August 2014].
• WebMD. 6 August 2012. Complete Blood Count (CBC).
[online]. Available from: http://www.webmd.com/a-to-z-
guides/complete-blood-count-cbc [Accessed 25 August
2014].
• https://www.youtube.com/watch?v=pP0xERLUhyc
• https://www.youtube.com/watch?v=WWS9sZbGj6A
• Google images
Hemoglobin
Function, Structure, Formation, Destruction,
Tests
Functions of Hemoglobin
- Carried to all tissues of the body by erythrocytes
• Transport
- Has a major role in oxygen transport to the
tissue
- CO2 transport back to the lungs
-Designed to take up oxygen in areas of high
oxygen tension and release them in areas of low
oxygen tension
• Major buffering system of the body
• Regulator of iron metabolism
Structure of Hemoglobin
• Large, spherical, complex
protein molecule (approx.
64,000 MW)
• Has 2 main parts:
- Heme (4 units) – 3%
- Globin (2 pairs) – 97%

Structure of Adult Hemoglobin (Hemoglobin A)

(http://www.nutralegacy.com/blog/wp-
content/uploads/benefits-of-iron-to-
hemoglobin1.jpg)
Heme
• Comprises a porphyrin
ring with iron chelated in
the center
• Iron – the site of
reversible Oxygen
attachment
• Enclosed in a globular
protein, globin
Structure of Adult Hemoglobin (Hemoglobin A) and
Heme
Bishop, M. (2010), Clinical Chemistry:
Techniques, Principles, Correlations, 6th
Edition
Globin
• Each chain consists of 141 or more amino acids
• Has 6 types: (based on the variation of AA sequence)
- α Alpha
- β Beta
- δ Delta
- γ Gamma
- δ Zeta (in embryonic hemoglobin)
- ε Epsilon (in embryonic hemoglobin)
• Combinations of globin subunits determine the type
of hemoglobin
Globin
In the Embryo Subunit 1 Subunit 2
Gower I 2 δ Zeta 2 ε Epsilon
Gower II 2 α Alpha 2 ε Epsilon
Portland I 2 δ Zeta 2 γ Gamma
Portland II 2 δ Zeta 2 β Beta

In the Fetus Subunit 1 Subunit 2

Hemoglobin F 2 α Alpha 2 γ Gamma

In Postnatal People Subunit 1 Subunit 2

Hemoglobin A1 2 α Alpha 2 β Beta
Hemoglobin A2 2 α Alpha 2 δ Delta
Hemoglobin F 2 α Alpha 2 γ Gamma
Globin
In Embryos,
• Hemoglobin F is the main component, and is
about 60% normal hemoglobin at birth.
• Production of embryonic chains stop by week 8
of gestation
In Adults,
• Hemoglobin A1 is the most common
• Hemoglobin A2 is makes up 3% of normal adult
hemoglobin
• Hemoglobin F makes up the remainder.
Hemoglobin F
• Hemoglobin F has greater affinity for oxygen
than Hemoglobin A.
• Thus, more efficient carrier of oxygen for the
fetus.
• Differentiation of these two can be done by
using the alkali denaturation test, wherein
Hemoglobin F is more resistant to alkali.
• Alpha-fetoprotein – is a component of
Hemoglobin F that may serve as a marker for
liver cancer
Hematopoiesis and
Hemoglobin
• Mesoblastic Period
• Hepatic Period
• Myeloid Period
Mesoblastic Period
• It should be remembered that blood and its
precursors are mesodermal in origin
• Primary Hematopoietic Organ: Blood island
of the yolk sac
• Secondary Hematopoietic Organ: ventral
aspect of aorta in preumbilical area
• Cells are primitive hematopoietic cell lines.
They are not found after the 3rd month.
• Mainly embryonic hemoglobin
• Cells produced are nucleated and easily deformed
Hepatic Period
• Primary site of hematopoiesis: Liver
• Secondary Site: Spleen
• Hemoglobin F (α2γ2) is the main hemoglobin
• Thymus, Spleen and Lymph Nodes becomes
sites for the development of WBC
(lymphocytes)
Myeloid Period
• Primary site of hematopoiesis: Bone Marrow of
Long Bones
• As we age, red marrow is replaced by yellow (fatty)
marrow such that only the marrow of pelvis, ribs,
sternum, skull, and proximal extremities of the
long bones produce the blood cells
• Extramedullary hematopoiesis – production of
blood cells outside bone marrow (e.g. liver and
spleen), happens when there is decreased bone
marrow function.
• Hemoglobin A1 (α2β2) is the main hemoglobin
-HbA1C (glycosylated Hemoglobin A1) – marker
for high blood glucose levels/diabetes
HbA1C test
• Glycosylated HbA1
• Uses Whole Blood with Anticoagulant; 0.5 mL
fed to HPLC to analyze the glycosylated
Hemoglobin
• Normal Values: 4-6% (non-diabetic adults)
• Values greater than 7%: diabetes mellitus
Synthesis of Hemoglobin
• Occurs in the immature RBCs in the bone
marrow: 65% in the nucleated cells; 35% in
reticulocytes
• Heme – synthesized in the mitochondria of cells.
Iron is transported to the developing RBCs by
transferrin. Iron + PROTO ring = Heme
• Globin – protein synthesis occurs in the
cytoplasmic polyribosomes.
• Heme (leaving the mitochondria) + Globin (in
the cytoplasm) = HEMOGLOBIN
Fate of Hemoglobin
• After the death of RBCs (120 days – average life
cycle), heme and globin are separated and iron is
salvaged for reuse
• Globin is metabolized into amino acids and is
released into the circulation
• Heme is degraded to a yellow pigment called
bilirubin
• The liver secretes bilirubin into the intestines as bile
• The intestines metabolize it into urobilinogen
• This degraded pigment leaves the body in feces, in a
pigment called stercobilin
Hemoglobin Determination
(Cyanmethemoglobin/
Hemiglobincyanide Method)
Principle:
• Based on the determination of the
cyanmethemoglobin that has been adopted as a
standard method
• Hemoglobin (released from erythrocytes) →
(oxidized by ferricyanide) methemoglobin →
(cyanide) stable cyanmethemoglobin
Hemoglobin Determination
(Cyanmethemoglobin/
Hemiglobincyanide Method)
Principle:
• Absorbance of cyanmethemoglobin is measured
at 540 nm.
• It is directly proportional to the hemoglobin
concentration in the sample
• Hb is measured as g (Hb) / dL (deciliter of blood)
Hemoglobin Determination
(Cyanmethemoglobin/
Hemiglobincyanide Method)
Procedure:
1. 20µl of blood is diluted in a 5mL solution of
potassium ferricyanide and potassium cyanide
(Drabkin reagent).
2. Mix the solution well and allow to stand at room
temperature for at least 3 minutes.
3. The absorbance is measured against the reagent
blank in the photoelectric colorimeter at 540 nm.
Normal Ranges for Hemoglobin
Newborns 17-22 g/dL
1 week 15-20 g/dL
1 month 11-15 g/dL
Children 11-13 g/dL
Women after
11.7-13.8 g/dL
middle age
Adult women 12-16 g/dL
Men after middle
12.4-14.9 g/dL
age
Adult men 14-18 g/dL
< 10 g/dL = severe anemia (both sexes)
↑ age, Hb values ↓
 Factors Causing HIGH Hemoglobin
Levels
• Erythrocytosis
• Polycythemia
• Poor heart or lung function (lung and heart
diseases, poor cardiac output)
• Taking drugs or hormones, like Erythropoietin
(EPO)
• Smoking
• Cancers or tumors
• Living in high altitudes
 Factors Causing LOW Hemoglobin
Levels
• Vitamin deficiency that may cause anemias
(iron, folate, B12)
• Other forms of anemia (sickle cell anemia)
• Blood loss, Excessive bleeding, hemorrhages
• Pregnancy & Menstruation (Women)
• Problems with bone marrow
Common Disorders
• Can result from the abnormalities in subunits,
functions, and chemical structures

• Sickle Cell Disease/Anemia

• Thalassemia
Sickle Cell Anemia
• Autosomal recessive disorder
wherein the body makes
“sickle-shaped” RBCs due to
hemoglobin S or sickle
hemoglobin
• Caused by the substitution of
AA Valine (at position 6) to
AA Leucine → precipitation of
Hb → sickle RBCs (dies after
10-20 days)
• Sickle cells are stiff and sticky.
This may block blood flow and
cause pain and organ damage,
as well as increase the risk for
infection.

• http://www.nhlbi.nih.gov/health/health-
topics/topics/sca/
Thalassemias
• A family of autosomal recessive blood disorder
in which the body makes an abnormal form of
hemoglobin.
• This disorder results in the excessive destruction
of RBC, which leads to anemia
References
• Bishop, M., Fody, E. & Schoeff, L. (2010). Clinical
Chemistry: Techniques, Principles, Correlations. Lippincott
Williams & Williams.
• Marieb, E. N. & Hoehn, K. (2011). Human Anatomy and
Physiology. Pearson Benjamin Cummings.
• Blackburn, S. (2013). Maternal, Fetal and Neonatal
Physiology. Elsevier.
• Robbins and Cotran: Pathologic Basis of Disease, 8th ed.
• Henry’s Clinical Diagnosis and Management by Laboratory
Methods, 22th ed.
• http://www.nlm.nih.gov/medlineplus/ency/article/000587.htm
• http://www.nhlbi.nih.gov/health/health-topics/topics/sca/
• http://www.mayoclinic.org/symptoms/high-hemoglobin-
count/basics/causes/sym-20050862
Hematocrit
• Hematocrit measures the volume of RBCs compared to
the total blood volume.
• Can be used as a test for anemia.
• Hematocrit is used for other test.

What is it and why is it

important?
• Microsampling is used for blood collection
• Blood is collected in microhematocrit tube (heparinized)
• Tubes are then centrifuged for 3-5 mins
• The tubes are then measured in a hematocrit reader
• Values are presented as percentages

Procedure and
Computation
• Low hematocrit:
• Blood loss due to trauma
• Nutritional deficiencies
• Bone marrow problems
• High hematocrit
• Chronic smokers
• Dehydration
• Living in high altitudes

Reasons for abnormal

values
Mean Corpuscular
Volume
• The average volume of each RBC in a specimen
• Basically, it measure the size of the RBCs
• Can indicate microcytic, normocytic, or macrocytic RBCs
• Helps to classify anemia based on red cell morphology

What is it and why is it

important?
• Whole blood collected by venipuncture.
• Collected in EDTA tube
• Hematocrit and red blood cell count are
determined

• Equation

Procedure and
Computation
• Microcytic anemia
• Iron deficienecy anemia
• Anemia of chronic disease
• Thalassemia
• Sideroblastic anemia

• Macrocytic anemia
• Folate deficiency anemia
• Vit B deficiency anemia

Reasons for abnormal

values
• Liver disease
• Hemolytic anemias
• Hypothyroidism
• Excessive alcohol intake
• Aplastic anemia
• Myelodysplastic syndrome
Mean Corpuscular
Hemoglobin
Concentration
• It is the measure of hemoglobin in each re blood cell
• Can indicate hypochromic, normochromic or
hyperchromic RBCs
• To asses anemia based on hemoglobin levels

What is it and why is it

important?
• Venipuncture collected in EDTA tube
• Hemoglobin values and hematocrit are obtained

• Equation:

Procedure and Equation

Normal Values
RBC (M/MCL)
AGE MALE FEMALE
> 65 years 3.8 - 5.8 3.8 - 5.2
45 - 64 years 4.2 - 5.6 3.8 - 5.3
18 - 44 years 4.3 - 5.7 3.8 - 5.1
12 - 17 years 4.1 - 5.2 3.8 - 5.0
1 - 11 years 3.8 - 4.8 3.8 - 4.8
6 - 11 months 3.9 - 5.5 3.9 - 5.5
1 - 5 months 3.3 - 5.3 3.3 - 5.3
7 - 30 days 4.0 - 5.6 4.0 - 5.6
0 - 6 days 4.4 - 5.8 4.4 - 5.8

Red Blood Cell Count

WBC(K/MCL)
AGE MALE FEMALE
> 21 years 4.5 - 11.0 4.5 - 11.0
16 - 20 years 4.5 - 13.0 4.5 - 13.0
8 - 15 years 4.5 - 13.5 4.5 - 13.5
6 - 7 years 5.0 - 14.5 5.0 - 14.5
4 - 5 years 5.5 - 15.5 5.5 - 15.5
2 months - 3 years 6.0 - 17.5 6.0 - 17.5
2 days - 1 month 5.0 - 21.0 5.0 - 21.0
0 - 1 day 9.0 - 34.0 9.0 - 34.0

White Blood Cell Count

MCV (FL)
AGE MALE FEMALE
> 65 years 80 - 100 80 - 100
45 - 64 years 80 - 100 80 - 100
18 - 44 years 80 - 99 81 - 100
12 - 17 years 77 - 95 73 - 98
9 - 11 years 75 - 87 75 - 87
6 month s -8 years 68 - 85 68 - 85
4 - 5 months 76 - 97 76 - 97
2 - 3 months 84 - 106 84 - 106
1 month 91 - 112 91 - 112
0 - 30 days 88 - 140 88 - 140

Mean Corpuscular Volume

MCHC (G/DL)
AGE MALE FEMALE
> 65 years 31 - 36 32 - 36
45 - 64 years 32 - 36 31 - 36
18 - 44 years 32 - 37 32 - 36
12 - 17 years 32 - 37 32 - 36
6 months - 11 yrs 32 - 37 32 - 37
4 - 5 months 29 - 37 29 - 37
2 - 3 months 28 - 35 28 - 35
1 month 28 - 36 28 - 36
14 - 30 days 28 - 35 28 - 35
0 - 13 days 30 - 36 30 - 36

Mean Corpuscular Hemoglobin

Concentration
HEMOGLOBIN (G/DL)
AGE MALE FEMALE
> 65 years 12.6 - 17.4 11.7 - 16.1
4 5 - 64 years 13.1 - 17.2 11.7 - 16.0
18 - 44 years 13.2 - 17.3 11.7 - 15.5
12 - 17 years 11.7 - 16.6 11.5 - 15.3
9 - 11 years 12.0 - 15.0 12.0 - 15.0
6 months - 8 years 11.2 - 14.1 11.2 - 14.1
4 - 5 months 10.3 - 14.1 10.3 - 14.1
2 - 3 months 9.4 - 13.0 9.4 - 13.0
1 month 10.7 - 17.1 10.7 - 17.1
14 - 30 days 13.4 - 19.8 13.4 - 19.8

Hemoglobin
HEMATOCRIT (%)
AGE MALE FEMALE
> 65 years 37 - 51 35 - 47
45 - 64 years 39 - 50 35 - 47
18 - 44 years 39 - 49 35 - 45
12 - 17 years 35 - 45 34 - 44
9 - 11 years 34 - 43 34 - 43
6 months - 8
31 - 41 31 - 41
years
4 - 5 months 32 - 44 32 - 44
2 - 3 months 28 - 42 28 - 42
1 month 33 - 55 33 - 55
14 - 30 days 41 - 65 41 - 65
0 - 13 days 41 - 73 41 - 73

Hematocrit
 PLATELETSK/MCL
AGE MALE FEMALE
All ages 150 - 440 150 - 440

Platelet Count
Billet, H.H.(1990) Hemoglobin and Hematocrit. Retrieved
http://www.ncbi.nlm.nih.gov/books/NBK259/
Curry, C.V. (2012) Mean Corpuscular Volume. Retrieved
http://emedicine.medscape.com/article/2085770-overview#aw2aab6b3
Epstein, J. (2012) RBC Indices. Retrieved
http://www.healthline.com/health/rbc-indices#Overview1
McHill Physiology Vitual Lab (nd) MCV and MCHC. Retrieved
http://www.medicine.mcgill.ca/physio/vlab/bloodlab/mcv-mchc_n.htm
Mercy North Iowa (nd) CBC Normal Ranges. Retrieved
http://www.mercynorthiowa.com/cbc-normal-ranges
Shiel, W.C. (2014) Hematocrit. Retrieved
http://www.medicinenet.com/hematocrit/article.htm

References