Red Blood Cell (RBC)

and
White Blood Cell (WBC)
Counts
Blood
 Important Blood Terminology
• Whole blood: Composed of plasma (fluid portion of blood) and
formed elements (blood cells and platelets).
• Formed Elements: Red blood cells, white blood cells, and
platelets.
• Plasma: Fluid portion of blood that contains proteins, ions,
nutrients, hormones, antibodies, metabolites, enzymes, etc.
• Serum: Similar to plasma, but without clotting factors such as fibrin.
• Hematocrit: Percentage of total blood volume that is occupied by
red blood cells; also called packed red cell volume.
• Red cell count: The number of red blood cells per cubic milliliter
(mm3) of whole blood. 1 mm3 = 1 µL.
• Total white cell count: The total number of all white blood cells per
cubic milliliter (mm3) of whole blood. This number does not
distinguish between different white cell types.
• Differential white cell count: The relative numbers of the five
types of white blood cells. The numbers are reported as
percentages.
Taken from Diggs, L.W., Sturm, D., and Bell, A. The
Morphology of Human Blood Cells, 5th Edition, Abbot
Laboratories, Abbot Park, Illinois.
 Composition of Blood
• Plasma: Fluid portion of blood (~55%)
– Straw-colored
– Contains water, ions, nutrients, hormones,
antibodies, enzymes, metabolites, and
proteins.
– Na+ and Cl– are very high in concentration
and contribute most to plasma osmolality.
• Formed Elements: Cellular component of
blood (~45%)
– Red blood cells (erythrocytes)
– White blood cells (leukocytes)
– Platelets (thrombocytes)
Plasma Proteins (7–9% of plasma)
• Albumins (60–80% of all plasma proteins)
– Smallest in size
– Produced by liver
– Also important in contributing to plasma
osmotic pressure
– Many hormones and ions are bound to
albumin
Plasma Proteins (7–9% of plasma)
• Globulins
– Alpha globulins
– Beta globulins
• Both alpha and beta globulins are produced by the
liver and function to transport lipids and fat-soluble
vitamins.
– Gamma globulins
• Antibodies produced by lymphocytes
• Fibrinogens
– Produced by the liver
– Function in blood clotting
 Red Blood Cells
• Synonym: Erythrocytes
• Most numerous cells in blood
– Males: 4.5–6.0 million/mm3
– Females: 4.0–5.5 million/mm3
• Flattened biconcave discs
• Lack nucleus and mitochondria
• Each RBC has about 280 million
hemoglobin molecules
• Lifespan: 120 days
 White Blood Cells
• Synonym: Leukocytes
• 4,500 – 11,000 / mm3
• Contain nucleus and mitochondria
• Capable of amoeboid movement
– Diapedesis: the movement of leukocytes
through capillary walls.
• Classified based on stained appearance:
– Granular leukocytes
– Agranular leukocytes
– Lymphocytes
 Leukocytes
• Granular leukocytes
– Eosinophils (1–3%)
– Basophils (<1%)
– Neutrophils = Polymorphonuclear neutrophils
(PMN) (50–60%)
– Lifespan 12 hours to 3 days
 Leukocytes Continued
• Agranular leukocytes
– Monocytes (3–9%)
• Largest of leukocytes
• Kidney-shaped nuclei
• Lifespan of 100–300 days
– Lymphocytes (25–33%)
• Small cells
• Round nuclei
• Small cytoplasm
• T Lymphocytes
• B Lymphocytes
– Plasma cells (produce antibodies)
 Platelets
• Synonym: Thrombocytes
• 130,000–360,000 /mm3
• Smallest of formed elements
• Cytoplasmic fragments of megakaryocytes
in bone marrow
• Lack nuclei
• Capable of amoeboid movement
• Important in the formation of bulk of the
clot during blood clotting
• Lifespan of 5–9 days
Basic Procedure for RBC and WBC
• Obtain whole blood in a tube that contains
EDTA (Na+ or K+ salt).
• EDTA
– Ethylenediaminetetraacetic acid
• EDTA chelates Ca2+ and thus prevents the
formation of blood clots.
• This blood sample may be used as the
starting point for both the red blood cell
(RBC) count and white blood cell (WBC)
count procedures.
 Red Blood Cell Count
• Mix blood in tube well.
• Dilute whole blood in Hayem’s solution.
– 1:200 dilution
• Hayem’s Solution
– Isotonic with blood
– Contains
• 0.5% NaCl
• 2.5% Na2SO4 (to prevent clump formation)
• 0.25% HgCl2 (mercuric chloride) (preservative)
• Apply a small volume (~10 µl) of diluted
blood to one side of a hemacytometer.
Hemacytometer
Hemacytometer
Hemacytometer
Hemacytometer
Boundary Rule
 Calculation of RBC
N Count × Depth Factor × Dilution Factor
=
mm3 A

• N/mm3: Number of cells per cubic mm

• Count: Total number of cells counted
• Depth Factor: 10
• Dilution Factor: 200
• A: Area used for count in mm2 = 0.2 mm2
• Thus, for RBC: Multiply your total count
by 10,000.
Reference Ranges for Human RBC
Counts

• Adult Male: 4.5–6.0 x 106 /mm3

• Adult Female: 4.0–5.5 x 106 /mm3
• Newborn: 5.0–6.3 x 106 /mm3
 White Blood Cell Count
• Mix blood in tube well.
• Dilute whole blood in Turk’s solution.
– 1:100 dilution
• Turk’s Solution
– Red blood cells burst in this solution, but
white blood cells remain intact and are
stained.
– Contains
• 1% Acetic acid
• 0.01% Gentian violet (crystal violet) (stain)
• Apply a small volume (~10 µl) of diluted
blood to one side of a hemacytometer.
 Calculation of WBC
N Count × Depth Factor × Dilution Factor
=
mm3 A

• N/mm3: Number of cells per cubic mm

• Count: Total number of cells counted
• Depth Factor: 10
• Dilution Factor: 100
• A: Area counted in mm2 = 4 mm2
• Thus, for WBC: Multiply your total count
by 250.
 Reference Ranges for Human
WBC Counts
• Newborn: 9,000–30,000 /mm3
– Average: 18,000 /mm3
• 1 Year: 6,000–14,000 /mm3
– Average: 11,000 /mm3
• 6 Years: 4,500–12,000 /mm3
– Average: 8,000 /mm3
• Adult: 4,500–11,000 /mm3
– Average: 7,400 /mm3