Hematology

Biology 213.03/04
Dept. of Biology
San Francisco State University
Anthony Xavier Lopez
AIMS
! Learn about the basic composition of blood
! Learn typical hematology techniques:
! Hematocrit Determination
! RBC counting
! Hemoglobin Determination

Whole Blood
! Whole blood is composed of two elements: plasma and
formed elements
Plasma
! Extracellular fluid portion of the blood. Has osmolarity of
0.300 Osm or 300 mOsm.
! Contains 90% water and other dissolved solutes:
! Plasma proteins: albumin, globin, fibrinogen
! Nutrient materials: Glucose, lipids, amino acids
! Metabolic waste products: urea, lactic acid
! Gasses: Oxygen, carbon dioxide, nitrogen
! Electrolytes: Sodium, potassium, chloride
Formed Elements
! Formed elements consists of: Erythrocytes, Leukocytes,
Platelets
! Erythrocytes (Red Blood Cells): Carry oxygen and carbon
dioxide
! Most abundant cells in the blood
! Flattened, circular, biconcave shape
! Leukocytes (White Blood Cells): Defend body against
invading microorganisms and other foreign materials
! Platelets (Thrombocytes): Functions in blood clotting
Hematocrit
! Percentage of RBCs in whole blood (packed cell volume)
Sample is subjected to centrifugation
WBCs Buffy Coat
Red Blood Cell Count
! We will be counting the number of RBCs per cubic mm of
sheep blood by using a hemacytometer
! The average number of RBCs:
! Men 5.4 million cells/mm
3
! Woman 4.8 million cells/mm
3
! Sheep 8-14 million cells/mm
3

Counting RBCs Using a
Hemacytometer
! Hemacytometer is a specialized (and expensive)
microscope slide used to count cells.
! W: used in counting White
blood cells

! R: used in counting RBCs
We are to count 5 Rs (five
quadrants)

1 2
3
4
5
Counting RBCs Using a
Hemacytometer
! Each R square contains 5x5 counting
grids (total 25); we count these grids.
Hemoglobin Determination
! Hemoglobin (Hb) is a quaternary protein, each
containing one heme group that contains iron.
! A single Hb can carry a max of 4 oxygen molecules
! Hb also carries carbon dioxide
! We will discuss this further when we get into respiratory
physiology

Cyanmethemoglobin Method
! Common way to determine [Hb] is by cyanmethemoglobin
method
! [Hb] in whole blood is determined by chemically oxidizing the
ferrous iron (Fe
2+
) within the Hb molecule with potassium
ferricyanide (K
3
Fe(CN
6
) to methemoglobin (Fe
3+
)
! Potassium cyanide (KCN) then converts the methemoglobin to
cyanmethemoglobin.
! [cyanmethemoglobin] is measured using a spectrophotometer
Hb(Fe
2+
) MetHb(Fe
3+
) CNMetHb(Fe
3+
)
[K
3
Fe(CN)
6
]
Potassium Ferricyanide
(KCN)
(Potassium Cyanide)
Clinical Relevance
! Anemia is the most common blood disorder and roughly
affects more than 3 million Americans
! Anemia is a condition in which the body does not have
enough healthy red blood cells
! Anemia has many subtypes (listed are a few):
! Iron deficiency
! B12 deficiency
! Sickle Cell anemia
Q: How would an anemic individuals [Hb] and Hematocrit levels differ
from a non-anemic individual?