Introduction to

Hematology
OVERVIEW
LAB SAFETY
► Hematology is the branch of medical science that deals with the study of
the clinical, morphologic and laboratory disorders of the blood and the
blood forming organs.
► This study includes the analyses of the concentration, structure and
function of cells in the blood; their precursors in the bone marrow;
chemical constituents of plasma or serum intimately linked with blood cell
structure and function; and function of platelets and proteins involved in
hemostasis and blood coagulation.
► Blood is vital, life–sustaining fluid circulating constantly in a closed system of
blood vessels; it is pumped from the heart into arteries; from the arteries
into the capillaries, and from the capillaries into the veins to return back to
the heart.

► Blood is highly complex liquid connective tissue in which the cellular

elements are suspended in a liquid supporting or ground substance.
FUNCTIONS OF THE BLOOD

► 1. It transports oxygen from the lungs to the tissues and carbon dioxide from the
tissues to the lungs to be eliminated.
► 2. It serves as a vehicle for transport of food materials to the different tissues of
the body.
► 3. It picks up metabolic end products and delivers to other organ for use or
elimination.
► 4. It distributes the heat produced in active muscles and thus aids in the
regulation of body temperature.
► 5. It transports hormones from the glands in which they are produced to the
target organs.
► 6. Through the presence of buffer system, it helps maintain optimal pH of
the acid – base equilibrium.
► 7. It regulates the water balance through the effects of blood in the
exchange of water between circulating fluid and tissue fluid.
► 8. It serves in the mobilization of defense mechanism by protecting the
body against bacterial invasion and disease through the activities of
certain leukocytes and immune bodies in the blood stream.
► 9. It maintains a degree of irritability of the tissue cells so that functional
activity can be carried on satisfactorily.
COMPOSITION OF THE BLOOD

► A. Fluid portion – approximately 55%

► Blood plasma is the fluid portion of the blood. It is a straw colored fluid containing
a variety of substances and with complex chemical composition. It is the fluid
portion that remains when coagulation is prevented.
*** serum.
► Constituents of the liquid part:
► a. Water
► b. Inorganic constituents – sodium, calcium, potassium, magnesium, chloride,
bicarbonate, phosphate, sulfate
Plasma component…

► c. Organic constituents
► 1. Plasma proteins
► (a) Albumin – its primary function is to cause osmotic pressure at the capillary
membrane. This pressure called colloid osmotic prevents the fluid of the plasma from
leaking out of the capillaries into the interstitial spaces.
► (b) Globulin – alpha, beta and gamma globulins
► Alpha and beta globulins – transport substances by combining with them, acting as substrates
that react with other substances, transporting protein itself from one part of the body to
another.
► Beta and gamma globulins – protects the body against infection.
► Gamma globulins – antibodies that resist infection and toxicity thus providing the body with
immunity.
► (c) Fibrinogen – plays a role in blood clotting
Plasma proteins….

► d. Internal secretions (hormones), antibodies and various enzymes as

amylases, proteases, lipases, etc.
► e. Gases – oxygen, carbon dioxide, nitrogen
Buffy coat
layer
► B. Solid portion – approximately 41-45%
► The solid portion consists of the cellular elements:
► a. Red blood cells or erythrocytes
► b. White blood cells or leukocytes
► c. platelets or thrombocytes
RED BLOOD CELLS

► Anucleate, biconcave discoid cells

► Hemoglobin
► Abnormalities: Anemia, Polycythemia
► Tests:
► Hemoglobin (Cyanmethemoglobin method-Drabkin’s reagent)
► Hematocrit count/packed cell volume
► RBC count
► RBC indices (MCV, MCH, MCHC)
► RDW
► Reticulocyte count
White Blood Cells

► WBC count ( 4,500-11,500/uL)

► Leukocytosis - Leokocytopenia
► Leukemia – uncontrolled proliferation of WBC
► WBC differential count
► Neutrophils
► Bands
► Eosinophils
► Basophils
► Lymphocytes
► Monocytes
Platelets

► Platelet count
► Thrombocytosis - Thrombocytopenia
Complete Blood Count (CBC)

► RBC parameters: WBC parameters:

► RBC count *WBC count
► Hgb *Neut count % and absolute
► Hct *Lymph count % and absolute
► MCV *Mono count % and absolute
► MCH *EO and BASO count % and absolute
► MCHC Platelet parameters
► RDW *PLT count
► RETIC * MPV
Blood Film Examination
WBC differential count
Band/stab neutro vs
hypersegmented neutro
Other Hematological procedures:
Quality Assurance and Quality Control
Hematology Laboratory Safety
Applicable safety practices required
by the OSHA standard

► Proper Handwashing:
Applicable safety practices required by
the OSHA standard

► The following standards are applicable in a hematology laboratory and must be

enforced
► Hand washing is one of the most important safety practices. Hands must be washed
with soap and water. If water is not readily available, alcohol hand gels (minimum 62%
alcohol) may be used. Hands must be thoroughly dried. The proper technique for hand
washing is as follows:
► Wet hands and wrists thoroughly under running water.
► Apply germicidal soap and rub hands vigorously for at least 15 seconds, including
between the fingers and around and over the fingernails.
► Rinse hands thoroughly under running water in a downward flow from wrist to fingertips.
► Dry hands with a paper towel. Use the paper towel to turn off the faucet handles.
PPE

► Laboratory gown
► Gloves
► Eyewear
► Hairnet
► Face Mask
► Face shield
► Googles
► Closed flat shoes
Types of Hazard:

► Chemical
► The use of chemicals in research laboratories is inevitable, and the potential for harm
or injury could be significant if they are misused or mishandled
► OSHA Lab Standard,” requires laboratories to identify hazards, determine employee
exposures, and develop a chemical hygiene plan (CHP) including standard operating
procedures. The “lab standard” applies to the laboratory use of chemicals and
mandates written SOPs addressing the particular hazards and precautions required for
safe use.
► It requires providing material safety data sheets and employee training.
► Physical
► Included here are electrical safety hazards, ergonomic hazards associated with
manual material handling and equipment use, handling sharps, and basic
housekeeping issues.
► Biological
► Biological hazards encompass microbes, recombinant organisms, and viral
vectors. They also include biological agents introduced into experimental
animals
► Electrical
► Equip all electrical power outlets in wet locations with ground-fault circuit
interrupters, or GFCIs, to prevent accidental electrocutions. GFCIs are designed
to “trip” and break the circuit when a small amount of current begins flowing to
ground.
► Examples of sharps disposal systems. A
► , Molded foot pedal cart with hinged or slide top lid. B,
In-room system wall enclosures. C, Multipurpose container
with horizontal drop lid. D, Phlebotomy containers. Source:
(Courtesy Covidien, Mansfield, MA.)
Removal of gloves
Others:

► Housekeeping
► Laundry
► Vaccination
► Training and documentation
► Regulated medical waste management
► Occupational hazards
► Fire hazard – no smoking policy, fire extinguisher, alarms, smoke detectors,
sprinklers, manual fire alarm boxes near the exit doors.
► Drills, fire response plan, arrangement of combustible
ssifications and Use

Fire Extinguisher Classification and Use

Class/Type of Extinguisher Type of Fire
A Ordinary combustibles such as
wood, cloth, or paper.
B Flammable liquids, gases, or
grease.
C Energized (plugged-in) electrical
fires. Examples are fires involving
equipment, computers, fuse boxes,
or circuit breakers.

ABC Multipurpose for type A, B, and C

► The end
Lecture 2.1
Blood specimen collection
Blood Specimen Collection:

► Safety
► Standard precautions in blood collection (OSHA)
► Handwashing
► Responsibility of the phlebotomist in the infection control
► Quality Control (pre-ana, ana, post ana)
► Physiological factors (pre-ana)
► Posture * stress
► Diurnal rhythm * diet
► Exercise * smoking
Methods of Blood Collection

► Venipuncture
► Tourniquet
► Syringe with needle
► Vacutainer technique
► Butterfly technique

► Skin puncture
Capillary puncture

► Skin puncture or finger puncture

► Obtained from under the surface of the skin
► Ideal for tests requiring small amount of blood

► Ideal sites:
► Ring finger
► Great finger
► Later sides of the heels of infant ( heelstick)
Venipuncture

► Surgical puncture of a vein, also called phlebotomy

Site selection

► Sites for Venipuncture

► Most common: the inner arm at the
► bend of the elbow
► Back of the palm ( for difficult draw)
Materials used in VP

► Tourniquet
► Vacuum tubes and needle ( gauge 21-23)
► Syringe and needle
► Butterfly ( for difficult draws)
► Lavender-stoppered tubes ( EDTA)
► For whole blood collection
► Gray stoppered tubes
► Used in blood glucose test, contains oxalate
► Blue-stoppered tubes
► For testing prothrombin time, contains sodium citrate
► Green-stoppered tubes
► Contains heparin
► To determine several chemical constituents
Quality Assurance in Blood Collection

► Technical Competence
► Collection procedures
► Anticoagulants and other additives
► (Clot activators, antiglycolytic agent)
Requirements for a quality specimen are
as follows:
► 1. Patient properly identified
► 2. Patient properly prepared for draw
► 3. Specimens collected in the correct order and labeled correctly
► 4. Correct anticoagulants and other additives used
► 5. Specimens properly mixed by inversion, if required
► 6. Specimens not hemolyzed
► 7. Specimens requiring patient fasting collected in a timely manner
► 8. Timed specimens drawn at the correct tim
Reasons for specimen rejection

► 1. Test order requisition and the tube identification do not match.

► 2. Tube is unlabeled, or the labeling, including patient identification number,
is incorrect.
► 3. Specimen is hemolyzed.
► 4. Specimen was collected at the wrong time.
► 5. Specimen was collected in the wrong tube.
► 6. Specimen was clotted, and the test requires whole blood or plasma.
► 7. Specimen was contaminated with intravenous fluid.
► 8. Specimen is lipemic.
►Thank you for
listening!
 CELL
The Fundamental Unit Of Life
What is Cell?

Cell is the basic Structural and functional

unit of living organisms.

In other words, cells make up living things

and carry out activities that keep a living
thing alive.
Cell
Theory
Cell theory is a collection of ideas
and conclusions from many different
scientists over time that describes
cells and how cells operate.
1 A ll k n own livin g t h in g s a re
ma d e u p o f o n e o r mo re c e lls.
2 A ll livin g c ells arise f rom
p re - exist in g c e lls by d ivisio n .
3 Th e c e ll is t h e b a sic u n it o f
st ru c t u re a n d f u n c t io n in a ll
livin g o rga n isms.
 Cell Theory Timeline

1674
Anton Van Leeuwenhoek
Observed living cell

1665 1883
Robert Hooke Robert Brown
Discovered cell Discovered nucleus
 Cell Theory Timeline

1835 1839
Felix Dujardin J. E. Purkinje
Discovered fluid Named fluid content of
content of cell cell as protoplasm

1838
Matthias Schleiden
Proposed all plants are
made up of cells
 Cell Theory Timeline

1845
Carl Heinrich Braun
Proposed cell is the basic
unit of life

1839 1855
Theodor Schwann Rudolf Virchow
Proposed all animals Proposed all cells arise
are made up of cells from pre-existing cells
 Unicellular Organisms
An organism that is made up of only one cell is called as
unicellular organism.

Euglena Paramecium Yeast

 Multicellular Organisms
An organism that is made up of more than one cell is called as
multicellular organism.

Plants Animals Fungus

Multicellular Organisms Under Microscope

Leaf cells Muscle cells

Size of • Smallest cell

Cells
• Mycoplasma
• Size: 0.1 µm

Cells vary in size.

Most cells are very small

(microscopic), some may be
very large (macroscopic).
• Largest cell
The unit used to measure size • Ostrich egg
of a cell is micrometer.
• Size: 18 cm
1 µm = 1 / 1 0 00 m illi mete r
 Size of Cells in Humans

Smallest cell Largest cell Longest cell

Sperm cell Ovum cell Nerve cell
Size: 5 µm Size: 120 µm Size: 1 m

Shape of
Cells
Cells vary in shape.
Variation depends mainly upon
the function of cells.
Some cells like Euglena and
Amoeba can change their shape,
but most cells have a fixed shape.
Human RBCs are circular
biconcave for easy passage
through human capillaries.
Nerve cells are branched to
conduct impulses from one
point to another.
Human WBCs can change
their shape to engulf the
microorganisms that enter
the body.
Structure Compound
Of Cell microscope
Magnification 2000X

The detailed structure of a cell has

been studied under compound
microscope and electron microscope.

Certain structures can be seen only

under an electron microscope.
Electron
The structure of a cell as seen under microscope
an electron microscope is called
ultrastructure. Magnification 500000X
Animal Cell
12
11 1
10

1. Nucleus
2. Golgi body 9

3. Vesicle
4. Plasma membrane 8
5. Mitochondria
6. Cytoskeleton
7. Centriole
8. Lysosome 2

9. Cytoplasm 7 3
10. Rough endoplasmic reticulum
11. Smooth endoplasmic reticulum 4
12. Nucleolus 6
5
Plant Cell
12
11
1
10

1. Nucleus
2. Golgi body
9
3. Vesicle 2
4. Lysosome
5. Plasma membrane
8 3
6. Mitochondria
7. Chloroplast
8. Cell wall
4
9. Vacuole
10. Smooth endoplasmic reticulum 5
11. Rough endoplasmic reticulum 7

12. Nucleolus 6
Bacterial 9
10

Cell 8

1. Capsule
7
2. Cell wall
3. Plasma membrane 6
1
4. Cytoplasm
5. Flagellum
6. Food granule 2
7. Plasmid (DNA)
3
8. Ribosomes 4
5
9. Nucleoid
10. Pili
Structure
1. Pl asma Membrane
2. Nucleus

Of Cell
3. Cy toplasm

A. Cy tosol
B. Cell Organelles
If we study a cell under a microscope,
we would come across three features a) Endoplasmic reticulum
in almost every cell: plasma membrane, b) Go lgi bo dy
nucleus and cytoplasm. c) Lysosomes
All activities inside the cell and d) Vacuoles
interactions of the cell with its e) Mi tochondria
environment are possible due
f) Pl astids
to these features.
g) Centrosome
h) Cy toskeleton
Plasma Membrane • Extremely delicate, thin , elastic, living
and semi-permeable membrane
• Made up of two layers of lipid molecules
Carbohydrates in which protein molecules are floating
• Thickness varies from 75-110 A˚
• Can be observed under an electron
microscope only

Functions:
• Maintains shape & size of the cell
• Protects internal contents of the cell
• Regulates entry and exit of substances
Proteins in and out of the cell
Lipids
• Maintains homeostasis
 Cell wall • Non-living and outermost covering of a cell
(plants & bacteria)
• Can be tough, rigid and sometimes flexible
Pectin Cellulose
• Made up of cellulose, hemicellulose and
pectin
• May be thin or thick, multilayered structure
• Thickness varies from 50-1000 A˚

Functions:
• Provides definite shape, strength & rigidity
• Prevents drying up(desiccation) of cells
• Helps in controlling cell expansion
Plasma membrane Hemicellulose • Protects cell from external pathogens
 Nucleus
• Dense spherical body located near the
centre of the cell
Nucleus
• Diameter varies from 10-25 µm
• Present in all the cells except red blood
cells and sieve tube cells
• Well developed in plant and animal cells
• Undeveloped in bacteria and blue-green
algae (cyanobacteria)
• Most of the cells are uninucleated (having
only one nucleus)
• Few types of cells have more than one
nucleus (skeletal muscle cells)
 Nucleus
• Nucleus has a double layered covering called
Nuclear nuclear membrane
Nucleolus
pores
• Nuclear membrane has pores of diameter
about 80-100 nm
• Colourless dense sap present inside the
nucleus known as nucleoplasm
• Nucleoplasm contains round shaped
nucleolus and network of chromatin fibres
• Fibres are composed of deoxyribonucleic
acid (DNA) and protein histone
Chromatin • These fibres condense to form chromosomes
during cell division
Nuclear
envelope Nucleoplasm
 Nucleus
Gene DNA • Chromosomes contain stretches of DNA
called genes
• Genes transfer the hereditary information
from one generation to the next

Chromatin
Functions:
• Control all the cell activities like metabolism,
protein synthesis, growth and cell division
Histone
• Nucleolus synthesizes ribonucleic acid (RNA)
to constitute ribosomes
• Store hereditary information in genes
Chromatin fibre

Chromosome
Cytoplasm • Jelly-like material formed by 80 % of water
• Present between the plasma membrane
Organelles and the nucleus
• Contains a clear liquid portion called
cytosol and various particles
• Particles are proteins, carbohydrates,
nucleic acids, lipids and inorganic ions
• Also contains many organelles with distinct
structure and function
• Some of these organelles are visible only
under an electron microscope
• Granular and dense in animal cells and thin
in plant cells
Cytoplasm
 Endoplasmic • Network of tubular and vesicular structures
which are interconnected with one another
Reticulum • Some parts are connected to the nuclear
membrane, while others are connected to
the cell membrane
• Two types: smooth(lacks ribosomes) and
rough(studded with ribosomes)

Functions:
• Gives internal support to the cytoplasm
• RER synthesize secretory proteins and
membrane proteins
Rough ER
• SER synthesize lipids for cell membrane
Smooth ER • In liver cells SER detoxify drugs & poisons
• In muscle cells SER store calcium ions
Ribosomes
 Golgi body • Discovered by Camillo Golgi
• Formed by stacks of 5-8 membranous sacs
Incoming • Sacs are usually flattened and are called
Cis face transport the cisternae
Cisternae vesicle
• Has two ends: cis face situated near the
Lumen endoplasmic reticulum and trans face
situated near the cell membrane

Functions:
• Modifies, sorts and packs materials
synthesized in the cell
• Delivers synthesized materials to various
targets inside the cell and outside the cell
Outgoing • Produces vacuoles and secretory vesicles
Newly Trans face
forming transport • Forms plasma membrane and lysosomes
vesicle vesicle
 Nucleus
Smooth ER

Lysosomes

Golgi Body
At Work
Rough ER

Golgi body

Plasma
Vesicles
membrane
 Lysosomes • Small, spherical, single membrane sac
• Found throughout the cytoplasm
• Filled with hydrolytic enzymes
Hydrolytic enzymes Membrane • Occur in most animal cells and in few
type of plant cells

Functions:
• Help in digesting of large molecules
• Protect cell by destroying foreign
invaders like bacteria and viruses
• Degradation of worn out organelles
• In dead cells perform autolysis
Vacuoles • Single membrane sac filled with liquid
or sap (water, sugar and ions)
• In animal cells, vacuoles are temporary,
small in size and few in number
Tonoplast
• In plant cells, vacuoles are large and more
in number
• May be contractile or non-contractile

Functions:
• Store various substances including waste
products
• Maintain osmotic pressure of the cell
Vacuole • Store food particles in amoeba cells
• Provide turgidity and rigidity to plant cells
 Mitochondria • Small, rod shaped organelles bounded by two
membranes - inner and outer
• Outer membrane is smooth and encloses the
Ribosomes contents of mitochondria
Matrix • Inner membrane is folded in the form of shelf
like inward projections called cristae
Cristae
• Inner cavity is filled with matrix which contains
many enzymes
• Contain their own DNA which are responsible
for many enzymatic actions
DNA
Functions:
• Synthesize energy rich compound ATP
Outer membrane • ATP molecules provide energy for the vital
Inner membrane activities of living cells
Plastids
Plastids are double membrane-bound
organelles found inside plants and
some algae.

They are responsible for activities

related to making and storing food.

They often contain different types of

pigments that can change the colour
of the cell.
 Carrot
Pigment: Carotene

Chromoplasts

Chromoplasts are plastids that Mango

produce and store pigments Pigment: Xanthophyll

They are responsible for different

colours found in leaves, fruits,
flowers and vegetables.

Tomato
Pigment: Lycopene
 Potato tubers
Food: Starch

Leucoplasts

Leucoplasts are colourless Maize grains

plastids that store foods. Food: Protein

They are found in storage organs

such as fruits, tubers and seeds.

Castor seeds
Food: Oil
 Chloroplasts • Double membrane-bound organelles found
mainly in plant cells

Outer
• Usually spherical or discoidal in shape
Inner
membrane
membrane • Shows two distinct regions-grana and stroma
• Grana are stacks of thylakoids (membrane-
bound, flattened discs)
Thylakoid
• Thylakoids contain chlorophyll molecules
which are responsible for photosynthesis
• Stroma is a colourless dense fluid

Functions:

Stroma
• Convert light energy into chemical energy
in the form of food
Granum • Provide green colour to leaves, stems and
vegetables
 Centrosome
• Centrosome is the membrane bound organelle
Centrosome
present near the nucleus
matrix
• Consists of two structures called centrioles
• Centrioles are hollow, cylindrical structures
Microtubules
made of microtubules
• Centrioles are arranged at right angles to each
other

Functions:
Centrioles • Form spindle fibres which help in the movement
of chromosomes during cell division
• Help in the formation of cilia and flagella
 Cytoskeleton
• Formed by microtubules and microfilaments
Cell membrane
• Microtubules are hollow tubules made up of
protein called tubulin
• Microfilaments are rod shaped thin filaments
made up of protein called actin

Functions:
• Determine the shape of the cell
• Give structural strength to the cell
• Responsible for cellular movements

Microtubules
Microfilaments
Prokaryotic cell Eukaryotic cell

1. Nucleus is undeveloped 1. Nucleus is well developed

2. Only one chromosome is present 2. More than one chromosomes are present

3. Membrane bound organelles are absent 3. Membrane bound organelles are present

4. Size ranges from 0.5-5 µm 4. Size ranges from 5-100 µm

5. Examples: Bacteria and blue green algae 5. Examples: All other organisms
Animal cell Plant cell

1. G e n erall y s m a l l i n s i ze 1. G e n erall y l a rge i n s i ze

2. Ce l l wa l l i s a bs ent 2. Ce l l wa l l i s p resent

3. P l a st ids a re a bs ent 3. P l a st ids a re p re sent

4. Va c uo les a re s m a l l er i n s i ze 4. Va c uo les a re l a rger i n s i ze

and less in number a n d m o re i n n u m b e r

5. Ce nt r i oles a re p res ent 5. Ce nt r i oles a re a bs ent

THANK YOU...
Cell Cycle
• Interaction of Cdk’s & different cyclins triggers the stages of the cell
cycle