FORM NO.

F/ TL / 018
Rev.01 Date 20.10.2022
Page 1 of 2

Dr.MGR
EDUCATIONAL AND RESEARCH INSTITUTE
(Deemed to be University)

FACULTY OF PHARMACY

SUBJECT : HUMAN ANATOMY AND PHYSIOLOGY I

DEPARTMENT : PHARMACOLOGY

ACADEMIC YEAR : 2024 -2025

COURSE & YEAR : B.PHARM & I YEAR

SEMESTER : I SEM

FACULTY INCHARGE: Mrs. M. DEVI

Prepared by Reviewed by Approved by

Prepared by Reviewed by Approved by
 FORM NO. F/ TL / 018
Rev.00 Date 20.03.2020

INDEX

EXP. STAFF PAGE

DATE NAME OF THE EXPERIMENT
NO SIGN NO.
1 Study of Compound Microscope 01

2 Microscopic Study of Epithelial and 11

Connective tissues
3 Microscopic Study of Muscular and 23
Nervous tissues
4 Identification of Axial bones 27

5 Identification of Appendicular bones 33

6 Introduction to Hemocytometry 53

Enumeration of White Blood cell (WBC) 59

7
Count
8 Enumeration of Red Blood Corpuscles 63
(RBC) Count
9 Determination of Bleeding time 68

10 Determination of Clotting time 72

11 Estimation of Hemoglobin content 76

12 Determination of Blood Group 82

13 Determination of Erythrocyte 85
Sedimentation Rate (ESR) Determination
14 Determination of Pulse Rate/ Heart Rate 90,9
4
15 Recording of Blood Pressure 97

16 Determination of Differential Count 103

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LABORATORY GENERAL INSTRUCTIONS

1. All must bring your laboratory manual while coming to the laboratory
without fail, no one is allowed inside the lab without manual.
2. Before starting a new lab, you must always read the laboratory notes for
that experiment and the instructor will discuss the experiment with you.
If you don’t know the experimental procedure, the instructor may not let
you proceed with the experiment.
3. Notes must be made during each laboratory exercise.
4. Report to the instructor if you find equipment that is out of order or you
break something.
5. If you run out of some reagent, let the instructor know.
6. Do not smoke, eat, or drink in the laboratory.
7. Before beginning to work in the laboratory, you should be familiar
with the procedure you will be following, as well as with any special
precautions or changes that the instructor may note. Report any
unexpected events to the instructor.
8. No unauthorized experiments may be performed. Violators will be
subject to severe disciplinary action.
9. Before leaving the laboratory, wash your hands carefully.
10. Never taste or smell a chemical unless instructed to do so.
11. When smelling a chemical, fan vapors toward your face and
inhale cautiously.
12. Never pour water into concentrated acid; slowly add the acid to the
water with constant stirring in a Pyrex beaker or flask, not in a
graduated cylinder.
13. Never pipet liquids by mouth; use a suction bulb.
FORM NO. F/ TL / 018
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 Human Anatomy and Physiology I- Lab manual
EXP. NO.: 1 DATE:
STUDY OF COMPOUND MICROSCOPE

AIM: To study the different parts of a microscope.

OBJECTIVE: Microscopes are employed as a basic tool to observe microorganisms and to study
the structural details of the cells. Nowadays, highly sophisticated microscopes such as phase
contrast, dark field and electron microscopes are available but simple and compound microscopes
remain as a basic tool in routine microbiology research work.
The microscope is used in physiology to study the morphology of blood cells for making
different cell counts. A microscope magnifies the image of an object.
REQUIRMENT: Compound Microscope

DEFINITION:

A single magnified lens is known as a simple microscope. The compound microscope consists of
two magnifying lenses. They are the objectives and eyepiece. When the magnified image is
further magnified by one or more lens then it is known as a “Compound microscope”. A
compound microscope is a high power (high magnification) microscope that uses a compound
lens system.
PARTS OF THE COMPOUND MICROSCOPE

There are two commonly used compound microscopes. They are ‘monocular’ and ‘binocular’.
The monocular microscope has one eyepiece, whereas the binocular microscope has two eyepieces.
The structure of the compound microscope can be discussed under four main categories.

1. Support System (The framework)

2. The illumination system,

3. The Magnification system and

4. The Adjustment system.

1. THE SUPPORT SYSTEM

The support system is the framework of the microscope that holds the components of the
microscope.

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STRUCTURE OF COMPOUND MICROSCOPE

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The framework of the microscope consists of several units,

 BASE: The base supports the microscope and is horse – shoe shaped to give the maximum

stability to the microscope.

 PILLARS: Two upright pillars project upwards from the base and the handle of the

microscope is hinged to the pillars.

 HANDLE (ARM): The arm supports the magnification and adjusting systems.

 BODY TUBE: This is the tube that actually conducts the image.

 STAGE: The fixed stage is the horizontal platform on which the object being observed is

placed. Most microscopes have a mechanical stage which makes it much easier to

manipulate the objects being observed.

 NOSE PIECE: The fixed nose piece is attached to the lower end of the body tube and the

revolving nose piece carries the objective lens of different magnifying powers.

2. THE ILLUMINATION SYSTEM

The illumination system provides an uniform and soft –bright illumination of the entire

field viewed under the microscope.

 LIGHT SOURCE: The illumination system begins with a source of light. The

light source may be internal or external. The rays of light are reflected by a mirror towards

the object. The mirror is placed at the base of the microscope. It has two surfaces, ‘plane’

and ‘concave’. The plane mirror is used for the oil immersion objective whereas the

concave mirror is used for the low and high- power objectives.

 CONDENSER: The condenser focuses the rays of light reflected from the mirror onto the

object under examination and also helps in resolving the image. The position of the

condenser must be adjusted with each objective used to get the maximum focus of light and

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accurate resolving power of microscope. The condenser can be raised and lowered beneath

the stage by means of an adjustment knob.

o When a low power objective is used, the condenser is positioned at the lowest level;

with a high power objective, it is raised optimally and with an oil immersion

objective it is raised fully.

 IRIS DIAPHRAGM: The iris diaphragm regulates the amount of light that passes through

the material under observation. It is located at the bottom of the condenser. It has a central

aperture, which can be opened for more light or closed for less light according to necessity,

by means of a lever provided with the shutter.

3. THE MAGNIFICATION SYSTEM

The microscope magnifies the image of the object under view. The compound microscope

consists of two magnifying lenses, the eyepiece, and the objective. The total magnification

caused by a compound microscope is the product of the magnification caused by the

objective and that of the eyepiece.

i) EYEPIECE: The eyepiece or the ocular is the lens that magnifies the image formed

by the objective. It fits into the top of the body tube.

ii) OBJECTIVES: Objectives are the most important part of the magnification system.

The three objectives are,

a) 10 x : Low power objective,

b) 40 x or 45 x: High power objective, and

c) 90 x or 100 x: Oil immersion objective.

a) Low Power Objective: The low power objective is usually 10 x, which magnifies

the image 10 times.

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b) High Power Objective: It is usually a 40 x or 45 x magnification lens. It

magnifies the image 40 or 45 times. This objective is used for more detailed study

as the total magnification (with a 10 x eyepiece) is usually 400 or 450 times.

c) Oil Immersion Objective: The oil immersion objective is generally a 90 x or 100

x lens, which magnifies the image 90 or 100 times. The objective lens almost

rests on the slides when in use. It requires a special type of oil called ‘immersion

oil’, which is place between the objective and the slide. The most commonly used

immersion oil is cedar wood oil. Oil is used to increase the numerical aperture and

the resolving power of the objective.

4. THE ADJUSTMENT SYSTEM

The adjustment system consists of two adjustments:

i) Coarse adjustment and

ii) Fine adjustment.

i) COARSE ADJUSTMENT: Two coarse adjustment screws are used to makethe

coarse adjustment. These screws are mounted at the top of the handle by a

double-side micrometer mechanism, one on each side.

ii) FINE ADJUSTMENT: Two fine adjustment screws are used to make the fine

adjustment. Usually these screws are mounted in the handle below the coarse

adjustment screws by double side micrometer mechanism with one on each side.

These screws are operated for fine adjustment and exact focusing of the object.

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OTHER TYPES OF MICROSCOPES

1. Dark- Field microscope,

2. Fluorescence microscope,

2. Polarising microscope,

3. Phase- Contrast microscope,

4. Interference – Contrast microscope,

5. Electron microscope.

REPORT

Reference:

CL GHAI., A textbook of Practical Physiology, 8th Edition, JAYPEE Brothers.,Pg no: 2

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EPITHELIAL TISSUES

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EXP. NO.: 2 DATE :

MICROSCOPIC STUDY OF EPITHELIAL AND CONNECTIVE TISSUES

AIM
To study the microscopic structure, types and functions of human epithelial tissues and
Connective tissues.

REQUIREMENT: Permanent slides and Microscope

EPITHELIAL TISSUES
SQUAMOUS EPITHELIUM
Structure:
1. It is composed of a single layer of flattened cells
2. The cells are closely fitted together and arranged on a basement membrane
Sites:
It provides thin, smooth, inactive lining for the structures such as heart, blood vessels,
alveoli of lungs and lymph vessels.

Functions:
Its main function is to
1. Cover the structures
2. Protect the underlying organs
3. Absorb water
4. Secrete mucus
COLUMNAR EPITHELIUM
Structure:
1. This forms a single layer of cells shaped like elongated cubes.
2. Cells are column like closely fitted arranged on basement membrane.
Sites:
It lines the organs of the alimentary tract.
Functions:
1. It absorbs the product of digestion.
2. It is secret mucus.

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CUBOIDAL EPITHELIUM
Structure:
It consists of cube-shaped cells fitting closely together on the basement membrane
Sites:
It forms tubules of the kidney and the lining of glands.
Functions:
It involves in secretion and absorption.

STRATIFIED EPITHELIUM
Structure:
1. This is composed of several layers of cells of different shapes.
2. In the deepest layer, the cells are mainly columnar in shape.
3. The surface layer is composed of flat cells.
Sites:
It is found in
a) Conjunctiva of eye b) Lining of mouth
c) Lining of pharynx d) Lining of esophagus
Functions:
1. It protects the underlying structures.
2. It prevents drying of the cells in the deeper layer.

CILIATED EPITHELUM
Structure:
1. These are formed by columnar cells.
2. Cilia beats in the forward direction to propel the contents of the tube.
Sites:
1. It is found in the Fallopian tube of the female reproductive system, respiratory tract and
digestive system.
Functions:
1. In respiratory passage it propels mucus towards the throat. In the Fallopian tube, it propels
the ova towards the uterus.

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CONNECTIVE TISSUE

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TRANSITIONAL EPITHLEIUM
Structure:
1. It consists of 3 or 4 layers of cells and there occupies an intermediate position betweenthe
single-layered simple epithelium and many layered stratified epithelium.
2. The cells in the superficial layer are large, flat and irregularly quadrilateral.
Sites:
1. It is found in the pelvis , kidney, ureter and urinary bladder.
Functions:
1. Protective and prevents re absorption of excreted back to the excretory system.
2. Prevents drawing of water from blood and tissues.

CONNECTIVE TISSUES
ADIPOSE TISSUE
Structure:
1. It is a loose connective tissue.
2. It is composed of flat cells, containing fat globules.
3. These cells are put in varying numbers in matrix of aerolar tissue.
Sites:
1. These cells are put in the supporting organs such as kidney and eyes.
2. It is also found between bundles of muscle fibers.
Functions:
1. It reduces heat loss through skin.
2. It gives shape to limbs and body.
3. It serves as an energy reserve.
4. It protects the underlying organs from injury.

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CONNECTIVE TISSUE

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AREOLAR TISSUE
Structure:
1. It is a fibrous connective tissue.
2. Fibrocytes are embedded in the matrix which is semisolid.
3. Yellow elastic fibers appear singly and always branched.
Sites:
1. It is found in almost every part of the body in connecting and supporting other tissues, such as
a. Beneath the skin
b. Between the muscles
c. Supporting blood vessels and nerves
d. In the alimentary canal and
e. In gland supporting secretory cells
Functions:
1. It connects and gives mechanical support.

RETICULAR TISSUE
Structure:
1. Reticular tissue is a mesh-like, supportive framework for soft organs
Sites:
1. These are found in lymphatic tissue, the spleen, and the liver.
Functions:
1. Reticular cells produce the reticular fibers that form the network onto which other cells
attach. It derives its name from the Latin reticulus, which means “little net.”

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DENSE CONNECTIVE TISSUE

Dense connective tissue contains more collagen fibers than does loose connective tissue.
As a consequence, it displays greater resistance to stretching. There are two major categories of
dense connective tissue: regular and irregular. Dense regular connective tissue fibers are parallel
to each other, enhancing tensile strength and resistance to stretching in the direction of the fiber
orientations. Ligaments and tendons are made of dense regular connective tissue, but in
ligaments not all fibers are parallel. Dense regular elastic tissue contains elastin fibers in addition
to collagen fibers, which allows the ligament to return to its original length after stretching. The
ligaments in the vocal folds and between the vertebrae in the vertebral column are elastic.

ELASTIC CONNECTIVE TISSUE

The main fibers that form this tissue are elastic in nature. These fibers allow the tissues to
recoil after stretching. This is especially seen in the arterial blood vessels and walls of the
bronchial tubes

ELASTIC CARTILAGE
Structure:
1. It is yellow in colour and contains many elastic fibres.
2. It differs from hyaline cartilage only by the presence of its enormous elastic fibres in the
matrix.
3. The matrix, in addition to elastic fibres, contains collagen fibres.
Sites:
It is found in
a. External ear (pinna)
b. Eustachian tube and
c. In epiglottis
Functions:
1. To maintain rigidity and shape.
2. It maintains and strengthens the attached organs.

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CARTILAGE

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HYALINE CARTILAGE
Structure:
1. It appears as a smooth, bluish white tissue.
2. Under a microscope, the cell appears in a group of two or more and where they are in
contact with one another.
3. The matrix is solid and smooth.
Sites:
1. On the surface of parts of one which forms joints.
2. It forms the coastal cartilage which attaches the ribs and sternum.
Functions:
1. It covers and protects the joints of the body.

FIBRO CARTILAGE
Structure:
1. It consists of dense masses of white fibres in a solid matrix with the cells widely
dispersed.
2. It is tough and highly flexible tissue.
3. The matrix is solid and smooth.
Sites:
1. As a pad between the bodies of vertebrae, called “Inter vertebral disc”
2. Between the articulating surfaces of the bones of the knee joint known as “Semilunar
cartilages”.
3. Surrounding rims of bony sockets of the high and shoulder joints deepen the cavities.
Functions: Covers and protects the bony structure of the body.
Report

Reference: Ross and Wilsons., Human Anatomy and Physiology for health and illness, 18 th
Edition
S.R.Kale., R.R.Kale., Human Anatomy and Physiology., Nirali prakashan., Pg no: 47-60

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MUSCULAR TISSUES

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EXP. NO.: 3 DATE:

MICROSCOPIC STUDY OF MUSCULAR AND NERVOUS TISSUES

AIM
To study the microscopic structure and functions of human nervous and muscular tissues.

REQUIREMENTS: Permanent slides and Compound microscope

STRIATED MUSCLE
Structure:
1. The cells are cylindrical in shape.
2. Fibers are parallel to each other.
3. Each fibre has several nuclei and shows alternate dark and light bands i.e., striation.
Hence they are named as “Striated”.
4. These muscle fibers lines the skeletal muscles, hence they are also called “Skeletal
muscles”
Sites:
1. It is found in the lines of a skeleton.
Functions:
1. It involves contraction and relaxation.
SMOOTH MUSCLES
Structure:
1. The cells are spindle shaped.
2. Each fibre cell is mono nucleated.
3. Striations i.e., alternate dark and light bands are absence hence called “Smooth or non-
striated”
Sites:
1. It is found in
a. Walls of blood vessels
b. Walls of lymph vessels
c. Alimentary tract and respiratory tract
d. Urinary bladder and Uterus
Functions: It involves in contraction and relaxation of muscle fibers

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NERVOUS TISSUE

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CARIDAC MUSCLE
Structure:
1. This type of muscle tissue is found excessively in the walls of the heart.
2. Each fibre is parallel to each other but branched.
3. Each fibre has multiple nucleated.
4. Each fibre shows line which are thicker, darker
Sites:
1. Located exclusively in the walls of heart
Functions:
1. During contraction, the cardiac muscle propels blood from the auricle to the ventricle and
from theventricle to the pulmonary artery and aorta respectively.

NERVOUS TISSUE
Structure:
1. It consists of a vast number of units called “neurons”.
2. Each neuron is supported by a special type of connective tissue called “Neruoglia”
3. Each neuron consists of nerve cells, axons and dendrites.
4. Each nerve cell has an axon which carries nerve impulses.
5. A short and branched process called “dendrites” carries impulses towards the nerve cells.
Sites:
1. It is found in the brain and spinal cord.
Functions:
1. In co-ordination and controlling of nervous activities.
2. It involves the reception, discharge and transmission of stimuli.
Report

Reference: Ross and Wilson., Anatomy and Physiology in health and Illness., 12 th Edition.,
International edition
S.R. Kale., R.R. Kale., Practical Human Anatomy and Physiology.,Nirali Prakashan., Pg No: 47-
60.
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SKELETAL SYSTEM

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EXP. NO.: 4 DATE:

IDENTIFICATION OF AXIAL BONES

STUDY OF SKELETAL SYSTEM
AIM:
To study human skeletal system.

REQUIREMENT: Bone models and Chart

The skeletal system consists of two important divisions, namely

1. Axial skeleton

2. Appendicular skeleton

AXIAL SKELETAL SYSTEM: It consists of the skull, vertebral column and thoracic cage.

Skull: Two types of bones present in it are cranial bones and facial bones
Cranial bones: Facial bone:

 Frontal bone – 1  Zygomatic bones – 2

 Parietal bone – 2  Maxilla – 2

 Temporal bone – 2  Nasal bones – 2

 Occipital bone – 1  Lacrimal bones – 2

 Sphenoid bone – 1  Vomer – 1

 Ethmoid bone – 1  Palatine – 2

 Inferior nasal concha – 2

 Mandible -1

Vertebral column: The Vertebrmed by 5

types of bones

 Cervical vertebrae – 7
 Thoracic vertebrae – 12
 Lumbar vertebrae – 5

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BONES OF THE SKULL

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 Sacrum – 5 fused bones to form a single bone

 Coccyx – 4 fused bones to form a single bone

Thoracic cage:

It consists of two parts, namely Sternum and ribs

STUDY OF SKULL

The skull consists of two types of bone, namely Facial bones and cranial bones

Cranial bones:

Cranial bones are eight in number. They are

1. Frontal (1): It is the bone of fore head.

2. Parietal (2): These form the sides and roof of the skull which articulates with each other
at the sagittal sutures.

3. Occipital (1): This bone forms the back of the head and part of the base of the skull.

4. Temporal (2): These bones lie on each side of the head and forms fibrous immovable
joints with the parietal, occipital, sphenoid and zygomatic bones.

5. Sphenoid bone (1): This bone occupies the middle portion of the base of the skull and it
articulates with occipital, temporal, parietal and frontal bones.

6. Ethmoid bone (1): This bone occupies the anterior part of the base of the skull and
helps to form the orbital cavity.

There are four sutures present in the cranial bone

 Saggital sutures – between parietal bones

 Coronal sutures – between parietal and frontal

 Squamous sutures – between parietal and temporal

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BONES OF THE FACE

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 Lamboidal sutures – between parietal and occipital

Facial bones:

These are 14 in numbers which form the structure of the face.

 Zygomatic bones – 2

 Maxilla – 1

 Nasal bones – 2

 Lacrimal bones – 2

 Vomer – 1

 Palatine – 2

 Inferior conchae – 2

 Mandible -1

Report

Reference: Ross and Wilson., Anatomy and Physiology in health and Illness., 12 th Edition.,
International edition.

S.S.Randhawa., Atul Kabra., Human Anatomy and Physiology-I Practical Page no-21-23

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Exp. No.: 5 Date:

IDENTIFICATION OF APPENDICULAR BONES

AIM
To study the Human Appendicular Skeleton.

REQUIREMENTS: Appendicular bone models and charts.

APPENDICULAR SKELETAL SYSTEM:

The skeletal system consists of two parts

1. Shoulder girdle and upper limb

2. Pelvic girdle and lower limb

Shoulder girdle:

It forms two parts: Clavicle (Collar bone) and a Scapula (Shoulder bone).

Upper limb:

It consists of

 Humerus

 Ulna and Radius

 Carpals (Wrist bones) – 8

 Meta carpal bones – 5

 Phalanges – 14

Pelvic girdle:

It consists of Ilium, Pubis and Ischium

Lower limb:

It consists of

 Femur

 Tibia

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SCAPULA

CLAVICLE
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 Fibula

 Patella

 Tarsals (7)

 Meta tarsals (5)

 Phalanges (14)

The total number of bones present in our body is 206.

STUDY OF UPPER LIMB

Scapula:

1. Located in the posterior lateral region of the thorax.

2. It is a large flat triangular bone.

3. The Glenoid cavity for the articulation of the head of the humerus forms the shoulder joint.

4. There are two surfaces i.e., anterior and posterior, which provide fossa for attachment of
muscles

5. There are 3 angles and 3 borders.

Functions:

It provides articulation to the arm bones. It also provides attachment to the muscles of the
Arm.

Clavicle:

1. It is also known as “Collar bone”.

2. It is a long and curved bone of shoulder.

3. The sternal end articulates with sternum whereas the acromial end articulates with
acromion process of scapula.

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HUMERUS

RADIUS AND ULNA

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STUDY OF HUMERUS

 This is the bone of the upper arm. The head sits within the glenoid cavity of the scapula,
forming the shoulder joint.

 Distal to the head are two roughened projections of bone, the greater and lesser tubercles
and between them there is a deep groove, the bicipital groove or inter tubercular sulcus,
occupied by one of the tendons of the biceps muscle.

 The distal end of the bone presents two surfaces that articulate with the radius and ulna to
form the elbow joint.

 The Capitulum articulates with radius.

STUDY OF RADIUS AND ULNA

Ulna:

 It is the inner bone of the fore arm. The shaft of ulna gives attachment to the muscles of
fore arm.

 The lower part of ulna contains head and styloid process.

Radius:

 It is the outermost bone of the fore arm.

 The upper part of radius contains a head, neck and biceps tuberculi.

 The shaft gives attachment to muscles.

 The lower end of the radius forms the wrist joint with 3 carpal bones.

 It also forms radio-ulnar joint.

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BONES OF THE HAND

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CARPAL BONES

There are 8 carpal bones arranged in two rows of 4 forms outside inwards.

Proximal row: There are 4 bones

 Scaphoid bone

 Lunate bone

 Triquetral bone

 Pisiform bone

Distal row: There are 4 bones

 Trapezium

 Trapezoid

 Capitates

 Hamate

Features:

 Trapezium, trapezoid, capitates, hamate are closely fitted together and held in position by
ligaments which allow a certain movement.

 The proximal row is associated with the wrist , while the distal row forms joints with
metacarpal bones and the muscles of the fore arm move the wrist joints.

Meta carpal bones:

 These are 5 in number and form the structure of the palm of the hand.

 They are long, slender bones. The proximal ends of which articulate with the carpal
bones and the distal end with phalanges.

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Phalanges

 There are 14 phalanges arranged so that 3 in each finger two in the thumb.

 The proximal phalanges of each finger is the largest and articulate with corresponding
Meta carpal bone at another end with middle phalanx of other end.

 The distal phalanx is the smallest and forms the top of the finger.

 The thumb which is shortest of the digits has only two phalanges.

STUDY OF LOWER LIMB

Pelvic bone:

The pelvic girdle is formed from two innominate (hip) bones. The pelvis is the term given
to the basin-shaped structure formed by the pelvic girdle and its associated sacrum.

Innominate (Hip) bone:

 Each hip bone consists of three fused bones namely

 Ileum

 Ischium

 Pubis

On its lateral surface is a deep depression the acetabulum, which forms the hip joint with the
almost spherical head of femur.

 The ileum: The upper flattened part of the bone and its presents the iliac crest.
The anterior curve of which is called “Anterior superior iliac spine”.

 The ileum forms a synovial joint with the sacrm, the sacro-iliac joint, a strong joint
capable of absorbing the stresses of weight bearing and which tends to become fibrosed
in later life.

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FEMUR

PATELLA

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 The ischium: It is the inferior and posterior part. The rough inferior projections of the
Ischia, the ischial tuberositis bear the weight of the body when seated.

 The pubis: It is the anterior part of the bone and it articulates with the pubis of the other
hip bone at a cartilaginous joint, the symphysis pubis.

 The union of the three parts takes place in the acetabulum.

The pelvis:

 The pelvis if formed by the hip bones, sacrum and coccyx.

 It is divided into upper and lower part by the brim of the pelvis.

 Brim of the pelvis consisting of the promontory of the sacrum and ilio-pectineal
lines of the innominate bones.

 The greater or false pelvis is above the brim and the lesser or true pelvis is below the
brim.

STUDY OF FEMUR

Each leg contains 30 bones in it. They are

 Femur (Thigh bone) – 1

 Patella (Knee bone) – 1

 Tibia and Fibula (Leg bone) – 2

 Tarsals (Aukle) – 7

 Metatarsals (sole) – 5

 Phalanges (Toes) – 14

Femur:

1. It is thigh bone. It is the longest and strongest bone of the body.

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TIBIA & FIBULA

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2. The head is round and articulates with the acetabulum of innominate bone.

3. The head has body ridges called “Trochanters”.

4. The shaft which is smooth, cylindrical and rounded in front and the sides.

5. It contains linea aspera which is a ridge on the poseteior.

6. Gluteal ridges extended from linea aspera to the back of greater trochanter.

Patella:

1. It is a sesmoid bone.

2. The posterior surface is smooth.

3. The anterior surface is covered with bursa.

4. It forms the knee joint with the femur.

STUDY OF TIBIA AND FIBULA

Tibia:

 It is the innermost bone of the leg.

 The upper end possesses a head which articulates with condyles of femur.

 The shaft is triangular in shape

 The lower end takes part in the formation of ankle joint.

Fibula:

 It is the outer bone of the leg.

 The upper end contains head and styloid process.

 The shaft gives attachment to muscles.

 The lower end contains lateral malleolus and malleolar fossa.

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BONES OF THE FOOT

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STUDY OF BONES OF FOOT

Tarsal bones:

They include calcaneum, talus, navicular, cuboid and three cuneiform bones.

Metatarsal bones:

 They are 5 in number.

 They correspond with the five toes. All of them are long bones.

 They contain a head shaft and base.

 The first metatarsal is thick and stout. The second metatarsal is longer than others. The
fifth one has a projection at the lateral side of the base.

Phalanges:

 There are 14 bones. Two for the first two toes and three for the rest.

 All of them are long bones.

STUDY OF VERTEBRAL COLUMN

There are 26 bones in the vertebral column, of which 24 are separate vertebrae and one
sacrum and coccyx.

The vertebral column is divided into different regions.

 Cervical vertebrae – 7

 Thoracic vertebrae – 12

 Lumbar vertebrae – 5

 Sacrum formed from 5 fused vertebrae

 Coccyx formed from 4 fused vertebrae.

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SINGLE VERTEBAL BONE

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Each vertebrae is identified by the first letter of its region in the spine, followed by a
number indicating its position.

For example, the top most vertebrae is called as C1 and the third lumar vertebrae is
called as L3.

STUDY OF CERVICAL VERTEBRAE

Cervical vertebrae

 These are the smallest vertebrae. The transverse process has a foramen through which a
vertebral artery passes upwards to the brain. The first two cervical vertebrae, the atlas and
the axis are typical.

 The first cervical vertebrae, the atlas, is the bone on which the skull rests. Below the atlas
is the axis, the second cervical vertebrae C2.

 The atlas is essentially a ring of bone with no distinct body orspinous process although it
has two short transverse processes. It possesses two flattened facets that articulate with
the occipital one these are condyloid joints and they permit nodding of the head.

 The axis sits below the atlas, and has a small body with a small superior projection called
the “Odontoid process”. This occupies part of the posterior foramen of the atlas above
and is held securely within it by the transverse ligament.

 The seventh cervical C7 is also known as the “Vertebral prominens”.

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STUDY OF THORACIC AND LUMBAR VERTEBRAE

Thoracic vertebrae:

 The thoracic vertebrae are larger than the cervical vertebrae because this section of the
vertebral column has to support more body weight.

 The bodies and transverse processes have facets for articulation with the ribs.

Lumbar vertebrae

 These are the largest of the vertebrae because they have to support the weight of the
upper body.

 They have substantial spinous processes for muscle attachment.

STUDY OF SACRUM AND COCCYX

Sacrum

1. This consists of five rudimentary vertebrae fused to form a triangular or wedge shaped
bone with a concave anterior surface.

2. The upper art or base articulates with the 5th lumbar vertebrae.

3. On each side it articulates with the ilium to form a “Sacro-iliac joint” and at its inferior
tip it articulates with the coccyx.

4. The anterior edge of the base, the promontory, protrudes into the pelvic cavity.

5. The vertebral foramina are present and on each side of the bone there is a series of
foramina for the passage of nerves.

Coccyx:
This consists of 4 terminal vertebrae fused to form a small triangular bone. The broad
base of which articulates with the tip of the sacrum.
Report

Reference: Ross and Wilson., Anatomy and Physiology in health and Illness., 12 th Edition.,
International edition.
S.S.Randhawa, Atul Kabra, Human Anatomy and Physiology I Practical., Page no: 24-26

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EXP. NO.: 6 DATE:

INTRODUCTION TO HEMOCYTOMETRY
Aim

To study about Haemocytometry

OBJECTIVE

Haemocytometry is the study of counting the number of cells in a blood sample. Since, the whole

blood contains a large number of blood cells; it is difficult to count them under a microscope. This

difficulty can be overcome by diluting blood with suitable diluting fluid and then counting them.

The whole process of counting blood cells involves.

1. Collection of blood sample from the finger prick.

2. Filling the blood in a pipette.

3. Dilution of blood with suitable diluting fluid.

4. Filling counting chamber with diluted blood.

Counting cells and reporting the result in terms of the number of cells per cubic millimetre.

Blood cell counting is performed using specific apparatus called haemocytometer consists of

1. Diluting pipettes

2. Counting chamber/ Neubauers Chamber

3. Cover slip.

DILUTING PIPPETTES

Two different glass capillary pipettes having a bulb are provided for diluting the blood for RBC

and WBC counting. The pipette has a long narrow stem with a capillary bore. It is divided into ten

equal parts and only two markings are numbered ie., 0.5 in the middle of the stem, 1.0 at the

junction of the stem and bulb.

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Differentiation of RBC and WBC pipettes are shown in the table.

RBC pipette WBC pipette

The Calibration is 0.5 and 1.0 below the bulb The Calibration is 0.5 and 1.0 below the bulb and
and 11above the bulb.
101 above the bulb.
The capillary bore is narrow, thus it is a The capillary bore is narrow, thus it is a fast
slowspeed pipette. speed
pipette.
The bulb is larger and has a red bead. The bulb is smaller and has a white bead.
The volume of the bulb is 100 times to The volume of the bulb is 10 times to the
the volume contained in stem. volume contained in stem.

Dilution can be 1: 100 or 1 : 200 Dilution can be 1: 10 or 1 : 20

COUNTING CHAMBER

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The stem widens into a bulb containing a free rolling bead which helps in mixing blood with

diluting fluid. The color of bead is read in RBC pipette and white in WBC pipette.

The bulb narrows again into short stem to which a rubber tube is attached with wearing a mouth

piece (Red mouth piece for RBC pipette and white mouth piece for WBC pipette).

NEUBAUER’S CHAMBER OR COUNTING CHAMBER

The counting chamber is a single solid heavy glass slide. There are three parallel platforms or pillar

separated from each other by shallow trenches extending across middle third portion of slide. The

central platform is wider and exactly 0.1mm lower than the two lateral platforms. It is divided into

two equal parts by a short transverse trench in its middle. Thus, there is H shaped trench enclosing

the two platforms. The lateral platforms support the coverslip and central platforms contains

counting grid.

THE COUNTING GRID

Each counting grid measures 9 mm2 (3mm x 3mm) and divided into 9 squares each of 1mm 2

(1mm x 1mm).

Out of these 9 squares four large corner squares are used for counting WBC’s. Each large square is

divided by single lines into 16 medium sized squares. Each of these medium sized squares has a

side of 1/4mm and a area of 1/16mm2 (1/4mm x 1/4mm).

The central large square is divided into 25 medium sized squares each is having aside of 1/5mm.

These medium squares are separated by double or triple lines. These lines extend in all direction

beyond the boundaries of 9mm square i.e., in between all the WBCs squares around the central

RBCs square. Each of the 25 medium squares is further divided into 16 smallest squares by single

lines. These smallest squares are having a side of side of 1/20 mm and an area of 1/400 mm2. RBC

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are counted in four corner and one central medium square each containing 16 smallest squares i.e.,

in a total of 80 smallest squares.

Report

Reference: CL GHAI., A textbook of Practical Physiology, 8th Edition, JAYPEE Brothers.,

Pg no: 23-30

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W.B.C. COUNT

CALCULATION

Dilution factor = 1:20 (i,e.20)

Depth factor = 10

Area counted = 4 Sq.mm

Number of cells counted= N

RBC Count = Number of cells counted X Dilution factor X Depth factor

-----------------------------------------------------------------------
Area counted

RBC Count = N X 20 X10

----------
4
= N X 50 Cells/ Cu.mm of blood

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EXP. NO.: 7 DATE:

ENUMERATION OF WHITE BLOOD CELL (WBC) COUNT

AIM

To enumerate the number of white blood cells in one cubic millimeter of blood of your own blood.

PRINCIPLE

Blood is diluted with suitable solution, which removes the red cells by hemolysis and accentuates

the nuclear of white cells. The WBC diluting fluid is Turk’s fluid. Counting is done under low

power and with the knowledge of the volume of dilution fluid obtained; the number of white blood

cells per cubic millimeter per mm3 of undiluted blood is calculated.

REQUIREMENTS

APPARATUS

 Microscope,

 Hemocytometer (WBC diluting pipette and counting chamber),

 Equipment for a sterile finger prick,

 Watch glass,

 Cover slip.

WBC DILUTING FLUID (TURK’S FLUID)

Composition

 1 per cent of glacial acetic acid solution (destroys RBC)

 Gentian Violet or aqueous Methylene blue 0.3 per cent W/V (stains the nuclei of the

leucocytes).

 Distilled water (solvent).

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PROCEDURE

 Clean and dry the necessary apparatus.

 Prick the finger under aseptic conditions.

 Take necessary precautions and suck blood up to 0.5 marks in WBC pipette.

 Immediately suck diluting fluid up to mark 11.

 Keep the pipette in horizontal position and thoroughly mix the contents for about 5 to

10 minutes.

 Discard first two drops and charge the Neubauer’s chamber.

 Allow time for the cells to settle down.

 Count the WBCs in the four corner squares under low power.

 WBCs are identified as clear nucleated and refractile bodies.

DILUTION OBTAINED

Blood is drawn up to 0.5 and dilution fluid up to 11. Volume of the bulb is 10 (11-1). Dilution

takes place only in the bulb and the fluid in the stem (from tip of the pipette to mark 1) does not

take part in dilution and is discarded before charging the chamber. So when 0.5 in 10 is doubled we

get 1 in 20.

RESULT

The total WBC count of my blood was found to be .

Reference: CL GHAI., A textbook of Practical Physiology, 8 th Edition, JAYPEE Brothers.,Pg no:

60-67

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R.B.C. COUNT CALCULATION

Dilution factor = 1:200 (i,e.200)

Depth factor = 10
80 1
Area counted = ---------- = -------- Sq.mm
400 5
Number of cells counted= N

RBC Count = Number of cells counted X Dilution factor XDepth factor

-----------------------------------------------------------------------
Area counted

RBC Count = N X 200X10

----------
1/5
= N X 10,000 Cells/ Cu.mm of blood

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EXP. NO.: 8 DATE:

ENUMERATION OF TOTAL RED BLOOD CELLS (RBC) COUNT

AIM

To enumerate the number of red blood cells in one cubic millimeter of my own blood.

PRINCIPLE

The blood specimen is diluted (usually 200 times) with the diluting fluid, which does not remove

the white cells, but allows the red cells to be counted in a known volume of fluid. Finally, the

number of cells in undiluted blood is calculated and reported as the number of red cells per cubic

millimeter of whole blood.

REQUIREMENTS

APPARATUS
 Microscope,
 Hemocytometer (RBC diluting pipette and counting chamber),
 Equipment for a sterile finger prick.
 Watch glass,
 Cover slip.

RBC DILUTING FLUID

The RBC diluting fluid is isotonic therefore it prevents hemolysis. Of the different diluting fluids
used for red cell count the most frequently used one is Hayem’s fluid.
 Hayem’s fluid Composition:
 Sodium Chloride: 0.5 grams (maintains osmolarity )
 Sodium Sulfate: 2.5 grams (prevents aggregation of RBC)Mercuric Chloride: 0.25
grams (acts as a preservative, it is anti-fungal and anti-bacterial)
 Distilled water: 100 ml (act as a solvent)

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PROCEDURE

 Assemble equipment needed for practical and ensure they are clean and dry.

 Prick the finger under aseptic condition, suck blood into the pipette upto the 0.5

mark, and suck the diluting fluid upto mark 101.

 Hold the pipette horizontally, close both ends, and gently mix the contents of the

bulb until the blood is thoroughly mixed with the diluting fluid.

 Discard the first two drops of fluid from the pipette.

 Charge the Neubauer’s chamber and allow two minutes for the cells to settle down.

 Focus the cells under low power and then under high power.

 Count the red blood cells in 80 small squares. Care should be taken not to count the

same cells again.

DILUTION OBTAINED

Blood is drawn up to 0.5 and dilution fluid drawn upto 101. Volume of blood is 100 (101-1).

Dilution takes place only in the bulb, and the fluid in the stem (from the tip of the pipette to mark

1) does not take part in dilution and is discarded before charging the chamber. So when 0.5 in 100

is double what we get the dilution as 1 in 200.

RESULT

The RBC count of my blood was found to be .

Reference:
CL GHAI., A textbook of Practical Physiology, 8th Edition, JAYPEE Brothers.,Pg no: 45- 52

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EXP. NO.: 9 DATE:

DETERMINATION OF BLEEDING TIME

Aim

To determine the bleeding time of my own blood.

METHODS OF DETERMINATION

Bleeding time is generally determined by two methods:

1) Duke method,

2) lvy method.

DUKE METHOD

The Duke method is more frequently used method to determine bleeding time in clinical laboratory

as it is easy to perform and requires minimal equipment and laboratory skill.

PRINCIPLE

A deep skin puncture is made and the length of time required for bleeding to stop is recorded. It

determines the function of the platelet and integrity of the capillary. The bleeding time is increased

in conditions such as scurvy, aplastic anemia, multiple myeloma and allergic conditions, infectious

mononucleosis and glannzman’s disease. The bleeding time is increased because of shortage of

platelets, deficiency of plasma factors and inadequate functions of platelets.

EQUIPMENTS

 Equipment for a sterile finger prick,

 Blotting paper or filter paper,

 Stop watch.

PROCEDURE

 Assemble all necessary materials.

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 Clean the fingertip with alcohol and allow the skin to dry completely,

 Make a deep puncture with the help of a sterile lancet.

 Immediately start the stopwatch.

 Blot the drop of blood coming out of the incision every 30 seconds by using the blotting

paper or filter paper. Place a subsequent drop of blood a little further along the side of

the filter paper.

 Stop the stopwatch as soon bleeding ceases.

 Count the number of drops on the filter paper and multiply it by 30 seconds.

NORMAL VALUES:
 1-9 minutes: Normal
 Normal bleeding time:
 Duke method – Less than 3 minutes
 IVY method – Less than 8 minutes
 Prolonged bleeding times greater than 5 minutes in the Duke method and
10 minutes in the IVY method are concerning for coagulopathy.
Abnormalities would require further evaluation with a focus on the
coagulation pathway of interest.
 9-15 minutes: Platelet dysfunction

REPORT

The bleeding time of my own blood was found to be .

Reference: S.R. Kale., R.R. Kale., Practical Human Anatomy and Physiology.,Nirali Prakashan.,
Pg No: 45-46.

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EXP. NO.: 10 DATE:

DETERMINATION OF CLOTTING TIME

Aim

To determine the clotting time of my own blood.

Requirements

Sterile needle, Capillary tube, cotton, spirit and stop watch.

Principle

Whenever a blood vessels rupt ure, bleeding is continuous for a few minutes. Blood
loses its fluidity and sets into a semisolid mass. This mass is referred to as “Clot” and the
phenomena is known as “Coagulation”.

Clotting time is the time interval between onset of bleeding time and the appearance of a
semisolid mass that is “Clot’. The method is called “Capillary glass method”.
Method: Capillary tube method

Procedure:

1. Sterilize the finger tip. Take a blood prick to have a free flow of blood.

2. Suck blood in a capillary glass tube of 15 cm long.

3. A small bit of glass tube of 1 cm is carefully broken for every 30 sec until a fine thread of
clotting blood appears while the capillary tube is being broken.
4. The period of in between appearance of blood in finger and formation of clot is taken as

“Clotting time”.

The normal value is 3 to 7 min.

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Anticoagulants

It is a substance that po st po ne s the process of coagulation. The commonly used

anticoagulants are sodium citrate, sodium oxalate and others are heparin and potassium fluoride.

Report:

My own clotting time according to Wright’s capillary method is min.

Reference: S.R. Kale., R.R. Kale.,Practical Human Anatomy and Physiology.,Nirali Prakashan.,
Pg No: 33-34.

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EXP. NO.: 11 DATE:

ESTIMATION OF HEMOGLOBIN CONTENT

Aim

To estimate the concentration of hemoglobin (Sahli’s acid haematin method) of my own blood.

PRINCIPLE

Hemoglobin is converted to acid haematin by addition of N/10 HCI. The acid haematin solution is

further diluted until its color matches with that of permanent standard of the comparator box.

Hemoglobin concentration is read directly from the calibration tube.

DIFFERENT METHODS

Different methods of estimation of hemoglobin can be classified into the following categories:

1) VISUAL METHODS:

a) Sahli’s method

b) Dare’s method

c) Haden’s method

d) Wintrobe’s method

e) Haldane’s method

f) Tallquist’s method

2) GASOMETRIC METHOD

3) SPECTROPHOTOMETER METHOD

4) AUTOMATED HEMOGLOBINOMETRY

5) NON-AUTOMATED HEMOGLOBINOMETRY,

6) OTHER METHODS.

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Significance
The variance in normal value of Hb concentration is observed during the pregnancy and
disease conditions and it also depends upon age, sex attitude. The new born infant having higher
Hb value than adults. At high altitude, the 0.2 content of earth decreases the RBC count and Hb
value increases. In pregnancy, body gains fluid and red blood cells become less count and Hb.The
determination of Hb content it shows some diagnostic appearance. The value drops below normal
in anaemia, leukemia, while its value rises below normal in dehydration conditions and
polycythemia.
SAHLI’S ACID HAEMATIN METHOD

REQUIREMENTS

Sahli’s haemoglobinometer. It contains a comparator, hemoglobin tube, hemoglobin pipette, and

stirrer.

PROCEDURE

 Fill haemoglobinometer tube with N/10 HCI upto its lowest mark (2 grams %)

 Prick the finger under aseptic conditions, suck blood into 20 cubic millimeter mark of

pipette and transfer it into tube containing N/10 HCI.

 Mix it thoroughly with a stirrer.

 Leave the solution in the tube for 10 minutes for maximum conversion of hemoglobin

to acid haematin which occurs in the first 10 minutes.

 After 10 minutes dilute the acid haematin with distilled water drop by drop, mix it with

stirrer.

 Match the color of the solution in the tube with the standard in the comparator.

 While matching hold the stirrer above the level of the solution.

 Note the reading when the color exactly matches with the standard.

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Formula

% of Hb = Colour index = Total % of Hb X 100

Total % of RBC

(or) standard formula = 14.5 gm of Hb = 100%

Test value

Reading of Hb= X 100

Standard value

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 Express the hemoglobin concentration as grams per 100 ml of blood.

Report

The hemoglobin content of my blood is gm / 100 ml

Reference: S.R. Kale., R.R. Kale., Practical Human Anatomy and Physiology.,Nirali Prakashan.,
Pg No: 05-09.

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ANTI-A SERUM ANTI-B SERUM AGGLUTINOGENS BLOOD GROUPS

(in RBC)

+ - A A

- + B B

+ + AB AB

- - Nil O

+ : Agglutination - : No Agglutination

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EXP. NO.: 12 DATE:

DETERMINATION OF BLOOD GROUP

Aim

To determine the blood group of my own blood sample.

Apparatus Requirement

Porcelain tiles, antiseptic solution, antiserum A, antiserum B, anti Rh serum, needle and
cotton.

Principle

The ABO and Rh blood grouping system is based on agglutination reaction. When red blood
cells carrying one or both the antigens are exposed to the corresponding antibodies, they interact
with each other to form visible agglutination or clumping. The ABO blood group antigens
are O-linked glycoproteins in which the terminal sugar residues exposed at the cell surface of
the red blood cells determine whether the antigen is A or B. Blood group A individuals have A
antigens on RBCs and anti-B antibodies in serum. Similarly, blood group B individuals have B
antigens on RBCs and anti-A antibodies in the serum. Blood group AB individuals have both A
and B antigens on RBCs and neither anti-A nor anti-B antibodies in serum. Whereas, blood
group O individuals have neither A antigens nor B antigens, but possess both anti-A and anti-B
antibodies in serum. The Rh antigens are trans membrane proteins in which the loops exposed on
the surfaceof red blood cells interact with the corresponding antibodies

Procedure

Sterilize your finger tip. Give one bold prick. Discard the first drop of blood. Place the
second, third and fourth on the previously cleaned tiles, which are already divided into 3 portions
with the help of a glass marking pencil. Now immediately add anti serum A, anti-serum B and
anti Rh serum. Mix thoroughly with 3 separate sticks. Wait for some time. Watch the
development of agglutination formed, the group of blood is determined. If the agglutination is
seen in antiserum

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‘A’, it denotes the blood group is “A”. If it is seen in antiserum ‘B’, then it is “”B group. If both
‘A’ and ‘B’ has agglutination it denotes “AB” group. If there is no agglutination seen in both ‘A’
and ‘B’ then it is “O” group. With the anti Rh presence of agglutination confirms Rh + ve and
absence confirms Rh – ve.

Report

My blood group is

Reference: S.R. Kale., R.R. Kale., Practical Human Anatomy and Physiology.,Nirali Prakashan.,
Pg No: 34-35.

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WESTERGREN PIPETTE WITH STAND

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EXP. NO.: 13 DATE:

DETERMINATION OF ERYTHROCYTE SEDIMENTATION RATE (ESR)

AIM

To determine the erythrocyte sedimentation rate of your own blood.

METHODS OF DETERMINATION

There are two traditional methods for determining ESR (Westergren’s and Wintrobe’s) methods

WESTERGREN METHOD

PRINCIPLE

Anti- coagulated blood is taken in a pipette and left undisturbed in a vertical position. The

erythrocytes settle at the bottom due to density and specific gravity as compared to blood plasma

which accumulates above it. The level of the column of red cells is noted in the beginning (0 h) and

after 1 and 2 hours. The distance in (mm) the column moves is noted as the ESR (mm/h).

REQUIREMENTS

APPARATUS REQUIRED

Westergren pipette is an open-ended tube. It is 300 mm in length with an internal bore of 2.5

mm. It is graduated from 0 to 200 mm along the lower two thirds of its length. The graduated

volume of the pipette is 1.0 ml. The pipettes are held vertically in the Westergren rack after being

filled with blood and the rack is provided with rubber pads at the lower end and metal clip at the

upper end.

The Westergren Rack is a special rack designed to hold the Westergren pipette in a vertical

position. It is constructed in such a way that the rubber stoppers attached to the springs close the

open ends of thetubes when they are placed in the rack.

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BLOOD SAMPLE

Anti-coagulated blood is taken for study. Sodium citrate solution (3.8%) is the preferred anti

coagulant. The ratio of the blood to the anti coagulant is 4.1 i.e. 4 parts of blood (say 2 ml) is

mixed with one part of anti-coagulant (0.5 ml). EDTA can also be used.

PROCEDURE:

 Mix the blood thoroughly by inversion or swirling.

 Fill the citrated blood into the Westergren pipette upto the 0 mark making sure that no

air bubbles in the blood column drawn through the tube.

 Immediately close the upper end of the Westergren tube to prevent the blood from

running down.

 Place the Westergren tube in the rubber pad of the Wetergren rack and fix it vertically

with the metal clips provided on the rack.

 Note the time and allow the tube to stand for 1 hour.

 Record your observation (note the level to which the red cell column has fallen) after 1

hour.

NORMAL VALUE

In male: 0-9mm

In female: 0-20mm

REPORT
The erythrocyte sedimentation rate of my own blood was found to be .

Reference: CL GHAI., A textbook of Practical Physiology, 8 th Edition, JAYPEE Brothers.,Pg no:

93-97
S.S.Randhawa, Atul Kabra, Human anatomy and Physiology I., Practical, Page no: 44

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EXP. NO.: 14 DATE:
a. DETERMINATION OF PULSE RATE

Aim
To determine my own pulse rate.

Principle

Pulse can be defined as the wave of distension which moves along the artery in such a way
that it coincides with the beat of the heart. The elastic walls of the artery expand when blood flows
through it with pressure. This pressure is exerted when the blood is forced out of the left ventricle.
With each beat, an additional amount of blood is forced out of the heart into the arteries. It is a type
of wave that passes along the artery, therefore it is also called pulse wave. The expansion and
contraction of the artery with each wave can be felt with finger tips. The rate of contractions and
expansions of the pulse not only indicates the condition of the heart but also the general condition
of the patient. One can feel the pulse in the radial, brachial, carotid, femoral, axillary and temporal
arteries.
Significance

The examination of the pulse is of a great clinical importance to the condition of the heart, the
condition of arteries, the extent of blood pressure etc., can be known.

The following features are to be noted during the examination of the pulse.

a) Rate: It means frequency of pulse per minute.

b) Rhythm: It indicates whether the beats are equidistant or not.

c) Volume: It means rise of pulse way above diastolic level.

d) Tension: It is an approximate measure of systolic pressure.

Procedure

Palpate the superficial arteries by pressing them against the underlying bone. Usually the
radial arteries at the wrist level are taken. The pulse rate is recorded for 1 minute.
Normal value

1. In new born babies, the pulse rate – 135 to 140 beats/min.

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OBSERVATION TABLE
S.no Observation Pulse rate (Pulse/min)

No.

1. 1st reading

2. 2nd reading

3. 3rd reading

4. Mean

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2. In children (5 years) – 90 to 110 beats/min.

3. In adult humans – 60 to 80 beats/min.

An increased pulse rate above the normal level is called as “Tachycardia”, where as
thedecrease in pulse rate is called as “Bradycardia”.

PROCEDURE

 Locate the radial artery at the own wrist level.

 Palpate the radial artery by pressing them with your finger against the underlying bones

(thepulse will be felt).

 Record the pulse for 1 minute.

 Take three readings at the intervals of 5 minutes and calculate the mean pulse rate.

Report

The pulse rate was found to be pulses per minute.

Reference:

1. S.R. Kale., R.R. Kale., Practical Human Anatomy and Physiology.,Nirali Prakashan.,

2. https://labmonk.com/determination-of-heart-rate-and-pulse-rate-of-the-patient

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b. DETERMINATION OF HEART RATE

AIM

To determine the Heart rate.

REQUIREMENTS

Sthethoscope is an instrument used for listening to sound produced inside the body with

amplification. The commonly used stethoscope consists of:

Ear frame: It consists of two curved metallic tubes joined together with a flat U shaped spring

which keeps them pulling together.

Conducting tubes: Simple flexible and soft [pressure tubes of rubber or latex material.

Chest piece: It consists of a bell and flat diaphragm which causes an amplification of the body

sound.

PRINCIPLE

The number of heart beats recorded per minute is called as heart rate. The heart rate is normally

initiated by impulses generated in the sinoatrial (SA) node. The rhythm is determined by the route

of impulse transmission through the conducting system. It is usually measured by a stethoscope

but, more specifically, determined by an electrocardiogram (ECG).

An increased heart rate above normal is tachycardia and a decreased heart rate below normal is

bradycardia.

PROCEDURE

 Select the subject and make him/her to sit comfortably on the chair.

 Place the chest piece of stethoscope against the thoracic wall.

 Record the heart beat for the period of 1 minute.

 Take three readings at the interval of 5 minutes and calculate the mean heart rate.

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OBSERVATION TABLE

S.no Observation Heart rate (Beats/min)

No.

1. 1st reading

2. 2nd reading

3. 3rd reading

4. Mean

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Note:

Normal values of heart rate/minute:

1. Foetus: 140-160 beats.

2. Children: 140 beats.

3. Adult males: 64-74 beats.

4. Adult Females: 72-80 beats.

RESULT

The heart rate was found to be beats per minute .

Reference:

1. S.S.Randhawa, Atul Kabra, Human anatomy and Physiology I., Practical, Page no: 46

1. S.R. Kale., R.R. Kale., Practical Human Anatomy and Physiology.,Nirali Prakashan.,

2. https://labmonk.com/determination-of-heart-rate-and-pulse-rate-of-the-patient

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Observation

1. Name

2. Age

3. Gender

4. Date of recording

5. Time of recording

Observation Table

S.No Systolic BP Diastolic BP

1.

2.

3.

4. Mean

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EXP. NO.: 15 DATE:

DETERMINATION OF BLOOD PRESSURE

AIM

To determine the blood pressure of the subject.

METHODS OF MEASUREMENT

Blood pressure is measured by two methods

1. Direct Method

2. Indirect Method

INDIRECT METHOD (SPHYGMOMANOMETRY)

The instrument used in this method is the sphygmomanometer.

PRINCIPLE

The cuff of the sphygmomanometer is wrapped around the arm of the subject. The bag is then

inflated until the air pressure in the cuff overcomes the arterial pressure and obliterates the arterial

lumen. This is confirmed by palpating the radial pulse that disappears when the cuff-pressure is

raised above the arterial pressure. The pressure is then raised further by about 20 mm Hg and then

slowly reduced. When the pressure in the cuff reaches just below the arterial pressure, blood

escapes beyond the occlusion into the peripheral part of the artery and pulse starts reappearing.

This is detected by the appearance of sounds in the stethoscope and is taken as the systolic

pressure. Subsequently, the quality of the sound changes and finally, disappears. The level where

sound disappears is taken as the diastolic pressure. The sound disappears because the flow in the

blood vessels becomes laminar.

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Blood Indication/ Factor responsible

Pressure

Increased  Obesity, Ageing, Emotional stress, Exercise.

 Hypertension.

 Heart disease.

 Renal disease.

 Diabetes Mellitus.

Decreased  Hypotension

 Severe blood loss

 Severe water and electrolyte imbalance.

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REQUIREMENTS

Sphygmomanometer, Stethoscope.

PALPATORY METHOD

PROCEDURE

 Check the level of the mercury column in the sphygmomanometer.

 Expose the arm up to the shoulder.

 Wrap the cuff around the middle of the arm in such a way that the lower edge of the

cuff remains at the minimum distance of one inch above the cubital fossa.

 Palpate the radial artery at the wrist by placing the middle three fingers over it.

 Hold the rubber bulb in the other hand in such a way that your thumb and index finger

remain free to manipulate the scram.

 Raise the pressure and simultaneously feel the pulse.

 Note the level of mercury where the pulse disappears.

 Raise mercury level 10 mm Hg above the point of disappearance of pulse.

 Reduce pressure gradually, 2-4 mm Hg per second. Note the level of mercury where

thepulse reappears. Pulse reappears at the same level where it had disappeared.

 Note your observation and express result in mmHg

AUSCULTATORY METHOD

PROCEDURE

 Record blood pressure by the palpatory method as described.

 Raise the pressure 30 mm Hg above the palpatory level

 Place the diaphragm of the stethoscope lightly on the brachial artery in the cubital fossa.

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 Lower the pressure at the rate of 2 to 4 mm Hg per second.

 Note the level where the sounds are first heard as the systolic pressure and the levelwhere the

sound ceases as the diastolic pressure.

Normal BP Value is

 Systolic BP-120 mm Hg.

 Diastolic BP-80 mm Hg

Report

The blood pressure of the selected subject was found to be mm Hg.

Reference: CL GHAI., A textbook of Practical Physiology, 8th Edition, JAYPEE Brothers.,Pg no: 168-173.

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WBC DIFFERENTIAL COUNT

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EXP NO: 16 DATE:

DIFFERENTIAL LEUKOCYTE (WBC) COUNT

AIM: To Determine the differential leukocyte for own blood

Apparatus Required: 4-5 clean glass slides, Leishman’s stain, blood lancet, dropper,

glass rod, compound microscope, cedar wood oil, buffered water and staining tray.

Principle:

White Blood Cell Differential Count determines the number of each type of white blood

cell like neutrophils, lymphocytes, monocytes, eosinophils and basophils are present in the

blood. It can be expressed as a percentage (relative numbers of each type of WBC in

relationship to the total WBC) or as an absolute value (percentage x total WBC). A blood

film appropriately prepared and stained is seen under the microscope in oil immersion.

Different WBCs are seen which are counted in percentage. The total of 100 leukocytes are

studied, identified and recorded from a blood smear.

Procedure:

1A. Blood film preparation by Wedge technique.

 Place 3 x 1 inch glass slide on a flat surface (Stationary slide

 A drop of blood is placed on the writing hand side of the slide.

 It is spreaded by a haemocytometer cover glass by pushing slide forward withsmooth

and rapid stroke so that the blood will be pulled behind the spreader.

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OBSERVATION TABLE:

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1B. Staining of the blood film by Rack Method.

 Place the slide on glass rods overlying a sink or dish that keeps the glass slide in a

horizontal position.

 The glass slide is covered with chemically pure methanol (CH3COOH).

 The slide is allowed to dry and the blood film becomes fixed.

 The surface of the blood film is flooded with Leishmann’s stain with the help of adropper.

 After 10 minutes the film is flushed with the stream of distilled water till all stain isremoved.

 The residual stain on the back of the slide is cleaned off with cotton gauze or tissuepaper

and the specimen is dried in the ‘air.

2. Draw one hundred squares on a paper.

3.The differential cells are counted manually. An area where the film is made quite clear under the
low power magnification is selected. The best area for cell counting in a slide is the body of the film

and not the head or tail.

4. The two drops of cedar wood oil are placed over the slide and the oil immersion lens
is shifted in contact with cedar wood oil.

5. Identify various types of leukocyte and enter them by hrst letter (NNeutrophil, EEosinophil,
Basophil, Lymphocyte, M-Monocyte) in all 100 squares on the paper. At least 100 WBCs are noted.

6. Report the result of 100 cells classified as a percentage.

7. Count the cells in a specific sequence to avoid repeated counting. The track method is usually used to
count WBCs. (Longitudinal pattern method or lengthwise method is also used) in which cells are
countered in consecutive fields in strips or rows from the tail end straight back towards the thick end of
film i.e. Cells are counted in one direction and then the field is shifted and counting is done in

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opposite direction. The field is again shifted in the same direction and counting is done in the same

direction as in the beginning. .

8. The percentage of various types of white cells is calculated by counting the different cells into

various squares.

Normal Values: Normal Leukocyte count

1. Granulocytes.

Neutrophil

Eosinophil 1- 4%

Basophil O- 1%

2. Agranulocytes
Lymphocyte 20- 40%

Monocyte 2- 8%
RESULT: The Differential leukocyte count was found for the own blood

1. Neutrophil: _

2. Eosinophil:

3. Basophil:

4. Lymphocyte: _

5. Monocyte:

Reference
1. Dr. Ramesh K Goyal. Dr. N.M. Patel Practical Anatomy & Physiology. B.S.ShahPrakashan.
15th Edition.

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