HUMAN ANATOMY

AND
PHYSIOLOGY
An Experimental Handbook

For
B. Pharm Students

Vishnu N. Thakare
Ph.D.
Assistant Professor
Department of Pharmacology
Sinhgad Institute of Pharmaceutical Sciences
Kusgaon (Bk), Lonavala-Pune 410401,
India.

Price ` : 160.00

N 1656
HUMAN ANATOMY AND PHYSIOLOGY ISBN 978-93-83750-85-6
Fourth Edition : April 2018
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Preface
It is with great pleasure that I introduce the book "Human Anatomy and Physiology –
An Experimental Handbook". The book allows for the lucid understanding of human
anatomy and physiology, which is extremely necessary for the clear understanding of the
pathophysiology of various disorders/diseases and effects or actions of drugs. This book is a
sincere attempt to explain the basics of experimental human anatomy and physiology in a
simple and interesting manner and as per the syllabus prescribed for the first year B. Pharm
students by the University of Pune. This book imparts insight into each experiment along
with its clinical and pathological significance. Questions that could be asked for viva-voce are
also included at the end of each experiment.

All efforts have been made to keep the text error-free and to present the subject in a
student friendly and easy to understand manner. However, any suggestions and constructive
comments would be highly appreciated and incorporated in the next edition.

Vishnu N. Thakare
Acknowledgements
I feel indebted to Hon. Prof. M. N. Navale (Founder President), Dr. (Mrs.) Sunanda M.
Navale (Secretary) Sinhgad Technical Education Society, Pune for their continuous
encouragement and support.

I am sincerely grateful to Dr. S. R. Naik (Head of Department, Pharmacology), Dr. S. B.

Bhise, Principal, Sinhgad Institute of Pharmaceutical Sciences, Lonavala, for their support,
help and guidance.

I express my sincere gratitude to Dr. K. S. Jain, Sinhgad Institute of Pharmaceutical

Sciences, Lonavala; Dr. (Mrs.) Bhoomika M. Patel, Nirma Institute of Pharmacy, Ahmedabad;
Dr. Niraj Vyawahare, D. Y. Patil College of Pharmacy Pune; Dr. M. M. Ghaisas, lndira College of
Pharmacy Pune; Dr. Sohan Chitlange, Dr. D.Y. Patil Institute of Pharm. Sciences, Pune;
Dr. R. V. Patil and Dr. N. S. Bhajipale, College of Pharmacy Akola and Mrs. Anupama Suralkar
for their continuous help, valuable guidance and advice.

I also express my gratitude to my colleagues Dr. Valmik Dhakane, Mr. R. R. Patil,

Mr. R. R. Wadekar, Mr. M. K. Aswar, Mr. Y. P. Kulkarni, Ms. D. K. Ingawale, and Mr. P. G.
Pandhare, for their help and support during writing of this book.

My special thanks to my wife Vaishali and my students, Ms. Malvika and Ms. Sanyogita
Navale for their kind help.

I express my thanks to Mr. S. B. Gokhale and staff of Nirali Prakashan.

I am also thankful to Mr. Jignish Furia of Nirali Prakashan for publishing this book.

Vishnu N. Thakare
Contents
1. STUDY OF MICROSCOPE 1-4
EXPERIMENTS ON BLOOD
2. DETERMINATION OF HAEMOGLOBIN CONTENT AND OXYGEN CARRYING CAPACITY
OF BLOOD SAMPLE 5-10
3. DETERMINATION OF THE NUMBER OF RBCs AND COLOUR INDEX OF BLOOD 11-16
4. STUDY OF OSMOTIC FRAGILITY OF RBCs 17-20
5. DETERMINATION OF THE NUMBER OF WBCs IN BLOOD SAMPLE 21-24
6. DETERMINATION OF DIFFERENTIAL LEUKOCYTE COUNT OF BLOOD 25-28
7. DETERMINATION OF PLATELET COUNT OF BLOOD 29-32
8. DETERMINATION OF RETICULOCYTE COUNT OF BLOOD 33-36
9. DETERMINATION OF ARNETH COUNT OF BLOOD 37-40
10. DETERMINATION OF ERYTHROCYTE SEDIMENTATION RATE 41-42
11. DETERMINATION OF BLOOD GROUP 43-46
12. DETERMINATION OF HEMATOCRIT CONTENT OF BLOOD 47-48
13. DETERMINATION OF BLEEDING TIME AND CLOTTING TIME OF BLOOD 49-50
14. DETERMINATION OF BLOOD PRESSURE 51-54
EXPERIMENTS ON CVS, RESPIRATORY PARAMETERS
15. RECORDING OF PULSE RATE 55-56
16. RECORDING OF ELECTROCARDIOGRAM 57-58
17. MEASUREMENT OF BODY TEMPERATURE 59-60
18. DETERMINATION AND RECORDING OF RESPIRATORY VOLUME 61-64
19. DETERMINATION OF BREATH HOLDING TIME AND HYPERVENTILATION TIME 65-68
EXPERIMENTS ON CELL, TISSUES AND BODY SYSTEMS
20. STUDY OF HUMAN CELLS, TISSUES AND HISTOLOGY OF ORGANS 69-78
21. STUDY OF THE CARDIOVASCULAR SYSTEM 79-86
22. STUDY OF THE DIGESTIVE SYSTEM 87-94
23. STUDY OF THE RESPIRATORY SYSTEM 95-98
24. STUDY OF THE URINARY SYSTEM 99-102
25. STUDY OF THE HUMAN SKULL 103-106
26. STUDY OF THE PECTORAL GIRDLE AND BONES OF THE UPPER LIMB 107-112
27. STUDY OF BONES OF THE VERTEBRAL COLUMN 113-116
28. STUDY OF THE PELVIC GIRDLE AND THE LOWER LIMB 117-120
29. STUDY OF VARIOUS JOINTS 121-126
30. STUDY OF THE CENTRAL NERVOUS SYSTEM 127-132
31. STUDY OF THE LYMPHATIC SYSTEM 133-135
EXPERIMENTS ON REPRODUCTIVE SYSTEM
32. STUDY OF THE MALE REPRODUCTIVE SYSTEM 136-138
33. STUDY OF THE FEMALE REPRODUCTIVE SYSTEM 139-142
34. PREGNANCY DIAGNOSTIC TEST 143-144
35. STUDY OF FAMILY PLANNING METHODS AND DEVICES 145-148
EXPERIMENTS ON SENSE ORGANS
36. STUDY OF THE HUMAN EYE 149-151
37. DETERMINATION OF VISUAL ACUITY FOR NEAR AND DISTANCE VISION 152-155
38. DETERMINATION OF DOMINANCE OF EYE 156-156
39. DETERMINATION OF SPECIAL SENSES 157-159
40. STUDY OF THE HUMAN EAR 160-162
41. STUDY OF THE HUMAN SKIN 163-165
BIOCHEMICAL ANALYSIS OF URINE
42. BIOCHEMICAL ANALYSIS OF URINE 166-169
EXPERIMENTS ON NERVE MUSCLE PREPARATION (OF FROGS)
43. STUDY OF THE SIMPLE MUSCLE TWITCH 170-172
44. EFFECT OF TEMPERATURE ON SIMPLE MUSCLE TWITCH 173-174
45. STUDY OF FATIGUE ON GASTROCNEMIUS SCIATIC NERVE MUSCLE
(OF FROG) 175-176
• APPENDIX 177-178
• GLOSSARY 179-185
• BIBLIOGRAPHY 186-186




 EXPERIMENT NO. 1
STUDY OF MICROSCOPE

Aim
To study the microscope

Microscope
Microscope is an optical instrument which uses visible light and a system of lenses to
view magnified images of very small objects. This microscope also called light microscope.

Types
(a) Simple microscope – has one set of lens and low magnification.
(b) Compound microscope – has two sets of lenses and objectives with higher
magnifications.
(c) Electron microscope – This microscope differs from optical microscope; in that the
electrons interact with the sample to generate image instead of light acting as an
illuminating source.
In general, experiment involving the study various cells, the compound microscope is
used for the study of morphological characteristics of cells. Compound microscope works
on the principle of formation of an enlarged image of sample / object in the plane of focus.

Physical terms
(a) Compound microscope : It is an arrangement of objective and eyepiece used to
magnify an object to the point where it can be seen with the human eye.
(b) Resolution : The resolution of an optical microscope is defined as the shortest
distance between two points on a specimen that can still be distinguished by the
observer as separate entities. Consequently it describes how small objects can lie
close to each other and can still be recognisable. Resolution with human eye is
around 0.25 mm, with light microscope it is around 0.25 µm and with the electron
microscope it is 0.5 nm.
(c) Working distance : It is the distance between the objective lens and the specimen.
The magnifications increase with decrease in working distance. The ideal working
distance is 0.15-1.5 mm for the oil immersion objective, 0.5-4 mm for high power
objective and 5-15 mm for low power objective.

(1)
 Eye piece

Coarse adjustment knob

Fine adjustment knob

Objective
Arm

Glass slide

Condenser Stage

Mirror
Base

Figure 1.1 : A Compound Microscope

Parts of a Compound Microscope
(1) The support system
(2) The illuminating system
(3) The magnification system
(4) The adjustment system.
1. The Support system: The support system consists of various parts which are described
below.
(a) Base of microscope: It is horse shoe shaped and is the base on which the
microscope rests.
(b) Pillars: Projecting upward from the base, it is joined to the handle of microscope
(c) Handle (arm) : Handle is curved shaped or ‘c’ shaped and supports the magnifying
and the adjusting system.
(d) Body tube : It is the tube which houses the lenses and through which light passes to
form an image.
(e) Stage : It is a horizontal platform on which the specimen under observation is
placed. Fixed and mechanical stage microscopes are generally used. The specimen
is mounted on a slide which is held in place by clips that are present on the stage.
The screws enable the movement of the slide sideways or vertically.
Nosepiece : It is the part of the microscope that holds the objective lenses. It is also
called as a revolving nosepiece or turret. Nosepiece is attached to the lower end of the
body tube and consists of objective lenses of different magnifications attached to it. The

Human Anatomy and Physiology 2 An Experimental Handbook

objective of required magnitude can be focused over the specimen by moving the nose
piece.
2. The illuminating system : A good illuminating system is the one which provides
uniform and bright illumination of the entire field viewed under the microscope.
There are six types of illuminating systems based on which the microscope functions.
(a) Bright field microscope –the source of illumination used is white light, either
external sunlight or internal tungsten filament lamp.
(b) Dark field microscope – dark field condenser is used to block scattered light
(c) Fluorescent microscope – UV lamp is used as light source.
The illuminating system of a compound microscope is composed of a light source, a
condenser, and a diaphragm.
Mirror is fixed at the base and is used to reflect light from an external light source up
through the bottom of the stage. As the rays of light are reflected by mirror through the
condenser on the object, the object on the stage appears clear.
Condenser – the rays of light reflected by the mirror pass through the condenser
located in between the mirror and stage of the microscope, then fall onto the object under
examination and thus help in resolving the image. The position of the condenser needs to
be adjusted with each object used in order to alter the light and to improve the resolving
power of the microscope.
3. Magnification system: As the name indicates, it directly implies magnifying the
image of the object under observation and comprises of the eyepiece and objectives.
(a) Eye piece: The eyepiece is a lens that fits into the top of the body. It magnifies the
image formed by the objectives; generally 5x and 10x eyepieces are used. Monocular
microscope uses one eyepiece; where as binocular microscope has provision for fitting 2
eyepieces. The magnification formed by the eyepiece multiplied by objective
magnification gives the total magnification of the object being viewed.
Objectives : Generally these objectives are screwed into the resolving nosepiece in a
compound microscope. The nosepiece is a pivot that ensures quick changes of objectives.
(1) 10X - low power objective- magnifies the image ten times greater.
(2) 40 or 45X - high power objective- magnifies images 40 or 45 times . It is used for a
broad view of blood films or histological sections prior to their examination under
oil-immersion objectives.

Human Anatomy and Physiology 3 An Experimental Handbook

 (3) Oil-immersion objective (100X) - This requires immersion oil viz, cedar wood oil.
Oil is employed to increase the numerical aperture (is a ratio of the diameter of
lens to its focal length) and the resolving power of the objectives. Light passes
through glass at same speed as it travels through the immersion oil. Hence, the ray
of light that passes through oil undergoes minimum diffraction when it passes
through glass. As a result the resulting image is much clearer and sharper.
4. Adjusting system: Two adjustment systems are used
(a) Coarse adjustment-Two coarse adjustment screws are employed for coarse
adjustment. These screws are mounted at the top of the handle by a double side
micrometer mechanism, one on each side.
(b) Fine adjustment-Two fine adjustment screws are mounted on the handle below the
coarse adjustment screws by double side micrometer mechanism, one on each
side.

EXERCISE
(a) Give the principle of microscope.
(b) Enlist the various parts of a compound microscope.
(c) What is resolution? Give its significance.
(d) State the importance of condenser and iris diaphragm
(e) Give the role of illuminating system in a microscope.
(f) Give the significance of oil used in oil immersion objectives.


Human Anatomy and Physiology 4 An Experimental Handbook

 EXPERIMENT NO. 2
DETERMINATION OF HAEMOGLOBIN CONTENT AND
OXYGEN CARRYING CAPACITY OF BLOOD SAMPLE

Aim
To determine the haemoglobin (Hb) content and oxygen carrying capacity of one's
own blood sample.

Apparatus
Sahli’s Hellige haemoglobinometer (Figure 2.1), stirrer, micropipette (200 cubic
millimeter), disposable needle (24 gauge), Pipette-having single mark 0.02 ml (20 cu nm)
without any bulb, 0.1N HCL, 70% alcohol or spirit, cotton, distilled water.
Sahli’s haemoglobinometer consists of comparator, tube, pipette and stirrer.
Comparator-the haemoglobinometer at the center point is provided with opening
which holds the haemoglobin tube. Two non fading standard brown tubes are provided on
both sides of the central haemoglobin glass tube for colour matching.
Tube- Gram% markers on one side up to 30 and % mark (20-140) on another side for
easy reading.

Principle
Reaction of blood with hydrochloric acid (HCl) causes the formation of hematin acid
by hydrolysis of haemoglobin. Acid hematin is reddish brown in colour. This is then
diluted with distilled water until its colour matches exactly with that of the permanent
standard of the comparator block. Matching of sample with standard tubes gives exact
concentration of haemoglobin of sampled blood.

Theory
Haemoglobin (Hb) is a protein comprising of heme and globin present in RBCs, which
carries oxygen and carbon dioxide. Heme portion is involved in transportation of oxygen
from lungs to the tissue. Oxyhaemoglobin is the combination of oxygen (4) with one
molecule of haemoglobin.

Human Anatomy and Physiology 5 An Experimental Handbook

 1 gram of Hb carries 1.34ml of oxygen. Haemoglobin also acts as a buffer by
maintaining blood pH.

Normal Range
Adult male : 14-18 gm% of blood
Adult female : 12-16 gm% of blood
In newborn : 16-22 gm% of blood
Infants : 12-40 gm% of blood

Procedure
1. Take Sahli’s haemoglobinometer and pipette and make sure that it is dry.
2. Fill the haemoglobinometer up to its lowest mark 10% (2 gram %) by adding 0.1N
HCL with the help of a dropper.
3. Sterilise the finger tip with spirit or 70% alcohol and prick the finger tip with a
sterile needle to allow free flow of blood.
4. Allow a large drop of blood to form on the finger tip, dip the tip of pipette on to the
blood drop and suck up to 0.02 ml mark taking care to avoid the formation of an
air bubble.
5. Transfer immediately 0.02 ml blood into the haemoglobinometer (containing
0.1N HCL) by blowing pipette.
6. Leave the solution in the tube haemoglobinometer for about 10min.
7. After 10 min, dilute the solution with distilled water, drop by drop and mix it with a
stirrer. Keep adding water until colour of solution in the tube matches with
standard of the comparator (while matching the two colours, take care to hold the
stirrer above the level of the solution).
8. Note the reading when the colour of solution matches to the standard and express
the haemoglobin concentration as gram%.

Precaution
Following precautions to be taken while performing this experiment.
1. Blood should be immediately transferred from pipette into haemoglobinometer
tube to prevent clotting of blood in the tube.

Human Anatomy and Physiology 6 An Experimental Handbook

 2. 10 min should be given after addition of blood into haemoglobinometer tube, for
complete conversion of haemoglobin to acid hematin.
Result : The haemoglobin content of the blood sample was found to be …… gm%.

Haemoglobinometer

Dropper

Pipette

Figure 2.1 : SAHLI’S Haemoglobinometer

Calculation
Observed reading of haemoglobin ------ gm%.
Percentage of haemoglobin
14.5 gms Hb = 100%.
Observed Hb value = ------ gm%.
= 100 × observed value / 14.5
= Y%

Oxygen carrying capacity of haemoglobin

100% Hb = 18.5 cc of oxygen
Z = Y% × 18.5 / 100 Zcc of blood.

Human Anatomy and Physiology 7 An Experimental Handbook


Other various methods of haemoglobin content estimation
1. Dare’s method

2. Harden’s method

3. Wintrobe’s method

4. Halden’s method

5. Tallquist’s method

6. Gasometric method

7. Spectrophotometric oxyhaemoglobin method - A cyanmethaemoglobin method

8. Specific gravity method

Significance
Physiological

RBC contains Hb that forms nearly about 90% of dry weight of the cell.
RBC is called so, due to presence of red coloured haemoglobin. Reduced supply of
haemoglobin to cell/ tissues leads to hypoxic condition, since haemoglobin carries oxygen
to various cell/ tissues.

Clinical
Determination of haemoglobin content in blood will give an idea about whether the
person is suffering from anaemia (a condition in which the oxygen carrying capacity of
blood is reduced) causing weakness, muscle cramp or not. A person is said to be anemic if
the Hb content falls below the normal range. Symptoms of anemic person are weakness
muscle cramp, nausia, abdominal pain etc.
Hb content increases in the following conditions.

• High altitude

• Excessive sweating

• New born/ infants

• Diarrhoea

Human Anatomy and Physiology 8 An Experimental Handbook

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