HUMAN ANATOMY AND PHYSIOLOGY LAB December 30, 1899

LAB 2:
Study of microscope and its use
__________________________ __________________________
Student Name Date Lab Performed
Submission Date/Time: ______________________________
Note: Do the task/make report according to your understanding and use plane A4 sheets
Note: It should be handwritten and submitted physically.

Lab objectives

At the conclusion of this lab, the student should be able to

 Identify when a compound microscope would be used in the lab

 Describe the proper way to handle a microscope
 Identify the parts of a compound microscope
 Determine the total magnification of an object being viewed by a compound light
microscope if given the ocular lens and
 Objective lens magnification

Key points to remember:

 Microscope is a costly equipment. Therefore, it should be handled carefully. Always

keep the microscope in an upright position while taking it from one place to another. As
far as possible don’t tilt the arm.
 Clean the lenses of the microscope with the lens paper or muslin cloth, never with the
filter or any other kind of paper.
 If you are using the high power objective lens then after the observation is over, turn
the nose piece and bring low power objective lens in line with the hole in the stage.
 Objective lens should be kept at least 1 cm above the stage.
 After using the microscope always keep it in the box.
 Take care to see that the stage of microscope, the eye piece, the objective lens are dry
and clean. No chemical should stick to these.

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 Adjustment knobs and joints should be protected from rusting by applying Vaseline.

Background:

Many organisms (bacteria) and parts of organisms (cells) that biologists study are too small to
be seen with the human eye. We use microscopes to enlarge specimens for our investigation.
The word microscope means “to see small” and the first primitive microscope was created in
1595

Compound microscope

A compound microscope is an instrument that is used to view magnified images of small

specimens on a glass slide. It can achieve higher levels of magnification than stereo or other low
power microscopes.

It achieves this through the use of two or more lenses in the objective and the eyepiece. The
objective lens or objectives located on the nosepiece have a short focal length and are close to
the target specimen where it collects light and focuses the image of the object into the
microscope. The second lens, in the eyepiece, has a longer focal length and further enlarges the
image.

Parts of a compound microscope

Each part of the compound microscope serves its own unique function, with each being
important to the function of the scope as a whole. The individual parts of a compound
microscope can vary heavily depending on the configuration & applications that the scope is
being used for. Common compound microscope parts include

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1. Ocular lens/Eye piece (10x)- the magnifying lens closest to your eye.
2. Arm-Support the microscope head and attaches it to the base
3. Revolving Nose Piece - holds the objective lens and allows objectives to be rotated for
differing magnifications.
4. Objective lenses are the lenses closest to the object (specimen).
5. Scanning objective (4x) is lowest power and shortest. Used to locate portion of
specimen to observe.
6. Low power objective (10x) - the middle length lens.
7. High power objective (40x)- the longest length lens.

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8. Stage - supports the object (specimen).

9. Stage Clips hold the object slides in place.
10. Illuminator/light - shines light upwards through the slide. Low voltage Halogens bulb
are mostly used as source of illumination
11. Base - supports the microscope.
12. Diaphragm/Aperture - controls the amount of light striking the slide.
13. Mirror-Reflects light into the base of the microscope. Earlier microscopes used mirrors
that reflected light into the base of the microscope instead of halogen bulbs as their
source of illumination.
14. Coarse adjustment knob - used to focus with scan objective.
15. Fine adjustment knob - used to focus on low or high power objective
16. Bottom lens/Brightness adjustment- Knob used to adjust the amount of light that
reaches through the specimen

Total magnification

Magnification is the ratio of the image size with the microscope to the actual size of the object.
When you say that the magnification is 10, the image you see using the microscope is ten times
bigger than viewing the specimen with the naked eye. Remember with a compound light
microscope you are magnifying with two lenses, so to calculate the total magnification you
multiple the objective magnification by the ocular magnification.

Total magnifications= Objective lens magnification x Ocular magnification

Lab Questions
1. Generate a lab report on how to handle a compound microscope.
2. Discuss the different types of objective lens and their magnification power.
3. Calculate the total magnification for each lens in the table below

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Objective lens Ocular lens =Total magnification

Scan objective
Low objective
High objective

4. State the function of each part of microscope in your own words

Parts Function
Oculars

Arm

Nosepiece

Coarse adjustment

Fine adjustment

Scanning objective

Illuminator

Sphygmomanometer

Safety Notes:
1. When inflating the cuff pressure should not exceed 180 mm Hg.
2. Rate of deflation of cuff should be between 2 and 3 mm Hg per second (never leave the cuff
inflated on the subject for more than a few seconds) severe bruising may occur.
3. Cuff should never be inflated unless being used on a subject.

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4. The stethoscope is a sensitive instrument which dramatically magnifies sound. Once in place
the person using the stethoscope should never be exposed to loud noise, i.e. tapping, yelling,
etc.

Introduction:
1. Blood Pressure and Hypertension
The blood pressure, the force that the blood exerts in all directions in any given area, is
the basis for maintaining a stable movement of blood from the heart throughout the whole
body and back into the heart.
Because a fluid moves from an area of high pressure to an area of low pressure, the
pressures at different points in the circulatory system differ. Also, in any blood vessel the
pressure depends on the amount of blood present and the size of the vessel. When the
ventricles contract, more blood is forced into the arteries and the pressure increases; this is
called the systolic pressure. While the ventricles are relaxed, blood goes out of the arteries and
into the capillaries and venous system, lowering the pressure. The lowest pressure occurs just
before the ventricles contract and is called the diastolic pressure. The pressure at any given
point in an artery is not constant but is always changing between high pressure (systolic) and
low pressure (diastolic). Pressure along an artery is highest at the end closest to the ventricle
and lowest at the end farthest from the ventricle. The pressure exerted by the blood is
expressed as the height in millimeters that it will move a column of mercury (mm Hg). The
systolic pressure is written first and the diastolic pressure second, e.g. 120/80 mm Hg.
The difference between the systolic and diastolic pressure is called the pulse pressure
and represents the change in the arterial pressure between heartbeats. The pulse pressure is
an approximation of the stroke volume of the ventricle.
The mean blood pressure in the larger arteries of a normal young adult is about 100 mm
Hg, in the arterioles about 60 mm Hg, in the capillaries about 18 mm Hg, in the veins about 7
mm Hg, and at the entrance to the right atrium about 3 mm Hg. While the arterial pressures
change throughout the cardiac cycle, the mean blood pressure is usually fairly constant.
When we refer to the blood pressure medically we usually are referring to the pressure
in the arterial system. Unless otherwise indicated, blood pressure readings are the pressures in
the brachial arteries in the arms.

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Name of component Explanation

Anerold Manometer Enables the observer to observe reading

Gauge

Hose / Tube These provide a connection among anerold manometer gauge, cuff and
Connectors inflation bulb.

Inflation Bulb This part is pressed gradually with small intervals due to which the needle of
anerold manometer gauge rises that results in the inflation of cuff.

Air Release Valve The valve connected with inflation bulb that is used to release air so that the
needle in the anerold manometer gauge falls and diastolic pressure can be
noted.

Cuff A part of BP apparatus that is placed around the arm on the brachial artery
and then inflated with the help if inflation bulb to observe blood pressure.

Artery position Indicates the direction of BP Apparatus on right and left arms.
indicator lebel (Right

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and Left arms)

Cuff Size Tells the size whether it can be worn only by adults or not.

Velcro Fastening Used to tie, stick and hold the cuff tightly around the arm.

2. Measuring Blood Pressure

Blood pressure can be most accurately measured by inserting a needle directly into the
artery and connecting it to a mercury manometer or electronic transducer. Because this
method is not practical, a less precise, indirect method using a sphygmomanometer consists of
an inflatable cuff, an inflation pump, a gauge to register pressure, and a controlled exhaust
valve. The cuff is wrapped around the upper arm just above the elbow and inflated. When the
pressure in the cuff exceeds that in the artery, the artery collapses and stops blood flow. As
pressure is gradually released from the cuff, blood flows through the artery only when its
pressure exceeds the pressure of the cuff. Sounds made by the blood as it spurts through the
artery under the cuff can be heard using a stethoscope. These sounds are called Korotkov's
sounds. The first sound heard corresponds to the highest or systolic pressure, while the
muffling or disappearance of the sounds corresponds to the lowest or diastolic pressure.
3. Stethoscope
The stethoscope has a head (or bell) containing a diaphragm and is connected to two
ear pieces by tubing. The ear pieces are inserted into the ears and the end of the stethoscope
is placed on the bare chest. The strongest sounds are found to the patient's left of his or her
chest midline in the space between the fifth and sixth ribs. Heart sounds can also be heard at
other points on the chest and back.
The electronic stethoscope is much the same in appearance as a regular stethoscope,
but it greatly amplifies the sounds of the heart and allows one to listen to heart sounds even
through clothing. It also lets you amplify and hear faint sounds not readily discernible with the
simple stethoscope.

4. Sphygmomanometer
The sphygmomanometer is an instrument which gives an indirect measurement of
blood pressure. It consists of a gauge (manometer) to register pressure, an inflatable
compression bag with an unyielding cloth cover (cuff), an inflating device (inflation bulb or
pump), and a means of controlled exhaust.
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The fitting of the compression bag and cuff are very important to obtain correct
readings. The bag should be about 20% wider than the diameter of the arm and long enough to
go at least halfway around the arm. A standard adult inflation bag is usually about 14 cm wide
and 60 cm long. For youths below the age of 14 and adults with thin arms, a smaller cuff and
bag should be used. Using too small a cuff and bag gives high blood pressure reading, while
using too large a cuff and bag gives low readings.
Basically, there are three types of gauges used in sphygmomanometers; the mercury
gravity manometer, the aneroid manometer, and the electronic manometer.
5. Normal Blood Pressure
As mentioned earlier, when we medically refer to blood pressure we are talking about
the arterial pressure as measured in the brachial artery of the arm.
Normal blood pressure varies with age and from person to person. It also varies
depending on whether a person is lying, sitting, or standing, and is affected by such factors as
time of day, fullness of bladder, amount of recent exercise, and recent intake of caffeine,
nicotine, or alcohol. Some average blood pressures at different ages are:

Age Pressure (sys/dias)

4 99/66

6 100/56

12 113/56

16 120/61

18 120/74

30 126/79

It must be remembered that these figures are just averages and that blood pressures
normally vary above and below these figures for a given age group. For instance, at age 16 the
average given was 120/61 mm Hg, while the range for tested youths in this group was 102 to
140/51 to 71 mm Hg.

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High blood pressure is a pressure reading of 140 systolic or higher and 90 diastolic or
higher. If either or both of systolic or diastolic pressures continually exceed the 140/90 mm Hg
readings, the person is considered to be hypertensive. These hypertensive values should be
thought of as the person's continuing blood pressure; single high readings from stress or
exercise normally exceed these limits for short periods of time.

Korotkoff Sounds:

 Korotkoff sounds are produced underneath the distal half of the blood pressure cuff.
 The sounds appear when cuff pressures are between systolic and diastolic blood
pressure, because the underlying artery is collapsing completely and then reopening
with each heartbeat.
 Consist of four phases on the basis of intensity and quality of sound.

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Objectives:
1. The student will demonstrate a competence in the correct and responsible usage of the
stethoscope and sphygmomanometer.
2. The student will take blood pressure readings from a human subject.
3. Students should compare this blood pressure readings to the normal readings listed in the
introduction.

Materials:
stethoscope
sphygmomanometer

Procedure:
Exercise: Taking Blood Pressure
1. Have the subject lying supine or seated comfortably and relaxes with the forearm on a
smooth surface at heart level.
2. The mercury meniscus or the needle of the manometer should be at zero with the cuff
deflated and the exhaust valve open.
3. Locate the brachial artery with your fingers just as you do for taking the pulse. The brachial
artery is located on the inside of the upper arm just above the bend in the elbow on the body
side.
4. The deflated cuff should be wrapped snugly (but not tightly) around the upper arm 2 to 3 cm
above the bend in the elbow. Be sure to position the inflatable bag so about half of it is on each
side of the brachial artery. When using an electronic sphygmomanometer or a cuff with a built-
in stethoscope head, be sure the microphone pick up or stethoscope head is directly over the
brachial artery.
5. While feeling the radial pulse in the wrist with one hand, close the exhaust valve and rapidly
inflate the cuff with the other hand. Inflate to a pressure about 30 mm Hg above the point
where the radial pulse ceases.
6. Without undue pressure, place the stethoscope over the brachial artery just below the cuff.
Exhaust the cuff at a rate of 2 to 5 mm Hg per second. As the cuff exhausts, listen for the first
clear tapping sound; the manometer reading at this point is the systolic pressure. Continue to
listen until the sound disappears; this is the diastolic pressure.

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7. Before taking the blood pressure again, allow the subject to relax for 30 to 60 seconds with
the cuff deflated. Never leave the cuff inflated on the subject for more than a few seconds.

Evaluation:

Name:_____________________ ______________________
_____________________ ______________________

Title: Blood Pressure

Objectives:
You are to demonstrate a competence in the correct and responsible usage of the
stethoscope and sphygmomanometer, take blood pressure readings from a human subject, and
be able to compare this blood pressure readings to the normal readings listed in the
introduction.

Procedure:
Refer to previous procedure

Results:
___________________________________________________________
Student Trial #1 Trial #2 Trial #3 Normal Reading
(sys/dias)

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Task:
1. Compare your blood pressure with the normal values listed in the introduction.

2. Calculate the pulse pressure for each reading and also the mean pulse pressure of all three
readings. Pulse pressure is the difference between the systolic and the diastolic readings and is
a crude estimate of the stroke volume of the heart (pulse pressure = systolic pressure - diastolic
pressure).

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