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English Language and Foundation Studies Centre

EPBIOL259/EPCHEM314 Life Sciences


Laboratory Session

Part A: Using a Microscope


Part B: BMR and Cardiovascular Physiology
Student Name:
Student Number:
Course:
(please circle)

EPBIOL259

EPCHEM314

Marking:
./10
Demonstrator
Signature:

Life Sciences Laboratory Session


Marks from this assessment will contribute towards your final grade for this
course. Please complete all working and answers on this sheet. This assessment
must be submitted before leaving the laboratory.
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Conducting an experiment in a laboratory involving glassware, equipment and other


people requires skill and patience. The most important thing is for you to be fully
prepared. Always read carefully ahead before beginning, try to visualise the steps, write
down any questions and clear them with the instructor before you begin.
Safety Issues and Requirements:
1. All students must wear covered footwear; entry to the laboratory will not be
permitted otherwise. Long hair must be tied back, no bare midriffs.
2. Eating, drinking or smoking is not permitted in the laboratory.
3. Any sharps, broken glass must be placed in the special containers provided.
4. Only supervised work is permitted in the lab.
5. Any spills or accidents must be reported immediately.
6. Consult the MSDS for unfamiliar chemicals.
7. Wash your hands immediately after leaving the laboratory.
8. If in doubt always ask the demonstrator.
Human Derived Products:
All human derived products and equipment contaminated by the products such as
blood and saliva must be considered as potential health hazards to you and to others
in the laboratory.
The student who has donated the product must be solely responsible for
the handling of the product.
All glassware and contaminated disposable material are to be disposed of
as instructed by the demonstrator in charge.
Pre-lab Questions:
Read and complete the following before entering the laboratory.
As you read the descriptions provided, label the parts on the diagram on the following
page.

Ocular Lens

Also known as the eyepiece


May be one only or a set of two
Lens closest to your eye, usually the highest part of the microscope
Includes a magnification factor engraved on the barrel, e.g. 10x indicates the
image is magnified ten times
Should be cleaned only with lens cleaner and lens paper

Nosepiece

Revolving circular mechanism that holds the different objective


lenses
Rotation of this changes the objective lenses

Objective Lenses

Individual lenses attached to the nosepiece


Includes a magnification factor engraved on the barrel
Changed by rotating the nosepiece

Provides the safest way to hold a microscope. Use two hands.

Arm

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Stage

Also called the mechanical stage


A surface that supports the slide, with the help of stage clips

Stage controls

Usually located on the top or side of the


stage
Moves the slide around the
stage.

Condenser

Located under the stage


Focuses the light through the hole in the stage and onto the specimen
Adjusts the quality and the amount of light passing through the specimen
May be raised or lowered with the condenser-adjustment knob

Iris Diaphragm

Located under the condenser


Adjusts the intensity of light passing through the specimen
Use the iris diaphragm lever to open or close

Coarse focus adjustment knob

Large knob located on the arm


Adjusts the distance between the stage and the objective lens, in large
increments
Used initially to bring the specimen into focus. It is dangerous to use this
knob when the objective lens is already near the slide
Should be turned very slowly to avoid breaking the slide

Fine focus adjustment knob.


Small knob located inside the course adjustment knob
Adjusts in small increments
Typically used after the objective lens is already near the slide and the
specimen is almost in focus

Lamp

Small light source located under the condenser


May be turned on / off with switch on the base

Base

Square or horseshoeshaped support for the whole


microscope Usually quite heavy to prevent tilting

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I dentify and label the parts of the micros cope.

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Activity 1 - Preparing a Wet Mount Slide

Materials for this activity


Paper towel
Clean slide and cover slip
Dropper bottle of methylene blue stain. Dropper
bottle of physiological saline solution Flat
toothpick / paddle pop stick.
Microscope

Collect a microscope. It should always be carried in an upright position.


Never place the microscope close to the edge of the bench.
Never touch the lens with your hands.
Use the revolving nosepiece to change objective lenses.
Always locate the object to be studied with the scanning-power objective in
position and then turn to the higher power objectives if a more detailed study
is required.
Do not use the coarse adjustment when the high power objective is in
position. It is difficult to control the movement of this objective through small
distances except by use of the fine adjustment knob.
Never focus upward while looking through the eyepiece. Rather, look at the
objective from the side to prevent moving the body tube too far and possibly
damaging the objective lens to say nothing of the possibility of smashing the
slide.

1. Place a drop of saline solution on the clean slide. Gently scrape the inner surface
of your cheek with the flat part of the toothpick, (Be careful not to draw blood. If
blood is drawn, tell the instructor.) Transfer the cells from the toothpick to the
saline solution by carefully stirring the end of the toothpick with the cheek cells in
the saline (Figure a).
2. Place a small drop of the methylene blue stain directly on the saline/cell smear
(Figure b). Stir it again with the toothpick and immediately dispose of the
toothpick in the autoclave bag.
3. Hold the cover slip by the sides to avoid getting fingerprints on it. Place one edge
of the cover slip next to one edge of the smear, and then lower it slowly into the
smear (Figure c). By lowering it slowly you will avoid forming air bubbles. Absorb
any excess fluid around the edges of the cover slip using paper towel.

(a) Stir cheek cells into saline solution b) Add drop of stain to the saline/cell smear (c) Lower cover slip onto smear

4. Place the cheek smear slide onto the stage. Focus with the scanning power lens
first. Scan for some individual cheek cells first. Notice that each cell has a
distinctive dark centre called the nucleus. Draw one of these cells in results table
on the following page. Be sure to include the proportion of the cell to the whole
visual field, including details like the shape of the cell and nucleus and if the
cytoplasm appears cloudy, clear or grainy.
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5. Return to the scanning-power lens, and scan for a cluster of cheek cells. Again
look for a cluster that is separate from the others. Draw the cluster of cells in your
results table.

Results:
Individual cheek cell:
Scanning power

Low power

High power

Low power

High power

Grouped cheek cells:


Scanning power

10
Questions:
1.

What is the total magnification if the ocular lens is 10x and the objective lens is
100x? Show the calculations.

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2.

Why should you scan a specimen under scanning power before using higher
magnifications?

3.

How is poor light quality affected by raising the condenser?

4.

How is poor light quality affected by opening the iris diaphragm?

5.

What is the shape of the individual cheek cells?

6.

What does the thickness and the shape of the cells tell you about their function?

7.

Was the nucleus centrally or peripherally located in the cells?

8.

How do you think cheek cells connect to each other?

9.

Why did you make the slide using physiological saline solution? Could you just
use water instead?

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Activity 2 - Basal Metabolic Rate


Basal Metabolic Rate (BMR) is the minimum amount of energy required to
sustain life and its calculation forms the basis of estimating daily energy
requirements. The BMR does not account for any activity the individual may
participate in during the day. It simply estimates the energy needed to sustain a
heartbeat, breathing and normal body temperature with the body at rest (not
sleep) at room temperature.
Calculate your own and the class averages for BMR using one of the following
formulae:
Male:

BMR = 66 + (13.7 x weight) + (5 x height) (6.8 x age)

Female:

BMR = 655 + (9.6 x weight) + (1.7 x height) (4.7 x age)

For these equations, use weight in kilograms and height in centimetres.

Calculations:
My height = cm
My weight =
Results:
My
calories
TheBMR
classisaverage for males
is per day.
The class average for females is

calories per day.


calories per day.

Questions:
1. Which was higher male or female?
2. Explain the results.

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kg

Activity 3 Cardiovascular Physiology


Many of the functions of the cardiovascular system can be measured using a few
simple instruments. These give an indication of the condition of various structures.
In this activity you will investigate, measure and record the following:
1. heart sounds using stethoscopes
2. pulse rate from two superficial pulse points (three repetitions) by palpation and
timing with a watch
3. blood pressure (using a sphygmomanometer) and pulse (radial)
(a) at rest sitting
(b) after 3 minutes of intense activity (immediately and 5 minutes after you
cease)

1. Heart Sounds
To hear the heart sounds you need to use a stethoscope.
The earpieces need to be cleaned with alcohol swabs before use by each student.
Once the alcohol has dried, place the ear pieces in your ears and the bell of the
stethoscope on your partners thorax just to the medial side of the left nipple at the
5th intercostal space. A characteristic lup dup sound can be heard with each beat of
the heart, representing the closure of the atrio-ventricular and semi-lunar valves
respectively.
2. Pulse
Pulse rate can be elicited by palpating one of the major superficial pulse points on
the body. The common carotid artery (neck), radial artery (in the wrist on the thumb
side) and femoral artery (groin) are frequently used.
Take 3 readings of your partners pulse at two pulse points over a 1-minute period,
and compare the averages.
3. Blood Pressure
Blood pressure is measured in millimetres of mercury using a sphygmomanometer.
Firstly determine the blood pressure in (a) a relaxed sitting position.
Collect a stethoscope, a sphygmomanometer and some alcohol swabs from the
front bench. Clean the earpieces as before and remove any excess air from the cuff
of the sphygmomanometer.
Wrap the cuff of the sphygmomanometer around your partners upper arm just
above the elbow. Find the arrow marked on the cuff and adjust the cuff so that the
arrow points to the brachial artery. Secure the cuff using the Velcro wrap.
Palpate (find by touch) the brachial artery and place the bell of the stethoscope on
the artery. Inflate the cuff to approx 160 mm pressure. Keeping the stethoscope in
place, slowly release the pressure valve on the sphygmo and listen for the first
thudding sounds of blood moving through the partially blocked (occluded) artery.
This is the systolic pressure. Record this on the results table.

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Continue to release the pressure until the sound muffles or disappears and again record
the result. This is the diastolic pressure. Repeat once.
Have your partner repeat the procedure to find your blood pressure. Blood pressure is
recorded as the systolic pressure divided by the diastolic pressure.
Now determine the blood pressure and pulse of your partner and yourself after 3
minutes of intense activity (immediately and at 5 minutes).

Results:
1. Pulse rate:
Pulse point 1

Pulse point 2

Reading 1
Reading 2
Reading 3
Average
Question: How could you explain the difference between the average results for
the two different pulse points?

2. Blood pressure:
After 3 minutes of activity
At rest
0 min
Blood pressure
Pulse
Blood
pressure
Question: Why did the blood pressure readings Pulse at rest
change depending on your activity?
(average
from above)
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5 min

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When you are exercising your muscles need more oxygen. The heart then pumps more
blood around your body with more powerful contractions which will make your blood
pressure increase. Your systolic blood pressure increases during exercise as the
cardiovascular system delivers more blood to the working muscles and your diastolic
blood pressure stays roughly the same or decrease slightly.

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