Practical Manual: Name of Course: Course Code: Compiled By: The Manual Belongs To

Programme: Bachelor of Medical Laboratory Science
Practical Manual
Name of Course: Introduction to Medical Laboratory Science
Course Code: IML511S
Compiled by: Ms E van der Colf
The Manual belongs to:__________________
2020
NAMIBIA UNIVERSITY OF SCIENCE AND TECHNOLOGY DEPARTMENT OF HEALTH SCIENCES
IML511S INTRODUCTION TO MEDICAL LABORATORY SCIENCE
PRACTICAL MANUAL
CONTENT
Objective
13 February 7:00-8:30 NUST Clinic Hepatitis B vaccination
14 February WILSIM lab Hand out of white coats
20 February Lecture on laboratory safety – hand out of safety manuals
27 February Practical 1: Safety rules. Daily maintenance in the laboratory. Microscope
5 March Practical 2: Phlebotomy Technique
12 March Practical 3: Specimen collection
19 March Presentations on specimen collection
26 March Practical 4: Microscopy
2 April Practical 5: Centrifuges. Balances – make up normal saline
16 April Practical 6: Serial dilution, pipetting and spectrophotometry
23 April Practical 7: Pipetting skills, Precision
30 April Practical 8: Units of measurement, Levey-Jennings chart
2
IML511S Introduction to Medical Laboratory Science
Objective
The objective of practical classes is to introduce students to basic techniques and equipment
used in the medical laboratory. Basic skills like phlebotomy and pipetting are developed.
At the end of the practical training students should be able to:
• Perform daily maintenance of microscopes and decontaminate benches.

• Perform the phlebotomy technique on a manikin
• Identify appropriate Vacutainers needed for specimen collection in order to perform
specific tests in the clinical laboratory
• Use a microscope effectively to correctly identify all the types of white blood cells in a
normal blood smear
• Balance, set and start a centrifuge
• Use a balance and laboratory glassware to make up normal saline, and biocide
solution according to instructions on the package
• Use a micropipette to make serial dilutions of a blue coloured solution, record
absorbance at the correct wavelength on a spectrophotometer and draw a standard
curve.
• Demonstrate accurate pipetting skills as illustrated on a standard curve. This
practical will be repeated till each student has reached the set standard of pipetting
skill.
• Demonstrate understanding of units of measurement, basic interpretation of lab
results and calculations pertaining to the Levey-Jennings chart.
Please note:
Before commencement of practicals for this semester students should familiarize themselves
with the Safety Manual which was handed out at the beginning of the year. They have
signed that they are familiar with safety rules in the medical laboratory as spelled out in the
Safety Manual and they are responsible to see that they adhere to these rules at all times
while in the laboratory.
Students who have failed to register as Medical Laboratory Scientist Students with the
Health Professions Councils of Namibia will not be allowed to do practicals where patient
samples are handled. They will forfeit any marks allocated for such practical.
Practical sessions are compulsory for all students while some practicals have marks
allocated towards the full period mark. Students are encouraged not to miss a practical since
no practical session will be repeated.
Students will have to use a scientific calculator for these practicals.
3
Practical 1: Observe safety rules in the laboratory. Perform daily maintenance

of microscopes and decontaminate benches.
Objective:
Students will be held responsible to adhere to safety rules in the laboratory and should know
where to find information pertaining to safety. Students should be able to perform daily
maintenance on the microscopes and at all times adhere to the laboratory rules pertaining to
this. Whenever patient samples are handled, proper bench decontamination should be done
afterwards.
A. Safety Manual and Standard Operating Procedures
Students should be familiar with the content of the Safety Manual, know where to find it, as
well as know where Standard Operating Procedures pertaining to laboratory safety are kept.
B. Maintenance of Microscopes
Instrumentation:
Microscopes
Reagents and Consumables:
• Ethanol
• Lens wipes
• Maintenance log sheet
Method:
Before a practical with microscopes, proper cleaning should be done according to

instructions. After using the microscope, each microscope should be cleaned again and the
maintenance sheet should be filled in by the student and signed by the supervisor.
C. Biocide
Objective:
Students should observe a demonstration of how to make up a biocide solution used

commonly in the clinical laboratory to disinfect working benches.
Reagents:
• Sachet of Biocide
• Beaker of water
• Squirt bottles
Method:
Students are to disinfect bench tops before they leave. Squirt biocide solution over bench
tops and wipe down with paper towels. Students should fill the decontamination sheet and
have the supervisor sign it.
4
Practical 2: Phlebotomy Technique
Objective:
At the end of the practical each student should be able to perform the phlebotomy technique
on a manikin. The technique will be demonstrated by an instructor, after which each student
will get the opportunity to draw artificial blood from a manikin.
Instrumentation:
Manikin
• Artificial blood
• Sealed, unused needle
• Needle cap
• Cuff
• Vacutainer tubes
• Sharps container
• Cotton wool swabs
• Ethanol
Method:
Follow instructions in Study Notes. Students should prepare for the practical by studying the
Study Notes. They should be familiar with the correct sequence of steps, how they follow
upon each other.
Lab report:
Sign attendance register
5
Practical 3: Specimen collection
Objective:
At the end of the practical each student should be able to identify appropriate Vacutainers
needed for specimen collection in order to perform specific tests in the clinical laboratory.
One of each type of Vacutainer per group:
Yellow top; Red top; Light blue top; Green top; Purple top; Grey top; PPT tube (viral load)
One blood culture bottle per group (four groups)
Method
Each group to prepare a poster with the following information:
Example of each type of colour coded Vacutainer, plus a blood culture bottle
For each container, indicate:
Stopper colour Additive Tests Minimum draw Special remarks

volume
Refer to Namibia University of Science and Technology IML511S Study Notes
Lab report:
Group Presentation
Each group to present as follows:
Group Topic
1 Additives
2 Tests performed
3 Minimum draw volume
4 Special remarks / procedures
Marks will be allocated per group. Since everybody in the group will get the same mark,
participation is expected from each and every group member.
6
Practical 4: Microscopy: ID Leukocytes in normal blood smear
Objective:
At the end of the practical each student should be able to use a microscope effectively to
correctly identify all the types of white blood cells in a normal blood smear
Instrumentation:
Microscope
Covered slide with normal blood smear
Immersion oil
Lens paper
Ethanol
Xylene to clean slides
Industrial wiper roll
Method:
Use the microscope as instructed in the lesson.
1. Mount the slide in the specimen holder, ensuring that it fits tightly.
2. Select the area which you are going to investigate – it should be at the round end of
the slide
3. Use the 40X objective to select an area where red blood cells are evenly distributed
4. Move the objective away
5. Insert a small drop of oil on the area
6. Position the 100X objective over the oil and carefully adjust the mechanical stage.
The objective should just touch the oil.
7. Select an area where red blood cells are evenly distributed, and don’t overlap.
8. Identify the five different types of leukocytes. Have the instructor sign your lab report
when you correctly identify a type of leukocyte.
7
Lab Report
IML511S: Laboratory Practical on Microscopy: ID of leukocytes
Refer to Hole’s Human Anatomy and Physiology
The following leukocytes were identified: [5]
Cell identified Tutor signature
Neutrophil
Eosinophil
Basophil
Monocyte
Lymphocyte
8
Practical 5: Centrifuges. Balances – make up normal saline
Centrifuge
Objective:
At the end of the practical each student should be able to balance, set and start a centrifuge.
An instructor will be available at each centrifuge to supervise students. Students may work
in pairs.
A. Instrumentation: Centrifuge
Ten Vacutainer tubes containing different volumes of artificial blood (one set for each
centrifuge)
Method:
1. Load the Vacutainer tubes into the centrifuge, balancing equal volumes opposite
each other.
2. Call the supervisor to check the loading.
3. Carefully close the lid firmly.
4. Set the time and speed as indicated by the instructor
5. Press the start button
6. When the centrifugation starts, check for imbalance.
7. Wait for the centrifuge to stop, open the lid and remove Vacutainers.
Lab report: Sign attendance register
B. Normal (Physiological) Saline
Objective: At the end of the practical each student should be able to use a balance and
laboratory glassware effectively to correctly make up normal saline. The effect of hypotonic
solution on red blood cells will be demonstrated. Before the practical, students should refer
to the reference to calculate the correct amount of NaCl to be weighed out, and to complete
the lab report.
Instrumentation:
Balance, spatula
Reagents and Consumables for each student:
• NaCl 2 test tubes with screw caps

• Beaker with DDW anticoagulated whole blood
• Weighing boat timer
• Plastic pasteur pipette fine markers
• 200 ml volumetric flask with cap pipette 20-200ul
• Funnel pipette tips x 2
• 200 ml bottle with label to store normal saline
9
Method:
1. Weigh out the correct amount of NaCl to make up 200 ml of normal

(physiological) saline.
2. Use the funnel to fill the volumetric flask two thirds full with DDW
3. Add the NaCl to the DDW in the volumetric flask
4. Rinse the weighing boat with DDW using the plastic pasteur pipette
5. Add to the volumetric flask
6. Cap the volumetric flask and swirl gently till all the NaCl is dissolved
7. Fill the volumetric flask with DDW up to the line on the neck. You may use the
pasteur pipette to add the last few drops.
8. Remember the fault of Parallaks when aligning the bottom of the meniscus with
the engraving on the neck of the flask
9. Transfer the solution to the bottle and label correctly for storage
Test for red blood cell haemolysis:
1. Mark one test tube “DDW” and add 5 ml DDW using a Pasteur pipette.
2. Mark the other tube “Normal saline” and add 5 ml of the solution you have made up.
3. Add 50ul of anticoagulated blood to each tube and mix gently.
4. Leave the tubes upright for 15 minutes.
5. Centrifuge for 5 minutes at 3000 rpm
6. Record the colour of the solution in each tube.
Lab report:
Complete the questions and hand in before you leave the practical
10
Lab Report
IML511S: Introduction to Medical Laboratory Science practical on Physiological

(Normal) saline
DATE: ___________ NAME: _______________________________
MARKS: [46]
Reference: http://faculty.etsu.edu/currie/solutions.htm
1. We often use the terms “normal” or “physiological” saline to describe a

…………………. (%) solution of ……………. (2)
2. This means that 100 ml of normal saline contains …………. g of …………….. (2)
3. Show your calculations to determine the molarity of normal saline. (5)
4. Show your calculations to determine how 200 ml of normal saline must be made up.
(2)
5. Define “osmolarity” of a solution
(3)
6. Define an “electrolyte” and give an example of an electrolyte found in the human

body.
(2)
7. Define the following solutions:
Hypotonic:
11
Isotonic:
Hypertonic
(3)
8. Describe the following as hypotonic, isotonic or hypertonic:
• Seawater in comparison to plasma _____________________

• Plasma in comparison to interstitial (intercellular) fluid _________________
• Intercellular fluid in comparison to intracellular (inside cells) fluid
______________ (3)
9. What happens to a red blood cell (RBC) in DDW?
Why?
What happens if the cell swells too much?
What is this called? (5)
10. What happens to a red blood cell in normal saline?
Why?
(2)
11. What happens to a red blood cell in hypertonic solution?
Why?
12
What is this called? (4)
12. Record the colour of the solutions in the test tubes:
Saline: ___________________
Distilled water: ______________________________ (4)
13. Marks for label on bottle of saline: (7)
Reagent: ………………………………… Date prepared: ………………..…

Use for: ……………………………………… Expiry date: …………………..…
Store at: …………………………………….. QC checked by: ……….……..…
Lot Number: ………………………………... Date QC checked: ………….……
Prepared by: ……………………………….
Reference: (2)
13
Practical 6: Serial dilution, pipetting and spectrophotometry
Objective:
At the end of the practical each student should be able to use a micropipette to make serial
dilutions of a blue coloured solution, record absorbance at the correct wavelength on a
spectrophotometer and draw a curve.
Demonstrate accurate pipetting skills as illustrated on a curve.
A. DILUTIONS
Instrumentation:
Automated pipette 1000 µl adjustable volume
Consumables for each student: Four tips, waste bucket, 4 test tubes with screw caps (4ml)
in a rack
Reagents for each student: Sample (1.5 ml of water mixed with blue dye)
Double distilled water (5 ml)
Fine marker
Method:
1. Pipette into 4 test tubes: 1 ml water each (use same tip)

Remember to push to first stop to aspirate and push to second stop to release the
volume.
Release the stopper between tubes.
2. Transfer one ml of sample to tube 1. (Use a new tip)
3. Mix well
4. Use same tip and transfer 1 ml from tube 1 to tube 2.
5. Repeat 3 and 4 through tube 4.
6. Discard 1ml from tube 4.
7. Calculate the final dilution in each tube.
1. ___________
2. ___________
3. ___________
4. ___________
8. Complete the sentence: “This is called a ………. fold dilution.”
14
B. SPECTROPHOTOMETRY
Instrument: Spectrophotometer
Consumables:
• 4 cuvettes per student (to be washed and reused after the practical)
• squirt bottle with DDW
• one cuvette for blank at each spectrophotometer
• Waste bucket
• Paper towel
Reagents: Dilutions of the sample (blue dye) prepared above
Method:
If the sample is observed by the eye as blue, it means that the colour absorbed in the
spectrophotometer is ……………………. (Refer to study notes).
The instrument wavelength should be set at …………………. nanometer.
Taking a reading on the spectrophotometer:
1. Fill the blank cuvette with water.

2. Wipe with paper towel. Don’t touch the clear sides
3. Insert the cuvette into the spectrophotometer
4. Take the reading and record it or zero the instrument if possible.
5. Pipette 1ml of each dilution into a set of four cuvettes
6. Repeat 2 – 4 for each cuvette
7. Rinse each cuvette with DDW
15
Spectrophotometer readings:
TUBE Absorbance Absorbance
Lab report:
1. Prepare a graph:
a. X-axis: absorbance at …………. nanometer
b. Y-axis: dilution e.g. 1:2 at the top and indicating higher dilutions nearer
to zero
2. Plot the absorbance for each dilution
3. Draw a line through the points
4. Ideally the graph should go through 0.
16
17
Practical 7: Pipetting skills - precision
Objective:
Students are to practice pipetting skills.
Instruments
Weighing balance
Pipettes: 200 – 1000 µl and 20-200 µl
Reagents / consumables
Pipette tips
Beaker with water
Weighing boat
Wiper paper
Method:
Calibrate the balance to zero with the weighing boat.
Pipette any volume of water into the boat and weigh it. Record the weight.
Discard the water and wipe the boat dry.
Repeat with the same volume and take five measurements in total.
Repeat with other pipette.
Weight pipette 1 Volume µl Weight pipette 2 Volume µl
Plot the weights obtained with each pipette on the graph. Use left Y-axis for one pipette’s
weights and the right Y-axis for the other pipette’s weights. Alternatively you could draw two
separate graphs.
Evaluate the precision obtained with each pipette. [20]
18
19
Practical 8: Units of measurement, Levey-Jennings chart
20
21
22
Lab Report: Interpret lab results
Full blood count results:
Use your study notes and indicate whether all tests are resulted in SI units.
YES/NO _______________
If NO, which one is not in SI units? ________________________________________
LFT results:
Use your study notes and indicate whether all tests are resulted in SI units.
YES/NO _______________
If NO, which one is not in SI units? ________________________________________
Total protein and glucose result:
What is wrong with the reporting of these results?
_________________________________________________________________________
CRP result:
What is your diagnosis of this patient? ________________________________________
BHCG result:
What is your diagnosis of this patient? ________________________________________
What does the Dl hCG indicate? _____________________________________________
Give the reason why it was performed.
________________________________________________________________________
________________________________________________________________________
(9)
23
Levey-Jennings Chart
How the manufacturer calculates the Standard Deviation (SD) for a commercial
glucose control sample:
The factory ran a glucose test on a commercial glucose control sample over ten consecutive
days, and obtained the following results in mmol/L:
Day number Test result Mean - value Square the

difference
1 10
2 6
3 12
4 13
5 7
6 10
7 9
8 8
9 14
10 11
SUM SQUARES:
1. Compute the mean __________
2. Subtract each value from the mean (ignore negatives)
3. Square the difference
4. Add the squares together _____________
5. Subtract 1 from n _______________
6. Divide n – 1 into the sum of squares ____________________
7. Take the square root of the number from step 6 ____________________
The answer in step 7 equals one SD.
This means that in this set of data there is a spread of 2.58 (1 SD) around the mean of ___
• So how do you determine 2 SD and 3 SD?

2 SD = 1 SD X 2 = ____________________
3 SD = 1 SD X 3 = ____________________
• Calculate one SD above the mean ___________________
• Calculate one SD below the mean ____________________
• Calculate two SD above the mean ____________________
• Calculate two SD below the mean _____________________ (7)
24
Plotting the Levey-Jennings chart for a haemoglobin control sample
The WILSIM lab bought three haemoglobin control samples from the manufacturer. You will
be shown the control results obtained in the WILSIM lab over several days using the
haematology analyser. Let us say you worked in the WILSIM lab and you tested the normal
haemoglobin control sample each morning over ten days. You obtained the following
haemoglobin results in g/dl:
Day number Test result

1 14.5
2 13.9
3 12.9
4 13.5
5 14.0
6 15.1
7 14.9
8 14.1
9 14.0
10 15.1
Refer to the package insert from the manufacturer and insert the SD’s of the normal control
in the Levey-Jennings chart. Plot the haemoglobin value obtained on each of the ten days.
Levey-Jennings chart – haemoglobin test done on normal control (17)
25
• 65% of the values will be +/- 1SD
For this LJ chart, __________% of the values is within +/- 1SD
• 95% of the values should fall between +/- 2SD (confidence limits)
• 99% of the values should fall between +/- 3 SD (repeat analysis)
TOTAL [36]
26
Semester marks for practicals (for student’s own records):
Practical Student mark Total mark Percentage
Specimen collection 30
ID leukocytes 5
Physiological saline 46
Pipetting 20
Levey Jennings 36
Mean Percentage
27

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Practical Manual: Name of Course: Course Code: Compiled By: The Manual Belongs To

Uploaded by

Copyright:

Available Formats

Programme: Bachelor of Medical Laboratory Science

Compiled by: Ms E van der Colf

The Manual belongs to:__________________

IML511S INTRODUCTION TO MEDICAL LABORATORY SCIENCE

13 February 7:00-8:30 NUST Clinic Hepatitis B vaccination

14 February WILSIM lab Hand out of white coats

20 February Lecture on laboratory safety – hand out of safety manuals

27 February Practical 1: Safety rules. Daily maintenance in the laboratory. Microscope

5 March Practical 2: Phlebotomy Technique

12 March Practical 3: Specimen collection

19 March Presentations on specimen collection

26 March Practical 4: Microscopy

2 April Practical 5: Centrifuges. Balances – make up normal saline

16 April Practical 6: Serial dilution, pipetting and spectrophotometry

23 April Practical 7: Pipetting skills, Precision

30 April Practical 8: Units of measurement, Levey-Jennings chart

At the end of the practical training students should be able to:

• Perform daily maintenance of microscopes and decontaminate benches.

Students will have to use a scientific calculator for these practicals.

Practical 1: Observe safety rules in the laboratory. Perform daily maintenance

A. Safety Manual and Standard Operating Procedures

Reagents and Consumables:

Before a practical with microscopes, proper cleaning should be done according to

Students should observe a demonstration of how to make up a biocide solution used

Practical 2: Phlebotomy Technique

Reagents and Consumables:

Sign attendance register

Practical 3: Specimen collection

Reagents and Consumables:

One of each type of Vacutainer per group:

One blood culture bottle per group (four groups)

Each group to prepare a poster with the following information:

For each container, indicate:

Stopper colour Additive Tests Minimum draw Special remarks

Refer to Namibia University of Science and Technology IML511S Study Notes

Each group to present as follows:

Practical 4: Microscopy: ID Leukocytes in normal blood smear

Reagents and Consumables:

Covered slide with normal blood smear

Xylene to clean slides

Industrial wiper roll

Use the microscope as instructed in the lesson.

IML511S: Laboratory Practical on Microscopy: ID of leukocytes

Refer to Hole’s Human Anatomy and Physiology

The following leukocytes were identified: [5]

Cell identified Tutor signature

Practical 5: Centrifuges. Balances – make up normal saline

Reagents and Consumables:

Lab report: Sign attendance register

B. Normal (Physiological) Saline

Reagents and Consumables for each student:

• NaCl 2 test tubes with screw caps

1. Weigh out the correct amount of NaCl to make up 200 ml of normal

Test for red blood cell haemolysis:

IML511S: Introduction to Medical Laboratory Science practical on Physiological

DATE: ___________ NAME: _______________________________

1. We often use the terms “normal” or “physiological” saline to describe a

5. Define “osmolarity” of a solution

6. Define an “electrolyte” and give an example of an electrolyte found in the human

7. Define the following solutions:

8. Describe the following as hypotonic, isotonic or hypertonic:

• Seawater in comparison to plasma _____________________

DATE: _ NAME: _____________________