INTERNATIONAL MEDICAL SCHOOL

BACHELOR’S IN MEDICAL SCIENCE

SUBJECT/CODE: GENETIC DISORDER/QDS20101P

Lab 1: INTRODUCTION TO LAB INSTRUMENTS
LECTURER NAME: DR SITI ARFFAH KAMARULZAMAN

NAME: ILHAASHINI KRISHNAN

IC NUMBER: 010301-05-0546
ID NUMBER: 012021090323
Introduction
Laboratory Instrument is any implement, tool, or utensil used for laboratory tests. An

instrument is a device that measures a physical quantity, such as flow, concentration,

temperature, level, distance, angle, or pressure. Using laboratory equipment correctly is

necessary for working safely with hazardous substances. This activity's primary elements are

maintenance of equipment and regular examination. The incorrect usage or maintenance of

laboratory equipment is accountable for a large number of accidents that take place there.

The safe handling of equipment used often in laboratories is covered in this section. In many

different types of subjects, including biology, chemistry, physics, etc., lab equipment is crucial.

Various lab equipment may perform a similar position. Therefore, to execute the experiments

properly, it is our responsibility as students to know how to use these lab instruments. This will

reduce accidents, which frequently happen in laboratories, and improve the accuracy of the

results. In this practical, the students will be exposed to the ideas and theories behind the

usage of several devices that are widely used in laboratories. Students can thus distinguish

between how various types of apparatus are used. The pupils are then able to comprehend

which laboratory equipment needs to be used for each experiment.

Objective:

At the end of the practical student should be able to:

• Familiar with the instruments that will be used in the molecular lab

• To practice proper use and technique of using a pipette and a micropipette which is

essential in molecular biology.

Methods:

Exercise I

1. A permanent marker is used to label two 1.5 microcentrifuge tubes A and B.

2. The matrix below is used as a checklist while adding solutions to each tube.

Tube Sol.I Sol.II Sol.III Sol.IV

A 100µl 200µl 150µl 550µl

B 150µl 250µl 350µl 250µl

3. The micropipette was set and appropriate volumes of solution I was added to tubes A

and B.

4. A fresh tip is used to add the appropriate volume of Solution II on tubes A and B.

5. A fresh tip is used to add the appropriate volume of Solution IIl on tubes A and B.

6. A fresh tip is used to add the appropriate volume of Solution IV on tubes A and B.

7. Tops of the tubes are closed.

8. The reagents were mixed by placing the tubes in a centrifuge and applying a short pulse

of several seconds. Make sure the tubes are placed in a balanced configuration in the

microcentrifuge rotor. Spinning tubes in an unbalanced position will damage the

microcentrifuge rotor.

9. A total of 1000 microlitres of reactants was added to each tube.

10. To check the accuracy of the instruments, the micropipette was set to 1000 microlitters

and carefully the solution was withdrawn from each tube.

a. Is the tip barely filled?

Tips of Tube A and B are not barely filled.

b.Does a small volume of fluid remain in the tube?

In tube A there is a small volume of fluid remaining in the tube. Whereas, there is no volume

remaining in tube B.

c.After withdrawing all fluid, is an air space left at the end of the tip? If your measurements

were inaccurate, repeat the exercise to obtain nearly perfect results.

Yes, there is an air space left at the end of the tip for tubes A and B. This is due to human

error which is a wrong technique of withdrawing of solution.

Exercise II

A.Zeroing the spectrophotometer-cuvette with 1ml distilled water and press the zero

button.

B.Preparing a dilution of Colour dye

1. An appropriate micropipette to measure 100 microlitres was chosen

2.100 microlitres of dye was transferred to an empty microcentrifuge tube

3.p1000 used to transfer 1ml of water to microcentrifuge tube.

4. The tube was mixed vigorously.

C.Spectrophotometer reading

1.1ml of the dye was pipetted from a microcentrifuge tube into a cuvette.

2. Cuvette was inserted into the spectrophotometer

3. The reading was observed from a digital display on a spectrophotometer.

4. Reading was recorded.

D.Independent trials

1.100 microlitres of dilution repeated two more times as outlined in boxes A-C.
2. The values were recorded in the data collection sheet.

3. The values for the same dilution were compared.

Repeating with other dilutions

1. Procedures as outlined in boxes A-C were repeated.

2. New dilutions using the following quantities of color dye were prepared such as 75

microlitres and 50 microlitres.

3. The data was recorded on the table collection sheet attached.

4. Three trials of each dilution were conducted.

Results

Dilutions Trials Absorbance

1 0.227

100µl 2 0.221

3 0.215

1 0.365

75µl 2 0.349

3 0.349

1 0.354

50µl 2 0.341

3 0.344
Discussion

We carried out two exercises that helped to learn how to use the micropipette, centrifuge, and

spectrophotometer. In the first exercise, we learned the proper way to handle the micropipette

and centrifuge. There are techniques and ways to withdraw the solution accurately. In the

second exercise, we learned the proper way to use a micropipette and also

spectrophotometer.

In the first exercise, There are various types of micropipettes such as adjustable volume

micropipettes and fixed volume micropipettes. there are different volumes of micropipettes

and there are different colors and sizes of tips for each of them. In this exercise, our aim is to

the accuracy of the micropipette when transferring the solution from the microcentrifuge tubes.

From this exercise, the proper way of using a micropipette is important to get accurate results.

Based on our results, In tube A, when we withdrew 1000 microlitter solution from the tube

using a micropipette, there was a remaining solution in the tube. This indicates that there are

more than 1000 microliters of solution in the tube. In tube B, when we withdrew 1000 microlitter

solution from the tube, there was a lack of 1000 microlitter solution in the tube and it caused

the presence of air bubbles in the micropipette. This is because of several factors that affected

the results such as technical error and human error. For technical errors, we didn’t use the

pipette with the correct volume. Therefore we cannot withdraw the accurate amount of solution

at one time. This causes the inaccurate amount of the solution in tube A and tube B. Next the

human error is, that we used the wrong technique of withdrawing the solution which led to the

presence of air bubbles in the solution. For example, keeping the angle of the pipette at an

angle less than 20 degrees from the vertical position will give accurate results. Therefore, the

technique of using a pipette is important because it may affect the results during tests.

In the second exercise, We learned the proper way to handle the micropipette and

spectrophotometer. The spectrophotometer is an instrument that measures the amount of the

photo absorbed after it passes through the sample solution. We mixed blue dye solution in

three different concentration which is 100 microlitres,75 microlitres, and 50 microlitres with
1000 microlitres of distilled water. Then we prepared distilled water as a blank solution to zero

the spectrophotometer. Then we insert the solutions in cuvettes accordingly. We insert the

cuvette inside the spectrophotometer with the wavelength to obtain the result. For each

concentration, we inserted three times and recorded the readings. Absorbance is proportional

to the substance’s concentration. When the concentration is higher, it produces a high value

of absorbance. Based on our result, we had inconsistent findings that the higher concentration

which is 100 microlitres obtain the lowest absorbance value. Whereas, compared to 50

microlitre concentration,75 microlitre concentration is higher which is the same as estimated

results. This is due to the instrumental error where there are scratches on the cuvettes which

may produce inconsistent findings. And also can be due to the presence of air bubbles during

the withdrawal of the solution using a micropipette. When we compare the three times of trials

of each concentration, there is not much difference between the absorbance values. This

indicates that the spectrophotometer gives the same reading for all the trials.

Question

1. Describe the function of micropipette parts and tips and centrifuge.

Plunger: It primarily serves the following two functions: Volume change, liquid aspiration, and

liquid dispensing

Tip Ejector: Micropipettes come with a tip ejector that makes it possible to quickly, easily, and

safely expel tips.

Volume display: The volume of the liquid to be taken in or dispensed is displayed using the

volume display. In the range of micropipettes, for example, a micropipette with a capacity of

10 l to 100 l can be modified to any value between 10 l and 100 l according to the extensive

range of adjustments available with variable pipettes.

Tipcone: The tipcone helps the tips fit together. Because it improves the instrument's

compatibility with most common tips, a micropipette with a universal tip cone is advised.

Grip: It makes the micropipette easier to support and comfortably grasp.

Tip for the pipette: It is a piece of equipment made of virgin polypropylene or plastic that has

been molded into it and is in touch with the liquid. Tips are thrown away. There are several tip

sizes and colors for each type of micropipette.

Centrifuge: For spinning liquid samples at high speeds that are as small as 2 ml or even less,

microcentrifuges are used. The purpose of centrifuging samples that are contained in tiny

capillary tubes is to divide their components into two phases.

2. List precautions steps or rules when using a micropipette and microcentrifuge.

• Don't ever change the volume above what the micropipette can handle. No

micropipette should ever be set lower than zero L. Never adjust the P20 above 20 L,

the P200 over 200 L, or the P1000 over 1 mL.

• Never turn the volume dial with force. If turning the knob becomes difficult, the pipette

may be destroyed or you are using it beyond its capacity. Alert your instructor to the

problem.

• Don't let the micropipette drop.

• When using the micropipette, always move it smoothly. This will aid in providing

accurate measurements and prevent pipette breakage. It shouldn't make "snapping"

sounds.

• Always hold pipettes vertically.

• Never place a pipette on a bench with liquid in the tip.

• Always use a pipette of the correct size for the volume you are measuring.

• Always place tips in the proper rubbish container.

• To prevent contamination, use a separate tip for every sample.

• Avoid touching the tip with your hands.

 Microcentrifuge

Before spinning, check that the sample is stable and that the spindle is clean. For instance,

if we have two samples, we can place them in opposing positions to ensure that the

samples are stable. Also, check that the microcentrifuge's lid is closed before is on.

3. Describe the purpose of the two stops on a micropipette plunger.

The first stop is used to fill the micropipette tip, and the second stop is used to dispense

the contents of the tip. The operator depresses the plunger to the first stop, immerses the

tip into the sample then releases the plunger slowly to withdraw the sample. The second

stop is to dispense the contents of the tip into the test tube.

Conclusion

There are many equipment and instruments in the laboratory. For each instrument, there are

proper ways to handle it. This proper way is to reduce the errors and to get accurate results

without error. On the other hand, precautions when using laboratory instruments are to avoid

damage to the instruments and avoid laboratory accidents. In this lab, we learned about the

proper way to use the micropipette, microcentrifuge, and spectrophotometer. Some results we

didn’t get due to some errors. Therefore these techniques of handling laboratory instruments

are very important.

References

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Evidence: