COLEGIO SAN AGUSTIN-BACOLOD

College of Health and Allied Professions MLS 111A LABORATORY ACTIVITY SHEET
Medical Technology Program

LABORATORY ACTIVITY 1
Laboratory Resources (Pipette)

I. Desired learning outcomes

As future laboratory scientists, it is important that medical technology students learn
the basic supplies and equipment used in Clinical Chemistry. One of these is a pipette.
Although most clinical chemistry laboratories use semi-automated or fully automated
pipettes, there is still the need to master the manual operation of these pipettes to be
equipped with the complete skills of aspirating and transferring fluids from one receptacle
to another.
After performing this activity, students should be able to:
1. Identify correctly the different kinds of pipettes according to classification, description,
and function;
2. Choose the appropriate pipette to use and
3. Perform proper pipetting technique.
II. Materials
1 mL Serological Pipette Stirring rod/Stirrers
5 mL Serological Pipette Water
1000 uL Micropipette Serum
100 uL Micropipette Whole blood
Pipette tips Colored powder
Aspirator bulb Folded tissue paper
Beakers Freshly prepared 10% bleach solution

III. Procedure
A. SEROLOGICAL PIPETTE
1. Check the pipette to ascertain its correct size and check for broken delivery or suction tips.
2. Wearing protective gloves, hold the pipette lightly between the thumb and last three
fingers, freeing the index finger.
3. Place the tip of the pipette well below the surface of the liquid to be pipetted.
4. Using mechanical suction or an aspirator bulb, carefully draw the liquid into the pipette
until the liquid level is well above the calibration mark.
5. Quickly cover the suction opening at the top of the pipette with the index finger.
6. Wipe the outside of the pipette dry with a piece of clean tissue to remove excess fluid.
7. Hold the pipette vertically with the delivery tip against the inside of the original vessel.
Carefully allow the liquid in the pipette to drain by gravity until the bottom of the meniscus
is exactly at the calibration mark. To do this, do not entirely remove the index finger from
the suction hole end of the pipette; rather, by rolling the finger slightly over the opening,
allow slow drainage.
8. While holding the pipette vertically, touch the pipette’s tip to the inside wall of the
receiving vessel. Remove the index finger from the top of the pipette to permit free
drainage. Remember to keep the pipette in a vertical position for correct drainage. In TD
(to deliver) pipettes, a small amount of fluid will remain in the delivery tip.
9. To be certain that the drainage is as complete as possible, touch the delivery tip of the
pipette to another area on the inside wall of the receiving vessel

B. MICROPIPETTE
1. Select the appropriate micropipette to use. Turn the dial to the exact amount of sample to
be aspirated.
Note: Take note of the minimum and maximum limits of the micropipette. DO
NOT EXCEED THESE LIMITS. Exceeding these values will put the pipet out of calibration.

2. Hold the micropipette in your dominant hand, and gently but securely place the end of the
micropipette into the proper size tip. Once the tip is on, be careful not to touch the tip on
 COLEGIO SAN AGUSTIN-BACOLOD
College of Health and Allied Professions MLS 111A LABORATORY ACTIVITY SHEET
Medical Technology Program

anything! If your tip touches the bench, lab coat, etc., eject the tip into the waste
container and get a new clean pipet tip.
3. With your other hand, open the cap of the sample tube and bring the tube to eye level.
4. Push the micropipette plunger down to the first stop with your thumb and hold it in this
position.
5. Place the pipet tip into the tube and gently release your thumb from the plunger to draw
fluid into the tip. The micropipette should be upright while aspirating. Confirm that the tip
has liquid and that no bubbles are present within the tip.
6. Close the sample tube and place it back in the tube rack.
7. Gently touch the tip to the side of the tube and slowly push all the way down (to the
second stop) on the plunger to dispense the liquid.

IV. Guide Questions

1. Make a 3-column table to classify the different pipettes, their corresponding
specific or unique features, and their function. Then, with your own experience
while using them, make a graph to show their comparison according to the degree
of easiness to manipulate or use. (Criteria: 1- being the easiest to use, 2- being not
so easy and not so difficult to use, and 3- being the most difficult to use). If the
mode of learning is virtual, the comparison of the three pipettes will be on the
design and functionality only. Use a table.
2. Why do you have to carefully select the pipette that you use in your work?
3. When do you apply the law of “lower and upper meniscus levels”? How can
improper reading of the volume of specimens or reagents inside the pipette lead
to inaccuracy of result?
 COLEGIO SAN AGUSTIN-BACOLOD
College of Health and Allied Professions MLS 111A LABORATORY ACTIVITY SHEET
Medical Technology Program

NAME OF STUDENT: _00

I. Observation/Results

Figure 2. Receiving beaker and

Figure 1. Micropipette, Serological
source beaker with fluids
Pipette, and Aspirator bulb

Figure 3. Putting of aspirator bulb in the serological Figure 4. Drawing the liquid
pipette with its orifice tip immersed in the water using aspirator bulb
 COLEGIO SAN AGUSTIN-BACOLOD
College of Health and Allied Professions MLS 111A LABORATORY ACTIVITY SHEET
Medical Technology Program

Figure 5. Holding, draining to Figure 6. Blowing out

calibration, and dispensing

Figure 7. Sucking using the Figure 8. Transferring the

micropipette micropipette with liquid

Figure 9. Dispensing the

micropipette

II. Discussion
 COLEGIO SAN AGUSTIN-BACOLOD
College of Health and Allied Professions MLS 111A LABORATORY ACTIVITY SHEET
Medical Technology Program

Clinical chemistry delves into the biochemical examination of bodily fluids to aid in illness
diagnosis and therapy (Marshall et al., 2020). One crucial equipment in clinical chemistry is the pipettes,
which are used to move precise quantities of liquid for analysis. The appropriate pipette must be chosen
for accurate liquid handling during sample preparation, which is required for reliable findings.

Identifying various types of pipettes involves understanding their classifications, descriptions,

and functions. Based on their design classification, pipettes can be divided into two groups: "to contain"
(TC) and "to deliver" (TD). Turgeon (2015) says that TD pipettes accurately dispense the volume stated,
in contrast to TC pipettes, which hold a given volume but may not dispense the correct amount.
Additionally, pipettes can be further classified based on how they drain, including blowout and self-
draining varieties (Carreiro-Lewandowski, 2013). Measuring pipettes come in various types, including
micropipettes, Ball/Kolmer/Kahn, Mohr, bacteriologic, and serological pipettes. Volumetric, Ostwald-
Folin, Pasteur, and automated macro- and micropipettes are examples of transfer pipettes (Guzman,
2001). Every type has a specific purpose; self-draining pipettes allow gravity drainage, whereas blowout
pipettes have continuously etched rings for total liquid expulsion. Serological and Mohr pipets, which fall
under the classification of graduated/measuring pipets, are TD/blowout and TD/no blowout,
respectively, and may measure different liquid volumes—however, they cannot be a calibrator. Without
graduations but still equipped with a bulb, volumetric pipettes extremely precisely measure a single
volume of reagent or sample and can dispense diluent to lyophilized control (Church, 2018). Viscous
solutions are ideal for the Ostwald-Folin pipet but are not as common now. Micropipettes, also known
as TC, are devices that measure small amounts of liquid and have a variety of uses. The micropipette's
content is typically expelled using a small pipetting bulb, and the inner surfaces are cleansed by
aspirating the diluent from the receiving container with a minor aspirating bulb. While electronic
pipetters offer programmed modes with digital volume display for various pipetting activities,
semiautomated pipettes are more convenient and use plastic tips as they do not require pipetting bulb
and keep less inner surface film (Hawach, 2023). A multichannel pipette operates similarly to a single-
channel pipette but can accommodate multiple tips simultaneously. Lastly, unlike conventional pipettes,
the repeat pipette dispenser is designed for dispensing specific volumes into multiple receptacles
without aspirating between dispenses, thus saving time (Pipettes.com, 2022).

When measuring precise volumes, the appropriate pipette depends on the specific application.
TC and TD pipettes are for accurate measurements without or with discharge, respectively (Turgeon).
Blowout and self-draining pipettes are used when complete dispensing without residual volume or easy
one-handed operation is vital (Carreiro-Lewandowski). Measuring pipettes like Micropipettes are best
for small-volume measurements, while Ball/Kolmer/Kahn and Mohr pipettes are suitable for varying
dispensing needs, the latter without blowout (Guzman). Bacteriologic and serological pipettes address
specialized tasks, the former for bacterial cultures and the latter for large-volume titrations. Transfer
pipettes include volumetric, and Ostwald-Folin pipettes for viscous controlled transfers and fine
titrations, while Pasteur pipettes offer quick non-precise transfers (Church). For automation, electronic
pipetters and semiautomated options enhance efficiency. Multichannel pipettes are ideal for high-
throughput assays, whereas repeat pipette dispensers ensure consistent dispensing of the same volume.

Executing precise pipetting technique, especially with air-displacement pipettes, May (2018)
enumerates several crucial steps: firstly, set the desired volume and depress the plunger; subsequently,
submerge the tip to the correct depth, allowing smooth plunger release to its initial position, and wait
briefly for liquid intake; then, position the pipette at a 10-45 degree angle against the chamber wall,
press the plunger to the first stop, pause, and depress to the second stop; glide the tip along the vessel
wall while lifting to detach, and let the plunger return to rest. Notably, electronic pipettes automate
certain stages, streamlining the process for improved precision.
III. Guide Questions
 COLEGIO SAN AGUSTIN-BACOLOD
College of Health and Allied Professions MLS 111A LABORATORY ACTIVITY SHEET
Medical Technology Program

1. Make a 3-column table to classify the different pipettes, their corresponding specific or unique
features, and functions. Then, with your own experience while using them, make a graph to show
their comparison according to the degree of easiness to manipulate or use. (Criteria: 1- being the
easiest to use, 2- being not so easy and not so difficult to use, and 3- being the most difficult to use).
If the mode of learning is virtual, comparing the three pipettes will be on the design and functionality
only. Use a table.

Pipette Type Features Functions

TC and TD Color-coded bands, TC: used when the remaining liquid in the tip is not
Pipettes Different calibration to be dispensed. TD: used when the entire liquid,
including the droplet in the tip, is to be dispensed
(Turgeon, 2015).
Blowout Pipettes Presence of etched rings They are used when the entire liquid, including the
or bands droplet in the tip, is to be dispensed (Carreiro-
Lewandowski, 2013).
Self-Draining Absence of etched rings They are used when the remaining liquid in the tip is
Pipettes or bands not to be dispensed (Carreiro-Lewandowski, 2013).
Micropipettes Digital display, It is used to accurately and precisely transfer very
adjustment dial small volumes of liquid (Rodrigues & Curtis, 2010).
Ball/Kolmer/Kahn Presence of a ball or bulb They are used specifically for serological and
Pipettes in the middle bacteriological tests for mixing and dispensing
specific volumes of liquid (Guzman, 2001).
Mohr Pipettes Straight tube, graduation Used for accurately measuring and transferring a
marks stop before the tip; variety of liquid volumes, except the last drop
TD/No Blowout (Guzman, 2001).
Bacteriologic May have a cotton plug at They are used for transferring and mixing liquids in
Pipettes the top bacteriological applications to avoid cross-
contamination (Hewitt, 2012).
Serological Graduations go down to They are used for accurately measuring and
Pipettes the tip, TD/Blowout. transferring a variety of liquid volumes, including the
last drop (Guzman, 2001).
Volumetric Single graduation mark They are used for preparing solutions with precise
Pipettes volumes for quantitative analyses, like titrations
(Church, 2018).
Ostwald-Folin Larger bulb, no They are used specifically to accurately transfer
Pipettes graduation marks viscous or foamy liquids (Guzman, 2001).
Pasteur Pipettes Dropper pipette, usually Used for transferring small amounts of liquid, not for
plastic or glass; Thin stem, precise measurements. Commonly used in mixing
often comes with a solutions and liquid-liquid extractions (Liu et al.,
rubber bulb. 2019).
Automated Motorized, programmable They are used for high-throughput, repetitive tasks
Macro- and to avoid manual errors and fatigue (Henry, 2011).
Micropipettes
Electronic Electronic display, buttons They are used for precise, repetitive tasks, improving
Pipetters for programming efficiency, and reducing manual errors (Hawach,
2023).
Semiautomated Combination of manual Allows manual control for certain tasks while
Pipettes and electronic features automating others, often programmable for specific
tasks (Hawach, 2023).
Multichannel Multiple tips, manual or They are used simultaneously, transferring multiple
 COLEGIO SAN AGUSTIN-BACOLOD
College of Health and Allied Professions MLS 111A LABORATORY ACTIVITY SHEET
Medical Technology Program

Pipette electronic samples and increasing efficiency in high-throughput

applications (Pipettes.com, 2022).
Repeat Pipette Often larger, with a dial Enables rapid, repetitive dispensing of the same
Dispenser for volume selection volume of liquid without refilling (Pipettes.com,
2022).

-
Micropipette 10mL Serological pipette 1mL Serological pipette

Figure 10. Degree of ease to manipulate Micropipette, 10mL Serological pipette, and 1 mL Serological
pipette.

2. Why do you have to carefully select the pipette that you use in your work?

Bentivegna (2023) advises that carefully selecting the right pipette is vital for accuracy in work.
One must opt for the smallest one suitable for the volume to maintain precision near the pipette's
capacity limit. Using a larger pipette for a small volume can compromise accuracy. Consistently using the
same pipette for similar volumes ensures reproducibility. An oversized pipette might cause sample
carryover. Avoid extremes in volume relative to pipette capacity to prevent over-pipetting or under-
pipetting.

3. When do you apply the law of “lower and upper meniscus levels”? How can improper reading of the
volume of specimens or reagents inside the pipette lead to inaccuracy of result?

The law of “lower and upper meniscus levels” is applied when reading the volume of liquid in a
pipette. The meniscus is the curved surface of the liquid that forms at the top of the pipette (Dillon &
 COLEGIO SAN AGUSTIN-BACOLOD
College of Health and Allied Professions MLS 111A LABORATORY ACTIVITY SHEET
Medical Technology Program

Friedl, 2022). When reading the volume of liquid, one should read the bottom of the concave meniscus
at eye level to ensure an accurate measurement—particularly on transparent or lightly colored fluids
(Helmenstine). However, reading the upper convex meniscus level is advised when dealing with viscous
fluids or dark-colored solutions where the lower meniscus is not clearly visible.

Improper reading of the volume of specimens or reagents inside the pipette can lead to
inaccurate results and, worse, misdiagnosis, leading to adverse patient outcomes (Felder, 2014). As
Dillon and Friedl point out, one may have an incorrect measurement if one reads the volume from the
wrong part of the meniscus. Additionally, not reading the volume at eye level may introduce parallax
error, which can also lead to inaccurate measurements—reading while the pipette is above level will
falsely increase results and vice versa. Any false elevated reading of the sample will falsely increase the
result and vice versa, while a false elevated reading of the reagent can falsely decrease the result and
vice versa. Thus, following proper pipetting techniques and taking care when reading volumes is
important to ensure accurate and reliable results.

IV. Conclusion

In the laboratory, I have learned about the pivotal role of pipettes in clinical chemistry,
emphasizing the need for precise liquid handling through proper pipette selection. The objectives were
sufficiently achieved as I successfully chose the appropriate pipette and performed proper pipetting. The
diverse classifications, designs, and functions of pipettes provided me with options for delivering
tailored solutions for accurate measurements and transfers. When pipetting, meticulous technique,
including volume setting, plunger control, tip submersion, proper positioning, gliding along vessel walls,
and allowing the plunger to return, is crucial for reliable results. However, others seem to be difficult in
pipetting, and some are easy initially. Still, it is important to practice and carefully select the pipette that
one must use in their work to maintain precision and accuracy, such as by choosing the smallest one
suitable for the volume and liquid one is working on. Adhering to the proper reading method, such as
using the law of "lower and upper meniscus levels," ensures precise readings, particularly considering
viscosity and transparency, because mishandling or misreading volumes can introduce significant
inaccuracies, underscoring the importance of meticulous technique. Thus, it is vital to master the
selection and practice of the pipette—because this mere handheld device orchestrates groundbreaking
diagnostics, wielding the power to measure, transfer, and shape even the tiniest volumes of the very
essence of life itself.

V. References

Bentivegna, T. (2023). Are you using the right type of micropipette? [how-to]. INTEGRA.
https://www.integra-biosciences.com/china/en/stories/are-you-using-right-type-micropipette-
how
 COLEGIO SAN AGUSTIN-BACOLOD
College of Health and Allied Professions MLS 111A LABORATORY ACTIVITY SHEET
Medical Technology Program

Carreiro-Lewandowski ,E. (2013). Basic principles and practices. Clinical Chemistry: Principles,
Techniques, and Correlations, 2.

Church, R. (2018). Clinical biochemistry and Pathology. Scientific e-Resources.

Dillon, A., & Friedl, E. (2022). What is a Meniscus in Science? Study.com | Take Online Courses. Earn
College Credit. Research Schools, Degrees & Careers. https://study.com/learn/lesson/meniscus-
liquid-uses-examples-science.html

Felder, R. A. (2014). Automated specimen inspection, quality analysis, and its impact on patient safety:
beyond the bar code. Clinical Chemistry, 60(3), 433-434.

Glassware, V., Flasks, V., Pipettes, S., Pipettes, T. C., Pipettes, T. D., Pipettes, B., ... & Grade, C. P. (2015).
Systems of Measurement, Laboratory Equipment, and Reagents. Linne & Ringsrud's Clinical
Laboratory Science-E-Book: The Basics and Routine Techniques, 131.

Guzman, K. (2001). Pipetting: A practical guide. The American Biology Teacher, 63(2), 128-131.

Hawach. (2023, July 19). Classification of Pipettes. Hawach Scientific Co., Ltd.
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Helmenstine, T. (2021, May 2). How to read a meniscus of a liquid - what is a meniscus? Science Notes
and Projects. https://sciencenotes.org/how-to-read-a-meniscus/

Henry, K. (2011). How to use a micropipette - Sample Delivery with Variable Automatic Micropipettes.
Molecular, Cell and Developmental Biology - UCLA.
https://research.mcdb.ucla.edu/Goldberg/HC70AL_Su14/pdf/How%20to%20Use%20a
%20Micropipettor.pdf

Hewitt, W. (2012). Theory and application of microbiological assay. Elsevier.

Liu, S., Wei, M., Liu, R., Kuang, S., Shi, C., & Ma, C. (2019). Lab in a Pasteur pipette: low-cost, rapid, and
visual detection of Bacillus cereus using denaturation bubble-mediated strand exchange
amplification. Analytica Chimica Acta, 1080, 162-169.

Marshall, W. J., Lapsley, M., Day, A., & Shipman, K. (2020). Clinical chemistry. Elsevier Health Sciences.

May, M. (2018). A guide to proper pipetting. Lab Manager. https://www.labmanager.com/a-guide-to-

proper-pipetting-2235

Pipettes.com. (2022). Types of pipettes and how to use them. Pipettes.com.

https://www.pipettes.com/calibration-services/pipettes-university/accuracy-matters-blog/how-
to-use-various-types-of-pipettes

Rodrigues, G., & Curtis, R. (2010). Instrument performance verification: micropipettes. Practical
approaches to method validation and essential instrument qualification, 327-346.

Turgeon, M. L. (2015). Linne & Ringsrud's Clinical Laboratory Science-E-Book: The Basics and Routine
Techniques. Elsevier Health Sciences.

Yanase, I., Ohtaki, T., & Watanabe, M. (2002). Combinatorial study on nano-particle mixture prepared by
a robot system. Applied surface science, 189(3-4), 292-299.