Basic Laboratory Equipment and Good Practice Guide for

Laboratory general measurements

1. Objective

In laboratory, the correct weighing and liquid handling is essential to ensure precise, accurate
results. Scientific staff has a broad range of equipment to choose and selection of the most
appropriate type is crucial.

This is an introduction of basic laboratory equipment and practical guide to the proper use of
general measurements in laboratory - weighing and liquid handling for students and laboratory
workers. Their exercises mainly focus on practice guide for micropipettes with dispensing
volumes ranging from microliters to milliliters and for laboratory balances with proper weighing
from gram to milligram.

2. Learning outcome

The learning outcome includes recognize general equipment for measuring and liquid handling
in laboratory.

Student know proper use of micropipettes and weighing of balances.

Following experiments is mostly intended for students to develop their skills using
micropipettes and practice weighing in a laboratory setting. The aim of the practice including:

● Accuracy: Indication of how close a measurement is to the actual value.

● Precision: Indication of how reproducible the measurement is.

3. Basic Laboratory Equipment

In a laboratory, there are many equipment and tools with different size. Knowing equipment
and their use will ensure the safety and correct of laboratory practices. Details of common
laboratory equipment and their appropriate use are identified as table 1.
Table 1: Details of common laboratory equipment and their appropriate use

Beaker Normal container with wide mouth for holding or

heating substance

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Erlenmeyer flask Container with narrow mouth for transporting,
heating, or storing substance. Can add a stopper, if
need.

Wash bottle Squeeze container with nozzle cap. Use for

washing with cleaning liquids or keep materials
moist.

Test tubes Contain small volumes of liquids for mixing, heating

or testing a reaction.

Eppendorf tube Contain small volumes of liquids (normally <2.5 ml)

Use for running reactions, storage and centrifuging
samples.
Volumetric flask Calibrated flask for a precise volume and precise
dilution at particular temperature. They usually use
for creating standard solutions.

Graduated Use for normal measuring of specific amounts of

cylinder liquids (not as precise as volumetric flask)

Mortar and Use for grinding samples

pestle

Test tube rack Use for holding test tubes

Funnel Use for filtrating (with filter) or transferring liquids

into narrow mouth containers.

Alcohol burner Laboratory heating source.

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Spatulas and Use for removing amounts of solids or powders of
scoopulas substance.

Stirring rod Used for stirring or mixing.

Dropper Use for obtaining small volumes of liquids

(semi-quantify by drops).

Disposable Use for performing a semi-quantitative transfer of a

Pasteur Pipette small volume (usually 1–10 ml) of liquid.

Calibrated Use for measuring and dispensing precise small

Pipette amounts of liquids.

Pipet bulb/pump Use for drawing liquids into pipette (also known as
pipetor or pipet-aid).

Burette Use for dispensing an accurate volume of a liquid.

Ring Stand Use for holding burette or clamp laboratory

equipment in place.

Watch Glass Used to hold solids while they are being weighed or
to cover a beaker.

4. Reading a meniscus technique

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Reading right level of a liquid is necessary in
using volumetric glassware. A liquid meniscus is
a curvature present at the liquid surface of
small-diameter container (pipette, burette,
volumetric flasks, graduated cylinder, etc.). The
curve can be center-downward or
center-upward depend on their cohesive and
adhesive forces.
For reading, it is important to move the eyes to
the same level of liquid meniscus (Figure 1). The
Figure 1: Meniscus and the right level for
correct level of the liquid was read at the lowest reading
point of the liquid level.

5. Laboratory general measurements

5.1. Weighting

Weighing is a common and critical technique in laboratory. For precise balance of solid or liquid
substance, laboratory-specific analytical balance is developed (Figure 2). Analytical balances
usually require weighing technique including proper balance operation and a stable external
environment.

Figure 2: Standard Analytical Balance (Mettler-Toledo brand)

5.1.1. External influences

External influences are physical factors that affect laboratory balance accuracy, for examples:

● Airflow
● Environmental vibrations
● Evaporation
● Moisture uptake
● …

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Physical factors also effect sample because of their characteristics (liquid evaporation, solid
hygroscopicity) and could give a weighing mistake.

5.1.2. Proper balance operation

Each of analytical balance have slightly differences, manual should be read before starting to
use a new one. Besides, balance is need to be adjusted the sensitivity regularly for precision. An
analytical balance always has capacity and readability, don’t overload.

It is importance to avoid to put supernumerary of weighing substance back to original container

to keep substance pureness.

The following steps (using Mettler-Toledo balance as an example) is tips to obtain reliable
weighing results (Table 2).

Table 2: Weighing tips

Leveling The first thing to do is adjustment the air bubble to

and press the center of the level indicator before weighing.
Zero After adjustment, press Zero button to set a new zero
point on the balance.

Using Use suitable clean weighing vessel for a temporally

weighing substance container.
vessel Open the draft shield and place the weighing vessel
in the middle of the weighing pan.

Tare Using tare button to add tare point to the weighing.

It is used when the weight of the container is
important or when more than one sample will be
add into one single container.
Put Slowly put small amount of substance on weighing
substance vessel several times, until get desired weight.
and close The draft shield is closed after that for the constant
draft in weighing chamber.
shield

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Reading The result is records only after the showed number is
the results stable (not change).

Cleaning Take away the sample and weighing vessel out.

Clean the balance after use, especially weighing
chamber and weighing pan.

5.2. Liquid handling

Pipettes are used to transfer desired volume of liquid accurately to a container. In this part,
two common types of pipette are mentioned: graduated pipettes and micropipette.

5.2.1. Graduated pipettes

In the laboratory, graduated pipette normally uses for transferring volumes of liquid
from ranging from 0.1 mL to 25 mL. Depend on different applications, they are disposable
sterilized plastic and reusable sterilizable glass graduated pipette.

Figure 3: Graduated pipette structure and types

Structure of a graduated pipette (Figure 3) includes:

(1) Aspiration area with or without cotton filter to take liquid in

(2) Long graduated tube for liquid containing and measuring

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 (3) Conical outlet opening for liquid out
(4) Technical data will be marked on pipette. The labelling normally contents manufacture,
reference temperature, nominal volume, increment, volume unit, accuracy classification
(class A, AS, AW or B) and calibration (Ex/TD - “to deliver”). It may also content waiting
time, error limit, country of origin and designation of the standard (DIN EN ISO).

Pipet-aid

Pipette need pipet-aid (also call pipetor) for aspirating and dispensing. There are two common
types of pipet-aids can be used with pipette, pipette bulb and pipette pump (Table 3). Using
ancient mouth pipetting is not recommended because of serious adverse side effects.
Table 3: Common types of pipet-aids

Pipette bulb Classic and cheap bulb. It can be a simple bulb no valve or
and “triple “triple valve" bulb with (A) - displaces air from the bulb, (S)
valve" - take up liquid, (E) expel the liquid.

Pipette pump Easier to handle but more expensive. Its structure has a
suction roller to take up or slowly drop down liquid and
empty button to quick expel the liquid.

Pipette technique

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 (1) Firstly, chose suitable pipette for experiment
and always handle pipette with caution. While
pipetting, try not to let the conical outlet
opening contact with hands or outer surfaces.
(2) Fix pipet-aid tightly to the aspiration area.
Hold the pipette carefully
(3) Place conical outlet opening into liquid and
adjust pipet-aid slowly to aspirate liquid.
(4) Fill pipette with liquid to several millimeters
above the graduated mark (Figure 4).
(5) Control the liquid flow out slowly to obtain
meniscus at desired mark while hold the
pipette vertically and conical outlet opening
down.
(6) Remove liquid remaining outside pipette if
exists.
(7) Transfer pipette to new container and adjust
pipet-aid to delivery liquid in contact with the Figure 4: Graduated pipette liquid delivery
inner surface of new container. (4 mL for example): (A)drains completely
(8) Waiting until all the liquid drains completely and (B) point-to-point delivery. Black arrow
(Figure 4A) or until meniscus reach another indicate liquid moving direction from
desired mark (point-to-point delivery, Figure step-to-step: (1) aspirate, (2) adjust
4B). Take the pipette out. meniscus and (3) delivery.
(9) Lastly, remember to clean the pipette and
pipet-aid after use.

5.2.2. Micropipette

The micropipette uses for precise transferring small volumes of liquid from ranging from ~ 0.001
mL to 1 mL with microliters (µL) scales. There are two type of micropipette including air
displacement micropipette for standard applications and positive displacement pipette for
special applications (such as viscous or volatile samples) worked like a syringe. This part focus
on standard pipetting applications and air displacement micropipette. Depend on amount of
tests, it has single channel (Figure 5A) or multichannel micropipette (Figure 5B). They also have
different sizes of micropipette such as P2 for 0.2-2 μL; P10 for 1-10 μL; P20 for 2-20 μL; P200 for
20-200 μL; and P 1000 for 200-1000 μL. The Eppendorf tube is common container of
micropipette liquid transfer. Micropipette can has its accuracy and precision within ± 5% of
specifications.

Visual structure of a micropipette (Figure 5,6) includes:

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(1) Plunger or push button use for liquid aspiration and delivery. It may also use to adjust
transfer volume (adjustment knob). Plunger controls the piston to push the air out for liquid
aspirating. There are three positions including: (a) rest position: non-pressed position, (b)
first stop: pressed position for aspirate desired volume of liquid or dispense liquid in the tip
and (c) second stop: hard pressed position to blow-out liquid remains in the tip.

Figure 5: Plunger position

(2) Tip eject button use for eject used tip.

(3) Finger-hook use to comfort finger and avoid pipette falling.
(4) Volume indicator use for indicating transfer volume.
(5) Technical data will be marked on pipette. The labelling normally contents manufacture,
volume range, volume unit. It may also content autoclave information, country of origin and
designation of the standard.
(6) Tip ejector is a part use for tip pushing out controlled by tip eject button.
(7) Tip attachment is a part use to attach tips for pipetting.

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 Figure 6: Visual structure of single channel (A) or multichannel (B) micropipette

Micropipette tips

Micropipette tips are used with micropipette to contain

aspirated liquid. Micropipette tips normally made by
disposable and autoclavable clear plastic tips. They also
have different sizes to fix with micropipette such as
white tips for 0.5-10µL and 2-20µL; yellow tips for
20-200µL (Figure 7B); and blue tips for 200-1000µL
(Figure 7A). Micropipette tips normally set in a
autoclavable plastic tip-box which has similar color of
contained tips. Figure 7: Micropipette tips: blue tip
1,000µL (A), yellow tip 200µl (B)
Micropipette technique

(1) Firstly, chose suitable micropipette for experiment and always handle micropipette with
caution. While pipetting, try not to let the tip contact with hands or outer surfaces.
(2) Adjust the volume by turning adjustment knob slowly to reach the desired volume. Read
and adjust carefully the indicated number with the eyes at the same level of volume
indicator (Figure 8). It important to note that the setting over intended range can let
micropipette malfunctioning.

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 Figure 8: Tips for adjusting right intended volume
(3) Fit a tip on by slowly pressing down the tip attachment into a tip in tip-box while slightly
twist the micropipette to ensure a tight seal.
(4) Hold the micropipette vertically with the thumb is on the plunger and other fingers is under
finger-hook (Figure 9).

Figure 8: Tip for holding micropipette

(5) To aspirate liquid, a preparation step is set by press down the plunger to first stop position.
(6) Place the tip into liquid to a suitable depth (2–6 mm). release plunger slowly to the rest
position to draw in liquid without air bubbles.
Table 4: Tips for suitable tip immersion depth

Volume range (μL) Immersion depth (mm)

0.1 - 1 1-2

> 1 – 100 2-3

> 100 – 1000 2-4

> 1000 3-6

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(7) Transfer micropipette to new container. Hold the micropipette at an angle of less than 45°
and the tip is in contact with the inner surface of new container.
(8) Press down the plunger slowly to the first stop position and then to the second stop to
delivery all liquid inside the tip. Waiting until all the liquid drains completely. Take the tip
out.
(9) Release plunger slowly to the rest position.

Figure 9: Tip for aspirating and delivering liquid. White arrow indicate plunger moving
direction from step-to-step: (1) prepare, (2) draw in, (3) delivery and (4) back to rest.

(10) Press down tip eject button to discard the tip into a waste box.
(11) Lastly, remember to clean the micropipette after use.

* There is a reverse technique used for viscous and foaming liquid at slower pipetting speeds.
The principle of this technique is replacing first stop by second stop position for liquid aspirating
and replacing second stop by first stop position for liquid delivery. This may avoid the error
come from liquid viscosity and foam formation. (following guideline of Gilson, S. A. S. "Guide to
Pipetting”, 2015)

6. Materials and methods

6.1. Materials

Micropipette and tips

Graduated pipette

Balance

Eppendorf tube

Beaker

Filter paper

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 Parafilm

6.2. Regents

Food’s dye

Sugar

Glycerol

Alcohol

Water

6.3. Experimental designs

6.3.1. Weighing

Weighing granulated sugar with analytical balance for 0.01, 0.10, 1.00, 10 and 100 g.

Try not to take supernumerary of weighing substance back and put them to original container.

Results

Note the balance technical data and each step of weighing. This note reflects proper technique.

6.3.2. Graduated pipette

6.3.2.1. Pipetting practice

Using graduated pipette transfer following amounts of water: 10.0, 8.9, 7.8, 5.7, 3.6, 1.5, 0.4
and 0.2 mL to a beaker.

Results

Note the used pipette data and each step of pipetting. This note reflects proper technique.

6.3.2.2. Gravimetric practice

Weighing one empty dry beaker, using Tare button to set 0.

Using 1mL graduated pipette transfer 10 times of 0.5 mL of water (m1 –m10). Record each
experimental volume weight separately.

Results

Average of experimental weight of water is:

(𝑚1+𝑚2+…+𝑚10)
𝑚(𝑔) = 𝑛

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With n = total weighing times.

Using density of water 0.997 g/mL (at 25°C) to calculate average of experimental volume of
water:
𝑚
𝑣 [𝑚𝐿] = 0.997

The total theoretical water volume adding should be 5 mL (v).

Percentage of accuracy (A) is calculate using equation as follows:

𝑣− 𝑣
𝐴 (%) = 𝑣
×100

The standard deviation (SD) can determine as follows:

2 2 2
(𝑚1−𝑚) +(𝑚2−𝑚) +…+(𝑚10−𝑚)
𝑆𝐷 = 𝑛−1

Coefficient of variation here is defined as the percentage of standard deviation and mean
volume:
𝑆𝐷
𝐶𝑉 [%] = ×100
𝑣

The percentage of accuracy (A) represent how difference the average of experimental water
volume to theoretical water volume.

Coefficient of variation (CV) represent how close the repeat values in pipetting.

If there is a big error, how do you explain the error in your experiment? List at least three
possible causes.

6.3.3. Micropipette
6.3.3.1. Parafilm test

Prepare a dye solution by adding several drops of food’s dye into 50 mL of water and stir.

Take 5 pieces of parafilm (wax paper) and tape on a notebook paper.

Place each of volume of dye solution: 2, 4, 6, 10 and 20 μL on separated parafilm. Mark the
volume.

Using appropriate pipettes and try to transfer the entire amount in one load to have a single
drop.

Results:

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Try to define and to place side by side form small to big seen volume (without following marked
volume) and compare with marked volume (indicated volume).

Compare the results with others.

6.3.3.2. Filter paper test

Take a filter paper, place ten drops of 2 uL on one line using 2-20 uL pipette. It will make ten
colored dots on filter paper.

Repeat the test, with 4 and 6 uL.

Results

Compare ten drops of the same volume if there is any visible difference in dot sizes?

Compare the results with others.

6.3.3.3. Colored practice

Pipetting amounts dye solution as 40, 60 and 100 µL into one clean Eppendorf tube with a
20-200 µl pipette. Set the pipette to 200 µl and withdraw all the solution.

Results

Compare the results (before withdraw) with others.

Note the result after withdraw and try to explain.

6.3.3.4. Weighing practice

Repeat the above gravimetric test (6.3.3.2.) with micropipettes.

Results

Calculate the percentage of accuracy (A) and coefficient of variation (CV).

If there is a big error, how do you explain the error in your experiment? List at least three
possible causes.

Compare two types of pipettes.

6.3.3.5. Liquid practice

Redo the test 6.3.3.2, 6.3.3.3, replace dye solution with alcohol or glycerol.

7. Reports (requirements)

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Writing reports following each experiment above.

Try to explain the problems.

Correct your technique to get better results.

8. References
1. Blues J., Bayliss D., Buckley M., Measurement Good Practice Guide No. 69: The
Calibration and Use of Piston Pipettes. National Physical Laboratory, United Kingdom,
2004
2. Brand, “Volumetric Measurement in the Laboratory”, Brand GMBH + CO KG, Germany (.
3. Integra Biosciences, “Evolve Manual Pipettes” Hudson, New Hampshire, USA (2016)
4. Jim Wolf, College of Canyons. ‘Introduction to Biotechnology” Custom Lab Exercises
5. Lisa A. Seidman and Cynthia J. Moore, Basic Laboratory Methods for Biotechnology,
Textbook and Laboratory Reference Prentice-Hall, Inc. (2000)
6. Gilson, S. A. S. "Guide to Pipetting." Third edition (2015).
7. Sanders, Erin R. "Aseptic Laboratory Techniques: Volume Transfers with Serological
Pipettes and Micropipettors." Journal of visualized experiments: JoVE 63 (2012).
8. Toledo, Mettler. "Weighing the right way: proper weighing with laboratory balances."
Mettler-Toledo AG, Laboratory Weighing, Greifensee, Switzerland.

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