10 TIPS FOR

PROPER
PIPETTING
 1 Pipetting proficiency is
one of the most critical
elements to ensure
OPERATOR accuracy and precision,
TRAINING particularly in the face
of high personnel
turnover, growing
laboratory workloads
and increasingly
complex tests.
 Like all mechanical

2 instruments, pipettes
are subject to random
failure. Electronic rely
on internal high

PIPETE
precision components
to function properly.
Silent mechanical
CALIBRATION failures, which are
often not visible or
evident by the
pipette’s “feel,” can be
diagnostic tests and
experiments.
 3 Sample preparation
for most analytical
SAMPLE and
processes
production
requires
PREPARATION accurate, precise
liquid handling
 Every pipette shaft has
4 a specific taper angle
that is designed to fit
optimally to the
PIPETTE recommended tips.
Tips have to form a
TIP FIT tight seal with the
pipette shaft, as a leaky
seal contributes to
errors in pitted
volumes.
 5
Temperature and relative
humidity are usually
programmed to provide a
comfortable working
environment in the laboratory.

LABORATORY
Instruments also have a
comfort zone. Air displacement

ENVIRONMENT
pipettes use an air cushion to
couple the plunger to the
aspirated liquid in the tip. This
air cushion, or captive air
volumes, closely follows the
ideal gas law and is therefore
susceptible to changes in
temperature and pressure.
 6 PIPETTING MODES
STANDARD MODE REVERSE MODE

For pipetting most

aqueous solutions, Reverse mode is
forward mode should be recommended for
used. It typically yields viscous solutions,
better accuracy and because forward
precision than reverse
mode for this
mode and is used by
manufacturers to calibrate application can result
their pipettes. in underdelivery
 7 ASPIRATING
During aspiration, the pipette is held in the vertical position
and the tip is immersed to the proper depth (User should
avoid touching the side walls or bottom of the sample
container). Sample is dispensed with the tip touching the
receptacle wall at an angle and dragging it slightly along the
wall.

Pipetting should be slow and steady. Operators should pause

immediately after aspirating, with the pipette tip still in the
sample liquid. Failure to do so can lead to inconsistent
aspiration volumes because the liquid requires about a
second is acceptable, but anything longer than that may result
in underdelivery.
 8 DISPENSING
Consistency and care are critical also when
dispensing sample. Before dispensing, pipettes
should be checked for droplets on the outside of
the tip, with visible droplets removed very carefully
with a lint-free cloth.

Once clean, the tip should be placed against the

receptacle wall and the plunger pressed to the second
stop when using forward mode. The tip should then be
slightly dragged up the receptacle wall to allow all liquid to
be drawn from the tip. Repeated actions produce
repeatable results the plunger should always be pressed
and released with consistent speed and pressure.
9 TIP IMMERSION

Tip immersion depth Pipetted volume

is important and will determine the
requires subtlety : if correct immersion
it is too shallow, depth, 1 generally
there is a risk that are 2-4 mm below the
can be aspirated; too surface for
deep in the sample, volume from 1ul,
and droplets may and 3-4 mm for
volumes larger
collect on the outside
than 1 ml.
of the tip and more
sample might be
forced inside.
 10 The barrel of a pipette
will heat up after
prolonged handling due
AVOID HEAT to heat transfer from the
palm of the hand. This
TRANFER can lead to an expansion
FROM of the captive air volume
HANDS and ultimately decrease
the delivered sample
volume.