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BUENAFE
WHY?
Techniques among the users may vary with the users background,
preferences, and training.
The difference in technique and execution can affect accuracy
and precision of the experiment or the test.
TYPES OF PIPETTES
Volumetric Pipettes
Measuring Pipettes
- Mohr
- Serological
Automatic (Microliter) Pipettors
PIPETTE AIDS
Types of Suction Device:
Rubber Aspirator (Bulb)
VOLUMETRIC PIPETTES
Shaped like rolling pins with a large belly, one blunt end, the
neck, and one tapering end, the tip.
Used for accurate measurements, since it is designed to deliver
only one volume and is calibrated at that volume.
Should be used when accuracy and reproducibility are crucial,
because these can achieve accuracy to four significant figures.
(e.g. 100.0mL)
VOLUMETRIC PIPETTES
Specifications:
Indicate how much liquid will be transferred if the liquid is
drawn up to the calibration line on the neck
VOLUMETRIC PIPETTES
Helpful Hints: Using a Volumetric Pipette
- Examine each pipette you take
from the drawer because
occasionally you will find a
stowaway who got in and couldn't
get out.
- Examine the tips for breakage. Many
breaks are trivial, i.e., they show minor
chipping around the edge of the tips.
Here's one in which the entire wall has
fractured at one point. This pipette is
unusable because the liquid path has been
compromised.
VOLUMETRIC PIPETTES
Helpful Hints: Using a Volumetric Pipette
- Hold the pipette by the upper third of the tube
and keep the tip from touching anything.
- Squeeze the aspirator before pressing it against
the blunt end of the pipette. Make sure you have an
air-tight seal between the bulb and the pipette.
- Place the tip of the pipette into the solution to be drawn
up and release the bulb slowly. Do not allow the tip of the
pipette to break the surface of the liquid while drawing in
the solution. (The sudden decrease in viscosity at the tip will
cause a large amount of liquid to contaminate the inside of
the rubber bulb because of the entry of air pushing the
liquid up beyond the mouth of the pipette .)
- Draw up the solution until the meniscus is several
centimeters above the calibration line, then quickly put your
finger over the open hole of the pipette.
VOLUMETRIC PIPETTES
Helpful Hints: Using a Volumetric Pipette
- Making sure that the line of sight is perpendicular to
the calibration mark, allow a tiny amount of air in so that
the meniscus drops to the mark. When the bottom of
the meniscus coincides with the calibration mark, the
pipette contains a precisely measured volume.
-Remove the pipette from the reagent solution
and allow the liquid to drain into the receiving
vessel.
-Volumetric pipettes should NOT be "blown out"
to eject all liquid at the tip these pipettes are
calibrated in a manner that takes into account
the solution which remains at the tip due to
surface tension.
MEASURING PIPETTES
They are straight glass or plastic tubes with one tapering end.
Calibrated into small divisions so that various amounts of liquid
can be measured with the same pipette.
Usually used to measure any amount between 0.1mL and 25.0mL.
They are not as accurate due to the fact that any imperfection
in their internal diameter will have a greater effect on the volume
delivered.
MEASURING PIPETTES
Types of Measuring Pipettes:
- Mohr Pipettes
The graduations on these
pipettes always end before
the tip.
- Serological Pipettes
The graduations on these
pipettes continue to the tip.
MEASURING PIPETTES
Specifications:
Indicate the maximum volume of liquid that can be transferred.
Indicate the size of the divisions on the pipette.
Indicate the temperature at which calibrations were made.
Indicate whether the pipette is a to deliver (TD) or to
contain (TC) pipette.
MEASURING PIPETTES
Specifications:
MEASURING PIPETTES
Serological Pipettes:
If a serological pipette has a frosted band or two thin rings around
the neck, then it is a blow-out pipette.
(The frosted band should not be confused with
thicker colored rings or colored dots, which are a
manufacturers code for the maximum volume
of the pipette. Remember, only blow-out a
serological pipette if it has a frosted band
or two thin rings.)
This means that after all the liquid has been allowed to drain out by
gravity, the last drop remaining in the tip is also forced out.
In a 1000L Pipettor,
this reads 262L
Place a fresh tip tightly onto the barrel. (Tips are contained in a package or dispenser so that you
do not handle or contaminate the points with your fingers.)
Attaching tips to
the pipettor
Hold the pipette vertically and dip it 2 5mm into the sample liquid.
Slowly and smoothly release the plunger to aspirate the liquid, pausing about a second after
aspiration. Never let the plunger snap up!
receiver and gently depress to the first stop. Allow the liquid to
drain along the inside walls of the tip, then depress the plunger
beyond the first stop to squirt the last drop of liquid out of the tip.
Fig 1
Fig 2
Fig 3
Never lay the pipette on its side nor invert the pipette if liquid is in the
tip.
1.
2.
The smallest volume to be dispensed (position 0.5 on the Volume Selection Dial)
3.
The step sequence of volume to be dispensed if dial setting is increased from 0.5
4.
5.
Graduation scale
6.
7.
US Patent Numbers
AE [ x ]
where
standard value
2
(
x
x
)
( N 1)
where
s
%CV ( ) 100
x
xi