PIPETTES
Kathlene Joy Labrador-Limcumpao, RMT, MSMT
LEARNING OBJECTIVES
At the end of this activity, the student shall:
1. Identify the basic parts of a pipette.
2. Classify each pipette observed as to the calibration,
drainage characteristic, and type.
3. Differentiate one kind of pipette from another.
4. Demonstrate the proper use of pipettes.
5. Transfer the correct amount of liquid as instructed.
LEARNING OBJECTIVES
How to choose proper type of pipette
What the specifications on a pipette mean
Handling and disposal of pipettes
PIPETTES
These are instruments that are used to transfer
liquids from one container to another.
≤ 20 mL
Glass or plastic, reusable or disposable
It has parts:
The tip
The stem
The mouth
PIPETTES
Classification:
Calibration/Design
A. To Contain
B. To Deliver
Drainage Characteristic
A. Blow-out
B. Self draining
Type
A. Transfer pipette
B. Measuring/ Graduated pipette
C. Micropipette
D. Automatic pipette
PIPETTES
Classification as to Calibration/Design
To Contain
Holds a particular volume
Do not dispense the exact volume
To Deliver
Delivers the exact amount of volume it holds to a container
PIPETTES
Classification as to Drainage characteristic:
Blow-out
With etched ring/double ring
Exact volume is obtained when the last drop is blown out
Self draining
No etched ring/double ring
Liquid is allowed to drain by gravity
PIPETTES
Classification as to Type:
Transfer Pipettes
Designed to dispense one volume without further
subdivisions
Examples:
a. Ostwald Folin
Bulb like enlargement at the tip
Viscous fluid/liquid
Blow-out, both TC and TD
b. Volumetric pipette
Bulb like enlargement at the middle
Aqueous solution
Self-draining, TD
PIPETTES
Volumetric Pipette
Specifications
How much liquid will be transferred if the liquid is drawn
up to the calibration line on the neck
Temperature at which the calibration was made
Whether it is a TD or TC pipette
PIPETTES
Classification as to Type:
Measuring or Graduated pipette
Has graduation lines along the stem
Measure and transfer several fractional volume
Examples:
a. Serologic pipettes- calibrated up
to the tip
With graduation marks up to the tip
Blow-out, TD
b. Mohr pipettes- calibrated between
two mark
No graduation marks up to the tip
Self-draining, TD
c. Micropipettes
PIPETTES
Classification as to Type:
Measuring or Graduated pipette
Micropipette
≤ 1 mL, TC
Examples of Micropipette:
WBC pipette
RBC pipette
Sahli Helige pipette
Lang Levy pipette- TD/TC
Kirk and Overflow pipette
PIPETTES
PIPETTES
Examine pipette A and B.
Which is the serological and which is the Mohr?
PIPETTES
Measuring Pipette
Specifications
Maximum volume of liquid that can be transferred
Size of the divisions on the pipette
Temperature at which calibration were made
Is the pipette TD or TC
PIPETTES
Pipette is calibrated in
1/10 ml divisions and
will deliver up to 5.0
mL within published
tolerance levels at
20oC
PIPETTES
Pipette is calibrated in
1/100 mL divisions
and it will deliver up
to 1.00 mL within
published tolerance
levels at 20oC.
PIPETTES
Classification as to Type:
Automatic pipettes
Mechanism of aspirating and dispensing of fluid is
inherited to the pipet
According to mechanism, classification includes:
Air Displacement Pipette- piston for suction creation for
drawing sample
Disposable tips
Positive Displacement Pipette- moving piston in the pipet
tip or barrel
No requirement of different tip
Dispenser/Dilutor Pipette- obtaining liquid from a
common reservoir and dispense it repeteadly
PIPETTES
PIPETTES
PIPETTES
Transfer of uncalibrated volumes up to 2.5 mL can be
accomplished using glass “transfer” or “Pasteur” pipettes
shown below. These may be sterilized before use.
Roughly calibrated volumes of 1 and 2 mL can be
transferred with the one piece plastic transfer pipettes
which may be purchased as sterile or non-sterile units.
PIPETTES
PIPETTES
How to use pipette properly?
Materials:
PIPETTES- HOW TO USE A GLASS
PIPETTE?
1. Squeeze pipette bulb and touch to mouth of
pipette.
2. Hold pipette to upper third of the tube.
3. Place tip on the liquid, slowly releasing pressure
on the bulb.
PIPETTES- HOW TO USE A GLASS
PIPETTE?
4. If you already drawn liquid up past desired level,
replace bulb with the index finger.
5. Set to Zero (0), holding the meniscus level in
place, gently lift pipette out of solution, then
transfer the liquid.
PIPETTES- HOW TO USE A GLASS
PIPETTE?
6. In transferring, pipette should be in vertical
position , the tip is place against the side of the
receiving vessel.
7. Let the liquid drain in the vessel.
8. Just in case, its not the maximum volume that is
needed, return the remaining liquid on the
aspirating vessel.
PIPETTES- HOW TO USE A GLASS
PIPETTE?
PIPETTES
Notes to Remember:
Class A pipettes are available in the laboratory.
In calibrating pipettes, pipettes that are TD are
calibrated with distilled water whereas TC pipettes
are calibrated using mercury.
Gravimetric and Spectrometric methods –
accuracy and precision of pipettes
Acid dichromate (soapy water solution in a
tray)
PIPETTES
PIPETTES- HOW TO USE AN AUTOMATIC
PIPETTE?
Materials:
PIPETTES- HOW TO USE AN AUTOMATIC
PIPETTE?
PIPETTES- HOW TO USE AN AUTOMATIC
PIPETTE?
1. Set the volume of pipette.
2. Attached the correct tip to use.
3. Push the control button. You have to feel the first
resistance/first stop.
4. Immerse the tip and slowly release depression on the
control button.
5. Remove tip slowly from the liquid, wipe the tip against
the tube wall.
6. Dispense liquid, pipette in vertical position, place the tip
on the wall of the tube.
7. Press the control button slowly until the liquid is
dispense.
8. To completely dispense, press up to second
resistance/second stop the control button.
9. Eject tips by pressing the tip ejector.
PIPETTES- HOW TO USE AN AUTOMATIC
PIPETTE?
PIPETTES- HOW TO USE AN AUTOMATIC
PIPETTE?
PIPETTES- HOW TO USE AN AUTOMATIC
PIPETTE?
PIPETTES- HOW TO USE AN AUTOMATIC
PIPETTE?
PIPETTES- HOW TO USE AN AUTOMATIC
PIPETTE?
PIPETTES- HOW TO USE AN AUTOMATIC
PIPETTE?
NTR:
If pipettes are dirty, clean the outside with
ethanol/alcohol.
Tip and pipettes should be appropiate.
Do not lay down or invert pipette when its already
filled with liquid for it can contaminate the inside of
pipette.
Pre-wetting of pipette before sample aspiration is
sometimes recommended.
PIPETTES
END
“I’m not telling you it’s
going to be easy, but,
I’m telling you it’s
going to be worth it. “
Keep Faithing mga ka-
CC! Aja Aja!