5 Operation and Functional Verification of Automated Pipetor
5 Operation and Functional Verification of Automated Pipetor
5 Operation and Functional Verification of Automated Pipetor
1 Micropipette operation
Learning Objectives
2 Micropipette operation
Chapter Outline
Introduction
Operating procedure
Pipetting techniques
Pipette troubleshooting
General working remarks
Principles of inspection and calibration
3 Micropipette operation
Introduction to Automated Micropipette
A pipette, pipet, pipettor or chemical dropper is a
laboratory tool commonly used to transport a measured
volume of liquid.
4 Micropipette operation
Introduction to Automated Micropipette
6 Micropipette operation
Parts of Automatic Micropipette
7 Micropipette operation
Operating the Micropipette
Step 1: Set the Volume
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Step 2: Read the Volume
9 Micropipette operation
Setting Adjustable Volume
10 Micropipette operation
Micropipette operation
Step 3: Attach the Disposable Tip
Attaching the
disposable tip
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Step 4: Depress the Plunger to the First Stop
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Step 8: Withdraw the Tip
Remove the tip from the sample liquid.
No liquid should remain on the outside of the tip.
Wipe away any droplets on the outside of the tip with a lint-free tissue,
but only wipe droplets from the side of the tip.
Never touch the tip opening or you may absorb part of your sample.
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Step 9: Dispense the Sample
14 Micropipette operation
Step 10: Withdraw the Pipette
15 Micropipette operation
Pipetting Techniques
Commonly, there are two types of pipetting techniques;
Forward pipetting
Reverse pipetting
2. Dip the tip into the liquid to a depth of 1 cm, and slowly release
the operating button. Withdraw the tip from the liquid, touching it
against the edge of the reservoir to remove excess liquid.
3. Dispense the liquid into the receiving vessel by gently pressing the
operating button to the first stop. After one second, press the
operating button to the second stop. Remove the tip from the
vessel, sliding it along the wall of the vessel.
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Pipette Troubleshooting
20 Micropipette operation
General working Remarks
21 Micropipette operation
General working Remarks , Cont’d..
Maintain consistent angle of pipette when
aspirating (min. 30º)
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Principles of Preventive Maintenance
23 Micropipette operation
Routine Maintenance
24 Micropipette operation
Inspection and Calibration of Pipettes
25 Micropipette operation
Gravimetric Calibration
26 Micropipette operation
This table shows the accuracy and precision of pure
water dispensed that is allowed for micropipettes based
on their volumes.
For example, pipettes that dispense 100 µl should be accurate to 0.1 mg with
a standard deviation of up to 0.2 mg of pure water weighed with an analytic
balance
27 Micropipette operation
Testing Pipetting Accuracy
The following procedure can be used to test how accurately
the pipette measures volume (water is used because the weight
of water is 1g/mL):
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Testing Pipetting Accuracy, Cont’d….
The accuracy of pipetting should be evaluated periodically (i.e. every
6-12 months)
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Checking Pipetting Precision
Set the pipette to 50 µL (0.050mL)
30 Micropipette operation
Precision and Accuracy Calculation
Case scenario: A lab professional observed that when using the
pipette method for analysis, the volume of sample dispensed
into each tube seemed to look different.
Question:
Answer:
31 Micropipette operation
Example Data and Calculations
The following data were obtained after 10 replicate of
volume measurments was done. Calculate the mean, sd and
%cv of the data.
Weight
0.0495 Mean: ?
0.0506 sd: ?
0.0498 %CV: ?
0.0501
0.0506
0.0492
0.0498
0.0507
0.0502
0.0491
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Pipetting
0.0000 g
0.1499
0.0495
0.1001
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Gravimetric Measurements
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Calculations
35 Micropipette operation
Calculations
36 Micropipette operation
Calculations
37 Micropipette operation
Application 2:
Precision Calculation
In order to be precise for patient testing, the pipette is
required to have a %CV of <2% and accuracy was
required to have < 1% error rate.
A quick precision study is performed with the following
results:
0.048 mL
0.045 mL
0.051 mL
0.049 mL
Micropipette operation
0.053 mL
Application 2:
Precision Calculation
Arithmetic mean
x
x
N
x
2
Standard deviation SD
x 2
N
N 1
Micropipette operation
Application 2 Debrief: Precision Calculation
Questions Answers
1. What is the mean? 0.0492 mL or 49.2 uL
2. What is the coefficient of 6.1%
variation (%C.V.)?