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© 2016 Artel 1
Impact of Standardizing on Traceable QC
for Volume Transfers
© 2016 Artel 2
How is liquid handler performance evaluated in your lab?
What information do you obtain (accuracy and/or precision)?
How long does it take?
How much does it cost?
Who takes ownership?
Are you confident that every liquid handler within your organization
dispenses equal volumes?
Are you confident that your liquid handler QC method is performed
reproducibly across your organization?
The challenges
© 2016 Artel 3
“In the modern drug discovery, clinical, and forensic laboratories there
is an ever-increasing requirement for validation of every aspect of
laboratory operations …
A single error in a liquid transfer step can destroy a complete
assay, and in most cases, this error is not caught until the assay is
completed resulting in a loss of time and resources ....”
G.J. Wendel, J. Assoc. Lab. Autom., 2006, 11, 88-91
Liquid Handling Validation
© 2016 Artel 4
Goal:
Reproducibly deliver target volume
with accuracy & precision
Solution:
Rapidly verify transferred target
volumes with a standardized,
traceable platform technology
Tecan QC Kit
Artel MVS
© 2016 Artel 5
Automation-ready MVS
A Standardized QC Platform:
Traceable
Fast & Easy
Accurate
Precise
Known Performance
Data Integrity
© 2016 Artel 6
Process Work Flow:
Checking Volume Transfer Performance
© 2016 Artel 7
Increase Liquid Handling Quality – Simply By
Knowing Performance
Dispense Dye
Solutions and
Mix.
Read Plate. Know
Performance.
© 2016 Artel 8
Dual-Dye Photometric Method: Dispensing and
Reading
• Photometric measurement of liquid volume
• Two dyes measured at two wavelengths
• Ratiometric calculation of results per well
• Simultaneous measurement of accuracy & precision per tip
• Artel’s patented dual-dye ratiometric photometry is included in ISO 8655:
Part 7 and the measurement results are NIST-traceable
Mix
Photometric Measurements
Dispense Sample (quantitative)
Dispense Diluent (non-quantitative)
© 2016 Artel 9
Liquid handler
Reagents
Tips
Environment
Operator
contribute directly to the liquid
dispensing variability
affect the
measurement process
Mixing
Detector
Plate Type
Sources of Variability for Liquid Handler
Calibration Measurements (1 of 2)
© 2016 Artel 10
Sources of Variability for Liquid Handler
Calibration Measurements (2 of 2) A Standardized QC Method Removes Many Variables
Liquid handler
Reagents
Tips
Environment
Operator
contribute directly to the liquid
dispensing variability affect the
measurement process
Mixing
Detector
Plate Type
© 2016 Artel 11
Standardization Allows for Accuracy Comparisons:
Accuracy Matters
© 2016 Artel 12
Defining Accuracy & Precision: There is a Difference
5 µL target
volume; 8 µL
dispensed with
repeatability
5 µL target
volume; 5 µL
dispensed with
repeatability
5 µL target
volume; 5 µL
averaged
dispense without
repeatability
5 µL target
volume; 8 µL
dispensed
without
repeatability
precise,
not accurate
precise
& accurate
accurate,
not precise
not precise,
not accurate
© 2016 Artel 13
Example:
Liquid Handler* QC Volume Information
Albert & Bradshaw, J. Assoc. Lab. Autom., 2007, 12, 172 – 180.
All tests have similar precision, but varying accuracy
6.80 mL
9.97 mL
* not a Tecan instrument
© 2016 Artel 14
40 mL total working volume
% difference in compound concentration = 37.9%
(6.80 mL)(2 mM)/(40 mL) = 0.34 mM vs. (9.97 mL)(2 mM)/(40 mL) = 0.499 mM
Knowing Accuracy, and Therefore, Concentration
6.80 mL of a 2 mM
compound
9.97 mL of a 2 mM
compound
© 2016 Artel 15
40 mL total working volume (± 4 mL)
% difference in compound concentration = 56.8%
(6.80 mL)(2 mM)/(44 mL) = 0.309 mM vs. (9.97 mL)(2 mM)/(36 mL) = 0.554 mM
6.80 mL of a 2 mM
compound
9.97 mL of a 2 mM
compound
+ 4 mL - 4 mL
Knowing Accuracy, and Therefore, Concentration
© 2016 Artel 16
Standardizing on a QC Platform Allows for
Direct Comparisons….
© 2016 Artel 17
Standardization: Multiple User Types
Research and development
High throughput screening & compound management
Food & agriculture
Automated liquid handler manufacturers
Automation engineers
Service, production support, manufacturing
Clinical
Forensics
Government
Other
© 2016 Artel 18
A (Global) Standard Measurement Platform
5 mL, 8-tip LiHa 5 mL, 96-tip MCA
Images from: www.tecan.com
28 26
CAMBRIDGE, UK SAN DIEGO, CA
=
Compare performance regardless of user, model, date or
location…remove robot-to-robot differences
© 2016 Artel 19
Corrective actions are easier to implement
Comparability (local, off-site, global)
SAT, FAT, Process control
Transferability between methods, devices, assays, groups
Reduction in errors, re-work
Monitoring accuracy and precision with ability for trending analysis
Ability to pick the best device for the task
Confidence, take the guess-work out of liquid handling QC
Removing ‘blame game’ – improves communication(s)
Known uncertainties, shelf-life, etc. with measurement tool(s)
Impact of Standardizing on Traceable QC
for Volume Transfers
Tecan QC Kit
OVERVIEW AND OPERATION
Automated Quality Control with QC Kit
• Easy to use
• Fast
• Comprehensive data
• Traceable
21
22
Consumables
QC VERIFICATION PLATES
• Two Formats: 96-well and 384-well
• Lot characterized plates
• Extensive testing of every plate lot
• Well dimensions are characterized through certified
reference materials maintained by NIST; this ensures
traceability to SI
• 2D barcode on plate boxes carries necessary
information about performance, dimension, and lot
number
QC RANGE SOLUTIONS
• Kit contains two Dyes: red and blue
• Distinct absorbance maxima: 520 & 730 nm
• Different concentrations of red dye for
different volume ranges
• Stable and traceable to international standards
23
Calibrator Plate and Reader
24
• Sealed precision cuvettes filled with same dyes as sample solutions and diluent
• Barcoded absorbance traceable to international standards
• Calibration of reader ensures consistency before runtime
• Tecan Infinite reader:
• Infinite F50 for 96 well plates
• Infinite 200 PRO for 96 & 384 well plates
• Narrow bandwidth filters
Easy to Use
CONVENIENT AND SECURE
• Step by step software guidance with clear
instructions eliminates mistakes and reduces
training time to a minimum
• Pre-mixed solutions; open and use
• Verified hardware selected specifically for QC
• Built in reporting module generates automatic
reports
25
Fast
START, MEASURE, REPORT
• No need for sample preparation, standard curve
generation, and post-measurement calculation
• Testing of one volume takes:
• <15 min for standard liquid handling arm (1 plate)
• <15 min for MultiChannel Arm™ 96 (3 plates)
• <15 min for MultiChannel Arm™ 384 (3 plates)
• Frequency
26
Comprehensive Data
LIQUID HANDLING AND
PERFORMANCE DATA
• The automatically generated
printout lists overall accuracy
and precision (CV) and gives
volume information per tip
• Report shows the overall
outcome (Pass or fail)
• Color coded “heat map”
• Easy to identify if a tip
deviates from the expected
volume
27
Traceable
TRACEABLE RESULTS FOR REGULATORY
COMPLIANCE
• Consumables
• QC Kit test results are traceable to
international standards and can be compared to
the manufacturer's specification used for IQ/OQ
• Benefits include
• Traceability enables compliance with regulatory
standards for peace of mind for upcoming audits
• Comparison of liquid handling data between different
sites
• Compare instruments within the company;
mandatory for protocol transfer from R&D to quality
control and production
• QC Kit tests before running important assays will
remove any question of pipetting robot in case of
surprising results
28
Software
THE QC KIT SOFTWARE IS AN EVOWARE
ADD-ON
• EvoWare
• Automated with Freedom EVO liquid handler
• It functions as a wizard to help the user at run-
time
• Value
• Security and Convenience
• Time Savings
• Comprehensive Results
• Consistent
30
31
Automated workflow
32
33
34
35
36
37
38
39
Calibrator, Baseline, QC Plate
40
Pipetting & Volume Verification Success
Failure
TROUBLESHOOTING LIQUID HANDLER
41
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Results
Advantages
• Fast, Easy, comprehensive
• QC Kit is integrated with
EVOware and supported by
Tecan
• The software includes verified
pipetting templates for common
configurations
• The QC Wizard will automatically
execute the pipetting, shaking,
measuring, reporting
• Traceable components
• Traceable results
43
44
© 2016 Artel 45
Environmental Testing of Tecan EVO using the QC Kit
© 2016 Artel 46
Experiment - Environmental Conditions Tested
© 2016 Artel 47
Experiment - Volumes Tested
© 2016 Artel 48
Measured Volumetric Performance
© 2016 Artel 49
Trueness
© 2016 Artel 50
Conclusions Precision of the Tecan Freedom EVO was found to be insensitive to environmental conditions Trueness does depend on environmental conditions. Trueness exhibited a linear correlation with the “evaporation potential” Although the Tecan QC Kit is intended to be used for checks such as IQ/OQ, this study shows that the Tecan QC Kit can also be used for performance checks under environmental conditions that deviate from the manufacturer’s specified operating conditions. note 1: Tecan specifies the pipetting behavior of the Tecan Freedom EVO under controlled environmental conditions (T = 20 °C to 27 °C; relative humidity at 25 °C between 30% and 60%). Volumetric performance specifications apply to this controlled limit. For operation outside these limits, meeting the manufacturer’s specifications for volumetric performance is not guaranteed.