Emily SalmonAdvisors: Norman Purvis, PhD, Esoterix
Dr. Paul King, Vanderbilt University
Development, Simulation and Optimization of Oncology Laboratory Sample Processing Automation
Introduction•Esoterix Oncology performs Immunophenotyping of Leukemia/Lymphoma Conditions
•Assays are Performed manually by Certified Technicians
•Purpose of Study to examine effects of complete automation of the initial screening process
Goals:–Develop automation protocols–Model operating procedures through simulations–Show benefits (time or cost savings) through optimization
Current Statistics
• Decrease Cost
• Reduce Staff
• Increase Throughput
• Decrease Time
• Increase Efficiency
Why Automate?
• 60 Blood/Bone Marrow Specimens, 40 Tissue Specimens, Daily• 8 test tubes per Blood/Bone, 9 test tubes per Tissue Patient Basic Screening• 6-10 Certified FTE’s processing Blood/Bone Marrow samples, 3-4 Certified FTE’s processing
Tissue samples
• Conservative Projected Growth of 100%, 3 years
• Increase Flexibility
• Increase Quality, Accuracy • Technicians freed for complex
analysis
• More time for further panels
Flow Cytometry
• Leukemia/Lymphoma Cells express lineage associated antigens• Cells Labeled with fluorescent Antibodies• Flow Cytometer uses Laser Beam• Sorts and quantifies populations according to light scatter and
Fluorescence
Lymphs
Blast
10 20 30 40 50 60 70 80 90 100 120
SSC -->
1020
3040
5060
7080
9010
012
0C
D45
-->
FAB-M1
PMN
ProGrn
Immat._PMN
Blast
Mono
10 1 10 2 10 3 10 4
SIDE SCATTER -->
101
102
103
104
CD
45-P
ER
CP
-->
Normal
LymphsBlast
10 1 10 2 10 3 10 4
SSC-Height -->
101
102
103
104
CD
45-P
ER
CP
-->Lymphs
Blast
10 20 30 40 50 60 70 80 90 100 120
SSC -->
1020
3040
5060
7080
9010
012
0C
D45
-->
FAB-M3 FAB-M4
The Laboratory
Blood/Bone Marrow Screening Lab
Tech adding antibodies
Tissue Screening Lab Tech at sink, Centrifuge
ORCANT ROBOT
Six Axis of Motion:•movement along rail•height•reach•bend•twist•grip
Sample Manual Process
ReceiveSamples
To FlowCytometer
Add Reagent#3
Decant
Centrifuge
Add Reagent#2
Incubate
AddAntibodies
Decant
Centrifuge
Add Reagent#2
Decant
Centrifuge
Add Reagent#2
Decant
Centrifuge
Incubate
Mix
Add Reagent#1
Add PatientSpecimen
Corresponding Automated
ProcessSet-up
Automation,Start Assay
Run,
FlowCytometer
ReagentDispenseStation
VacuumFiltrationStation
ReagentDispenseStation
PipettingStation
VacuumFiltrationStation
ReagentDispenseStation
VacuumFiltrationStation
ReagentDispenseStation
VacuumFiltrationStation
Incubator
ReagentDispenseStation
VacuumFiltrationStation
Incubator
PipettingStation
ReagentDispenseStation
Design-Choosing a Procedure to ImplementProcess of adding lyse, shaking wells, centrifuging at
1300 rpm for 5 min, then decanting wells best automated by three different options:
• “No Wash” Procedure– Pros: Fastest, Easiest to Perform, Least # Steps– Cons: Least Chemically Feasible
• Wash, Centrifuge, Aspirate– Pros: Most like Manual Procedure
– Cons: Most Steps, Most Time, Manual Interface
• Wash, Filter– Pros: Fast, Easy to Implement, Easy to Automate– Cons: Filters and Lyse may not Work
InstrumentationBeckman Coulter Instruments:
– Biomek Fx Liquid Handling System
– Multidrop
– ORCA Robot
– Filtration Station
– Carousel/Incubation for 96 well plate
– Flow Cytometer
Simulation Software
SAGIANTM Core System• SAMI Automated Method Development and
Scheduling softwareBuild methods by connecting and configuring nodes, sources, and stations.
• SILAS Integration softwareRun SAMI methods, gather resource use statistics and time values
• BIOMEK FX Automation WorkstationSeparate module for complicated pipetting schemes
• ORCA NT Motion Editor SoftwareUsed for framing and “teaching” of transportation robot
Creating a SAMI Method
Biomek FX
Custom Pipetting
The Biomek FX
Biomek FX Method Editor
Scheduling and Summary Outputs
Allocation of time per task, each plate Resource Utilization Bar Graph
Plate Location Summary Text timing Details
Design Aspects• Choosing a lysing strategy, Choosing Basic Labware• Dimension Configurations• Viewing complex log files to verify/forecast actual machine
behavior• Specifying patterns through special transfer file• Analysis of Filtration Simulation• Adding Rotating ALP to Rotate plates
– Saved time of ~30 sec on Tissue samples, Reduce time of Patient Antibody, Patient Sample additions by ~5 min each,
• Well-Pattern “Minor Software Bug” Assessment• “If” Statement Programming to increase flexibility
Series of Revisions, Trouble-Shooting, and Improvements
Simulation Times
Patient per Plate
Time per Plate
12 46 min
10 69 min
12 40 min
10 35 min
Blood/Bone Marrow 12 46 min
10 43 min
Screen Type
Tissues
standard multidrops, outdated filtration stations, no Rotation.
Deepwell plates, Biomek methed revision
Deepwell plates, Biomek Rotating ALP
Description of Revision Aspects
Revision I
Revision II
Revision III
Blood/Bone Marrow
Blood/Bone Marrow
Tissues
Tissues
SAMI Simulation Times vs. Manual Model Times, Projected Volume
Current 6 Months 1 Year 3 Years
Growth (Proj) 1 16% 33% 100%
Bl/BMarrow no. samples 60 67 83 120
Tissues no. samples 40 45 53 80
Total Simulation Time 2:30:31 3:17:36 3:56:24 4:44:48
Blood/Bone Marrow Current 6 Months 1 Year 3 Years
Simulated Time 1:54:16 2:13:20 2:38:49 3:36:46
Manual Proj Time 5:43 6:12 7:39 11:40
Tissues Current 6 Months 1 Year 3 Years
Simulated Time 1:21:20 1:36:04 1:50:49 2:10:17
Manual Proj Time 3:32 3:51 4:10 6:04
Conclusion• Protocols created are basis for future
implementation• Automation can be shown to save time and
money
• Still need to verify Filtration Method works
• Collect More Experimental Data to Model Manual Times; Re-Evaluate Costs & Proj. Growth
• Explore Integration with LIMS, Variable Work Files
Recommendations
References
References:The Purdue Cytometry CD-ROM Volume 4, J. Watson, Guest Ed., J.Paul Robinson, Publisher. Purdue University Cytometry Laboratories, West Lafayette, IN 1997 http://www.ctyo.purdue.edu 18 April 2003Beckman Coulter “Cytomics FC 500 Series Flow Cytometry Systems” © 2002 http://www.beckman.com/products/instrument/flowcytometry/fc500series.asp 21 April 2003Beckman Coulter “Orca Robot” © 2002 http://www.beckman.com/products/instrument/automatedsolutions/integsystems/orcarobot_dcr.asp 21 April 2003Beckman Coulter “SAGIAN Core Systems” © 2002 http://www.beckman.com/products/instrument/automatedsolutions/integsystems/sagian_coresystems_inst_dcr.asp 21 April 2003