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Development, Simulation and Optimization of Oncology Laboratory Sample Processing Automation Emily Salmon Advisors: Norman Development, Simulation and Optimization of Oncology Laboratory Sample Processing Automation Emily Salmon Advisors: Norman Purvis, Ph. D, Esoterix Dr. Paul King, Vanderbilt University

Introduction • Esoterix Oncology performs Immunophenotyping of Leukemia/Lymphoma Conditions • Assays are Performed manually 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 • 60 Blood/Bone Marrow Specimens, 40 Tissue Specimens, Daily • 8 test Current Statistics • 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 Why Automate? • • • Decrease Cost Reduce Staff Increase Throughput Decrease Time Increase Efficiency • 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 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 Normal FAB-M 1 FAB-M 3 FAB-M 4

The Laboratory Blood/Bone Marrow Screening Lab Tech adding antibodies Tissue Screening Lab Tech at 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 ORCANT ROBOT Six Axis of Motion: • movement along rail • height • reach • bend • twist • grip

Sample Manual Process Corresponding Automated Process Sample Manual Process Corresponding Automated Process

Design-Choosing a Procedure to Implement Process of adding lyse, shaking wells, centrifuging at 1300 Design-Choosing a Procedure to Implement Process 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

Instrumentation Beckman Coulter Instruments: – Biomek Fx Liquid Handling System – Multidrop – ORCA Instrumentation Beckman 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 software Build Simulation Software SAGIANTM Core System • SAMI Automated Method Development and Scheduling software Build methods by connecting and configuring nodes, sources, and stations. • SILAS Integration software Run SAMI methods, gather resource use statistics and time values • BIOMEK FX Automation Workstation Separate module for complicated pipetting schemes • ORCA NT Motion Editor Software Used for framing and “teaching” of transportation robot

Creating a SAMI Method Creating a SAMI Method

Biomek FX Custom Pipetting Biomek FX Method Editor The Biomek FX Biomek FX Custom Pipetting Biomek FX Method Editor The Biomek FX

Scheduling and Summary Outputs Allocation of time per task, each plate Plate Location Summary Scheduling and Summary Outputs Allocation of time per task, each plate Plate Location Summary Resource Utilization Bar Graph Text timing Details

Design Aspects • Choosing a lysing strategy, Choosing Basic Labware • Dimension Configurations • 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 Simulation Times

SAMI Simulation Times vs. Manual Model Times, Projected Volume Current 6 Months 1 Year 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 2: 30: 31 3: 17: 36 3: 56: 24 4: 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 5: 43 6: 12 7: 39 11: 40 Tissues Current 6 Months 1 Year 3 Years Simulated Time 1: 20 1: 36: 04 1: 50: 49 2: 10: 17 3: 32 3: 51 4: 10 6: 04 Total Simulation Time Manual Proj Time

Conclusion • Protocols created are basis for future implementation • Automation can be shown Conclusion • Protocols created are basis for future implementation • Automation can be shown to save time and money Recommendations • 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

References: The Purdue Cytometry CD-ROM Volume 4, J. Watson, Guest Ed. , J. Paul 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 2003 Beckman Coulter “Cytomics FC 500 Series Flow Cytometry Systems” © 2002 http: //www. beckman. com/products/instrument/flowcytometry/fc 500 series. asp 21 April 2003 Beckman Coulter “Orca Robot” © 2002 http: //www. beckman. com/products/instrument/automatedsolutions/integsystems /orcarobot_dcr. asp 21 April 2003 Beckman Coulter “SAGIAN Core Systems” © 2002 http: //www. beckman. com/products/instrument/automatedsolutions/integsystems /sagian_coresystems_inst_dcr. asp 21 April 2003