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

2. 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

3. 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

4. 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 FAB-M1 NormalFAB-M3FAB-M4

5. The Laboratory Blood/Bone Marrow Screening Lab Tech adding antibodies Tissue Screening LabTech at sink, Centrifuge

6. ORCA NT ROBOT Six Axis of Motion: movement along rail height reach bend twist grip

7. Sample Manual Process Corresponding Automated Process

8. 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

9. Instrumentation Beckman Coulter Instruments: –Biomek Fx Liquid Handling System –Multidrop –ORCA Robot –Filtration Station –Carousel/Incubation for 96 well plate –Flow Cytometer

10. Simulation Software SAGIAN TM 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

11. Creating a SAMI Method

12. Biomek FX Custom Pipetting The Biomek FX Biomek FX Method Editor

13. Scheduling and Summary Outputs Allocation of time per task, each plateResource Utilization Bar Graph Plate Location SummaryText timing Details

14. 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

15. Simulation Times Revision I Revision II Revision III Revision I Revision II Revision III Revision I Revision II Revision III Revision I Revision II Revision III Revision I Revision II Revision III Revision I Revision II Revision III

16. SAMI Simulation Times vs. Manual Model Times, Projected Volume Current6 Months1 Year3 Years Growth (Proj)116%33%100% Bl/BMarrow no. samples606783120 Tissues no. samples40455380 Total Simulation Time2:30:313:17:363:56:244:44:48 Blood/Bone MarrowCurrent6 Months1 Year3 Years Simulated Time1:54:162:13:202:38:493:36:46 Manual Proj Time5:436:127:3911:40 TissuesCurrent6 Months1 Year3 Years Simulated Time1:21:201:36:041:50:492:10:17 Manual Proj Time3:323:514:106:04

17. 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

18. 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 2003http://www.ctyo.purdue.edu Beckman Coulter “Cytomics FC 500 Series Flow Cytometry Systems” © 2002 http://www.beckman.com/products/instrument/flowcytometry/fc500series.asp 21 April http://www.beckman.com/products/instrument/flowcytometry/fc500series.asp 2003 Beckman Coulter “Orca Robot” © 2002 http://www.beckman.com/products/instrument/automatedsolutions/integsystems /orcarobot_dcr.asp 21 April 2003 http://www.beckman.com/products/instrument/automatedsolutions/integsystems /orcarobot_dcr.asp Beckman Coulter “SAGIAN Core Systems” © 2002 http://www.beckman.com/products/instrument/automatedsolutions/integsystems /sagian_coresystems_inst_dcr.asp 21 April 2003 http://www.beckman.com/products/instrument/automatedsolutions/integsystems /sagian_coresystems_inst_dcr.asp