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