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Optimization and Implementation of the Sysmex SP-1000i in a Multispecies Laboratory
Samantha Wildeboer, MT(ASCP), MSPfizer Global Research and Development
Groton, CT
Sysmex User GroupOctober 16, 2012
Outline
• Pfizer Groton Clinical Pathology Laboratory
• Former Process
• Evaluation
• Optimization– Initial Set-up
– Troubleshooting
• Implementation
• Project Outcomes
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Pfizer Drug Safety Research and DevelopmentGroton Clinical Pathology Laboratory
• Multispecies Clinical Pathology Laboratory Supporting:– Multispieces preclinical GLP and non-GLP safety studies
– Multi-site colony health monitoring
– Research and Business Unit exploratory studies
– Phase I/II Clinical Support
– Veterinary Medicine Research and Development
– Average two species, 50-100 samples/day
– Species: Monkey, Dog, Rat, Mouse, Rabbit, Cat, Pig, Horse, Cow, and Goat
• Platforms include:– Siemens:120/2120, 1800/2400, Centaur, Atlas, Immulite 2000,
CentraLink Middleware
– Stago Compact
– Sysmex SP-1000i, XT-2000iV
– Cerner Millennium LIS
Former Process –Hematology Sample Processing
• Pre-label slides with required information
• After Samples received, slides would be made manually– Large animal – average 30.36 seconds/slide
– Rodent – average 33.76 seconds/slide
– Average of 20 min/day (based on average sample number)
– 83.3 hours/year
• Samples sent to instrumentation for analysis
• Slides left to dry
• Load slide carousels and stain using a Wescor 7120 Aerospray
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Former Process – WescorEosin/Thiazin/Azure B/Methylene Blue Stain
Canine
Non-Human Primate• Advantages– Staining time
< 5 minutes
– Prepared reagents
• Disadvantages– 12 slide carousel
– Very little customization (Lighter/Darker)
– No staining of rabbit basophils(basophils in general)
– Pre-fix used for bone marrow staining
RabbitBasophils
Inspiration!
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Evaluation of Available Technology
• Gathered our requirements:– Stand alone– Read barcode– Interface to LIS (Cerner)– Print study important information on slide– Able to support our tube size(s)– Different settings for variable hematocrit– Several stain protocols available– Make a smear and stain, or just stain
• Contacted Vendors
Sysmex SP-1000i Highlights
• Stand alone
• Can interface with LIS
• Prints up to three lines (3D barcode available)
• No premixing of stain
• Makes smears, stains, or stains premade slides
• Supports 13 x 75 (2 mL) and Microtainer (500 uL)
• Variable angle settings (hematocrit)
• Multiple stain protocols available
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Optimization – Initial Settings
• On-site technical specialist provided:– Training– Evaluation of dog, non-human primate, and rat
samples– Smear angles– Initial optimization of available stain (Wright-
Giemsa)– pH testing?– NERL Water
Optimization – Initial Settings (cont.)
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Optimization – Initial Settings (cont.)
Optimization – The Next Steps
Stain Optimization
Date: __________________
Species/Slide #: _______________
Settings
Stain 1 prefix: ___________
Stain 1 Dry Time: ___________
Stain Cycle: ___________
Stain 1 Dilution: ___________
Stain 1 Dilution Time: ___________
Stain 2 Dilution Time: ___________
Rinse: ___________
Heater: ___________
Dry Time: ___________
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Optimization – Artifact Crenation
Low grade, appearing on 85% of slides
Optimization – Troubleshooting
• Probably not the stain
• Slide preparation or drying
• Slides (try a new lot)
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Optimization – May Grüenwald/Giemsa Stain
• Feedback to Sysmex– Contacted SP-1000i users in Germany
– Provided May Grüenwald/Giemsa staining protocol
• SP-1000i was converted to the two stain system– We loved the stain, but the crenation was still
present
Optimization - Troubleshooting
• Sysmex provided two SP-1000i Experts to help troubleshoot the issue– Back to a single stain system (PROTOCOL)– Determined to be a combination of things
o Different slide typeo Adjustment to the smear settings
Wright-Giemsa - Rodent
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The final battle: Wrights-Giemsa vs. May Grüenwald-Giemsa
Unanimous vote for May-Grüenwald!
May-Grüenwald-Giemsa - Rodent
May Grüenwald-Giemsa
Human
Non-Human Primate
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May Grüenwald-Giemsa
Canine
Rodent
Wescor Stain
Rabbit
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May Grüenwald-Giemsa
Rabbit
Basophils!
Implementation - Validation
• Acceptable Stain Quality
– 5 samples from four selected representative species (Human, Rat, Dog, and Monkey) will be stained and evaluated for acceptable stain quality.
– Cells should be easily identified, including Red Blood Cell and platelet morphology. Manual differentials performed will match automated differentials from the Siemens Advia 120 Hematology analyzer.
• Sample Identification
– At least 10 samples will be placed on the SP-1000iand slides prepared by the unit will be visually checked to ensure correct sample identification.
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Implementation - Metrics
• < 1 min to load the SP-1000i with 40 specimens
– Average of 0.825 min/day (based on average sample number)
– 3.4 hours/year
• Pre-labeling/post-labeling
• Stain Time Loss 5 min vs. 40 min (hands off)
• Shorter turn around to get the specimens to the hematology analyzer
• Approximately 80 Technologist hours saved/year!
Additional Utilization
• Bone marrow staining– Cytospin preps
– Histology brush preps
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Rat Bone Marrow Cells
SP-1000i Project Outcomes
• Improvement in stain quality
• Standardization of peripheral blood smears
• Efficiency gains
• Allows for utilization of cell imaging and classification technology (CellaVision)
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Questions?
Back-Up Slides
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May-Grüenwald-Giemsa Stain Protocol
Stain 1 prefix Use
Stain 1 Fix time 00’03
Stain 1 Dry time 00’05
Stain Cycle 5
Stain 1 Staining 04’30
Stain 1 dilution USE
Stain 1 dilution time 05’00
Stain 2 dilution time 20’00
Rinse Use
Heater Not use
Dry time 03’00
All pictures included in this presentation were taken using
CellaVision DM96
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The final battle: Wrights-Giemsa vs. May Grüenwald -Feline
The final battle: Wrights-Giemsa vs. May Grüenwald -Mouse
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The final battle: Wrights-Giemsa vs. May Grüenwald –Non-human Primate
The final battle: Wrights-Giemsa vs. May Grüenwald -Canine
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The final battle: Wrights-Giemsa vs. May Grüenwald -Rat
