1. 1. Confocal Imaging of Spheroids Using Corning ULA Plates with the Yokogawa CV7000 Platform
2. 2. Thick Sample Imaging Tissue slices SpheroidsS Key Feature
3. 3. Yokogawa CV7000 Time-lapse imaging of spheroid growth on Corning Ultra-Low Attachment Surface 96-well and 384-well Plates
4. 4. Spheroid Imaging with the CV7000 Platform Spheroid models have become common for research applications such as drug discovery and toxicity assays. Corning 96-well round bottom ultra low attachment spheroid microplates enable cells to form a spheroid centered in each well. This typicall requires 2-3 days incubation. The 96-well and 384-well formats are useful formats for high- throughput screening. The challenge has been imaging the spheroids. The CV7000 Platform solves this challenge
5. 5. Spheroid Imaging with the CV7000 Platform 1 hour 127 hours 11 z-slices at 10 micron intervals 100 microns total
6. 6. Spheroid Imaging with the CV7000 Platform Timepoint (hr) 50000 45000 40000 25000 30000 35000 Area(m2) 0 6545 55 60504035302520155 10 Spheroid Growth based on image area
7. 7. Cell Voyager 7000 Configuration
8. 8. Simultaneous Imaging with 1 - 3 Cameras sCMOS Cameras -20C 5.5MP = 2,560 x 2,160 6.5m pixel
9. 9. Objective Choices
10. 10. Choose Up To 6 Lenses Dry: 2x, 4x NA .16, 10x NA .4, 20x NA .75, 40x NA .95, 20x NA .45 LWD Water Immersion: 40xNA 1.15, 60x NA 1.2 Phase Contrast: 10X, 20X
11. 11. Plate Type Well Size Lens 96 well 6.5mm 2x objective 384 well 3.25mm 4x objective 1536 well 1.5mm 10x objective Complete Well FOV
12. 12. Imaging Modes Confocal Epifluorescence Phase Contrast Bright Field
13. 13. Illumination Choices Lasers: 405, 488, 561, 640nm UV LED: 365nm Halogen Lamp
14. 14. Kinetics ModuleKey Capability
15. 15. Visualize dynamic events in live cells Loader for Reagent plates Loader for Disposable tip racks 5l - 20l (384-well) 10l - 100l (96-well) Single tip dispensing Kinetics Module - Live imaging
16. 16. Reagents added with disposable tips Choice of 96-well, 384-well Multiple dispenses possible Begin imaging before dispensing Continue imaging during dispense Imaging speed 8 fps (no binning) Cut away view of Dispenser with CV7000 case removed Kinetics Module - Live imaging
17. 17. 2x binning: 15 fps at 1 color 7.5 fps at 2 color. 3x binning: 38 fps at 1 color 20 fps at 2 color Exceptional Image Quality Kinetics Module - Live imaging Imaging Speeds
18. 18. Confidential CV7000 Update January 2015 - AstraZeneca SearchFirstTM Automated Hit Finding Save Valuable Time Reduce Screening Costs Improve Imaging Results Key Capability
19. 19. SearchFirst quickly finds specific fields of interest (hits) throughout a plate, and then re-scans those specific fields of interest - automatically. 1. Low resolution scan of all wells (4X, 10X) 2. Images sent to analysis platform, hits returned 3. High resolution scan of hits (40X, 60XW) SearchFirst
20. 20. SearchFirst uses the full flexibility of the CV7000 interface enabling choice of colors, objectives, FOV, binning, and time lapse settings within a plate and even within a well (different fields within a well can have different settings). SearchFirst
21. 21. Most Flexible Imaging Live cell imaging for days Unmatched speed Proven reliability Yokogawa CV7000 www.wakoautomation.com