The document discusses imaging spheroids using the Yokogawa CV7000 confocal imaging platform. Key points include: 1) The CV7000 platform can perform time-lapse imaging of spheroid growth in Corning ultra-low attachment 96-well and 384-well plates over multiple days. 2) It images spheroids in thick tissue slices with various objectives, allowing high-throughput screening. 3) The platform measures spheroid growth over time, obtaining area measurements from images taken at regular intervals over 127 hours.

1. Confocal Imaging of Spheroids
Using Corning ULA Plates with the
Yokogawa CV7000 Platform

2. Thick Sample Imaging
Tissue slices
SpheroidsS
Key Feature

3. Yokogawa CV7000
Time-lapse imaging of spheroid growth
on Corning Ultra-Low Attachment Surface
96-well and 384-well Plates

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. Spheroid Imaging with the CV7000 Platform
1 hour
127 hours
11 z-slices at 10 micron intervals
100 microns total

6. Spheroid Imaging with the CV7000 Platform
Timepoint (hr)
50000
45000
40000
25000
30000
35000
Area(µm2)
0 6545 55 60504035302520155 10
Spheroid Growth based on image area

7. Cell Voyager 7000
Configuration

8. Simultaneous Imaging with
1 - 3 Cameras
sCMOS Cameras
-20˚C
5.5MP = 2,560 x 2,160
6.5µm pixel

9. Objective Choices

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. 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. Imaging Modes
Confocal
Epifluorescence
Phase Contrast
Bright Field

13. Illumination Choices
Lasers: 405, 488, 561, 640nm
UV LED: 365nm
Halogen Lamp

14. Kinetics ModuleKey Capability

15. Visualize dynamic events
in live cells
Loader for Reagent plates
Loader for Disposable tip racks
5µl - 20µl (384-well)
10µl - 100µl (96-well)
Single tip dispensing
Kinetics Module - Live imaging

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. 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. Confidential CV7000 Update January 2015 - AstraZeneca
SearchFirstTM
Automated Hit Finding
Save Valuable Time
Reduce Screening Costs
Improve Imaging Results
Key Capability

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. 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. Most Flexible Imaging
Live cell imaging for days
Unmatched speed
Proven reliability
Yokogawa CV7000
www.wakoautomation.com