Widefield Observer
Multi-well Imaging Setup
This guide covers the setup process for multi-well imaging using the Zeiss Observer microscope with Zen-blue software.
Initial Setup
- Launch Zen-blue software
- Calibrate the stage
- Navigate to acquisition tab
- Activate the tiles module
Sample Carrier Configuration
- In the tiles menu, select "Sample Carrier"
- Choose your desired carrier type
- Use Smart Setup to configure imaging conditions
At least one channel must be set up for carrier calibration
Calibration Process
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Stage Calibration - Option to calibrate using transmitted light (recommended) - Choose between re-calibrating or continuing with existing calibration
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Calibration Points - Select calibration method (7 points recommended) - Position points with high accuracy - More points generally yield better results
Advanced Setup
Region Setup Options
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Contour Method - Draw regions manually for imaging
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Predefined Method - Click on slide to add region - Use plus button to add setup region
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Carrier Method - Set fill factor for coverage - Configure columns/rows for custom layout - Click create to generate tile regions
Position Setup Options
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Location Method - Manual point placement - Click to add specific points
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Array Method - Add multiple points in random or grid pattern - Draw regions to define point distribution area
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Carrier Method - Add regions (columns/rows) in each well
Focus Settings
- Access Support Point Editor (circle with dot icon above calibrate button)
- Add support points for focus shift: - Manual addition via plus button - Add one per container - Set row/column distribution
- Adjust point positions as needed
Final Steps
- Verify all tile regions
- Review support point positions
- Confirm settings before starting acquisition
For optimal results, carefully verify focus points and tile regions before starting a long acquisition sequence.
Best Practices
- Use transmitted light for initial calibration when possible
- Add sufficient support points to maintain focus across the sample
- Verify all settings before starting long acquisitions
- Consider sample characteristics when choosing imaging regions