Molecular Devices ImageXpress Confocal HT.ai, User Manual

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ImageXpress Confocal HT.ai High-Content Imaging System User Guide
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To set up the ImageXpress Transmitted Light option for Koehler Illumination:
1. Load the sample (multi-well plate or slide).
2. Select a low magnification objective such as the 10x Plan Apo or a 4x objective.
3. Rotate the condenser turret (4) to select the A position (at the front). Open the condenser
aperture diaphragm by moving the field stop slider (5) above the A to the right.
4. In the MetaXpress software, do one of the following to display the Plate Acquisition Setup
dialog:
From the simplified menu, click Screening > Acquisition Setup.
From the standard menu, click Screening > Plate Acquisition Setup.
5. Move the stage to a well or slide area where there is visible sample.
6. Select one of the available Transmitted Light illumination settings.
7. Use laser-based autofocus, image-based autofocus, or live mode to focus on the sample.
8. Click Start Live.
9. Close the field aperture diaphragm (1) most of the way by moving the lever left or down if
you have the older model (1*).
10. Adjust the condenser height with the condenser focus control (2) until a bright polygon
surrounded by sharp dark edges is visible.
Figure 7-44: Condenser Height Focus Adjustment Comparison
11. If you cannot detect sharp edges, open the field aperture diaphragm (1) further, and try
adjusting the condenser height again.
12. Center the white polygon with the condenser adjustment screws (3), also known as field
diaphragm adjustment screws.
13. Open the field aperture diaphragm (1) by moving the lever right, or up if you have the older
model (1*), until the whole field is illuminated, and the dark edges are just outside of the
field of view. Opening this too far can increase glare in the images.
14. Close the condenser aperture diaphragm slightly by moving the field stop slider (5) above
the A to the left, to optimize the sharpness and contrast of the image.
15. Click Stop Live.