LSM980 Protocol: Fixed Cell Imaging

Setup

1. Turn on the power switch.
2. Login to the session and open ZEN Blue software.
3. Calibrate the system.
4. Place the sample on the stage. Make sure the objective lens is at the lowest magnification.

Sample Finding

1. In ZEN Blue, press AI Sample Finder.
2. Click Next, check DAPI, click Preview, then Finish.
3. A Navigation tab will appear.

Channel Configuration

1. Go to Smart Setup.
2. Add the channels you want to image: DAPI, AF647, AF488.
3. On the left panel of the Acquisition tab, press + Widefield to add a third track called TL Coherence.

Widefield Tile Imaging

1. Check only the WF (widefield) track.
2. Check Tile.
3. Double-click the area you want to image, then press Live.
4. Adjust focus until the image is in focus.
5. Move to the corner of the desired square.
6. Set up by Contour — draw a 2×2 square starting at the corner of the desired area. This defines the tile region to be collected.
7. Press Start Experiment.
8. Save the image as a .czi file.
9. The system will collect a 2×2 tile image of the drawn square.

Confocal Imaging

1. Uncheck the WF track.
2. Check the confocal tracks.
3. Press Live to adjust focus.
4. Press Start Experiment.
5. Save the image as a .czi file.

Moving to a New Area

1. Go back to the Navigation tab.
2. Double-click a new area to image and repeat the confocal imaging steps.