... | ... | @@ -99,31 +99,31 @@ Use the yellow marker pen slide with a coverslip (on the desk next to the lens c |
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### Channel Alignment
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1. Use the 63x 1.4 (SIM) or 1.46 (TIRF) objective, use your own bead slide, and focus on beads
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1. Use the 63x 1.4 (SIM) or 1.46 (TIRF) objective, use your own bead slide, and focus on beads. Use the same filters as for your experiment and set up the cameras and tracks the same way as you will use them in your experiment. Set the illumination time to 100ms and use 0.1-1% laser power (when pressing min/max you should have >1000 white/grey values, when ticking 'Show all' in the 'Display' panel). Use the optimal grid pattern.
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2. Take a large Z-stack (50 slices) using optimal sampling (double-click on [Opt])
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3. Go to processing – Channel alignment
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3. Go to Processing – Channel alignment
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{width=40%}
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4. Choose the input image, activate Fit and Affine, Click apply
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5. Save the results in order to correct your images later on
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6. You can apply your correction together with the SIM pocessing
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6. You can apply your correction together with the SIM processing
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## Shutdown
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Shut-down
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- Close Zen Black.
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- Press the Load position on the touch panel.
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- Clean immersion objectives with lens cleaning paper.
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- Close Zen Black
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- Press the Load position on the touch panel
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- Clean immersion objectives with lens cleaning paper
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{width=40%}
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Leaving system for the next user
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- Logout from the Windows session.
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- Logout from the Windows session
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Full shutdown (last user of the day)
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- Shut down the PC.
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- Switch off the microscope.
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- Shut down the PC
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- Switch off the microscope
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Always make sure that the system is back at 27°C and the CO2 is off when you have done any live cell imaging.
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