Capturing A M Ulti-­-Wavelength 3 D I Mage ( Z S Tack) - Leica SP5 confocal User Manual

Capturing a M ulti-­‐Wavelength 3 D I mage ( Z S tack)
1. Adjust scan parameters as for 2D image capture.
2. When optimising the image intensity for a z stack, move
through focus while live scanning to find the focal plane with
the highest signal intensity. Turn on the pixel saturation
indicator to aid in this process.
3. Ensure the Z stack panel is visible. View the live image and
move in one direction through focus using the joystick Z
control until you find the top/bottom of your sample. Click on
the Begin arrow on the diagram to record that position.
Then move to the opposite end of the sample and click the
End arrow. Stop scanning.
4. You can change the number of steps or step size by typing in the relevant boxes.
It is recommended that you use the System Optimised option. This will
calculated the step size and number of steps automatically according to the
Nyquist-Shannon sampling theorem.
5. Click Start button to acquire the Z stack.
6. Z stacks appear in the experiment folder with the name "series" by default.
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