Thermo Scientific EVOS M5000 User Manual page 40

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8. If you have opted to capture Z-Stack images, define the In Focus, Top, and
Bottom positions for the Z-Stack image set:
a. If not already on, click the Light button to illuminate the sample as you
locate the boundaries and the In Focus position of the Z-Stack.
b. To accept the Current Z as the In Focus position, click Set for In Focus.
To change the In Focus position, move the Coarse and Fine focus sliders
on the Capture tab to focus on the sample, then click Set.
c. Move the focus sliders up from the Current Z position to the desired Top
position (upper boundary of the Z-Stack), then click Set for Top.
d. Move the focus sliders down from the Current Z position to the desired
Bottom position (lower boundary of the Z-Stack), then click Set for Bottom.
Note: Click the Z number button for the Top, In Focus, and Bottom positions
to jump to that position along the Z-axis.
9. Define the Z-Stack parameters to determine the number of "optical sections"
captured in the Z-Stack image set:
•
Automatically compute Z-Stack Parameters: Select to calculate the
number of images and their Z-positions automatically based on the Z-Stack
boundaries and the default Step Size.
•
Manually Set Step Size: Select to enter the Step Size (Z-distance in µm
between focal planes) in the corresponding text box. The system calculates
the number and Z-positions of the images based on your entry.
•
Manually Set Number of Images: Select to enter the Number of Images
(Z-distance in µm between focal planes) in the corresponding text box.
The system calculates Step Size and the Z-positions of the images based on
your entry.
10. Select Generate Projection Image to generate a
projection image.
When selected, the system uses the most in-focus pixels from images captured
at different focal planes to generate a composite in-focus image. Each image in
the Z-Stack is also separately saved.
When unselected, the system only saves the individual Z-Stack images.
11. Click Next to proceed to create Run.
EVOS M5000 User Guide
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