Thermo Scientific EVOS M5000 User Manual page 46

For fluorescence and transmitted light applications
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3.
If not already on, click the Light button to illuminate the sample, then adjust
brightness and focus using the corresponding sliders on the Capture tab.
The focus position you set in this step is the initial
Current Z position.
4.
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.
5.
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.
6.
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: You can click the Z number button for the Top, In Focus, and Bottom
positions to jump to those positions along the Z-axis.
7.
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.
EVOS
M5000 User Guide

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