Invitrogen EVOS M5000 - Page 75

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
boundar
y 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 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 text box. The system calculates Step Size and the Z positions of
the images.
Note:
You can take up to 100 images (optical sections) for a Z-Stack.
8.
Click
Browse to open the Choose Save Folder dialog.
9.
Go to the destination folder for the Z Stack images, then click Select Folder.
10.
Select the
File Type: PNG or TIFF.
11.
Select
Generate Projection Image to generate a projection image.
When selected, the system uses the most in-focus pixels from images captured at dierent 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.
12.
Click
Start to capture the Z-Stack image sets based on your specifications.
The system provides information about the progress of the Z Stack protocol.
13.
When the Z-Stack routine is completed, the Review tab opens to the files captured in the routine.
Chapter8Capture Z-Stack
Capture Z-stack images
8
EVOS
™
M5000 Imaging System User Guide
75