Molecular Devices ImageXpress Confocal HT.ai, User Manual

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ImageXpress Confocal HT.ai High-Content Imaging System User Guide
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Option Description
Image-Based Available for the second or greater acquisition wavelengths and if you select
Enable Image-Based Focusing on the Acquisition tab. Specify the Z-Offset from
W1 in µm in the Offset field or click Calculate Offset to calculate the offset to use
for all subsequent wavelengths. If you select this option, specify the following
options:
Image-Based Range +/- µm specifies the range to use for the image-based
portions of autofocusing.
Note: Adjust the range based on the sample variability. A larger range
requires a longer time to focus.
Max. Step specifies the maximum step size in µm of a single Z move to use to
attain the correct focus position. This setting is dependent on the objective. Use
a smaller step size with higher NA objectives because the focus peak is
narrower.
Note: Smaller step sizes require more steps to arrive at the final focus
position. Increasing the number of image autofocus steps increases the
chances of photobleaching or phototoxicity.
If you select Custom Exposure Times on the Autofocus tab, and you select
Image-Based, specify a value for the following:
Table 5-8: W# Tab: Autofocus Options (options vary based on Acquisition tab selections)
(continued)
Option Description
Acquire Time
Series allows
Timelapse Specifies the image collection intervals for the wavelength.
At All Time Points allows acquisition of an image using the wavelength at each
time point in the experiment.
At Start of Experiment acquires an image using the wavelength at the first time
point only.
At Start/End of Experiment acquires an image using the wavelength at the first
time point and the last time point for the experiment.
Every Nth Timepoint acquires an image using the wavelength at the time point
interval, for example, every 5th time point, beginning with the first time point for
the experiment.
Table 5-9: W# Tab: Acquisition Options (options vary based on the Series option on the
Acquisition tab)