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Definite Focus 2
Definite Focus uses infrared light to keep your sample focused avoiding thermal and
vibrational drift, which will make your field-of-view change over time. It works by measuring
the refractive index mismatch between glass and your sample. Hence it only works is the
refractive index of your sample is different from glass, for example water-based media, such as
agarose pads or cell culture medium will work, whereas curing mounting media such as
Mowiol or Prolong Gold for example will NOT work once cured, intermediate mounting media,
such as Vectachield will be suboptimal unless diluted with an aqueous buffer.
To use this first select Definite Focus in the
Focus Devices and Strategy
tab, then in the
Definite Focus
tab click Store Focus and Periodic so the current focus is maintained.
Multidimensional Acquisition
All modes under Multidimensional Acquisition ca be used simultaneously. Eg: you can do
a time-lapse of a z-stack tile region in multiple areas of the sample.
Z-Stack
Since confocal imaging gives you optical sectioning acquiring images from different focal
planes may yield new information (as you will not collect out-of-focus light from the out-of-
focus imaging planes, effectively increasing the signal-to-noise ratio), selecting
Z-Stack
allows
you to do so.
