Zeiss LSM 880, Основное руководство пользователя

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Using the optimal distance between slices ensures the best coverage and 3D
reconstructions, while avoiding excessive bleaching and photo-toxicity on your sample (see
Nyquist sampling). Briefly, if you use a step size smaller than what the Nyquist sampling
theorem recommends you will not gain any more information and you will likely increase
bleaching and photo-toxicity on your sample by imaging the same “information” more times
than needed. You can choose the z-range to acquire by setting the first and last frames of the
stack or set the current frame as the central frame of the pile and choosing how many slices
you want to acquire. If you are using definite focus using Center might be the best option.
Also, when using Center insert an odd number of slices so the current frame is acquired.
Time Series
This allows you to perform time-lapse experiments, working best with Definite Focus in
longer time-lapses to ensure the sample stays focused. Number of cycles is the number of
pictures to take, if zero is selected the software will take pictures until manually stopped.
Interval is the time between each cycle, note that the interval timer resets when the picture
begins being taken, if the interval is shorter than the acquisition time the next cycle begins as
soon as the previous ends.
Tile Scan
Using Tile Scan you can have larger samples that do not fit in a single frame in the same
image with high magnification. This takes multiple images that overlap each other (at least
10% but optimally 30%) and then they can be stitched together into a single image. There are
three modes of tile scanning:
- Centered Grid - uses the current frame as a central point and acquires it and the
area around it;
- Bounding Grid – you set the top left corner and bottom right corner of the area you
want to acquire;
- Convex Hull – is used for irregular samples where a normal rectangular area would
have many blank frames that don’t need to be acquired, therefore reducing
acquisition time.