Zeiss LSM 880 Basic User's Manual

Also see for LSM 880: Operating manual Quick operation manual Quick start guide User guide

24Using the optimal distance between slices ensures the best coverage and 3Dreconstructions, while avoiding excessive bleaching and photo-toxicity on your sample (seeNyquist sampling). Briefly, if you use a step size smaller than what the Nyquist samplingtheorem recommends you will not gain any more information and you will likely increasebleaching and photo-toxicity on your sample by imaging the same “information” more timesthan needed. You can choose the z-range to acquire by setting the first and last frames of thestack or set the current frame as the central frame of the pile and choosing how many slicesyou 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 SeriesThis allows you to perform time-lapse experiments, working best with Definite Focus inlonger time-lapses to ensure the sample stays focused. Number of cycles is the number ofpictures 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 picturebegins being taken, if the interval is shorter than the acquisition time the next cycle begins assoon as the previous ends.Tile ScanUsing Tile Scan you can have larger samples that do not fit in a single frame in the sameimage with high magnification. This takes multiple images that overlap each other (at least10% but optimally 30%) and then they can be stitched together into a single image. There arethree modes of tile scanning:- Centered Grid - uses the current frame as a central point and acquires it and thearea around it;- Bounding Grid – you set the top left corner and bottom right corner of the area youwant to acquire;- Convex Hull – is used for irregular samples where a normal rectangular area wouldhave many blank frames that don’t need to be acquired, therefore reducingacquisition time.

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