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Leica AOBS Confocal Maximum and "Optimum" Zoom settings.
Why this matters:
Collecting optical information to the maximum resolution is often desired. "Oversampling" is known to cause sample destruction while not yielding additional information. Not sampling to the maximum resolution it is commonly called "Undersampling". While this does not destroy the sample, the maximum resolution is not attained. Should maximum resolution be necessary, optimal settings must be achieved.
Some individuals will not demand maximum resolution and should not concern themselves with this other than to note that they have no need to zoom beyond the maximum optimal zoom. These calculations are only important if you demand the optimum sampling size with minimal sample destruction.
Why not Oversample?
Oversampling will not yield increased resolution, and will simultaneously destroy the sample to a greater degree than is necessary. While oversampled images may sometimes look better, they are collected at the cost of excessive bleaching and without the gain of additional data. It is important to emphasize that oversampling with the confocal microscope is not beneficial to users demanding both high resolution and minimal sample degredation.
Is undersampling always bad?
Features that are clearly visible while undersampled may not necessitate the use of optimal zoom settings. Lower zoom settings may in fact be considered optimal even while the image is undersampled. The lower zoom subject samples to lower amounts of light and consequently less damage. If the lower setting achieve the desired image resolution then it is not necessary to increase the zoom in an effort to attain the maximum possible resolution.
Leica AOBS:
These conditions and approximate zoom settings were calculated for the Leica AOBS.
Click here for the Excel spreadsheet
1. Leica AOBS handbook
2. Pawley (1995). Handbook of Biological Confocal Microscopy (Second Edition)
Dustin Grzesik12/2003
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