Re: Z sections
Posted by
Fred Mast on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Z-sections-tp1142113p1142297.html
If the purpose of your z-sectioning is to 3D-reconstruct your image than the optimal z-section spacing is determined by the axial resolution of your objective and Nyquist's sampling theorem. As has been noted on this list in the past, there are a number of excellent websites and textbooks available that properly develop and explain these ideas further. I would recommend the Nyquist calculator on the SVI website:
The Handbook of Biological Confocal Microscopy by James Pawley is also an excellent resource in this regards. However, if time is of the essence and you can achieve your goals using a stereological approach then design a random sampling protocol and use statistics to extrapolate your data. The best protocol will be tailored to your needs and prior knowledge of your sample will help. ie. are you searching for a regular or irregular object... Your sample will determine the types of assumptions that can be made.
Hope that helps,
Fred
On 3-Oct-08, at 8:40 AM, Matthew Pearson wrote:
Hi all,
I have a question concerning Z sectioning. We have a Leica SP2 and when set the top and bottom range for a Z stack, the software automatically calculates the optimised number of sections, which is nearly always over 100! This is however not practical as the stacks take too long to acquire. Can anyone shed some light on how the software decides how many slices are necessary and if I haven't got time to take 100, how can I work out what will be a sufficient number.
Thanks for your help in advance.
Matt Pearson.
Fred D. Mast
Department of Cell Biology
Medical Sciences Building Room 5-14
University of Alberta
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Canada
Tel: 1-780-492-7407