Optimal z-step in confocal

Hi,

I’m trying to find out the formula for the optimal z-step in a confocal.
The only info I found is that most acquisition software do the calculation for the user.

Could someone give indicate to me what it is ?

Thank you

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https://svi.nl/NyquistCalculator

Most of the information you need will probably be on that page.

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The website gives a nice description. Of course it depends on what you’re trying to accomplish. If you want the best possible 3D reconstruction (if using a standard confocal), then in general, calculating the resolving capability of the objective in ‘Z’ and choosing Z-step sizes at half that value or smaller will give you good results. There will always be Z-stretch on conventional microscopes. There are exceptions that are discussed on that webpage, and sometimes reality will tell you that taking 85 slices, in 4 wavelengths, for tons of locations, at high quality scanning isn’t practical. Good luck!