Resolution formula for Second Harmonic generation microscopy

Hello everyone,

I would like to calculate the optimal pixel-size to meet the Nyquist-Shannon criterion for SHG microscopy but I can’t find the formula anywhere. Do you guys have anything related to it ?
We collect the signal with an epi configuration, does it change anything to its calculation ?

Thank you for your kind help


Hi Julien,

Since SHG requires emission collection at 1/2 the incident wavelength of excitation - we routinely use this wavelength to calculate sampling. So if you are using 900nm from your TiS laser for excitation - you can use 450nm to determine resolution using Rayleigh (0.61*lambda/NA). Then set your pixel size to 2.5-3x smaller than Rayleigh.

Hope this helps!


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Hi Jason,

Thank you very much for your input, and sorry for the delay in my answer.
It was my assumption as well to use the resolution formula of confocal microscopy, but I was wondering if it exists a formula dedicated to this technique and to its specific component.
Anyway, it wont be less than confocal microscopy, so having a bit of oversampling is not a big deal I guess !

Thank you again !