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
Julien
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!
-Jason
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 !