Hi @IR23 isabel.
The optical resolution of the 2 photon microscope is determined by the illumination wavelength and the numerical aperture of the objective lens
The image resolution depends on how the scan we set up.
ImageJ will try to read the image meta data and after opening the image will.tell you in image properties window what it read for the xy and z image resolutions.
In theory the optical resolution should be oversampled about 2X by the image resolution. But it’s some work to find out if that was the case.
If you tell us what imageJ reads the xy and z Pixel sizes (really it’s spacing not Size)
And the illumination wavelength and objective NA, we can calculate the optical resolution and see if it’s sampled nicely by the image resolution.
Read handbook of confocal microscopy by James Pawley for more details.