Schrader, M., Hell, S. W. and van der Voort, H. T. M.
Three-dimensional super-resolution with a 4pi-confocal microscope using image restoration. Journal of Applied Physics 84: 4033-4042 (1998).

The combination of two-photon excitation 4Pi-confocal fluorescence microscopy with image restoration leads to a fundamental improvement of three-dimensional resolution in the imaging of transparent, fluorescent specimens. The improvement is exemplified by randomly dispersed fluorescent beads and with actin filaments in a mouse fibroblast cell. For an illumination wavelength of 810 nm, we obtained lateral and axial full-width at half-maxima of point-like objects of 120-140 nm, and 70-100 nm, respectively. Fluorescent beads that are 150 nm apart are imaged with an intensity dip of ~25%. This amounts to a ~sixfold improvement of the axial resolution over standard two-photon confocal microscopy. In the cell, the 3D-images reveal details otherwise not resolvable with focused light.