Comparison of Widefield/Deconvolution and Confocal Microscopy for Three-Dimensional Imaging

The biggest limitation inherent in optical microscopy is its lateral spatial resolution, which is determined by the wavelength of the light used and the numerical aperture (NA) of the objective lens. Another important limitation is the resolution in the direction of the optical axis, conventionally called z, which is related to the depth of field. The presence of a finite aperture gives rise to undesirable and rather complicated characteristics in the image. In essence, the depth of field depends on the size of structure or spatial frequency being imaged.