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Resolution of oblique-plane images in sectioning microscopy

Imaging living biological specimens in three dimensions is particularly challenging because of their time-varying behaviour. Confocal microscopes can record three-dimensional images but usually at relatively low speeds since they need to acquire a series of through-focus (x-y) images at different focal (z) positions. To overcome this limitation, we have devised a system capable of acquiring diffraction-limited images over a user-defined sub region. Since the focal spot is not spherical the resolution is non-isotropic. We show, for example, that oblique plane images can contain spectral content that could not have been generated by specimen features lying wholly within the plane.