In scanned optical systems such as confocal laser microscopes wherein a
beam of light is focused to a spot in a specimen to excite a fluorescent
species or other excitable species in the spot, the effective size of the
excitation is made smaller than the size of the spot by providing a beam
of light of wavelength adapted to quench the excitation of the excitable
species, shaping this second beam into a pattern with a central intensity
minimum, and overlapping this central minimum with the central intensity
maximum of the focused spot, so that within the spot the intensity of
quenching light increases with distance from the center of the spot,
thereby preferentially quenching excitation in the peripheral parts of
the spot, and thereby reducing the effective size of the excitation and
thus improving the resolution of the system. In the preferred embodiment
of the preset invention, the central minimum of quenching light is
narrowed further by creating the pattern of quenching radiation in the
specimen by imaging onto the focal plane a plurality of pairs of sources
of quenching light, arrayed at the vertices of a regular, even-sided
polygon, the center of which is imaged in the specimen on the central
maximum of exciting radiation, and such that the two members of each pair
are on opposite vertices of the polygon and emit light mutually coherent
and out-of-phase, and the light emitted by different pairs is incoherent
with respect to each other.