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Nonlinear delayed fluorescence in confocal scanning microscopy. Point-spread-function analysis.

Kowalczyk M., Martinez-Corral M., Zapata Rodriguez C. J.

Optica Applicata

2000

Vol. 30, nr 2-3

231-238

Generation of the signal beam in the confocal scanning fluorescence microscopy by means of nonlinear delayed fluorescence phenomenon based on triplet-triplet annihilation is proposed. From the point of view of the spatial resolution of three-dimensional confocal imaging the superiority of the phenomenon proposed over the classical two-photon fluorescence is demonstrated. This is done in terms of axial and lateral point spread functions computed for the assumed model of nonlinearity.

Tunable axial resolution in confocal scanning microscopy by controlled symmetrical defocusing.

Martinez-Corral M, Kowalczyk M., Kowalczyk M., Andres P.

Optica Applicata

1998

Vol. 28, nr 3

213-225

There is presented a quite simple technique for improving the axial resolution capacity of confocal scanning microscopes without detriment to their transverse resolution. The technique, that is based in the equal contribution to the image of the illuminating and collecting lenses, consists in symmetrical defocusing of both parts of the imaging system.

Phase pupil filters for improvement of the axial resolution in confocal scanning microscopy.

Kowalczyk M., Zapata C., Silvestre E., Martinez-Corral M.

Optica Applicata

1998

Vol. 28, nr 2

127-137

Two phase pupil filters whose axial impulse response is asymmetric with respect to the focal point are proposed. It is shown that when either of the filters is placed both in the illuminating and collecting part of the system the impulse respon se becomes symmetric and the axial resolution increases. The filters are designed on the basis of the Fourier-transform properties of the Hermition functions. They consist of small number of zones (=3) whose amplitude transmittance is constant and its module is equal to unity. The width of all zones is adequately chosen. Several quantitative characteristic of the proposed filters which evidence the improvement of the axial resolution are presented.