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Fluorescence volume imaging with an axicon: simulation study based on scalar diffraction method

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Abstract

In a two-photon excitation fluorescence volume imaging (TPFVI) system, an axicon is used to generate a Bessel beam and at the same time to collect the generated fluorescence to achieve large depth of field. A slice-by-slice diffraction propagation model in the frame of the angular spectrum method is proposed to simulate the whole imaging process of TPFVI. The simulation reveals that the Bessel beam can penetrate deep in scattering media due to its self-reconstruction ability. The simulation also demonstrates that TPFVI can image a volume of interest in a single raster scan. Two-photon excitation is crucial to eliminate the signals that are generated by the side lobes of Bessel beams; the unwanted signals may be further suppressed by placing a spatial filter in the front of the detector. The simulation method will guide the system design in improving the performance of a TPFVI system.

© 2012 Optical Society of America

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