Quasi-spherical focal spot in two-photon scanning microscopy by three-ring apodization

Abstract : We present a beam-shaping technique for two-photon excitation (TPE) fluorescence microscopy. We show that by inserting a properly designed three-ring pupil filter in the illumination beam of the microscope, the effective optical sectioning capacity of such a system improves so that the point spread function gets a quasi-spherical shape. Such an improvement, which allows the acquisition of 3D images with isotropic quality, is obtained at the expense of only a small increase of the overall energy in the axial sidelobes. The performance of this technique is illustrated with a scanning TPE microscopy experiment in which the image of small beads is obtained. We demonstrate an effective narrowing of 12.5% in the axial extent of the point spread function, while keeping the 82% of the spot-fluorescence efficiency.
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Submitted on : Wednesday, October 20, 2010 - 2:42:33 PM
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Cristina Ibáñez-López, Genaro Saavedra, Karsten Plamann, Gilbert Boyer, Manuel Martínez-Corral. Quasi-spherical focal spot in two-photon scanning microscopy by three-ring apodization. Microscopy Research and Technique, Wiley, 2005, 67 (1), pp.22-26. ⟨10.1002/jemt.20175⟩. ⟨hal-00527854⟩

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