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Quasi‐spherical focal spot in two‐photon scanning microscopy by three‐ring apodization
Author(s) -
IbáñezLópez Cristina,
Saavedra Genaro,
Plamann Karsten,
Boyer Gilbert,
MartínezCorral Manuel
Publication year - 2005
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20175
Subject(s) - apodization , optics , point spread function , microscopy , optical sectioning , materials science , microscope , optical microscope , isotropy , focal point , fluorescence microscope , spherical aberration , beam (structure) , cardinal point , physics , fluorescence , lens (geology) , scanning electron microscope
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. Microsc. Res. Tech. 67:22–26, 2005. © 2005 Wiley‐Liss, Inc.