Quasi-spherical focal spot in two-photon scanning microscopy by three-ring apodization
✍ Scribed by Cristina Ibáñez-López; Genaro Saavedra; Karsten Plamann; Gilbert Boyer; Manuel Martínez-Corral
- Book ID
- 102334067
- Publisher
- John Wiley and Sons
- Year
- 2005
- Tongue
- English
- Weight
- 169 KB
- Volume
- 67
- Category
- Article
- ISSN
- 1059-910X
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✦ Synopsis
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.