6533b862fe1ef96bd12c6cba
RESEARCH PRODUCT
Quasi-spherical focal spot in two-photon scanning microscopy by three-ring apodization
Genaro SaavedraManuel Martínez-corralKarsten PlamannC. Ibáñez-lópezGilbert Boyersubject
Point spread functionHistologyMaterials scienceMicroscopeOptical sectioning02 engineering and technology01 natural scienceslaw.invention010309 opticsQuality (physics)OpticsTwo-photon excitation microscopyApodizationlaw0103 physical sciencesMicroscopyImage Processing Computer-AssistedInstrumentationtwo-photon excitation[PHYS.PHYS.PHYS-OPTICS]Physics [physics]/Physics [physics]/Optics [physics.optics]business.industry021001 nanoscience & nanotechnologybeam shaping3. Good healthMedical Laboratory TechnologyMicroscopy Fluorescence Multiphotonoptical sectioning effectAnatomy0210 nano-technologybusinessBeam (structure)Algorithmsdescription
International audience; 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.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2005-07-15 |