1. High-resolution mapping of the three-dimensional point spread function in the near-focus region of a confocal microscope
- Author
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Michael J. Nasse, Jorg C. Woehl, Serge Huant, Nano-Optique et Forces (NOF), Institut Néel (NEEL), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS), and Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)
- Subjects
Physics ,Point spread function ,[PHYS.PHYS.PHYS-OPTICS]Physics [physics]/Physics [physics]/Optics [physics.optics] ,Microscope ,Physics and Astronomy (miscellaneous) ,business.industry ,02 engineering and technology ,021001 nanoscience & nanotechnology ,01 natural sciences ,law.invention ,010309 optics ,Transverse plane ,Light intensity ,Optics ,Cardinal point ,law ,Optical transfer function ,0103 physical sciences ,Equidistant ,0210 nano-technology ,business ,Image resolution - Abstract
International audience; Fluorescent nanobeads with a diameter of 20 nm were used to map the three-dimensional point spread function in the near-focus region of a confocal microscope at high spatial resolution. Fluorescence images were taken in 109 equidistant planes (50 nm apart) parallel to the focal plane; postacquisition stacking of these images allows the reconstruction of the point spread function in the axial plane. The experimental distribution is compared to theoretical calculations based on an integral representation for the light intensity in the focus region that takes into account stratified media, polarization, the Gaussian illumination profile, and the finite exit pinhole size.
- Published
- 2007
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