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  • American Institute of Physics (AIP)  (2)
  • 1
    Electronic Resource
    Electronic Resource
    [S.l.] : American Institute of Physics (AIP)
    Journal of Applied Physics 84 (1998), S. 4033-4042 
    ISSN: 1089-7550
    Source: AIP Digital Archive
    Topics: Physics
    Notes: The combination of two-photon excitation 4Pi-confocal fluorescence microscopy with image restoration leads to a fundamental improvement of three-dimensional resolution in the imaging of transparent, fluorescent specimens. The improvement is exemplified by randomly dispersed fluorescent beads and with actin filaments in a mouse fibroblast cell. For an illumination wavelength of 810 nm, we obtained lateral and axial full-width at half-maxima of point-like objects of 120–140 nm, and 70–100 nm, respectively. Fluorescent beads that are 150 nm apart are imaged with an intensity dip of ∼25%. This amounts to a ∼sixfold improvement of the axial resolution over standard two-photon confocal microscopy. In the cell, the 3D-images reveal details otherwise not resolvable with focused light. © 1998 American Institute of Physics.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Woodbury, NY : American Institute of Physics (AIP)
    Applied Physics Letters 69 (1996), S. 3644-3646 
    ISSN: 1077-3118
    Source: AIP Digital Archive
    Topics: Physics
    Notes: We determine the resolution of high-performance confocal microscopes by measuring the three-dimensional point–spread function (3D-PSF) of an optimized confocal setup. The 3D-PSF is standardized by recording the scattered light of pointlike objects. For a wavelength of 543 nm and a specified numerical aperture of 1.4 (oil), we find an axial and lateral focal full width at half-maximum (FWHM) of 460±20 and 145±10 nm, respectively. A high signal-to-noise ratio is obtained by using recording times comparable to those of near-field scanning optical microscopy. We further reduce the effective PSF extent by means of a three-dimensional deconvolution technique exploiting the information gained from the measurement of the focus. We show that it is possible to obtain an axial and lateral FWHM of the far-field effective PSF after deconvolution of 80 and 40 nm, respectively. © 1996 American Institute of Physics.
    Type of Medium: Electronic Resource
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