Imaging of in vivo and ex vivo biological samples using full-field optical coherence tomography is demonstrated. Three variations on the original full-field optical coherence tomography instrument are presented, and evaluated in terms of performance. The instruments are based on the Linnik interferometer illuminated by a white light source. Images in the en face orientation are obtained in real-time without scanning by using a two-dimensional parallel detector array. An isotropic resolution capability better than 1 μm is achieved thanks to the use of a broad spectrum source and high numerical aperture microscope objectives. Detection sensitivity up to 90 dB is demonstrated. Image acquisition times as short as 10 μs per en face image are possible. A variety of in vivo and ex vivo imaging applications is explored, particularly in the fields of embryology, ophthalmology and botany.
We present results of studies in embryology and ophthalmology performed using our ultrahigh-resolution full-field OCT system. We also discuss recent developments to our ultrashort acquisition time full-field optical coherence tomography system designed to allow in vivo biological imaging. Preliminary results of high-speed imaging in biological samples are presented. The core of the experimental setup is the Linnik interferometer, illuminated by a white light source. En face tomographic images are obtained in real-time without scanning by computing the difference of two phase-opposed interferometric images recorded by high-resolution CCD cameras. An isotropic spatial resolution of ~1 μm is achieved thanks to the short source coherence length and the use of high numerical aperture microscope objectives. A detection sensitivity of ~90 dB is obtained by means of image averaging and pixel binning. In ophthalmology, reconstructed xz images from rat ocular tissue are presented, where cellular-level structures in the retina are revealed, demonstrating the unprecedented resolution of our instrument. Three-dimensional reconstructions of the mouse embryo allowing the study of the establishment of the anterior-posterior axis are shown. Finally we present the first results of embryonic imaging using the new rapid acquisition full-field OCT system, which offers an acquisition time of 10 μs per frame.
We have developed a white-light interference microscope as an alternative technique to conventional optical coherence tomography (OCT). The experimental setup is based on a Linnik interferometer illuminated with a tungsten halogen lamp. En face tomographic images are obtained in real-time without scanning by computing the difference of two phase-opposed interferometric images recorded by a CCD camera. The short coherence length of the source yields an optical sectioning ability with 0.7 μm resolution (in water). Transverse resolution of 0.9 μm is achieved by using high numerical aperture microscope objectives. A shot-noise limited detection sensitivity of 90 dB can be reached with ~ 1 s acquisition time. High-resolution images of mouse and tadpole embryos are shown.
We present a microscope set-up designed to produce three-dimensional images of the internal structures of various samples with high spatial resolution (a few tens of nanometers in axial and transverse directions). This level of resolution is reached by the use of nanometric sub-wavelength spherical gold beads as multiple local probes, dispersed in the hollow structures. The exploration, by Brownian motion, of the internal structures allows their three-dimensional reconstruction.
We present the new advances in full field optical coherence microscopy, an alternative method to conventional optical coherence tomography (OCT). The experimental setup is based on Linnik interferometer illuminated with a tungsten halogen lamp. En face tomographic images are obtained in real-time without scanning by computing the difference of two phase-opposed interferometric images recorded by a high-resolution CCD camera. The short coherence length of the source and the compensation of dispersion mismatch in the interferometer arms yield an optical sectioning ability with 0.8 μm resolution in water. Transverse resolution of 1.0 μm is achieved by using microscope objectives with a numerical aperture of 0.5. A shot-noise limited detection sensitivity of 86 dB can be reached with 2 s acquisition time. High-resolution images of the anterior segment of the rat eye are shown.
We present a high-resolution imaging technique using nanometric beads as multiple scattering local probes. The
positions of the beads are determined in three dimensions by white-light interference microscopy. The technique has
been applied to study the deformation of gels under mechanical constraint. The location of Brownian moving beads has
also been demonstrated with nanometer spatial precision and 10 μs acquisition time. High-resolution 3-D imaging of
hollow structures explored by the beads in relatively transparent materials should be possible.
KEYWORDS: Near field scanning optical microscopy, Particles, Near field optics, Near field, Scattering, Gold, 3D image processing, Optical spheres, Microscopy, Microscopes
Scanning near-field optical microscopy (SNOM) has proven to be very powerful in terms of both resolution and
efficiency. We report on new advances of this technique using metallic tips to scatter the optical field and induce
dramatic field enhancements. We also present a new technique under development using multiple nanometric beads as
scattering probes dispersed in the volume of the sample, rather than using a single tip. The bead positions are
determined in three dimensions (3-D) with a precision better than the diffraction limit, making possible high-resolution
3-D imaging of hollow structures in relatively transparent materials.
We present a white-light interference microscope designed to produce high-resolution three-dimensional images of biological media. This technique is an alternative to conventional optical coherence tomography (OCT). The experimental setup is based on a Linnik interferometer illuminated with a tungsten halogen lamp. En face tomographic images are obtained in real-time without scanning by computing the difference of two phase-opposed interferometric
images recorded by a high-resolution CCD camera. The short coherence length of the source and the compensation of dispersion mismatch in the interferometer arms yield an optical sectioning ability with 0.8 μm resolution in water. Transverse resolution of 1.0 μm is achieved by using high numerical aperture microscope objectives. A shot-noise limited detection sensitivity of 86 dB can be reached with 2 s acquisition time. High-resolution images of the Xenopus
Laevis tadpole are shown.
We present a new imaging modality using nanometric beads as multiple local probes. The positions of the beads are determined in three dimensions using white-light interference microscopy, by over-sampling and fitting the images. We measured the deformation of a Laponite gel with 100 nm resolution.
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