We present a simple and cost-effective wide-field, depth-sectioning, fluorescence microscope utilizing a commercial multimedia projector to generate excitation patterns on the sample. Highly resolved optical sections of fluorescent pollen grains at 1.9 microm axial resolution are constructed using the structured illumination technique. This requires grid excitation patterns to be scanned across the sample, which is straightforwardly implemented by creating slideshows of gratings at different phases, projecting them onto the sample, and synchronizing camera acquisition with slide transition. In addition to rapid dynamic pattern generation, the projector provides high illumination power and spectral excitation selectivity. We exploit these properties by imaging mouse neural cells in cultures multistained with Alexa 488 and Cy3. The spectral and structural neural information is effectively resolved in three dimensions. The flexibility and commercial availability of this light source is envisioned to open multidimensional imaging to a broader user base.
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http://dx.doi.org/10.1364/ao.46.007237 | DOI Listing |
Ptychography is a robust computational imaging technique that can reconstruct complex light fields beyond conventional hardware limits. However, for many wide-field computational imaging techniques, including ptychography, depth sectioning remains a challenge. Here we demonstrate a high-resolution three-dimensional (3D) computational imaging approach, which combines ptychography with spectral-domain imaging, inspired by optical coherence tomography (OCT).
View Article and Find Full Text PDFWide-field optical microscopy is efficient and robust in biological imaging, but it lacks depth sectioning. In contrast, scanning microscopic techniques, such as confocal microscopy and multiphoton microscopy, have been successfully used for three-dimensional (3D) imaging with optical sectioning capability. However, these microscopic techniques are not very suitable for dynamic real-time imaging because they usually take a long time for temporal and spatial scanning.
View Article and Find Full Text PDFUltramicroscopy
May 2014
School of Applied & Engineering Physics and Kavli Institute at Cornell for Nanoscale Science, Cornell University, Ithaca, NY 14853, USA.
To date, high-resolution (<1 nm) imaging of extended objects in three-dimensions (3D) has not been possible. A restriction known as the Crowther criterion forces a tradeoff between object size and resolution for 3D reconstructions by tomography. Further, the sub-Angstrom resolution of aberration-corrected electron microscopes is accompanied by a greatly diminished depth of field, causing regions of larger specimens (>6 nm) to appear blurred or missing.
View Article and Find Full Text PDFOpt Express
November 2010
Optical + Biomedical Engineering Laboratory, School of Electrical, Electronic & Computer Engineering, M018, The University of Western Australia, 35 Stirling Highway, Crawley, Western Australia 6009, Australia.
Fourier-holographic light scattering spectroscopy is applied to record complex angular scattering spectra of two- and three-dimensional samples over a wide field of view. We introduce a computational depth sectioning technique and, for the first time, demonstrate that a single-exposure hologram can generate a quantitative, three-dimensional map of particle sizes and locations over several cubic millimeters with micrometer resolution. Such spatially resolved maps of particle sizes are generated by Mie-inversion and could not be ascertained from the directly reconstructed intensity-distribution images.
View Article and Find Full Text PDFAppl Opt
October 2007
National Institute of Physics, University of the Philippines Diliman, Quezon City, the Philippines 1101.
We present a simple and cost-effective wide-field, depth-sectioning, fluorescence microscope utilizing a commercial multimedia projector to generate excitation patterns on the sample. Highly resolved optical sections of fluorescent pollen grains at 1.9 microm axial resolution are constructed using the structured illumination technique.
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