33,745 research outputs found
3D correlative single-cell imaging utilizing fluorescence and refractive index tomography
Cells alter the path of light, a fact that leads to well-known aberrations in
single cell or tissue imaging. Optical diffraction tomography (ODT) measures
the biophysical property that causes these aberrations, the refractive index
(RI). ODT is complementary to fluorescence imaging and does not require any
markers. The present study introduces RI and fluorescence tomography with
optofluidic rotation (RAFTOR) of suspended cells, quantifying the intracellular
RI distribution and colocalizing it with fluorescence in 3D. The technique is
validated with cell phantoms and used to confirm a lower nuclear RI for HL60
cells. Furthermore, the nuclear inversion of adult mouse photoreceptor cells is
observed in the RI distribution. The applications shown confirm predictions of
previous studies and illustrate the potential of RAFTOR to improve our
understanding of cells and tissues.Comment: 15 pages, 5 figure
Method for Assessing the Fidelity of Optical Diffraction Tomography Reconstruction Methods
We use a spatial light modulator in a diffraction tomographic system to
assess the accuracy of different refractive index reconstruction algorithms.
Optical phase conjugation principles through complex media, allows us to
quantify the error for different refractive index reconstruction algorithms
without access to the ground truth. To our knowledge, this is the first
assessment technique that uses structured illumination experimentally to test
the accuracy of different reconstruction schemes.Comment: 11 PAGES, 6 FIGURE
Magnetic resonance diffraction using the magnetic field from a ferromagnetic sphere
The theory of magnetic resonance diffraction is developed for the case of a crystal in close proximity of a ferromagnetic sphere. Distinct spectral peaks in the magnetic resonance signal are discovered for the specific ferromagnetic sphere and magnetic field configurations, and the appearance of the peaks is a direct signature of the presence of discrete atomic sites in the crystal lattice. The positions of the spectral peaks are sensitive to the crystal unit-cell size, thereby providing a method for determination of the basic parameters of the crystal at the atomic scale. The appearance of the spectral peaks is explained, and the dependence of the magnetic resonance spectra on the sphere size and the angle of the sphere magnetization with respect to the sample surface is analyzed. Applications to the studies of crystals, thin films, and crystallites are reviewed, and potential measurement methods for the confirmation of the diffraction theory are proposed. The analysis suggests that the long-desired goal of detecting atomic resolution magnetic resonance diffraction is well within reach of current experimental techniques
High-throughput intensity diffraction tomography with a computational microscope
We demonstrate a motion-free intensity diffraction tomography technique that enables direct inversion of 3D phase and absorption from intensity-only measurements for weakly scattering samples. We derive a novel linear forward model, featuring slice-wise phase and absorption transfer functions using angled illumination. This new framework facilitates flexible and efficient data acquisition, enabling arbitrary sampling of the illumination angles. The reconstruction algorithm performs 3D synthetic aperture using a robust, computation and memory efficient slice-wise deconvolution to achieve resolution up to the incoherent limit. We demonstrate our technique with thick biological samples having both sparse 3D structures and dense cell clusters. We further investigate the limitation of our technique when imaging strongly scattering samples. Imaging performance and the influence of multiple scattering is evaluated using a 3D sample consisting of stacked phase and absorption resolution targets. This computational microscopy system is directly built on a standard commercial microscope with a simple LED array source add-on, and promises broad applications by leveraging the ubiquitous microscopy platforms with minimal hardware modifications
Deep learning in computational microscopy
We propose to use deep convolutional neural networks (DCNNs) to perform 2D and 3D computational imaging. Specifically, we investigate three different applications. We first try to solve the 3D inverse scattering problem based on learning a huge number of training target and speckle pairs. We also demonstrate a new DCNN architecture to perform Fourier ptychographic Microscopy (FPM) reconstruction, which achieves high-resolution phase recovery with considerably less data than standard FPM. Finally, we employ DCNN models that can predict focused 2D fluorescent microscopic images from blurred images captured at overfocused or underfocused planes.Published versio
3D sub-nanoscale imaging of unit cell doubling due to octahedral tilting and cation modulation in strained perovskite thin films
Determining the 3-dimensional crystallography of a material with
sub-nanometre resolution is essential to understanding strain effects in
epitaxial thin films. A new scanning transmission electron microscopy imaging
technique is demonstrated that visualises the presence and strength of atomic
movements leading to a period doubling of the unit cell along the beam
direction, using the intensity in an extra Laue zone ring in the back focal
plane recorded using a pixelated detector method. This method is used together
with conventional atomic resolution imaging in the plane perpendicular to the
beam direction to gain information about the 3D crystal structure in an
epitaxial thin film of LaFeO3 sandwiched between a substrate of (111) SrTiO3
and a top layer of La0.7Sr0.3MnO3. It is found that a hitherto unreported
structure of LaFeO3 is formed under the unusual combination of compressive
strain and (111) growth, which is triclinic with a periodicity doubling from
primitive perovskite along one of the three directions lying in the
growth plane. This results from a combination of La-site modulation along the
beam direction, and modulation of oxygen positions resulting from octahedral
tilting. This transition to the period-doubled cell is suppressed near both the
substrate and near the La0.7Sr0.3MnO3 top layer due to the clamping of the
octahedral tilting by the absence of tilting in the substrate and due to an
incompatible tilt pattern being present in the La0.7Sr0.3MnO3 layer. This work
shows a rapid and easy way of scanning for such transitions in thin films or
other systems where disorder-order transitions or domain structures may be
present and does not require the use of atomic resolution imaging, and could be
done on any scanning TEM instrument equipped with a suitable camera.Comment: Minor fixes, especially in reference
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