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Register a new userCryptotomography: reconstructing 3D Fourier intensities from randomly oriented single-shot diffraction patterns
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We reconstructed the 3D Fourier intensity distribution of mono-disperse prolate nano-particles using single-shot 2D coherent diffraction patterns collected at DESYs FLASH facility when a bright, coherent, ultrafast X-ray pulse intercepted individual particles of random, unmeasured orientations. This first experimental demonstration of cryptotomography extended the Expansion-Maximization-Compression (EMC) framework to accommodate unmeasured fluctuations in photon fluence and loss of data due to saturation or background scatter. This work is an important step towards realizing single-shot diffraction imaging of single biomolecules.
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Speden is a computer program that reconstructs the electron density of single particles from their x-ray diffraction patterns, using a single-particle adaptation of the Holographic Method in crystallography. (Szoke, A., Szoke, H., and Somoza, J.R., 1997. Acta Cryst. A53, 291-313.) The method, like its parent, is unique that it does not rely on ``back transformation from the diffraction pattern into real space and on interpolation within measured data. It is designed to deal successfully with sparse, irregular, incomplete and noisy data. It is also designed to use prior information for ensuring sensible results and for reliable convergence. This article describes the theoretical basis for the reconstruction algorithm, its implementation and quantitative results of tests on synthetic and experimentally obtained data. The program could be used for determining the structure of radiation tolerant samples and, eventually, of large biological molecular structures without the need for crystallization.
Light field microscopy (LFM) uses a microlens array (MLA) near the sensor plane of a microscope to achieve single-shot 3D imaging of a sample without any moving parts. Unfortunately, the 3D capability of LFM comes with a significant loss of lateral resolution at the focal plane. Placing the MLA near the pupil plane of the microscope, instead of the image plane, can mitigate the artifacts and provide an efficient forward model, at the expense of field-of-view (FOV). Here, we demonstrate improved resolution across a large volume with Fourier DiffuserScope, which uses a diffuser in the pupil plane to encode 3D information, then computationally reconstructs the volume by solving a sparsity-constrained inverse problem. Our diffuser consists of randomly placed microlenses with varying focal lengths; the random positions provide a larger FOV compared to a conventional MLA, and the diverse focal lengths improve the axial depth range. To predict system performance based on diffuser parameters, we for the first time establish a theoretical framework and design guidelines, which are verified by numerical simulations, then build an experimental system that achieves $< 3$ um lateral and $4$ um axial resolution over a $1000 times 1000 times 280$ um$^3$ volume. Our diffuser design outperforms the MLA used in LFM, providing more uniform resolution over a larger volume, both laterally and axially.
The first experimental data from single-particle scattering experiments from free electron lasers (FELs) are now becoming available. The first such experiments are being performed on relatively large objects such as viruses, which produce relatively low-resolution, low-noise diffraction patterns in so-called diffract-and-destroy experiments. We describe a very simple test on the angular correlations of measured diffraction data to determine if the scattering is from an icosahedral particle. If this is confirmed, the efficient algorithm proposed can then combine diffraction data from multiple shots of particles in random unknown orientations to generate a full 3D image of the icosahedral particle. We demonstrate this with a simulation for the satellite tobacco necrosis virus (STNV), the atomic coordinates of whose asymmetric unit is given in Protein Data Bank entry 2BUK.
We represent three-dimensional Fourier transform light scattering, a method to reconstruct angle-resolved light scattering (ARLS) with extended angle-range from individual spherical objects. To overcome the angle limitation determined by the physical numerical aperture of an optical system, the optical light fields scattered from a sample are measured with various illumination angles, and then synthesized onto the Ewald Sphere corresponding to the normal illumination in Fourier space by rotating the scattered light signals. The method extends the angle range of the ARLS spectra beyond 90 degree, beyond the limit of forward optical measurements. Extended scattered light fields in 3D and corresponding ARLS spectra of individual microscopic polystyrene beads, and protein droplets are represented.
We derive a method to reconstruct Gaussian signals from linear measurements with Gaussian noise. This new algorithm is intended for applications in astrophysics and other sciences. The starting point of our considerations is the principle of minimum Gibbs free energy which was previously used to derive a signal reconstruction algorithm handling uncertainties in the signal covariance. We extend this algorithm to simultaneously uncertain noise and signal covariances using the same principles in the derivation. The resulting equations are general enough to be applied in many different contexts. We demonstrate the performance of the algorithm by applying it to specific example situations and compare it to algorithms not allowing for uncertainties in the noise covariance. The results show that the method we suggest performs very well under a variety of circumstances and is indeed qualitatively superior to the other methods in cases where uncertainty in the noise covariance is present.
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