Diffraction microtomography with sample rotation: influence of a missing apple core in the recorded frequency space

Diffraction microtomography in coherent light is foreseen as a promising technique to image transparent living samples in three dimensions without staining. Contrary to conventional microscopy with incoherent light, which gives morphological information only, diffraction microtomography makes it possible to obtain the complex optical refractive index of the observed sample by mapping a three-dimensional support in the spatial frequency domain. The technique can be implemented in two configurations, namely, by varying the sample illumination with a fixed sample or by rotating the sample using a fixed illumination. In the literature, only the former method was described in detail. In this report, we precisely derive the three-dimensional frequency support that can be mapped by the sample rotation configuration. We found that, within the first-order Born approximation, the volume of the frequency domain that can be mapped exhibits a missing part, the shape of which resembles that of an apple core. The projection of the diffracted waves in the frequency space onto the set of sphere caps covered by the sample rotation does not allow for a complete mapping of the frequency along the axis of rotation due to the finite radius of the sphere caps. We present simulations of the effects of this missing information on the reconstruction of ideal objects.Comment: 7 pages, 11 figures, presented at Focus On Microscopy 200

High-resolution ab initio three-dimensional X-ray diffraction microscopy

Coherent X-ray diffraction microscopy is a method of imaging non-periodic isolated objects at resolutions only limited, in principle, by the largest scattering angles recorded. We demonstrate X-ray diffraction imaging with high resolution in all three dimensions, as determined by a quantitative analysis of the reconstructed volume images. These images are retrieved from the 3D diffraction data using no a priori knowledge about the shape or composition of the object, which has never before been demonstrated on a non-periodic object. We also construct 2D images of thick objects with infinite depth of focus (without loss of transverse spatial resolution). These methods can be used to image biological and materials science samples at high resolution using X-ray undulator radiation, and establishes the techniques to be used in atomic-resolution ultrafast imaging at X-ray free-electron laser sources.Comment: 22 pages, 11 figures, submitte

Stain-Free Quantification of Chromosomes in Live Cells Using Regularized Tomographic Phase Microscopy

Refractive index imaging is a label-free technique that enables long-term monitoring of the internal structures and molecular composition in living cells with minimal perturbation. Existing tomographic methods for the refractive index imaging lack 3-D resolution and result in artifacts that prevent accurate refractive index quantification. To overcome these limitations without compromising the capability to observe a sample in its most native condition, we have developed a regularized tomographic phase microscope (RTPM) enabling accurate refractive index imaging of organelles inside intact cells. With the enhanced accuracy, we quantify the mass of chromosomes in intact living cells, and differentiate two human colon cancer lines, HT-29 and T84 cells, solely based on the non-aqueous (dry) mass of chromosomes. In addition, we demonstrate chromosomal imaging using a dual-wavelength RTPM, which shows its potential to determine the molecular composition of cellular organelles in live cells.National Institute of Biomedical Imaging and Bioengineering (U.S.) (9P41EB015871-26A1

Dielectric binary blazed gratings

Artificial index gratings, which are composed of binary microstructures of sizes less than the incident wavelength, are analyzed as functions of the filling factor or duty cycle of the microstructures. Different models for calculating the optimum duty cycles to produce high blazed diffraction efficiency are compared. Blazed binary grating designs in a material with a refractive index n = 2 show theoretical diffraction efficiencies as high as η = 80%. In the semiconductor material silicon, which has a refractive index n = 3.4, theoretical diffraction efficiencies as high as η = 70% are predicted.Not applicableab - kpw17/10/1

DESIGN OF AN OPTICAL DIGITAL COMPUTER

A possible implementation of the design of a digital optical computer is presented. A general technique for space-multiplexing arrays of beams is describe

Diffractive beam splitter for laser Doppler velocimetry

A miniaturized sensor head for the optical measurement of velocities of fluids based on laser Doppler velocimetry is demonstrated. Holographic optical elements mounted on a glass substrate are used for beam splitting and deflection. Volume holograms in dichromated gelatin exhibit good optical efficiency (75% transmission of a cascade of two holographic optical elements). With diffractive devices one can achieve achromatic behavior that makes the sensor insensitive to wavelength drifts or mode hopping of a semiconductor laser.Not applicabl