Quantitative phase measurements of human cell nuclei using X-ray ptychography.

The human cell nucleus serves as an important organelle holding the genetic blueprint for life. In this work, X-ray ptychography was applied to assess the masses of human cell nuclei using its unique phase shift information. Measurements were carried out at the I13-1 beamline at the Diamond Light Source that has extremely large transverse coherence properties. The ptychographic diffractive imaging approach allowed imaging of large structures that gave quantitative measurements of the phase shift in 2D projections. In this paper a modified ptychography algorithm that improves the quality of the reconstruction for weak scattering samples is presented. The application of this approach to calculate the mass of several human nuclei is also demonstrated.

Holography-guided ptychography with soft X-rays.

Ptychography is a lensless imaging technique that aims to reconstruct an object from a set of coherent diffraction patterns originating from different and partially overlapping sample illumination areas. For a successful convergence of the iterative algorithms used, the sample scan positions have to be known with very high accuracy. Here, we present a method that allows to directly encode this information in the diffraction patterns without the need of accurate position encoders. Our approach relies on combining ptychography with another coherent imaging method, namely Fourier-transform holography. We have imaged two different objects using coherent soft-X-ray illumination and investigate the influence of experimental and numerical position refinement on the reconstruction result. We demonstrate that holographically encoded positions significantly reduce the experimental and numerical requirements. Our ptychographic reconstructions cover a large field of view with diffraction-limited resolution and high sensitivity in the reconstructed phase shift and absorption of the objects.

Karyotyping human chromosomes by optical and X-ray ptychography methods.

Sorting and identifying chromosomes, a process known as karyotyping, is widely used to detect changes in chromosome shapes and gene positions. In a karyotype the chromosomes are identified by their size and therefore this process can be performed by measuring macroscopic structural variables. Chromosomes contain a specific number of basepairs that linearly correlate with their size; therefore, it is possible to perform a karyotype on chromosomes using their mass as an identifying factor. Here, we obtain the first images, to our knowledge, of chromosomes using the novel imaging method of ptychography. We can use the images to measure the mass of chromosomes and perform a partial karyotype from the results. We also obtain high spatial resolution using this technique with synchrotron source x-rays.

A general approach to obtain soft x-ray transparency for thin films grown on bulk substrates.

We present a general approach to thin bulk samples to transparency for experiments in the soft x-ray and extreme ultraviolet spectral range. The method relies on mechanical grinding followed by focused-ion-beam milling. It results in a uniformly thin area of high surface quality, suitable for nanoscale imaging in transmission. In a proof-of-principle experiment, nanoscale magnetic bits on a commercial hard drive glass disk are imaged with a spatial resolution below 30 nm by soft x-ray spectro-holography. Furthermore, we demonstrate imaging of a lithographically patterned test object via absorption contrast. Our approach is suitable for both amorphous and crystalline substrates and has been tested for a variety of common epitaxy growth substrates. Lateral thinning areas in excess of 100 µm2 and a remaining substrate thickness as thin as 150 nm are easily achievable. Our approach allows preserving a previously grown thin film, and from nanofocus electron diffraction, we find no evidence for morphological changes induced by the process, in agreement with numerical simulations of the ion implantation depth distributon. We expect our method to be widely applicable and especially useful for nanoscale imaging of epitaxial thin films.

Three-dimensional structure analysis and percolation properties of a barrier marine coating.

Artificially structured coatings are widely employed to minimize materials deterioration and corrosion, the annual direct cost of which is over 3% of the gross domestic product (GDP) for industrial countries. Manufacturing higher performance anticorrosive coatings is one of the most efficient approaches to reduce this loss. However, three-dimensional (3D) structure of coatings, which determines their performance, has not been investigated in detail. Here we present a quantitative nano-scale analysis of the 3D spatial structure of an anticorrosive aluminium epoxy barrier marine coating obtained by serial block-face scanning electron microscopy (SBFSEM) and ptychographic X-ray computed tomography (PXCT). We then use finite element simulations to demonstrate how percolation through this actual 3D structure impedes ion diffusion in the composite materials. We found the aluminium flakes align within 15° of the coating surface in the material, causing the perpendicular diffusion resistance of the coating to be substantially higher than the pure epoxy.