Numerical model for tomographic image formation in transmission x-ray microscopy.

We present a numerical image-formation model for investigating the influence of partial coherence, sample thickness and depth-of-focus on the accuracy of tomographic reconstructions in transmission x-ray microscopes. The model combines wave propagation through the object by finite difference techniques with Fourier methods. We include a ray-tracing model to analyse the origin of detrimental stray light in zone plate-based x-ray microscopes. These models allow optimization of x-ray microscopy systems for quantitative tomographic imaging of thick objects. Results show that both the depth-of-focus and the reconstructed local absorption coefficient are highly dependent on the degree of coherence of the optical system.

Laboratory soft-x-ray microscope for cryotomography of biological specimens.

Soft-x-ray cryotomography allows quantitative and high-resolution three-dimensional imaging of intact unstained cells. To date, the method relies on synchrotron-radiation sources, which limits accessibility for researchers. Here we present a laboratory water-window microscope for cryotomography. It is based on a λ=2.48 nm liquid-jet laser-plasma source, a normal-incidence multilayer condenser, a 30 nm zone-plate objective, and a cryotilt sample holder. We demonstrate high-resolution imaging, as well as quantitative tomographic imaging, of frozen intact cells. The reconstructed tomogram of the intracellular local absorption coefficient shows details down to ∼100 nm.

Imaging of vascular smooth muscle cells with soft X-ray spectromicroscopy.

Using X-ray microscopy and spectromicroscopy, vascular smooth muscle cells (VSMCs) were imaged, prepared without using additional embedding material or staining, but by applying simple, noncryo fixation techniques. The cells were imaged with a compact source transmission X-ray microscope and a scanning transmission X-ray microscope (STXM). With the STXM, spectromicroscopy was performed at the C K-edge and the Ca L(III,II)-edges. VSMCs were chosen because of their high amount of actin stress fibers, so that the actin cytoskeleton should be visible. Other parts of the cell, such as the nucleus and organelles, were also identified from the micrographs. Both in the spectra and the images, the effects of the different preparation procedures were observable. Furthermore, Ca hotspots were detected and their density is determined.

High-resolution computed tomography with a compact soft x-ray microscope.

Computed tomography based on high-resolution soft x-ray microscopy utilizes the natural contrast for biological specimens provided by the water window (lambda = 2.4 - 4.4 nm) and the high resolving power of zone plate objectives. It is capable of revealing the 3D structure of biological specimens at sub-visible-microscopic resolution. To date, the technique has only been available at synchrotron-based microscopes, which limits the researchers access. In the present paper we demonstrate high-resolution soft x-ray tomography with a laboratory zone-plate-based soft x-ray microscope. The specimen, a diatom mounted on a glass capillary, was reconstructed from a tilt series of 53 images covering 180 degrees using a filtered back projection algorithm. The resolution of the tomogram was estimated to a half period of 140 nm using a differential-phase-residual method. Cryo-fixation, increased source brightness and extended-depth-of-focus objectives are important for pushing the resolution of compact systems for biological samples.

Sub-25-nm laboratory x-ray microscopy using a compound Fresnel zone plate.

Improving the resolution in x-ray microscopes is of high priority to enable future applications in nanoscience. However, high-resolution zone-plate optics often have low efficiency, which makes implementation in laboratory microscopes difficult. We present a laboratory x-ray microscope based on a compound zone plate. The compound zone plate utilizes multiple diffraction orders to achieve high resolution while maintaining reasonable efficiency. We analyze the illumination conditions necessary for this type of optics in order to suppress stray light and demonstrate microscopic imaging resolving 25 nm features.

Theoretical development of a high-resolution differential-interference-contrast optic for x-ray microscopy.

In this paper, the theoretical background and development of a differential-interference contrast (DIC) x-ray optic is presented. The single-element optic is capable of high-resolution phase contrast imaging and is compatible with compact sources. It is shown that an understanding of the coherence requirements in this type of imaging is imperative and is explained in detail. The optic is capable of a wavefront separation equal to the resolution of the optic which places only minor constraints on the object illumination.

Laboratory arrangement for soft x-ray zone plate efficiency measurements.

We demonstrate a laboratory-scale arrangement for rapid and accurate measurements of the absolute and local efficiency of soft x-ray micro zone plates in the water window. This in-house instrument is based on a single-line lambda = 2.88 nm liquid-jet laser-plasma source. Measurements are performed by a simultaneous comparison of first diffraction-order photon flux with the flux in a calibrated reference signal. This arrangement eliminates existing source emission fluctuations. The performance of the method is demonstrated by the result from measurements of two approximately 55 microm diameter nickel micro zone plates, showing a groove efficiency of 12.9% +/- 1.1% and 11.7% +/- 1.0%. Furthermore, we show that spatially resolved efficiency mapping is an effective tool for a detailed characterization of local zone plate properties. Thus, this laboratory-scale instrument allows rapid feedback to the fabrication process which is important for future improvements.