X-Ray Microscopy: Preparations for Studies of Frozen Hydrated Specimens

X-ray microscopes provide higher resolution than visible light microscopes. Wet, biological materials with a water thickness of up to about 10 μm can be imaged with good contrast using soft X-rays with wavelengths between the oxygen and carbon absorption edges (at 24 and 43 Å). The Stony Brook group has developed and operates a scanning transmission X-ray microscope (STXM) at the National Synchrotron Light Source (NSLS) at Brookhaven National Laboratory. The microscope is used for imaging with a current resolution of 50 nm, and for elemental and chemical state mapping. Radiation damage imposes a significant limitation upon high resolution X-ray microscopy of room temperature wet specimens. Experience from electron microscopy suggests that cryo techniques allow vitrified specimens to be imaged repeatedly. This is due to the increased radiation stability of biological specimens in the frozen hydrated state. Better radiation stability has been shown recently with a cryo transmission X-ray microscope developed by the University of Gottingen, operating at the BESSY storage ring in Berlin, Germany. At Stony Brook, we are developing a cryo scanning transmission X-ray microscope (CryoSTXM) to carry out imaging and spectra-microscopy experiments on frozen hydrated specimens. This article will give an outlook onto the research projects that we plan to perform using the CryoSTXM

The history and future of X-ray microscopy

Abstract. We take a somewhat whimsical look at the history of X-ray microscopy, and extrapolate some trends into the future

The history and future of X-ray microscopy

Abstract. We take a somewhat whimsical look at the history of X-ray microscopy, and extrapolate some trends into the future. History This is a story of spies, heroes, villains, false starts, and a brush with real fame. We divide the history into seven epochs: 1) Ancient history 1895-1945; 2) The Classical Period 1946Period -1960 3) The Dark Ages 1961Ages -1971 4) The Renaissance 1972Renaissance -1982 5) Romanticism 1983 5) Romanticism -1993 6 Ancient History 1895-1945 This was the time of the pioneers who, following Röntgen's 1895 discovery of X-rays [1], established point-projection microscopy with a resolution of a few microns The Classical Period 1946-1960 Right after World War II several groups became interested in X-ray microscopy. Arne Engstrom in Sweden developed the technique of quantitative elemental imaging The Dark Ages 1961-1971 The sixities were a relatively quiet period in X-ray microscopy. The Cambridge group continued to be active, especially Theodore Hall and his collaborators Of course the seeds for the renaissance were planted during the dark ages: it was toward the end of the decade that Schmahl and Rudolph turned their attention to holographic fabrication of zone plates The Renaissance 1972-1982 The decade of the seventies saw the first lightsource-based X-ray microscopes: Horowitz and Howell demonstrated both transmission and fluorescence microscopy at the Cambridge Electron Accelerator Romanticism 1983-1993 Following the first uses of synchrotron light sources, a lot of new avenues were tried, some more successful than others. The King's College group built a STXM at Daresbury [28] (using DESY, 1976 ACO, 1983 ACO, 1983 Figure 4. The first zone plate TXMs (transmission x-ray microscopes) developed by Schmahl, Rudolph, and Niemann. At left is shown an instrument operated at DESY in Hamburg in 1976 [24], while an instrument operated at ACO in Orsay is shown in the middle image. At right is shown an image of a diatom obtained at Orsa

Quantitative Imaging of Single, Unstained Viruses with Coherent X-rays

Since Perutz, Kendrew and colleagues unveiled the structure of hemoglobin and myoglobin based on X-ray diffraction analysis in the 1950s, X-ray crystallography has become the primary methodology used to determine the 3D structure of macromolecules. However, biological specimens such as cells, organelles, viruses and many important macromolecules are difficult or impossible to crystallize, and hence their structures are not accessible by crystallography. Here we report, for the first time, the recording and reconstruction of X-ray diffraction patterns from single, unstained viruses. The structure of the viral capsid inside a virion was visualized. This work opens the door for quantitative X-ray imaging of a broad range of specimens from protein machineries, viruses and organelles to whole cells. Moreover, our experiment is directly transferable to the use of X-ray free electron lasers, and represents a major experimental milestone towards the X-ray imaging of single macromolecules.Comment: 16 pages, 5 figure

Development of Ground-testable Phase Fresnel Lenses in Silicon

Diffractive/refractive optics, such as Phase Fresnel Lenses (PFL's), offer the potential to achieve excellent imaging performance in the x-ray and gamma-ray photon regimes. In principle, the angular resolution obtained with these devices can be diffraction limited. Furthermore, improvements in signal sensitivity can be achieved as virtually the entire flux incident on a lens can be concentrated onto a small detector area. In order to verify experimentally the imaging performance, we have fabricated PFL's in silicon using gray-scale lithography to produce the required Fresnel profile. These devices are to be evaluated in the recently constructed 600-meter x-ray interferometry testbed at NASA/GSFC. Profile measurements of the Fresnel structures in fabricated PFL's have been performed and have been used to obtain initial characterization of the expected PFL imaging efficiencies.Comment: Presented at GammaWave05: "Focusing Telescopes in Nuclear Astrophysics", Bonifacio, Corsica, September 2005, to be published in Experimental Astronomy, 8 pages, 3 figure

An assessment of the resolution limitation due to radiation-damage in x-ray diffraction microscopy

X-ray diffraction microscopy (XDM) is a new form of x-ray imaging that is being practiced at several third-generation synchrotron-radiation x-ray facilities. Although only five years have elapsed since the technique was first introduced, it has made rapid progress in demonstrating high-resolution threedimensional imaging and promises few-nm resolution with much larger samples than can be imaged in the transmission electron microscope. Both life- and materials-science applications of XDM are intended, and it is expected that the principal limitation to resolution will be radiation damage for life science and the coherent power of available x-ray sources for material science. In this paper we address the question of the role of radiation damage. We use a statistical analysis based on the so-called "dose fractionation theorem" of Hegerl and Hoppe to calculate the dose needed to make an image of a lifescience sample by XDM with a given resolution. We conclude that the needed dose scales with the inverse fourth power of the resolution and present experimental evidence to support this finding. To determine the maximum tolerable dose we have assembled a number of data taken from the literature plus some measurements of our own which cover ranges of resolution that are not well covered by reports in the literature. The tentative conclusion of this study is that XDM should be able to image frozen-hydrated protein samples at a resolution of about 10 nm with "Rose-criterion" image quality.Comment: 9 pages, 4 figure

Nearfield Summary and Statistical Analysis of the Second AIAA Sonic Boom Prediction Workshop

A summary is provided for the Second AIAA Sonic Boom Workshop held 8-9 January 2017 in conjunction with AIAA SciTech 2017. The workshop used three required models of increasing complexity: an axisymmetric body, a wing body, and a complete configuration with flow-through nacelle. An optional complete configuration with propulsion boundary conditions is also provided. These models are designed with similar nearfield signatures to isolate geometry and shock/expansion interaction effects. Eleven international participant groups submitted nearfield signatures with forces, pitching moment, and iterative convergence norms. Statistics and grid convergence of these nearfield signatures are presented. These submissions are propagated to the ground, and noise levels are computed. This allows the grid convergence and the statistical distribution of a noise level to be computed. While progress is documented since the first workshop, improvement to the analysis methods for a possible subsequent workshop are provided. The complete configuration with flow-through nacelle showed the most dramatic improvement between the two workshops. The current workshop cases are more relevant to vehicles with lower loudness and have the potential for lower annoyance than the first workshop cases. The models for this workshop with quieter ground noise levels than the first workshop exposed weaknesses in analysis, particularly in convective discretization

Three-Dimensional X-ray Observation of Atmospheric Biological Samples by Linear-Array Scanning-Electron Generation X-ray Microscope System

Recently, we developed a soft X-ray microscope called the scanning-electron generation X-ray microscope (SGXM), which consists of a simple X-ray detection system that detects X-rays emitted from the interaction between a scanning electron beam (EB) and the thin film of the sample mount. We present herein a three-dimensional (3D) X-ray detection system that is based on the SGXM technology and designed for studying atmospheric biological samples. This 3D X-ray detection system contains a linear X-ray photodiode (PD) array. The specimens are placed under a CuZn-coated Si3N4 thin film, which is attached to an atmospheric sample holder. Multiple tilt X-ray images of the samples are detected simultaneously by the linear array of X-ray PDs, and the 3D structure is calculated by a new 3D reconstruction method that uses a simulated-annealing algorithm. The resulting 3D models clearly reveal the inner structure of the bacterium. In addition, the proposed method can easily be used for diverse samples in a broad range of scientific fields

Design Studies for a VUV--Soft X-ray Free-Electron Laser Array

Several recent reports have identified the scientific requirements for a future soft X-ray light source [1, 2, 3, 4, 5], and a high-repetition-rate free-electron laser (FEL) facility responsive to them is being studied at Lawrence Berkeley National Laboratory (LBNL) [6]. The facility is based on a continuous-wave (CW) superconducting linear accelerator with beam supplied by a high-brightness, high-repetition-rate photocathode electron gun operating in CW mode, and on an array of FELs to which the accelerated beam is distributed, each operating at high repetition rate and with even pulse spacing. Dependent on the experimental requirements, the individualFELs may be configured for either self-amplified spontaneous emission (SASE), seeded highgain harmonic generation (HGHG), echo-enabled harmonic generation (EEHG), or oscillator mode of operation, and will produce high peak and average brightness x-rays with a flexible pulse format ranging from sub-femtoseconds to hundreds of femtoseconds. This new light source would serve a broad community of scientists in many areas of research, similar to existing utilization of storage ring based light sources. To reduce technical risks and constructioncosts, accelerator research, development, and design studies at LBNL target the most critical components and systems of the facility. We are developing a high-repetition-rate low-emittance electron gun, high quantum efficiency photocathodes, and have embarked on design and optimization of the electron beam accelerator, FEL switchyard, and array of FELs. We continue our work on precision timing and synchronization systems critical for time-resolved experiments using pump-probe techniques