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A third-order aberration analytical analysis method of soft X-ray optical systems with orthogonal and coplanar arrangement of the main planes of elements is proposed. Firstly, the transfer equations of the aperture ray and the principle ray are derived; then, based on the third-order aberration theory with the aperture-ray coordinates on the reference exit wavefront of a plane-symmetric optical system, the aberration expressions contributed by the wave aberration and defocus of this kind of optical system are studied in detail. Finally, the derived aberration calculation expressions are applied to calculate the aberration of two design examples of such types of optical systems; the images are compared with ray-tracing results obtained using the Shadow software to validate the aberration expressions. The study shows that the accuracy of the aberration expressions is satisfactory. The analytical analysis method of aberration is helpful in the design and optimization of the soft X-ray optical systems with orthogonal and coplanar arrangement of the main planes of optical elements.

Carbon nanotubes (CNTs) have been promoted as nanodevices in the biomedicine areas of diagnosis and therapy. In terms of drug delivery, CNTs could be useful in the case of hydrophobic drugs and multiple co-deliveries of therapeutic and imaging molecules. Epithelial cells, being specialized in absorption and secretion, are the first barrier to overcome. At the same time, several frequent tumors are of epithelial origin (cervix, ovarian, liver). Synchrotron-based imaging and spectroscopic techniques are suitable tools to follow the fate of nanodrugs in epithelial cells. We present an example of multi-microscopy approach combining soft X-ray spectromicroscopy at the TwinMic beamline (Elettra Synchrotron, Trieste, Italy) with light and Scanning Electron (SEM) microscopies. In particular, we show that 2–4 h exposure to SWCNTs is not toxic (no viability changes) to a model of cervical epithelial cells, even though it causes temporary perturbation in the membrane and intracellular structures of exposed cells.

Soft X-ray microscopy coupled with low energy X-ray fluorescence is a powerful tool for investigating complex biological systems like cells and tissues. Due to certain characteristics of X-ray sources, sample stage motors, and detectors, the examination of large areas at high resolutions is very time consuming, often confining the analysis only to a restricted number of pre-selected representative regions. Here we propose and demonstrate a compressive sensing method that provides an alternative approach for overcoming such limitations and can be applied to different kinds of samples and other microscopy and analytical techniques.

Hydrophobic treatment is one of the most important interventions usually carried out for the conservation of stone artefacts and monuments. The study here reported aims to answer a general question about how two polymers confer different protective performance. Two fluorinated-based polymer formulates applied on samples of Cusa’s stone confer a different level of water repellence and water vapour permeability. The observed protection action is here explained on the basis of chemico-physical interactions. The distribution of the polymer in the pore network was investigated using scanning electron microscopy and X-ray microscopy. The interactions between the stone substrate and the protective agents were investigated by means of solid state NMR spectroscopy. The ss-NMR findings reveal no significant changes in the chemical neighbourhood of the observed nuclei of each protective agent when applied onto the stone surface and provide information on the changes in the organization and dynamics of the studied systems, as well as on the mobility of polymer chains. This allowed us to explain the different macroscopic behaviours provided by each protective agent to the stone substrate.

The overall CoCID concept is centred on providing virologists with a next-generation imaging device, which, through increased penetration and depth of focus, as well as through high natural contrast and sensitivity to organelle density (including virus-related organelles), will produce higher-fidelity ultrastructural images of whole intact cells. These insights will, in turn, help increase our understanding of the links between the structural reorganisation of cells and the mechanisms of viral entry, replication, assembly, and egress in cells. CoCID will provide this valuable imaging capability in the form of a compact lab-scale device that will greatly improve the accessibility of soft X-ray microscopy for the virologists' consortium within the CoCID project as well as the broader disease research community worldwide.

Progress in nanotechnology calls for material probing techniques of high sensitivity and resolution. Such techniques are also used for high-impact studies of nanoscale materials in medicine and biology. Soft X-ray microscopy has been successfully used for investigating complex biological processes occurring at micrometric and sub-micrometric length scales and is one of the most powerful tools in medicine and the life sciences. Here, we present the capabilities of the TwinMic soft X-ray microscopy end-station at the Elettra synchrotron in the context of medical and biological imaging, while we also describe novel uses and developments.

Male infertility is a worldwide clinical issue that increases the number of couples resorting to assisted reproductive technologies (ARTs) to achieve pregnancy. Photobiomodulation therapy (PBMT) is a promising technique that can biostimulate cells and tissues and it is currently successfully employed to enhance the sperm motility in vitro. Nevertheless, its use has been so far restricted to the research field. In the present work, we exploited two PBMT protocols at an 800 nm wavelength on sperm derived from infertile individuals, detecting an increase in sperm motility 1 hour after irradiation. Moreover, in order to add new information about the molecular effect of PBMT, the content of some light elements was evaluated using high resolution X-ray fluorescence imaging. Interestingly, an increase in Na content was detected in the irradiated samples, possibly suggesting a role of this element in sperm motility; indeed, a low Na content was previously correlated with a poor sperm quality, low semen volume, and modest fertilization rate. Amplifying the knowledge of PBMT in the ART field will expedite the translational potentiality of the PBMT use in clinical settings.

An Nd:YAG pulsed laser was employed to irradiate a nitrogen gas-puff target. The interaction gives rise to the emission of soft X-ray (SXR) radiation in the ‘water window’ spectral range (λ= 2.3÷4.4 nm). This source was already successfully employed to perform the SXR microscopy. In this work, a Silicon Carbide (SiC) detector was used to characterize the nitrogen plasma emission in terms of gas-puff target parameters. The measurements show applicability of SiC detectors for SXR plasma characterization.

The performance of MISTRAL is reported, the soft X-ray transmission microscopy beamline at the ALBA light source (Barcelona, Spain) which is primarily dedicated to cryo soft X-ray tomography (cryo-SXT) for three-dimensional visualization of whole unstained cells at spatial resolutions down to 30 nm (half pitch). Short acquisition times allowing for high-throughput and correlative microscopy studies have promoted cryo-SXT as an emerging cellular imaging tool for structural cell biologists bridging the gap between optical and electron microscopy. In addition, the beamline offers the possibility of imaging magnetic domains in thin magnetic films that are illustrated here with an example.

A method for the accurate evaluation of the organic carbon (OC) content of phytoplankton by soft X-ray microscopy (XM) was developed and applied to the picophytoplankton Synechococcus sp., whose cells are covered by extracellular polysaccharides (EPS). Based on the X-ray absorption coefficients and gray levels of the XM images, the OC content of EPS-covered cells of Synechococcus sp. (0.39–0.47 pg/cell) was found to be 2.0–2.4 times larger than that of EPS-removed cells (0.20 pg/cell). These findings suggest that soft XM could be a useful tool for evaluating the OC content of picophytoplankton and EPS without pretreatment steps.

The combination of high resolution X ray imaging with in situ mechanical testing, particularly the application of a specially designed indenter manipulator in the full field transmission X ray microscope at the U41 PGM1 XM beamline of the synchrotron radiation source BESSY II [1] see Fig. 1a , is used to study the fracture behavior of microchips. The X ray microscope was operated at a photon energy of 1200 eV for the experiments described here. The achieved spatial resolution of about 25 nm allows to image the metal interconnect structures in microchips manufactured in advanced technology nodes. The mechanical manipulation of the samples was performed using a Picoindenter PI95 Bruker Fig. 1b , equipped with a tungsten wedge indenter Fig. 1c [2]

Cryo-soft X-ray microscopy is an emerging imaging tool complementary to cryo-electron microscopy, allowing to image frozen hydrated specimens ten to hundred times thicker, but at lower resolution. We describe how the method, so far restricted to isolated small cells or cell monolayers, can be extended to large cells and tissue. We image the synapses of the Kenyon cells in frozen hydrated Drosophila brains combining cryo-soft X-ray microscopy of thick vitreous sections, and cryo-electron microscopy of ultrathin vitreous sections. We show how to obtain frozen hydrated sections of thicknesses ranging from 40 nm up to 2.5 μm, by tuning the sectioning speed of the cryo-microtome. A fluorescent stereo-microscope mounted on the cryo-microtome allowed us to target the regions of interest after GFP-labeling of synapses. Thick cryo-sections were imaged by cryo-soft X-ray microscopy at a resolution better than 25 nm, while ultrathin cryo-sections of the same regions were explored in parallel at the nanometre level of resolution by cryo-electron microscopy.

Imaging is one of the key drivers for new scientific insights – from the observation of distant stars in astronomy to microscopic studies of sub-cellular structures in biology. In the latter case, X-rays are a versatile probe due to their small wavelength and thus high spatial sensitivity. We give an overview of applicable lensless, coherent imaging approaches relying on scattering with a focus on ptychographic microscopy and discuss the experimental requirements for the soft X-ray scattering experiment HORST. Besides the experiment itself, we highlight the importance of sample environments, especially when biological specimens are investigated. Here, the water window is of central importance. In addition to exploitation of the contrast and resolution, resonant ptychography allows to distinguish chemical species at high spatial resolution with both phase and amplitude contrast.

The soft X-ray Microscopy beamline BL07W at National Synchrotron Radiation Laboratory is devoted to cryo nano-tomography for biological applications in the water window (284 - 530 eV) and for imaging of nanomaterials from 200 to 2500 eV. An ellipsoidal capillary used as condense to focus monochromatic light onto the sample. Two Ni zone plate (ZP) lenses made by Zeiss with 40 nm and 25 nm outer most zone widths, respectively, are available, giving spatial resolution in 2D of down to 40 nm and 30 nm, respectively. Hydrated biological specimens had been imaged in the water window photon energy range without chemical fixation, dehydration, chemical staining and physical sectioning. In addition, other applications such as nanomaterials imaging had been demonstrated.

Many signaling molecules involved in G protein-mediated signal transduction, which are present in the lipid rafts and believed to be controlled spatially and temporally, influence the potency and efficacy of neurotransmitter receptors and transporters. This has focus interest on lipid rafts and the notion that these microdomains acts as a kind of signaling platform and thus have an important role in the expression of membrane receptor-mediated signal transduction, cancer, immune responses, neurotransmission, viral infections and various other phenomena due to specific and efficient signaling according to extracellular stimuli. However, the real structure of lipid rafts has not been observed so far due to its small size and a lack of sufficiently sophisticated observation systems. A soft X-ray microscope using a coherent soft X-ray laser in the water window region (2.3–4.4 nm) should prove to be a most powerful tool to observe the dynamic structure of lipid rafts of several tens of nanometers in size in living cells. We have developed for the X-ray microscope a new compact soft X-ray laser using strongly induced plasma high harmonic resonance. We have also developed a time-resolved highly sensitive fluorescence resonance energy transfer (FRET) system and confirmed protein-protein interactions coupled with ligands. The simultaneous use of these new tools for observation of localization of G-protein coupled receptors (GPCRs) in rafts has become an important and optimum tool system to analyze the dynamics of signal transduction through rafts as signaling platform. New technology to visualize rafts is expected to lead to the understanding of those dynamics and innovative development of drug discovery that targets GPCRs localized in lipid rafts.

For the structural characterization of the polystyrene (PS)-based photonic crystals (PCs), fast and direct imaging capabilities of full field transmission X-ray microscopy (TXM) were demonstrated at soft X-ray energy. PS-based PCs were prepared on an O2-plasma treated Si3N4 window and their local structures and defects were investigated using this label-free TXM technique with an image acquisition speed of ~10 sec/frame and marginal radiation damage. Micro-domains of face-centered cubic (FCC (111)) and hexagonal close-packed (HCP (0001)) structures were dominantly found in PS-based PCs, while point and line defects, FCC (100), and 12-fold symmetry structures were also identified as minor components. Additionally, in situ observation capability for hydrated samples and 3D tomographic reconstruction of TXM images were also demonstrated. This soft X-ray full field TXM technique with faster image acquisition speed, in situ observation, and 3D tomography capability can be complementally used with the other X-ray microscopic techniques (i.e., scanning transmission X-ray microscopy, STXM) as well as conventional characterization methods (e.g., electron microscopic and optical/fluorescence microscopic techniques) for clearer structure identification of self-assembled PCs and better understanding of the relationship between their structures and resultant optical properties.

Transmission microscopy with soft X-rays (TXM) is applied to image in-situ polyelectrolyte assemblies in aqueous environment. The method is element specific and at this stage exhibits a lateral resolution of 20 nm. With the specific examples of hollow capsules and full spheres made of PAH/PSS polyelectrolyte multilayers, it is shown quantitatively that heat treatment irreversibly reduces the water content in the membrane. These experiments complement those reported recently on the polyion system PDADMAC/PSS, which shows a different glass-transition behavior. Finally, the potential and present limitations of TXM are discussed.

Radiation damage of biological samples remains a limiting factor in high resolution X-ray microscopy (XRM). Several studies have attempted to evaluate the extent and the effects of radiation damage, proposing strategies to minimise or prevent it. The present work aims to assess the impact of soft X-rays on formalin fixed cells on a systematic manner. The novelty of this approach resides on investigating the radiation damage not only with XRM, as often reported in relevant literature on the topic, but by coupling it with two additional independent non-destructive microscopy methods: Atomic Force Microscopy (AFM) and FTIR Microscopy (FTIRM). Human Embryonic Kidney 293 cells were exposed to different radiation doses at 1 keV. In order to reveal possible morphological and biochemical changes, the irradiated cells were systematically analysed with AFM and FTIRM before and after. Results reveal that while cell morphology is not substantially affected, cellular biochemical profile changes significantly and progressively when increasing dose, resulting in a severe breakdown of the covalent bonding network. This information impacts most soft XRM studies on fixed cells and adds an in-depth understanding of the radiation damage for developing better prevention strategies.

Magnetic materials with dimensions of a few tens of nanometers are important for the development of ultrahigh-density magnetic storage and sensor devices. Magnetic microstructure largely determines functionality, and imaging of magnetic domains and magnetization reversal behavior is an outstanding challenge. Magnetic X-ray microscopy makes it possible to investigate magnetization phenomena with elemental specificity and high spatial and temporal resolution.

The effect of poly(ethylene-co-vinyl acetate) or EVA on the hydration of a pure C3S was investigated by transmission soft X-ray microscopy in highly diluted systems. EVA particles are readily recognizable by their spherical shape and semitransparency to soft X-rays. Polymeric particles adsorb on the surface of hydrating C3S grains, and act as nucleation agents in the development of the composite microstructure. The formation of a polymeric film was observed after a few hours of hydration. Fourier-transform infrared spectroscopy supported the conclusions that a highly soluble phase is formed as a result of EVA-hydrating C3S chemical interactions.

Employing X-rays from the synchrotron radiation facility of NSRL and contact imaging technique, hydrated, intact bacterial cells in their states, named by XL 1-blue MRF', are studied. Some new information about the cells is acquired from their micrographs. The nucleiod containing DNA and proteins, and the mesosome has lower absorption to the X-ray with the wavelength 2.4nm. And the ends of the bacterial cell are great different from each other in structure. The causes resulted in these interesting phenomena and its importance biologically need to be further studied.

Laser plasma sources of soft x-rays an d extreme ultraviolet (EUV) developed in our laboratory for application in various areas of technology and science are presented. The sources are based on a laser-irradiated gas puff target approach. The targets formed by pulsed injection of gas under high-pressure are irradiated with nanosecond laser pulses from Nd:YAG lasers. We use commercial lasers generating pulses with time duration from 1ns to 10ns and energies from 0.5J to 10J at 10Hz repetition rate. The gas puff targets are produced using a double valve system equipped with a special nozzle to form a double-stream gas puff target which secures high conversion efficiency without degradation of the nozzle. The use of a gas puff target instead of a solid target makes generation of laser plasmas emitting soft x-rays and EUV possible without target debris production. The sources are equipped with various optical systems, including grazing incidence axisymmetric ellipsoidal mirrors, a “lobster eye” type grazing incidence multi-foil mirror, and an ellipsoidal mirror with Mo/Si multilayer coating, to collect so ft x-ray and EUV radiation and form the radiation beams. In this paper new applications of these sources in various fields, including soft x-ray and EUV imaging in nanoscale, EUV radiography and tomography, EUV materials processing and modification of polymer surfaces, EUV photoionization of gases, radiobiology and soft x-ray contact microscopy are reviewed. Keywords: laser plasmas, soft x-rays, extreme ultraviolet (EUV), laser plasma soft x-ray and EUV sources, soft x-ray microscopy, EUV radiography and tomography, EUV processing materials, radiobiology, soft x-ray contact microscopy, EUV photo-ionized plasmas

The wavelength diffraction limit, described by the Rayleigh criterion, can be overcome if short wavelength radiations are employed, thus it is possible to resolve smaller features by the use of radiation in the extreme ultraviolet (EUV) and soft X-ray (SXR) spectral ranges. In particular way, radiation from the “water window” spectral range, which extends between K-absorption edges of carbon and oxygen (280÷540 eV), could be used in order to obtain high-contrast biological imaging. Laser-plasma double stream gas puff target source is suitable for SXR microscopy in the “water window” spectral range, which recently allowed to develop a system, operating at He-like nitrogen spectral line λ=2.88 nm, which permits to obtain images with half-pitch spatial resolution of ∼ 60 nm, exposure time as low as a few seconds and represents an important alternative for high resolution imaging for biomedical applications, material science and nanotechnology using a very compact laser source. The goal of measurements, presented herein, is to show SXR images of various biological samples, proving high contrast in the “water window” and characterize in more detail such compact microscopy system, based on a laser plasma source with a double stream gas puff target and a Fresnel zone plate (FZP) objective. The influence of various acquisition parameters on the quality of the obtained SXR images, expressed in terms of a signal-to-noise (SNR) will be demonstrated. Moreover, because the measurements are performed on SXR images, similar measurements might be performed as a benchmark in order to characterize different imaging systems as well.

Molecular distribution in mammalian cells was studied by soft X-ray scanning transmission microscopy with respect to the quantitative aspect of analysis. NEXAFS profiles at the C, N and O K-absorption edges were combined and used for the analysis. For the estimation of quantity for nucleic acids and proteins, NEXAFS profiles of DNA and bovine serum albumin (BSA) at the N K-absorption edge were applied assuming that those were their representatives. The method has a potential to explore the other molecular components than nucleic acids and proteins.

X-ray microscopy is powerful in that it can probe large volumes of material at high spatial resolution with exquisite chemical, electronic and bond orientation contrast. The development of diffraction-based methods such as ptychography has, in principle, removed the resolution limit imposed by the characteristics of the X-ray optics. Here, using soft X-ray ptychography, we demonstrate the highest-resolution X-ray microscopy ever achieved by imaging 5â €..nm structures. We quantify the performance of our microscope and apply the method to the study of delithiation in a nanoplate of LiFePO 4, a material of broad interest in electrochemical energy storage. We calculate chemical component distributions using the full complex refractive index and demonstrate enhanced contrast, which elucidates a strong correlation between structural defects and chemical phase propagation. The ability to visualize the coupling of the kinetics of a phase transformation with the mechanical consequences is critical to designing materials with ultimate durability.

Short wavelength radiation, in the 1-20 nm wavelength range is discussed, with particular emphasis on applications to soft x-ray (SXR) microscopy and extreme ultraviolet (EUV) radiation. In this paper we bring attention to the fact that these two applications require a wide variety of SXR/EUV sources, some with good spatial coherence, others largely incoherent; some at fairly high average power (tens of watts), others at rather low (microwatts) power; some at fixed wavelength, others tunable. These provide great opportunities for attendees of this workshop who represent the world's body of knowledge in this area. We give reference to recent papers to illustrate these points, and call attention to recurring publications where further opportunities and progress can be monitored.

Nanoscale transmission x-ray microscopy bridges the resolution gap between light and electron microscopy, and a new laboratory soft x-ray source has extended its utility for imaging whole frozen hydrated unstained cells with

The chapter deals with main optical properties of Schwarzschild and Head objectives from the point view of geometrical optics. General approach in the study can be applied to both XUV and visible/IR optical instrumentation.

ةـصلاخلا : ة ينقتلا و ة يملعلا نيتيحا نلا نم ةر يبآ ة يمهأ اهل ونانلا مجحب ةبلص ةلاح ةمظنلأ ةيسيطانغملا صاوخلا نإ . و ةدحولا يه نور تكللإا ةنحش نأ نيح يف ةثيدحلا تاينورتكللإا يف ةيساسلأا ، ب ةيلبقتس ملا ي لزغملا نارود لا تا ينورتكلإ يف ي لزغملا نارود لا ة يرح ةجرد ذخأ نإف ني ع را بتعلاا حت ف ى لإ يدؤيس لبسلا نم ةريبآ ةعومجم . ةيللآل لضفأ يئايزيف مهف هاجتاب مدقتلا نإ ، و تاودأ مادختساب طقف ه قيقحت نكمي ةيس يطانغملا ونا نلا ةز هجلأ ةعجشملا تا قيبطتلا و ةنمضتملا ئدا بملا ةمدقتم ليلحت ، ثيح إ ا سايقمب ةيسيطانغملا بيآارتلل رشابملا ريوصتلا ن رمأ وه ةعيرسلا اهتيكيمانيد و ونانل لاعف . مدختسيو قيرط نع يو ق يس يطانغم نيا بت ة يلآ ليصحتل ة يرئاد ةبطقتسم سآإ ةعشأ ه لمع يف ة معانلا سآإ ةعشلأ سآاعلا يس يطانغملا رهجملا ةعشلأ يروشنملا يسيطانغملا نارودلا تاريثأت يتلا سآإ دنع ثدحت ا ز يمم دح د نع سآإ ةعشأ صاصتم . و مجي ةعشلأ سآاعلا يس يطانغملا رهجملا ع ىتح ايناكم لاصف ةمعانلا سآإ 10 يف يرطفلا تقولا ة بيآرتب تقؤم يئزج ماظن يف اتقؤم لاصف و ليسنرف ةقطنم ةحيفص تايئوض مادختساب ابيرقت من ة يلاحلا نورتوركنسلا رداصم . لكش ب حمس ي ة معانلا سآإ ةعشلأ سآاعلا يس يطانغملا رهجملا نإ ف يلا تلاب ة قيرط يف يئاوشعلا فرصتلا ةساردب د يرف ةديقملا ةيسيطانغمورفلا بيآارتلا يف لاجملا ناردج و ةيسيطانغملا تاماودلل ةعرسلا ةقئاف ايكيمانيدلا وأ ةقيقرلا تاقبطلل ةيسكعلا ةطنغملا . و ى لإ ةريصق سآإ ةعشأ تا نوتوف تابرض يطعت نأ نكمي ةيلبقتس ملا سآإ ةعشأ رداصم نأ ىلإ رظنلاب ة فيثآ و ة يناثوتميفلا ، رهجملا قرط نإ ف ةليصلأا ةيسيطانغملا لوطلا و تقولا سيياقم ىلإ لاوزن يسيطانغملا ريوصتلا دهع ىلع يقبتس ةمعانلا سآإ ةعشلأ سآاعلا يسيطانغملا .

Inner shell processes in the light elements provide the contrast mechanisms for soft X-ray microscopy. Bright X-ray sources and precision focusing elements have led to the development of high resolution instruments. Our efforts in scanning microscopy, holography and diffraction at the National Synchrotron Light Source are aimed at the development of microimaging techniques mostly for biological specimens.

Full-field magnetic transmission x-ray microscopy at high spatial resolution down to 20 nm is used to directly observe field-driven domain wall motion in notch-patterned permalloy nanowires. The depinning process of a domain wall around a notch exhibits a stochastic nature in most nanowires. The stochasticity of the domain wall depinning sensitively depends on the geometry of the nanowire such as the wire thickness, the wire width, and the notch depth. We propose an optimized design of the nanowire for deterministic domain wall depinning field at a notch.

The present status of the cement and concrete industry is not sustainable. The production of Portland cement is responsible for 7% of the CO2 emissions in the world and existing reinforced concrete infrastructure is deteriorating at a fast pace. The change in the existing technology requires new developments in our understanding of the nanostructure of hydration products and the complex deterioration reactions. We have been developing an elaborate research program to advance the existing cement and concrete science by characterizing its nanostructure by synchrotron radiation. A new generation of green cements is being studied using high‐resolution soft x‐ray microscopy at the nano‐level.