Synchrotron Beamlines Research Articles

Insights into Microscopic Crystal Growth Dynamics of CH3NH3PbI3 under a Laser Deposition Process Revealed by In Situ X-ray Diffraction

The process dynamics for the vacuum deposition of methylammonium lead iodide (MAPbI3) perovskite was analyzed by in situ X-ray diffraction using synchrotron radiation. MAPbI3 was fabricated by alternatingly supplying PbI2 and methylammonium iodide via a laser deposition system installed at the synchrotron beamline BL46XU at SPring-8, and in situ crystallization analysis was conducted. Microscopic insights into the crystallization were obtained, including observation of Laue oscillation during the PbI2 growth and octahedral unit (PbI6) rotation during the transformation into perovskite. On the basis of this analysis, conditions that favor the construction of atomically flat MAPbI3 perovskite films were deduced.

X-ray Phase-Contrast Computed Tomography for Soft Tissue Imaging at the Imaging and Medical Beamline (IMBL) of the Australian Synchrotron

The Imaging and Medical Beamline (IMBL) is a superconducting multipole wiggler-based beamline at the 3 GeV Australian Synchrotron operated by the Australian Nuclear Science and Technology Organisation (ANSTO). The beamline delivers hard X-rays in the 25–120 keV energy range and offers the potential for a range of biomedical X-ray applications, including radiotherapy and medical imaging experiments. One of the imaging modalities available at IMBL is propagation-based X-ray phase-contrast computed tomography (PCT). PCT produces superior results when imaging low-density materials such as soft tissue (e.g., breast mastectomies) and has the potential to be developed into a valuable medical imaging tool. We anticipate that PCT will be utilized for medical breast imaging in the near future with the advantage that it could provide better contrast than conventional X-ray absorption imaging. The unique properties of synchrotron X-ray sources such as high coherence, energy tunability, and high brightness are particularly well-suited for generating PCT data using very short exposure times on the order of less than 1 min. The coherence of synchrotron radiation allows for phase-contrast imaging with superior sensitivity to small differences in soft-tissue density. Here we also compare the results of PCT using two different detectors, as these unique source characteristics need to be complemented with a highly efficient detector. Moreover, the application of phase retrieval for PCT image reconstruction enables the use of noisier images, potentially significantly reducing the total dose received by patients during acquisition. This work is part of ongoing research into innovative tomographic methods aimed at the introduction of 3D X-ray medical imaging at the IMBL to improve the detection and diagnosis of breast cancer. Major progress in this area at the IMBL includes the characterization of a large number of mastectomy samples, both normal and cancerous, which have been scanned at clinically acceptable radiation dose levels and evaluated by expert radiologists with respect to both image quality and cancer diagnosis.

Sample Preparation and Transfer Protocol for In-Vacuum Long-Wavelength Crystallography on Beamline I23 at Diamond Light Source

Long-wavelength macromolecular crystallography (MX) exploits the anomalous scattering properties of elements, such as sulfur, phosphorus, potassium, chlorine, or calcium, that are often natively present in macromolecules. This enables the direct structure solution of proteins and nucleic acids via experimental phasing without the need of additional labelling. To eliminate the significant air absorption of X-rays in this wavelength regime, these experiments are performed in a vacuum environment. Beamline I23 at Diamond Light Source, UK, is the first synchrotron instrument of its kind, designed and optimized for MX experiments in the long wavelength range towards 5 Å. To make this possible, a large vacuum vessel encloses all endstation components of the sample environment. The necessity to maintain samples at cryogenic temperatures during storage and data collection in vacuum requires the use of thermally conductive sample holders. This facilitates efficient heat removal to ensure sample cooling to approximately 50 K. The current protocol describes the procedures used for sample preparation and transfer of samples into vacuum on beamline I23. Ensuring uniformity in practices and methods already established within the macromolecular crystallography community, sample cooling to liquid nitrogen temperature can be performed in any laboratory setting equipped with standard MX tools. Cryogenic storage and transport of samples only require standard commercially available equipment. Specialized equipment is required for the transfer of cryogenically cooled crystals from liquid nitrogen into the vacuum endstation. Bespoke sample handling tools and a dedicated Cryogenic Transfer System (CTS) have been developed in house. Diffraction data collected on samples prepared using this protocol show excellent merging statistics, indicating that the quality of samples is unaltered during the procedure. This opens unique opportunities for in-vacuum MX in a wavelength range beyond standard synchrotron beamlines.

Sample Preparation and Transfer Protocol for In-Vacuum Long-Wavelength Crystallography on Beamline I23 at Diamond Light Source

Long-wavelength macromolecular crystallography (MX) exploits the anomalous scattering properties of elements, such as sulfur, phosphorus, potassium, chlorine, or calcium, that are often natively present in macromolecules. This enables the direct structure solution of proteins and nucleic acids via experimental phasing without the need of additional labelling. To eliminate the significant air absorption of X-rays in this wavelength regime, these experiments are performed in a vacuum environment. Beamline I23 at Diamond Light Source, UK, is the first synchrotron instrument of its kind, designed and optimized for MX experiments in the long wavelength range towards 5 Å. To make this possible, a large vacuum vessel encloses all endstation components of the sample environment. The necessity to maintain samples at cryogenic temperatures during storage and data collection in vacuum requires the use of thermally conductive sample holders. This facilitates efficient heat removal to ensure sample cooling to approximately 50 K. The current protocol describes the procedures used for sample preparation and transfer of samples into vacuum on beamline I23. Ensuring uniformity in practices and methods already established within the macromolecular crystallography community, sample cooling to liquid nitrogen temperature can be performed in any laboratory setting equipped with standard MX tools. Cryogenic storage and transport of samples only require standard commercially available equipment. Specialized equipment is required for the transfer of cryogenically cooled crystals from liquid nitrogen into the vacuum endstation. Bespoke sample handling tools and a dedicated Cryogenic Transfer System (CTS) have been developed in house. Diffraction data collected on samples prepared using this protocol show excellent merging statistics, indicating that the quality of samples is unaltered during the procedure. This opens unique opportunities for in-vacuum MX in a wavelength range beyond standard synchrotron beamlines.

A microfluidic dosimetry cell to irradiate solutions with poorly penetrating radiations: a step towards online dosimetry for synchrotron beamlines.

Synchrotron radiation can induce sample damage, whether intended or not. In the case of sensitive samples, such as biological ones, modifications can be significant. To understand and predict the effects due to exposure, it is necessary to know the ionizing radiation dose deposited in the sample. In the case of aqueous samples, deleterious effects are mostly induced by the production of reactive oxygen species via water radiolysis. These species are therefore good indicators of the dose. Here the application of a microfluidic cell specifically optimized for low penetrating soft X-ray radiation is reported. Sodium benzoate was used as a fluorescent dosimeter thanks to its specific detection of hydroxyl radicals, a radiolytic product of water. Measurements at 1.28 keV led to the determination of a hydroxyl production yield, G(HO.), of 0.025 ± 0.004 µmol J-1. This result is in agreement with the literature and confirms the high linear energy transfer behavior of soft X-rays. An analysis of the important parameters of the microfluidic dosimetry cell, as well as their influences over dosimetry, is also reported.

SHELIXIR: automation of experimental phasing procedures using SHELXC/D/E

The program SHELIXIR represents a simple and efficient tool for routine phase-problem solution using data for experimental phasing by the single-wavelength anomalous dispersion, multiwavelength anomalous dispersion, single isomorphous replacement with anomalous scattering and radiation-damage-induced phasing methods. As indicated in its name, all calculation procedures are performed with the SHELXC/D/E program package. SHELIXIR provides screening for alternative space groups, optimal solvent content, and high- and low-resolution limits. The procedures of SHELXE are parallelized to minimize the computational time. The automation and parallelization of such procedures are suitable for phasing at synchrotron beamlines directly or for finding the optimal parameters for further data processing. A simple graphical interface is designed to make use easier and to increase efficiency during beam time.

Simultaneous ambient pressure x-ray photoelectron spectroscopy and grazing incidence x-ray scattering in gas environments

We have developed an experimental system to simultaneously measure surface structure, morphology, composition, chemical state, and chemical activity for samples in gas phase environments. This is accomplished by simultaneously measuring x-ray photoelectron spectroscopy (XPS) and grazing incidence x-ray scattering in gas pressures as high as the multi-Torr regime while also recording mass spectrometry. Scattering patterns reflect near-surface sample structures from the nano-scale to the meso-scale, and the grazing incidence geometry provides tunable depth sensitivity of structural measurements. Scattered x rays are detected across a broad range of angles using a newly designed pivoting-UHV-manipulator for detector positioning. At the same time, XPS and mass spectrometry can be measured, all from the same sample spot and under ambient conditions. To demonstrate the capabilities of this system, we measured the chemical state, composition, and structure of Ag-behenate on a Si(001) wafer in vacuum and in O2 atmosphere at various temperatures. These simultaneous structural, chemical, and gas phase product probes enable detailed insights into the interplay between the structure and chemical state for samples in gas phase environments. The compact size of our pivoting-UHV-manipulator makes it possible to retrofit this technique into existing spectroscopic instruments installed at synchrotron beamlines. Because many synchrotron facilities are planning or undergoing upgrades to diffraction limited storage rings with transversely coherent beams, a newly emerging set of coherent x-ray scattering experiments can greatly benefit from the concepts we present here.

Workflow and Tools for Crystallographic Fragment Screening at the Helmholtz-Zentrum Berlin.

Fragment screening is a technique that helps to identify promising starting points for ligand design. Given that crystals of the target protein are available and display reproducibly high-resolution X-ray diffraction properties, crystallography is among the most preferred methods for fragment screening because of its sensitivity. Additionally, it is the only method providing detailed 3D information of the binding mode of the fragment, which is vital for subsequent rational compound evolution. The routine use of the method depends on the availability of suitable fragment libraries, dedicated means to handle large numbers of samples, state-of-the-art synchrotron beamlines for fast diffraction measurements and largely automated solutions for the analysis of the results. Here, the complete practical workflow and the included tools on how to conduct crystallographic fragment screening (CFS) at the Helmholtz-Zentrum Berlin (HZB) are presented. Preceding this workflow, crystal soaking conditions as well as data collection strategies are optimized for reproducible crystallographic experiments. Then, typically in a one to two-day procedure, a 96-membered CFS-focused library provided as dried ready-to-use plates is employed to soak 192 crystals, which are then flash-cooled individually. The final diffraction experiments can be performed within one day at the robot-mounting supported beamlines BL14.1 and BL14.2 at the BESSY II electron storage ring operated by the HZB in Berlin-Adlershof (Germany). Processing of the crystallographic data, refinement of the protein structures, and hit identification is fast and largely automated using specialized software pipelines on dedicated servers, requiring little user input. Using the CFS workflow at the HZB enables routine screening experiments. It increases the chances for successful identification of fragment hits as starting points to develop more potent binders, useful for pharmacological or biochemical applications.

High-resolution ARPES endstation for in situ electronic structure investigations at SSRF

Angle-resolved photoemission spectroscopy (ARPES) is one of the most powerful experimental techniques in condensed matter physics. Synchrotron ARPES, which uses photons with high flux and continuously tunable energy, has become particularly important. However, an excellent synchrotron ARPES system must have features such as a small beam spot, super-high energy resolution, and a user-friendly operation interface. A synchrotron beamline and an endstation (BL03U) were designed and constructed at the Shanghai Synchrotron Radiation Facility. The beam spot size at the sample position is 7.5 (V) μm × 67 (H) μm, and the fundamental photon range is 7–165 eV; the ARPES system enables photoemission with an energy resolution of 2.67 meV at 21.2 eV. In addition, the ARPES system of this endstation is equipped with a six-axis cryogenic sample manipulator (the lowest temperature is 7 K) and is integrated with an oxide molecular beam epitaxy system and a scanning tunneling microscope, which can provide an advanced platform for in situ characterization of the fine electronic structure of condensed matter.

A facility for the research, development, and translation of advanced technologies for ion-beam therapies

This article describes the updated GSI radiotherapy research facility (Cave M) located at the GSI Helmholtz Center for Heavy Ion Research in Darmstadt, Germany. This facility was upgraded by modernizing the beamline that supported a pilot project in carbon ion cancer therapy in Europe from 1997 to 2008. Descriptions are provided of the modernized beamline, related hardware components and treatment delivery system. The performance specifications and general characteristics for each major component are described, along with example pre-clinical test results of selected components. These upgrades to Cave M allow for investigating novel therapy methods. The radiotherapy research facility is located on a beamline of the heavy ion synchrotron (Schwer-Ionen-Synchrotron, or SIS-18) accelerator complex, capable of delivering 0.1 to 2 GeV/u charged particle beams, ranging from protons to uranium. This beamline contains components for fast beam gating, aborting, focusing, scanning, monitoring, and shifting the range of the beam. The beam scanning magnets, position detectors, and beam monitors are described, along with tests of functionality and performance. A dose delivery system (DDS) was adapted from a clinical unit at the National Centre for Oncological Hadrontherapy (CNAO), Pavia, Italy, and consists of modular real-time hardware and software. The DDS was modified to enable research on adaptively-managed patient motion through the use of libraries of 4D-optimized radiation treatment plans, an unsolved problem of importance for treating moving tumors. The system is modular and is designed to support future research studies, such as high dose rate (Flash) radiotherapy and radioactive ion beams. A series of validation tests confirmed the functionality and performance of various key components and systems. For example, an end-to-end test revealed that dosimetric spatial homogeneity of over 95% was achieved for square treatment fields. More generally, all performance characteristics that were tested satisfied anticipated clinical requirements.

X-ray photoabsorption-induced processes within protonated rifamycin sodium salts in the gas phase

Up to now, the response of antibiotics upon ionizing radiation has been very scarcely reported. Here, we present the results of X-ray photoabsorption experiments on isolated rifamycin, a broad-range antibiotic against Gram-positive and Gram-negative bacteria. A mass spectrometer has been coupled to a synchrotron beamline to analyze cationic products of photoabsorption on protonated rifamycin dimer and monomer sodium salts. Absorption of a single photon in the 100–300eV energy range leads to ionization of the molecular system, followed by vibrational energy deposition and subsequent inter- and/or intramolecular fragmentation. Interestingly, we observe a proton transfer from sodiated rifamycin to rifamycin, a widely observed process in ionized molecular systems in the gas phase. Moreover, we show that another charge-transfer process occurs in both dimer and monomer: intramolecular sodium transfer, which has not been reported yet, to the best of our knowledge.

Effect of Combined Tempering and Aging in the Austenite Reversion, Precipitation, and Tensile Properties of an Additively Manufactured Maraging 300 Steel

Maraging 300 is an ultrahigh strength steel with significant alloy addition, resulting in a martensitic matrix hardened by precipitation through aging treatment. In these steels, intercritical tempering can provide reverted austenite and precipitation of intermetallic products, increasing the ductility of additively manufactured parts due to austenite presence. Studies deal with postprocessing of additive manufactured parts of maraging steel; however, few focused on phases evolution during the heat treatments and their mechanical response. In the present work, a maraging 300 steel processed by laser-based powder-bed fusion was studied with a focus on microstructural and mechanical properties after applying several postprocessing heat treatments. Tensile tests assessed the mechanical properties, and the microstructure was analyzed by scanning and transmission electron microscopy. A synchrotron beamline with x-ray diffraction was used to conduct in situ measurements of martensite and austenite evolution. The in situ phase evolution revealed that isothermal heat treatments were efficient in promoting martensite-to-austenite reversion. Likewise, the presence of austenite significantly enhanced the ductility, however, at some mechanical strength expense.

Automatic segmentation for synchrotron-based imaging of porous bread dough using deep learning approach.

In recent years, major capability improvements at synchrotron beamlines have given researchers the ability to capture more complex structures at a higher resolution within a very short time. This opens up the possibility of studying dynamic processes and observing resulting structural changes over time. However, such studies can create a huge quantity of 3D image data, which presents a major challenge for segmentation and analysis. Here tomography experiments at the Australian synchrotron source are examined, which were used to study bread dough formulations during rising and baking, resulting in over 460 individual 3D datasets. The current pipeline for segmentation and analysis involves semi-automated methods using commercial software that require a large amount of user input. This paper focuses on exploring machine learning methods to automate this process. The main challenge to be faced is in generating adequate training datasets to train the machine learning model. Creating training data by manually segmenting real images is very labour-intensive, so instead methods of automatically creating synthetic training datasets which have the same attributes of the original images have been tested. The generated synthetic images are used to train a U-Net model, which is then used to segment the original bread dough images. The trained U-Net outperformed the previously used segmentation techniques while taking less manual effort. This automated model for data segmentation would alleviate the time-consuming aspects of experimental workflow and would open the door to perform 4D characterization experiments with smaller time steps.

A theoretical study to focus a polychromatic synchrotron X-ray beam for microbeam radiation therapy

Microbeam radiotherapy (MRT) appears to target tumour cells while sparing normal healthy cells. However, it is in its preclinical stage at present. Clinical trials of MRT can be done in the Imaging and Medical Beamline (IMBL) of the Australian Synchrotron provided that the dose rate deliverable by the X-ray beam in Hutch 3B of the IMBL, where the required patient positioning equipment is located, is sufficiently high for MRT. At present the deliverable dose rate there is only 75Gys−1. However, a deliverable dose rate of between and including 375Gys−1 and 750Gys−1 is required for doing MRT clinical trials in Hutch 3B due to the irradiation profile inherent to MRT. The height of the focused beam in Hutch 3B must also be between and including 1.0 mm and 10 mm. A kinoform compound refractive lens (kinoform CRL) made of [3.5×1018Patomscm−3]-doped Si is proposed to be used in the Scenario where the X-ray beam, produced by a superconducting multipole wiggler source (SCMPW) operating at a maximum magnetic field of 4.000 T, is filtered by 0.6 mm of diamond before reaching the kinoform CRL. This is the Diamond-Optic Scenario. In this Scenario, the dose rate deliverable in Hutch 3B using the optic ranges from 380Gys−1 to 1300Gys−1 with beam heights ranging from 3.55 mm down to 1.0 mm, respectively. Also, the weighted average photon energies of the beam in Hutch 3B in this Scenario are almost entirely suitable for MRT in the IMBL. The X-ray beam in this Scenario can heat the P-doped Si optic to a few hundred degrees Kelvin above the hypothesised melting point of the P-doped Si due to its high power density. Therefore, the optic must be cooled lest it melt. The Diamond-Optic Scenario is not used routinely in the IMBL due to its excessive power. A variation of the Diamond-Optic Scenario, namely that the kinoform CRL be placed before sensitive components instead of after them is proposed for the routine use of this Scenario.

High-temperature deformation followed in situ by X-ray microtomography: a methodology to track features under large strain.

Metallic materials processing such as rolling, extrusion or forging often involves high-temperature deformation. Usually under such conditions the samples are characterized post mortem, under pseudo in situ conditions with interrupted tests, or in situ with a limited strain rate. A full in situ 3D characterization, directly during high-temperature deformation with a prescribed strain-rate scheme, requires a dedicated sample environment and a dedicated image-analysis workflow. A specific sample environment has been developed to enable highly controlled (temperature and strain rate) high-temperature deformation mechanical testing to be conducted while performing in situ tomography on a synchrotron beamline. A dedicated digital volume correlation algorithm is used to estimate the strain field and track pores while the material endures large deformations. The algorithm is particularly suitable for materials with few internal features when the deformation steps between two images are large. An example of an application is provided: a high-temperature compression test on a porous aluminium alloy with individual pore tracking with a specific strain-rate scheme representative of rolling conditions.

Versatile and high temperature spectroscopic cell for operando fluorescence and transmission x-ray absorption spectroscopic studies of heterogeneous catalysts.

A modular high-temperature cell consisting of a plug-flow microreactor with a fixed catalyst bed and long heating zone has been established for operando x-ray absorption/fluorescence spectroscopic and diffraction studies. The functionality of the cell is demonstrated for two important areas: emission control using 2 wt. % Pd/Al2O3 acting as a three-way catalyst and direct conversion of methane to olefins and aromatics on a 0.5% Fe/SiO2 catalyst. The performance has been determined by online infrared spectroscopy and mass spectrometry, respectively. In addition, the cell can be combined with optical spectroscopy, such as Raman spectroscopy. The catalyst, present as powdered/sieved samples, can be measured under reaction conditions at temperatures of up to 1050 °C. Another key aspect is a long isothermal heating zone with a small temperature gradient (<3 °C/mm at 1000 °C without reaction) including an inert zone for pre-heating of the reactant gas. Due to the small size of the microreactor and the heating system including a water cooling system, heating/cooling rates of up to 100 °C/min can be achieved. Moreover, due to the compact design and the autonomous control system, the high temperature operando setup fits to the space at the majority of synchrotron beamlines. In many cases, the concentration of the element of interest in the catalysts is low requiring x-ray absorption spectroscopy measurements in the fluorescence measurement mode. Hence, the microreactor was designed to fit such needs as well. More specifically, the case of Fe-containing catalysts was particularly considered by using iron-free materials for the reactor housing.

Integrated optical chip for a high-resolution, single-resonance-mode x-ray monochromator system.

An integrated optical chip that minimizes the size of the energy-tuning single-resonance-mode x-ray monochromator system into a 3cm×5cm silicon wafer is proposed. A Fabry-Perot x-ray resonator and two back-reflecting Si mirrors are employed on the wafer as the optical components, where Si(12 4 0) back reflection is used for both Fabry-Perot resonance and re-diffraction of the x-ray beams from the resonator in the incident direction. We can achieve an energy bandwidth of 3.4 meV in single-mode x rays and tune the energy by temperature variation. Such Si chips can be readily employed at the synchrotron beamlines and conventional x-ray laboratories for high-resolution investigations.

Measuring the atomic scattering factors near the iridium L-edges for the Athena silicon pore optics reflector

Athena, a future high-energy mission, is expected to consist of a large aperture x-ray mirror with a focal length of 12 m. The mirror surface is to be coated with iridium and a low Z overcoat. To define the effective area of the x-ray telescope, the atomic scattering factors of iridium with an energy resolution less than that (2.5 eV) of the x-ray integral field unit are needed. We measured the reflectance of the silicon pore optics mirror plate coated with iridium in the energy range of 9 to 15 keV and that near the iridium L-edges in steps of 10 and 1.5 eV, respectively, at the synchrotron beamline SPring-8. The L3, L2, and L1 edges were clearly detected around 11,215, 12,824, and 13,428 eV, respectively. The measured scattering factors were ∼3 % smaller than the corresponding values reported by Henke et al., likely due to the presence of an overlayer on the iridium coating, and were consistent with those measured by Graessle et al. The angular dependence of the reflectivity measured indicates that the iridium surface was extremely smooth, with a surface roughness of 0.3 nm.

Synchrotron radiation as a tool for macromolecular X-Ray Crystallography: A XXI century perspective

Intense X-rays available at powerful synchrotron beamlines provide macromolecular crystallographers with an incomparable tool for investigating biological phenomena on an atomic scale. The resulting insights into the mechanism’s underlying biological processes have played an essential role and shaped biomedical sciences during the last 30 years, considered the “golden age” of structural biology. In this review, we analyze selected aspects of the impact of synchrotron radiation on structural biology. Synchrotron beamlines have been used to determine over 70% of all macromolecular structures deposited into the Protein Data Bank (PDB). These structures were deposited by over 13,000 different research groups. Interestingly, despite the impressive advances in synchrotron technologies, the median resolution of macromolecular structures determined using synchrotrons has remained constant throughout the last 30 years, at about 2 Å. Similarly, the median times from the data collection to the deposition and release have not changed significantly. We describe challenges to reproducibility related to recording all relevant data and metadata during the synchrotron experiments, including diffraction images. Finally, we discuss some of the recent opinions suggesting a diminishing importance of X-ray crystallography due to impressive advances in Cryo-EM and theoretical modeling. We believe that synchrotrons of the future will increasingly evolve towards a life science center model, where X-ray crystallography, Cryo-EM, other experimental and computational resources, and knowledge are encompassed within a versatile research facility. The recent response of crystallographers to the COVID-19 pandemic suggests that X-ray crystallography conducted at synchrotron beamlines will continue to play an essential role in structural biology and drug discovery for years to come.

Evaluation of the effects of Azadirachtin on internal structures of Rhodnius prolixus head using low-energy X-ray microfluorescence

Rhodnius prolixus is the main triatomine insect vector of Chagas Disease, which affects between 6 and 8 million individuals, with approximately 12,000 deaths per year in Latin America. Azadirachtin is a drug that causes changes in several species of insects, including triatomines, thereby inhibiting their development.This work exploits Low-Energy X-Ray Microfluorescence (LEXRF) to evaluate the distribution of oxygen and sodium in the structures of the insect Rhodnius prolixus, and to understand the effects of azadirachtin treatment. Specimens of fifth-instar nymphs of Rhodnius prolixus were taken from the colony of the Laboratory of Biochemistry and Physiology of Insects (Oswaldo Cruz Foundation (FIOCRUZ), Brazil) and were divided in two groups. In the first group, the insects were fed with defibrinated rabbit blood containing azadirachtin and the other group only with simply defibrinated rabbit blood. Both groups were examined at the TwinMic soft X-ray microscopy synchrotron beamline, (Elettra Sincrotrone Trieste, Trieste, Italy), providing elemental map distribution of oxygen which was correlated to the high-resolution morphological information obtained by the simultaneous absorption and differential phase contrast images.Prior to Low-Energy X-Ray Microfluorescence (LEXRF) measurements two control samples were scanned at the SYRMEP beamline (Elettra Sincrotrone Trieste, Trieste, Italy) by microtomography for an overview of the structures that were subsequently studied at TwinMic. The results allowed us to compare the differences in structures of the head of Rhodnius prolixus providing new insights into the insect's development that may be of significance toward the control of the Chagas Disease.