Synchrotron Radiation-Based Three-Dimensional Visualization of Angioarchitectural Remodeling in Hippocampus of Epileptic Rats.

Characterizing the three-dimensional (3D) morphological alterations of microvessels under both normal and seizure conditions is crucial for a better understanding of epilepsy. However, conventional imaging techniques cannot detect microvessels on micron/sub-micron scales without angiography. In this study, synchrotron radiation (SR)-based X-ray in-line phase-contrast imaging (ILPCI) and quantitative 3D characterization were used to acquire high-resolution, high-contrast images of rat brain tissue under both normal and seizure conditions. The number of blood microvessels was markedly increased on days 1 and 14, but decreased on day 60 after seizures. The surface area, diameter distribution, mean tortuosity, and number of bifurcations and network segments also showed similar trends. These pathological changes were confirmed by histological tests. Thus, SR-based ILPCI provides systematic and detailed views of cerebrovascular anatomy at the micron level without using contrast-enhancing agents. This holds considerable promise for better diagnosis and understanding of the pathogenesis and development of epilepsy.

Investigation of inner ear anatomy in mouse using X-ray phase contrast tomography.

A thorough understanding of inner ear anatomy is important for investigators. However, investigation of the mouse inner ear is difficult due to the limitations of imaging techniques. X-ray phase contrast tomography increases contrast 100-1,000 times compared with conventional X-ray imaging. This study aimed to investigate inner ear anatomy in a fresh post-mortem mouse using X-ray phase contrast tomography and to provide a comprehensive atlas of microstructures with less tissue deformation. All experiments were performed in accordance with our institution's guidelines on the care and use of laboratory animals. A fresh mouse cadaver was scanned immediately after sacrifice using an inline phase contrast tomography system. Slice images were reconstructed using a filtered back-projection (FBP) algorithm. Standardized axial and coronal planes were adjusted with a multi-planar reconstruction method. Some three-dimensional (3D) objects were reconstructed by surface rendering. The characteristic features of microstructures, including otoconia masses of the saccular and utricular maculae, superior and inferior macula cribrosae, single canal, modiolus, and osseous spiral lamina, were described in detail. Spatial positions and relationships of the vestibular structures were exhibited in 3D views. This study investigated mouse inner ear anatomy and provided a standardized presentation of microstructures. In particular, otoconia masses were visualized in their natural status without contrast for the first time. The comprehensive anatomy atlas presented in this study provides an excellent reference for morphology studies of the inner ear.

Simultaneous Imaging of Cerebrovascular Structure and Function in Hypertensive Rats Using Synchrotron Radiation Angiography.

Hypertension has a profound influence on the structure and function of blood vessels. Cerebral vessels undergo both structural and functional changes in hypertensive animals. However, dynamic changes of cerebrovasculature and the factors involved in this process are largely unknown. In this study, we explored the dynamic changes of vascular structure in hypertensive rats using novel synchrotron radiation angiography. Twenty-four spontaneously hypertensive rats (SHR) and 24 Sprague-Dawley (SD) rats underwent synchrotron radiation (SR) angiography. Each group had 8 animals. We studied the cerebral vascular changes in SHR over a time period of 3-12-month and performed quantitative analysis. No vascular morphology differences between SHR and SD rats were observed in the early stage of hypertension. The number of twisted blood vessels in the front brain significantly increased at the 9- and 12-month observation time-points in the SHR compared to the SD rats (p < 0.01). The vessel density of the cortex and the striatum in SHR was consistently higher than that in SD rats at time points of 3-, 9-, and 12-month (p < 0.001). Vascular elasticity decreased both in SHR and SD rats with aging. There were statistically significant differences in the relative vascular elasticity of extracranial/intracranial internal carotid artery, middle cerebral artery, posterior cerebral artery and anterior cerebral artery between SHR and SD rats at 12-month (p < 0.01). We concluded that the dynamic vascular alterations detected by SR angiography provided novel imaging data for the study of hypertension in vivo. The longer the course of hypertension was, the more obvious the vascular differences between the SHR and the SD rats became.

In situ 3D topographic and shape analysis by synchrotron radiation X-ray microtomography for crystal form identification in polymorphic mixtures.

Polymorphism denotes the existence of more than one crystal structure of a substance, and great practical and theoretical interest for the chemical and pharmaceutical industries. In many cases, it is challenging to produce a pure crystal form and establish a sensitive detection method for the identification of crystal form in a mixture of polymorphs. In this study, an accurate and sensitive method based on synchrotron radiation X-ray computed microtomography (SR-µCT) was devised to identify the polymorphs of clopidogrel bisulphate (CLP). After 3D reconstruction, crystal particles were extracted and dozens of structural parameters were calculated. Whilst, the particle shapes of the two crystal forms were all irregular, the surface of CLP II was found to be rougher than CLP I. In order to classify the crystal form based on the quantitative morphological property of particles, Volume Bias Percentage based on Surface Smoothing (VBP) was defined and a new method based on VBP was successfully developed, with a total matching rate of 99.91% for 4544 particles and a lowest detectable limit of 1%. More important for the mixtures in solid pharmaceutical formulations, the interference of excipients can be avoided, a feature cannot achieved by other available analytical methods.

Visualization and quantification of deformation behavior of clopidogrel bisulfate polymorphs during tableting.

The deformation behavior of particles under pressure dominates the mechanical properties of solid dosage forms. In this study, the in situ 3D deformation of two polymorphs (I and II) of clopidogrel bisulfate (CLP) was determined to illustrate pressure distribution profiles within the tablet by the deformation of the crystalline particles for the first time. Synchrotron radiation X-ray computed microtomography (SR-µCT) was utilized to visualize and quantify the morphology of thousands crystalline particles of CLP I and CLP II before and after compression. As a result, the deformation was examined across scale dimensions from microns to the size of the final dosage form. Three dimensional parameters such as volume, sphericity, oblate and prolate of individual particle and distributions were computed and analyzed for quantitative comparison to CLP I and CLP II. The different degrees of deformation under the same compression conditions of CLP I and CLP II were observed and characterized quantitatively. The map of deformation degrees within the tablet illustrated the heterogeneous pressure distribution in various regions of the compacted tablet. In conclusion, the polymorph deformation behaviors demonstrated by SR-µCT quantitative structure analysis deepen understanding of tableting across dimensions from microns to millimeters for the macrostrcuture of tablet.

Ultra-high-resolution 3D digitalized imaging of the cerebral angioarchitecture in rats using synchrotron radiation.

The angioarchitecture is a fundamental aspect of brain development and physiology. However, available imaging tools are unsuited for non-destructive cerebral mapping of the functionally important three-dimensional (3D) vascular microstructures. To address this issue, we developed an ultra-high resolution 3D digitalized angioarchitectural map for rat brain, based on synchrotron radiation phase contrast imaging (SR-PCI) with pixel size of 5.92 µm. This approach provides a systematic and detailed view of the cerebrovascular anatomy at the micrometer level without any need for contrast agents. From qualitative and quantitative perspectives, the present 3D data provide a considerable insight into the spatial vascular network for whole rodent brain, particularly for functionally important regions of interest, such as the hippocampus, pre-frontal cerebral cortex and the corpus striatum. We extended these results to synchrotron-based virtual micro-endoscopy, thus revealing the trajectory of targeted vessels in 3D. The SR-PCI method for systematic visualization of cerebral microvasculature holds considerable promise for wider application in life sciences, including 3D micro-imaging in experimental models of neurodevelopmental and vascular disorders.

[An effective method for improving the imaging spatial resolution of terahertz time domain spectroscopy system].

Terahertz radiation is an electromagnetic radiation in the range between millimeter waves and far infrared. Due to its low energy and non-ionizing characters, THz pulse imaging emerges as a novel tool in many fields, such as material, chemical, biological medicine, and food safety. Limited spatial resolution is a significant restricting factor of terahertz imaging technology. Near field imaging method was proposed to improve the spatial resolution of terahertz system. Submillimeter scale's spauial resolution can be achieved if the income source size is smaller than the wawelength of the incoming source and the source is very close to the sample. But many changes were needed to the traditional terahertz time domain spectroscopy system, and it's very complex to analyze sample's physical parameters through the terahertz signal. A method of inserting a pinhole upstream to the sample was first proposed in this article to improve the spatial resolution of traditional terahertz time domain spectroscopy system. The measured spatial resolution of terahertz time domain spectroscopy system by knife edge method can achieve spatial resolution curves. The moving stage distance between 10 % and 90 Yo of the maximum signals respectively was defined as the, spatial resolution of the system. Imaging spatial resolution of traditional terahertz time domain spectroscopy system was improved dramatically after inserted a pinhole with diameter 0. 5 mm, 2 mm upstream to the sample. Experimental results show that the spatial resolution has been improved from 1. 276 mm to 0. 774 mm, with the increment about 39 %. Though this simple method, the spatial resolution of traditional terahertz time domain spectroscopy system was increased from millimeter scale to submillimeter scale. A pinhole with diameter 1 mm on a polyethylene plate was taken as sample, to terahertz imaging study. The traditional terahertz time domain spectroscopy system and pinhole inserted terahertz time domain spectroscopy system were applied in the imaging experiment respectively. The relative THz-power loss imaging of samples were use in this article. This method generally delivers the best signal to noise ratio in loss images, dispersion effects are cancelled. Terahertz imaging results show that the sample's boundary was more distinct after inserting the pinhole in front of, sample. The results also conform that inserting pinhole in front of sample can improve the imaging spatial resolution effectively. The theoretical analyses of the method which improve the spatial resolution by inserting a pinhole in front of sample were given in this article. The analyses also indicate that the smaller the pinhole size, the longer spatial coherence length of the system, the better spatial resolution of the system. At the same time the terahertz signal will be reduced accordingly. All the experimental results and theoretical analyses indicate that the method of inserting a pinhole in front of sample can improve the spatial resolution of traditional terahertz time domain spectroscopy system effectively, and it will further expand the application of terahertz imaging technology.

[Study of Terahertz Amplitude Imaging Based on the Mean Absorption].

A new method of terahertz (THz) imaging based on the mean absorption is proposed. Terahertz radiation is an electromagnetic radiation in the range between millimeter waves and far infrared. THz pulse imaging emerges as a novel tool in many fields because of its low energy and non-ionizing character, such as material, chemical, biological medicine and food safety. A character of THz imaging technique is it can get large amount of information. How to extract the useful parameter from the large amount of information and reconstruct sample's image is a key technology in THz imaging. Some efforts have been done for advanced visualization methods to extract the information of interest from the raw data. Both time domain and frequency domain visualization methods can be applied to extract information on the physical properties of samples from THz imaging raw data. The process of extracting useful parameter from raw data of the new method based on the mean absorption was given in this article. This method relates to the sample absorption and thickness, it delivers good signal to noise ratio in the images, and the dispersion effects are cancelled. A paper with a "THz" shape hole was taken as the sample to do the experiment. Traditional THz amplitude imaging methods in time domain and frequency domain are used to achieve the sample's image, such as relative reduction of pulse maximum imaging method, relative power loss imaging method, and relative power loss at specific frequency imaging method. The sample's information that reflected by these methods and the characteristics of these methods are discussed. The method base on the mean absorption within a certain frequency is also used to reconstruct sample's image. The experimental results show that this new method can well reflect the true information of the sample. And it can achieve a clearer image than the other traditional THz amplitude imaging methods. All the experimental results and theoretical analyses indicate that the method base on the mean absorption within a certain frequency can reflects sample absorb and thickness information, it can achieve good signal to noise ratio in the images. Because the absorption is mean absorption within in a certain frequency, so the method proposed in this article is especially suitable for samples with simple structure. And this new method can be a useful added tool for the other traditional THz amplitude imaging methods.

[In situ investigation to three dimensional structures of Chinese medicines seeds].

This paper is aimed to microscopic identification of traditional Chinese medicines (TCMs) using an in situ imaging method. In this study, two kinds of Zingiberaceae seeds, Amomi Rotundus Fructus and Alpiniae Katsumadai Semen, were investigated by synchrotron radiation in-line X-ray phase-contrast computed tomography (IXPCT) imaging method. The results showed that the microstructures of these Zingiberaceae seeds could be clearly obtained from the virtual slices information in different observing angles. It proves that IXPCT is an effective imaging method, which can provide the imaging information for the microscopic identification of the intact TCMs in situ and non-destructively.

3D online submicron scale observation of mixed metal powder's microstructure evolution in high temperature and microwave compound fields.

In order to study the influence on the mechanical properties caused by microstructure evolution of metal powder in extreme environment, 3D real-time observation of the microstructure evolution of Al-Ti mixed powder in high temperature and microwave compound fields was realized by using synchrotron radiation computerized topography (SR-CT) technique; the spatial resolution was enhanced to 0.37 µm/pixel through the designed equipment and the introduction of excellent reconstruction method for the first time. The process of microstructure evolution during sintering was clearly distinguished from 2D and 3D reconstructed images. Typical sintering parameters such as sintering neck size, porosity, and particle size of the sample were presented for quantitative analysis of the influence on the mechanical properties and the sintering kinetics during microwave sintering. The neck size-time curve was obtained and the neck growth exponent was 7.3, which indicated that surface diffusion was the main diffusion mechanism; the reason was the eddy current loss induced by the external microwave fields providing an additional driving force for mass diffusion on the particle surface. From the reconstructed images and the curve of porosity and average particle size versus temperature, it was believed that the presence of liquid phase aluminum accelerated the densification and particle growth.

In situ investigation of the 3D mechanical microstructure at nanoscale: nano-CT imaging method of local small region in large scale sample.

To investigate the local micro-/nanoscale region in a large scale sample, an image reconstruction method for nanometer computed tomography (nano-CT) was proposed in this paper. In the algorithm, wavelets were used to localize the filtered-backprojection (FBP) algorithm because of its space-frequency localization property. After the implementation of the algorithm, two simulation local reconstruction experiments were performed to confirm its effectiveness. Three evaluation criteria were used in the experiments to judge the quality of the reconstructed images. The experimental results showed that the algorithm proposed in this paper performed best because (1) the quality of its results had improved 20%-30% compared to the results of FBP and 10%-30% compared to the results of another wavelet algorithm; (2) the new algorithm was stable under different circumstances. Besides, an actual reconstruction experiment was performed using real projection data that had been collected in a CT experiment. Two-dimensional (2D) and three-dimensional (3D) images of the sample were reconstructed. The microstructure of the sample could be clearly observed in the reconstructed images. Since much attention has been directed towards the nano-CT technique to investigate the microstructure of materials, this new wavelet-based local tomography algorithm could be considered as a meaningful effort.

A synchrotron-based local computed tomography combined with data-constrained modelling approach for quantitative analysis of anthracite coal microstructure.

Quantifying three-dimensional spatial distributions of pores and material compositions in samples is a key materials characterization challenge, particularly in samples where compositions are distributed across a range of length scales, and where such compositions have similar X-ray absorption properties, such as in coal. Consequently, obtaining detailed information within sub-regions of a multi-length-scale sample by conventional approaches may not provide the resolution and level of detail one might desire. Herein, an approach for quantitative high-definition determination of material compositions from X-ray local computed tomography combined with a data-constrained modelling method is proposed. The approach is capable of dramatically improving the spatial resolution and enabling finer details within a region of interest of a sample larger than the field of view to be revealed than by using conventional techniques. A coal sample containing distributions of porosity and several mineral compositions is employed to demonstrate the approach. The optimal experimental parameters are pre-analyzed. The quantitative results demonstrated that the approach can reveal significantly finer details of compositional distributions in the sample region of interest. The elevated spatial resolution is crucial for coal-bed methane reservoir evaluation and understanding the transformation of the minerals during coal processing. The method is generic and can be applied for three-dimensional compositional characterization of other materials.

[Investigation of characteristic microstructures of adhesive interface in wood/bamboo composite material by synchrotron radiation X-ray phase contrast microscopy].

Third-generation synchrotron radiation X-ray phase-contrast microscopy(XPCM)can be used for obtaining image with edge enhancement, and achieve the high contrast imaging of low-Z materials with the spatial coherence peculiarity of X-rays. In the present paper, the characteristic microstructures of adhesive at the interface and their penetration in wood/bamboo composite material were investigated systematically by XPCM at Shanghai Synchrotron Radiation Facility (SSRF). And the effect of several processing techniques was analyzed for the adhesive penetration in wood/bamboo materials. The results show that the synchrotron radiation XPCM is expected to be one of the important precision detection methods for wood-based panels.

[Measuring the density of wood and bamboo using computed tomography].

CT is widespread non-destructive detection technique for wood materials, and the density measurement is a key role during this application. In the present report, the use of CT for air-dry density measurement of wood and bamboo is described. The authors found that there were marked linear correlations between air-dry density (0.303-1.061 g x cm(-3)) of 24 kinds of woods and their respective CT value, as well as 25 kinds of lignin materials (including 24 kinds of woods and 1 kind of bamboo) and the CT value, both with correlation coefficient of 0.99, which belonged to the CT technological breakthrough for wood quantitative detection These research results show that CT is an appropriate way to measure density for wood and bamboo, and would provide technical support for CT used in the field of wood science research and wood processing.

PITRE: software for phase-sensitive X-ray image processing and tomography reconstruction.

Synchrotron-radiation computed tomography has been applied in many research fields. Here, PITRE (Phase-sensitive X-ray Image processing and Tomography REconstruction) and PITRE_BM (PITRE Batch Manager) are presented. PITRE supports phase retrieval for propagation-based phase-contrast imaging/tomography (PPCI/PPCT), extracts apparent absorption, refractive and scattering information of diffraction enhanced imaging (DEI), and allows parallel-beam tomography reconstruction for conventional absorption CT data and for PPCT phase retrieved and DEI-CT extracted information. PITRE_BM is a batch processing manager for PITRE: it executes a series of tasks, created via PITRE, without manual intervention. Both PITRE and PITRE_BM are coded in Interactive Data Language (IDL), and have a user-friendly graphical user interface. They are freeware and can run on Microsoft Windows systems via IDL Virtual Machine, which can be downloaded for free and does not require a license. The data-processing principle and some examples of application will be presented.

[Detection of microvasculature in rat hind limb using synchrotron radiation].

OBJECTIVE: To detect deep-level microvascular structure in rat hind limb by synchrotron radiation and microangiographic technique. METHODS: Microangiography in vivo and ex vivo was performed by synchrotron radiation based absorption imaging and phase contrast imaging, with omnipaque and barium sulfate solution as contrast media, respectively, and synchrotron radiation-based micro-computed tomography (SRmCT) was also performed to reveal three-dimensional morphology of the blood vessel in rat hind limb. RESULTS: Using microangiographic technique in vivo and in vitro (with barium sulfate), blood vessels in the rat limb muscle could be visualized with high resolution, and the fourth branches of iliac artery in rat hind limb could be detected with the minimum visualized blood vessels about 40 µm and 9 µm in diameter, respectively. In addition, the vascular network could be defined and analyzed at the micrometer scale from the 3D renderings of limb vessel as shown by SRmCT. CONCLUSION: Synchrotron radiation-based microangiography and SRmCT thus provided a practical and effective means to observe the microvasculature of rat hindlimb, which might be useful in assessment of angiogenesis in lower limbs.

[Exploring the three-dimensional structure of dermal tissues of normal skin and scar in rat with synchrotron radiation X-ray imaging technology].

OBJECTIVE: To compare the morphological difference between dermal tissue of normal skin and that of scar in rat, and to explore its structural pattern. METHODS: The full-thickness skin and the scar tissue formed 3 weeks after wound healing from SD rats were harvested as samples, which were prepared appropriately afterwards. Samples were scanned and imaged with synchrotron radiation technology, micro-CT, and phase-contrast imaging technology. The images were rebuilt with three-dimensional software. RESULTS: The micro-CT was materialized by using X-ray generated by synchrotron radiation light source. The structure of dermal tissues was clearly shown with the assistance of phase-contrast imaging technology in the process. It was demonstrated that the dermal tissues of normal skin of rat were mainly composed of collagenous fibers, which twined together to form an olive-like structure. These olive-like structures as basic units were arranged randomly in a certain way. The collagenous fibers in dermal tissue of the scar were arranged in a parallel manner, while some fibers were crooked and arranged in a disorderly manner. CONCLUSIONS: Dermal tissue of normal skin in rat has stable three-dimensional structure, and its basic structure and manner of composition are obviously different from those of scar dermal tissue.

[Nondestructive imaging of elements distribution in biomedical samples by X-ray fluorescence computed tomography].

X-ray fluorescence computed tomography is a stimulated emission tomography that allows nondestructive reconstruction of the elements distribution in the sample, which is important for biomedical investigations. Owing to the high flux density and easy energy tunability of highly collimated synchrotron X-rays, it is possible to apply X-ray fluorescence CT to biomedical samples. Reported in the present paper, an X-ray fluorescence CT system was established at Shanghai Synchrotron Radiation Facility for the investigations of trace elements distribution inside biomedical samples. By optimizing the experiment setup, the spatial resolution was improved and the data acquisition process was obviously speeded up. The maximum-likelihood expectation-maximization algorithm was introduced for the image reconstruction, which remarkably improved the imaging accuracy of element distributions. The developed system was verified by the test sample and medical sample respectively. The results showed that the distribution of interested elements could be imaged correctly, and the spatial resolution of 150 m was achieved. In conclusion, the developed system could be applied to the research on large-size biomedical samples, concerning imaging accuracy, spatial resolution and data collection time.

[Quantitative analysis of nucleotide mixtures with terahertz time domain spectroscopy].

Adenosine, thymidine, guanosine, cytidine and uridine form the building blocks of ribose nucleic acid (RNA) and deoxyribose nucleic acid (DNA). Nucleosides and their derivants are all have biological activities. Some of them can be used as medicine directly or as materials to synthesize other medicines. It is meaningful to detect the component and content in nucleosides mixtures. In the present paper, components and contents of the mixtures of adenosine, thymidine, guanosine, cytidine and uridine were analyzed. THz absorption spectra of pure nucleosides were set as standard spectra. The mixture's absorption spectra were analyzed by linear regression with non-negative constraint to identify the components and their relative content in the mixtures. The experimental and analyzing results show that it is simple and effective to get the components and their relative percentage in the mixtures by terahertz time domain spectroscopy with a relative error less than 10%. Component which is absent could be excluded exactly by this method, and the error sources were also analyzed. All the experiments and analysis confirms that this method is of no damage or contamination to the sample. This means that it will be a simple, effective and new method in biochemical materials analysis, which extends the application field of THz-TDS.

Application of x-ray phase contrast imaging to microscopic identification of Chinese medicines.

In the last decade, x-ray phase contrast imaging (XPCI) has received considerable attention as a novel imaging technique, which has proved to be suitable for weakly absorbing materials such as biomedical samples and polymers. In this paper, the microstructures of traditional Chinese medicines (TCMs), which are used as judging criteria in the identification of TCMs, were investigated by XPCI based on a nano-focus x-ray tube. The results demonstrated that XPCI is a promising new method for the identification of TCMs, with advantages such as nondestructivity, no special sample preparation and suitability for thick samples.