Alberto Astolfo

Functional and Electrical Integration of Induced Pluripotent Stem Cell-Derived Cardiomyocytes in a Myocardial Infarction Rat Heart.

In vitro expanded beating cardiac myocytes derived from induced pluripotent stem cells (iPSC-CMs) are a promising source of therapy for cardiac regeneration. Meanwhile, the cell sheet method has been shown to potentially maximize survival, functionality, and integration of the transplanted cells into the heart. It is thus hypothesized that transplanted iPSC-CMs in a cell sheet manner may contribute to functional recovery via direct mechanical effects on the myocardial infarction (MI) heart. F344/NJcl-rnu/rnu rats were left coronary artery ligated (n = 30), followed by transplantation of Dsred-labeled iPSC-CM cell sheets of murine origin over the infarct heart surface. Effects of the treatment were assessed, including in vivo molecular/cellular evaluations using a synchrotron radiation scattering technique. Ejection fraction and activation recovery interval were significantly greater from day 3 onward after iPSC-CM transplantation compared to those after sham operation. A number of transplanted iPSC-CMs were present on the heart surface expressing cardiac myosin or connexin 43 over 2 weeks, assessed by immunoconfocal microscopy, while mitochondria in the transplanted iPSC-CMs gradually showed mature structure as assessed by electron microscopy. Of note, X-ray diffraction identified 1,0 and 1,1 equatorial reflections attributable to myosin and actin–myosin lattice planes typical of organized cardiac muscle fibers within the transplanted cell sheets at 4 weeks, suggesting cyclic systolic myosin mass transfer to actin filaments in the transplanted iPSC-CMs. Transplantation of iPSC-CM cell sheets into the heart yielded functional and electrical recovery with cyclic contraction of transplanted cells in the rat MI heart, indicating that this strategy may be a promising cardiac muscle replacement therapy.

A simple way to track single gold-loaded alginate microcapsules using x-ray CT in small animal longitudinal studies

UNLABELLED The use of alginate based microcapsules to deliver drugs and cells with a minimal host interaction is increasingly being proposed. A proficient method to track the position of the microcapsules during such therapies, particularly if they are amenable to commonly used instrumentation, would greatly help the development of such treatments. Here we propose to label the microcapsules with gold nanoparticles to provide a bright contrast on small animal x-ray micro-CT systems enabling single microcapsule detection. The microcapsules preparation is based on a simple protocol using inexpensive compounds. This, combined with the widespread availability of micro-CT apparatus, renders our method more accessible compared with other methods. Our labeled microcapsules showed good mechanical stability and low cytotoxicity in-vitro. Our post-mortem rodent model data strongly suggest that the high signal intensity generated by the labeled microcapsules permits the use of a reduced radiation dose yielding a method fully compatible with longitudinal in-vivo studies. FROM THE CLINICAL EDITOR The authors of this study report the development of a micro-CT based tracking method of alginate-based microcapsules by incorporating gold nanoparticles in the microcapsules. They demonstrate the feasibility of this system in rodent models, where due to the high signal intensity, even reduced radiation dose is sufficient to track these particles, providing a simple and effective method to track these commonly used microcapsules and allowing longitudinal studies.

Detectors for the imaging and medical beam line at the australian synchrotron

The Australian Synchrotron Imaging and Medical Beam Line (IMBL) began phased commissioning in late 2008 and was opened for Users this year (November, 2012). It will provide Australia with an unrivaled facility for x-ray imaging and radiotherapy research covering a wide range of applications in disease studies, treatments, and revealing physiological processes. The clinical research drivers for IMBL rely on the facilitys ability to support high spatial and contrast resolution imaging. The wide variety of demands for x-ray imaging with IMBL cannot be covered with any single detector system. A list of six detector categories was drawn up after assessing the techniques that are most likely to be used during our first years of operation. Detectors in this list will cover the fields of view, resolutions (both spatial and contrast), and frame rates required for a majority of the experiments. We present the six detectors within these categories. One detector system is the topic of a development project with the goal of producing a large field of view high aspect ratio system. Some initial design ideas are presented.

The first large area, high X-ray energy phase contrast prototype for enhanced detection of threat object in baggage screening

X-ray imaging is the most commonly used method in baggage screening. Conventional x-ray attenuation (usually in dual-energy mode) is exploited to discriminate threat and non-threat materials: this is essentially, a method that has seen little changes in decades. Our goal is to demonstrate that x-rays can be used in a different way to achieve improved detection of weapons and explosives. Our approach involves the use of x-ray phase contrast and it a) allows much higher sensitivity in the detection of object edges and b) can be made sensitive to the sample’s microstructure. We believe that these additional channels of information, alongside conventional attenuation which would still be available, have the potential to significantly increase both sensitivity and specificity in baggage scanning. We obtained preliminary data demonstrating the above enhanced detection, and we built a scanner (currently in commissioning) to scale the concept up and test it on real baggage. In particular, while previous X-ray phase contrast imaging systems were limited in terms of both field of view (FOV) and maximum x-ray energy, this scanner overcomes both those limitations and provides FOVs up to 20 to 50 cm2 with x-ray energies up to 100 keV.

Large field of view, fast and low dose multimodal phase-contrast imaging at high x-ray energy

X-ray phase contrast imaging (XPCI) is an innovative imaging technique which extends the contrast capabilities of ‘conventional’ absorption based x-ray systems. However, so far all XPCI implementations have suffered from one or more of the following limitations: low x-ray energies, small field of view (FOV) and long acquisition times. Those limitations relegated XPCI to a ‘research-only’ technique with an uncertain future in terms of large scale, high impact applications. We recently succeeded in designing, realizing and testing an XPCI system, which achieves significant steps toward simultaneously overcoming these limitations. Our system combines, for the first time, large FOV, high energy and fast scanning. Importantly, it is capable of providing high image quality at low x-ray doses, compatible with or even below those currently used in medical imaging. This extends the use of XPCI to areas which were unpractical or even inaccessible to previous XPCI solutions. We expect this will enable a long overdue translation into application fields such as security screening, industrial inspections and large FOV medical radiography – all with the inherent advantages of the XPCI multimodality.

Recent developments on techniques for differential phase imaging at the medical beamline of ELETTRA

Over the last decade different phase contrast approaches have been exploited at the medical beamline SYRMEP of the synchrotron radiation facility Elettra in Trieste, Italy. In particular special focus has been drawn to analyzer based imaging and the associated imaging theory and processing. Analyzer based Imaging (ABI) and Diffraction Enhanced Imaging (DEI) techniques have been successfully applied in several biomedical applications. Recently it has been suggested to translate the acquired knowledge in this field towards a Thomson Backscattering Source (TBS), which is presently under development at the Frascati National Laboratories of INFN (Istituto Nazionale di Fisica Nucleare) in Rome, Italy. Such source is capable of producing intense and quasi-monochromatic hard X-ray beams. For the technical implementation of biomedical phase imaging at the TBS a grating interferometer for differential phase contrast imaging has been designed and successfully tested at SYRMEP beamline.

A first investigation of accuracy, precision and sensitivity of phase-based x-ray dark-field imaging

In the last two decades, x-ray phase contrast imaging (XPCI) has attracted attention as a potentially significant improvement over widespread and established x-ray imaging. The key is its capability to access a new physical quantity (the ‘phase shift’), which can be complementary to x-ray absorption. One additional advantage of XPCI is its sensitivity to micro structural details through the refraction induced dark-field (DF). While DF is extensively mentioned and used for several applications, predicting the capability of an XPCI system to retrieve DF quantitatively is not straightforward. In this article, we evaluate the impact of different design options and algorithms on DF retrieval for the Edge-Illumination (EI) XPCI technique. Monte Carlo simulations, supported by experimental data, are used to measure the accuracy, precision and sensitivity of DF retrieval performed with several EI systems based on conventional x-ray sources. The introduced tools are easy to implement, and general enough to assess the DF performance of systems based on alternative (i.e. non-EI) XPCI approaches.

Compact and cost effective lab-based edge-illumination x-ray phase contrast imaging with a structured focal spot

We present a different implementation of the Edge Illumination (EI) X-ray Phase Contrast imaging method based on the use of multiple focal spots created through an additional x-ray mask. While this resembles directly inspired by the Talbot-Lau implementation of grating interferometry, the aim of the source mask and its effect on the acquired images are different. The individual “sourcelets” are much larger than in grating methods, and then still spatially incoherent; however, their use allows (a) exploiting cheap and large focal spot sources and (b) reducing the source spot size from the usual 70–100 μm typically used in EI to few tens of μm, which enables the realisation of more compact setups. However, in EI, multiple sources create images shifted by one detector pixel with respect to the other, imposing the use of an image restoration algorithm. Here, we show that the approach is feasible by deconvolving differential phase-contrast image profiles acquired with three separate sources, and comparing resu...

X-ray cell tracking: from ex-vivo to in-vivo experiments

The capacity to track cells (cell tracking) using x-rays on ex-vivo specimens of both malignant and non-malignant cell lines on small animals has been demonstrated recently. Gold nanoparticles have been used as a cellular contrast agent to render cells visible in x-ray microCT acquisitions. The limits of the technique proposed are basically driven by the imaging system used. Single cell resolution can be achieved using synchrotron radiation in-vitro or ex-vivo samples. Micro-focus x-ray tubes can be used to obtain high resolution cell tracking but with some limitations. However, the translation from ex-vivo to in-vivo experiments is not straightforward. The dose restrictions required for in-vivo longitudinal experiments set severe limitations on the technique. Here we present a detailed investigation showing a significant reduction of x-ray dose for the tracking of brain tumour cells. Monte Carlo simulations have been performed considering different spatial resolutions, photon fluence, number of projections, lesion dimension and cell contrast dilution. The findings are compared with real samples imaged using the same parameters. A pioneering in-vivo experiment conducted at the SYRMEP beamline (Elettra, Basovizza, Italy) is presented here as proof of principle of in-vivo longitudinal x-ray cell tracking experiments on small animals at low x-ray doses.

Large field-of-view asymmetric masks for high-energy x-ray phase imaging with standard x-ray tube

We report on a new approach to large field-of-view laboratory-based X-ray phase-contrast imaging. The method is based upon the asymmetric mask design that enables the retrieval of the absorption, refraction and ultra-small- angle scattering properties of the sample without the need to move any component of the imaging system. The sample is scanned through the imaging system, which also removes possible aliasing problems that might arise from partial sample illumination when using the edge illumination technique. This concept can be extended to any desired number of apertures providing, at the same time, intensity projections at complementary illumination conditions. Experimental data simultaneously acquired at seven different illumination fractions are presented along with the results obtained from a numerical model that incorporates the actual detector performance. The ultimate shape of the illumination function is shown to be significantly dependent on these detector-specific characteristics. Based on this concept, a large field-of-view system was designed, which is also capable to cope with relatively high (100 kVp) X-ray energies. The imaging system obtained in this way, where the asymmetric mask design enables the data to be collected without moving any element of the instrumentation, adapts particularly well to those situations in medical, industrial and security imaging where the sample has to be scanned through the system.