S. Matsuyama

MICROBEAM ANALYSIS SYSTEM AT TOHOKU UNIVERSITY

A microbeam analysis system has been developed at Tohoku University for biological applications. Spatial resolution of less than 1 μm has been achieved with a beam current of ~40 pA. In microbeam analysis of biological specimens, simultaneous measurement of structural and elemental properties is very important. Our system is applicable to simultaneous in-air/vacuum PIXE, RBS and STIM analyses. Typical results of biological application are shown.

MICRON-CT USING PIXE WITH MICRO-BEAMS

We developed a micron-CT consisting of micro-beam system and X-ray CCD camera (Hamamatsu photonics C8800X), whose element size is 8μm×8μm and a total number of image elements 1000×1000 gives an image size of 8mm×8mm. The sample is placed in a tube of a small diameter, which is rotated by a stepping motor. The transmission data through the sample are taken with characteristic Ti-K-X-rays (4.558 keV) produced by 3MeV proton and α particle micro-beams. After image reconstruction using an iteration method, 3D-images of small objects namely, hair and small ants were obtained with a spatial resolution of ~5μm. It is expected that our micron-CT can provide cross sectional images of in-vivo cellular samples with high resolution and can be applied to a wide range of research in biology and medicine.

Microbeam Analysis at Tohoku University for Biological Studies

A microbeam analysis system at Tohoku University has been improved in detection efficiency for application to single cell analysis. The system is applicable to STIM analysis and to simultaneous PIXE and RBS analysis. Sample preparation methods suitable for non-adhesive single cell analysis were developed and first results with the improved analysis system are shown.

MICROBEAM ANALYSIS OF SINGLE AEROSOL PARTICLES AT TOHOKU UNIVERSITY

A microbeam system has been developed for the analysis of single aerosol particles. Combination of PIXE, RBS and off-axis STIM methods enabled simultaneous analysis for hydrogen to metal elements. Aerosol particles were collected on thin polycarbonate film (~0.3 μm) resulting in good signal-to-noise ratio. Quantitative elemental correlation was measured for single aerosol particles. A total of 270 particles were analyzed and clustered into 4 groups. The analysis system reveals the chemical composition of aerosol particles and is a powerful tool for source identification.

DEVELOPMENT OF AN IMAGE RECONSTRUCTION METHOD FOR MICRON-CT USING PIXE

A prototype of micron-CT for biological research is being developed at Tohoku University. This micron-CT uses a point X-ray source with a spot size of 1μm and an X-ray CCD with 1000×1000 pixels of 8μm×8μm, achieving a spatial resolutions of the order of micro-meter. The event data obtained by the X-ray CCD is statistically poor and the 3 dimensional filtered back projection (3D FBP) algorithm, generally used in image reconstruction of X-ray CT, is not suitable because it is highly sensitive to statistical noise. Hence, we applied the expectation maximization (EM) algorithm for image reconstruction and developed an image reconstruction method using 3D EM algorithm. To confirm the validity of the reconstruction method, we irradiated two hairs inside a micro tube and reconstructed the CT image applying both EM and FBP algorithm on projection data. With 200×200×200 voxels of 4μm×4μm×4μm in the field of view, the computation time was less than 2 mins per iteration on a DELL Precision 650 Workstation 3.2GHz. The resulting EM image showed a better contrast than FBP image, and in the EM reconstructed CT image, we were able to reconstruct the micro tube of 270μm diameter and 45μm wall thickness and to visualize the two hairs inside.

Characterization of Si p-i-n diode for scanning transmission ion microanalysis of biological samples

The performance of a silicon p-i-n diode (Hamamatsu S1223-01) for the detection of charged particles was investigated and compared with the response of a standard passivated implanted planar silicon (PIPS) detector. The photodiode was characterized by ion beam induced charge collection with a micrometer spatial resolution using proton and alpha particle beams in the 1–3MeV energy range. Results indicate that homogeneity, energy resolution, and reproducibility of detection of charged particles enable the use of the low cost silicon p-i-n device as a replacement of conventional PIPS detector during scanning transmission ion microanalysis experiments. The Si p-i-n diode detection setup was successfully applied to scanning transmission ion microscopy determination of subcellular compartments on human cancer cultured cells.

In-Vivo Elemental Analysis by PIXE-μ-CT

We have developed “micron-CT”, using micro-PIXE for in-vivo imaging. This system comprises an X-ray CCD camera (Hamamatsu photonics C8800X9) with high resolution (pixel size: 8×8μm2, number of pixels: 1000×1000) and an X-ray-point-source with a spot size of 1.5×1.5μm2 which is generated by irradiation of a microbeam on a pure metal target. Thus we can acquire projection data with high resolution. The sample is placed in a small diameter tube and is rotated by a stepping motor. The 3D images were reconstructed from the obtained projection data by using cone-beam CT reconstruction algorithm. X-ray spectra produced by heavy charged particle bombardment, exhibit a much smaller continuous background compared to electron bombardment. Therefore, X-rays produced by ion beam can be used as a monochromatic and low energy X-ray source. The feature is very effective to investigate small insects. Moreover we can get elemental distribution image of object by choosing appropriate characteristic X-rays corresponding to the absorption edge. On the other hand, the conventional X-ray CT, in which continuous X-rays are used, provides images of the electron density in the object. Using this system, we were able to get 3D images of a living ants head with 6 μm spatial resolution. By using Fe-K-X-rays (6.40 keV) and Co-K-X-rays (6.93 keV), we can investigate the 3D distribution of Mn (K-absorption edge = 6.54 keV) in an ants head.

Development of an In-air on/off Axis STIM System for Quantitative Elemental Mapping

We have developed an in-air on/off axis STIM for simultaneous density mapping with PIXE and RBS, which will be useful for damage-monitoring in cell analysis and for yield correction based on the thickness distribution of X-ray self-absorption in samples. The in-air on/off axis STIM system provides a mass concentration map in the cell analysis. In the system, a thin scattering foil is placed downstream of the sample and scattered protons are detected by a Si-PIN photodiode set at 30 degrees with respect to the beam axis. These components are set in a He-gas-filled chamber to reduce energy loss, scattering and sample damage. Using this system, areal density mapping is carried out for RBL-2H3 cells simultaneously with PIXE and RBS. Correction for self-absorption is performed and areal density map of elements is converted into a mass-concentration map using the measured matrix density. The areal density distribution of P corresponds to that of matrix and mass concentration of P is uniform in the cell region. On the other hand, Br is concentrated in the nucleus, even in the mass concentration map. The Br accumulation in the nucleus is first confirmed in mass concentration using the on/off axis STIM and PIXE system. The in-air on/off STIM system will be effective for monitoring changes in cell density during beam irradiation.

BEAM DAMAGE OF CELLULAR SAMPLES IN IN-AIR MICRO PIXE ANALYSIS

The change of shape and elemental concentration of cellular samples in In-Air micro PIXE analysis was investigated. Cultured bovine aortic endothelial cells were analyzed in the atmosphere by using 2.6 MeV proton micro-beams. The shape of cross-sections of cells was not so much distorted by beam irradiation and the concentrations of trace elements did not change too, except for S. The concentration of S changed with strongly depending on the temperature rise due to beam irradiation.

CHARACTERIZATION OF AEROSOL PARTICLES IN A MECHANICAL WORKSHOP ENVIRONMENT

Indoor aerosols are directly affecting human lives. Especially aerosols in workshops, factories, and laboratories, where many chemical substances are used in treatment and production processes, might contain toxic elements: special care must be taken to alleviate air pollution and assure a clean breathing environment for the workers. For this study, size segregated aerosol particle sampling with a cascade impactor was performed in the machine workshop of Jožef Stefan Institute. The samples, collected during weekdays and weekend were analyzed with a microbeam facility at Tohoku University. Bulk PIXE analysis with scanning over the whole sample area was conducted along with multimodal microanalysis with microscopic scanning. Using bulk analysis, high concentrations of Pb and Ba were detected on weekend days, which was related to the removal of an old white paint from the furniture. On weekdays, concentrations of W and of soil origin elements increased, probably because of the machine operations and worker movements. At the same time high concentration of sulfur was detected. A microscopic multimodal analysis shows that it stems from a lubricant oil vapor. The combination of bulk and microanalysis of the size selected samples is an effective approach to aerosol characterization in the working environment.