0000000000024456
AUTHOR
Frank Watt
Objective improvement of the visual quality of ion microscope images
The need to operate with low ion beam fluences implies the images obtained using ion microscope (IM) are often grainy and have poor visual quality compared to what can be obtained using e.g. confocal microscopy. This results from the Poissonian distribution of counts in pixels. Here we report work on some different approaches for objectively improving the visual quality of IM images. In this work we present (i) dramatic improvement in the visual image quality of off-axis and direct-scanning transmission ion microscopy (STIM) images by suppression of zero-pixels; (ii) denoising of PIXE images using wavelet filtering and (iii) use of the feature preserving characteristics of wavelet filtering…
Adhesion of proton beam written high aspect ratio hydrogen silsesquioxane (HSQ) nanostructures on different metallic substrates
Abstract Hydrogen silsesquioxane (HSQ) behaves as a negative resist under MeV proton beam exposure. HSQ is a high-resolution resist suitable for production of tall (
WITHDRAWN: Measurement of beam focus quality in biomedical nuclear microscopy.
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Exploratory nuclear microprobe data visualisation using 3- and 4-dimensional biological volume rendering tools
Abstract The emergence of Confocal Microscopy (CM) and Atomic Force Microscopy (AFM) as everyday tools in cellular level biology has stimulated development of 3D data visualisation software. Conventional 2-dimensional images of cell (optical) sections obtained in a transmission electron or optical microscopes and more sophisticated multidimensional imaging methods require processing software capable of 3D rendering and mathematically transforming data in 3-, 4-, or more dimensions. The richness of data obtained from the different nuclear microscopy imaging techniques and often parallel information channels (X-ray, secondary electron, Scanning Transmission Ion Microscopy) is often not obviou…
Development of elastomeric lab-on-a-chip devices through Proton Beam Writing (PBW) based fabrication strategies
Abstract In recent years, one of the most exciting developments in fluidic device applications is the rapid evolution of miniaturized micro- and nanofluidic systems, the so called “lab-on-a-chip” devices. These devices integrate laboratory functions into a single chip, and are capable of various biochemical analysis and synthesis, such as sample injection and preparation, single cell/molecule observation, bioparticle sequencing and sorting etc. The evolvement of lab-on-a-chip concept implies the use of novel fabrication techniques for the construction of versatile analytical components in a fast and reproducible manner. Endowed with unique three-dimensional fabrication abilities, Proton Bea…
Nano-imaging of single cells using STIM
Scanning transmission ion microscopy (STIM) is a technique which utilizes the energy loss of high energy (MeV) ions passing through a sample to provide structural images. In this paper, we have successfully demonstrated STIM imaging of single cells at the nano-level using the high resolution capability of the proton beam writing facility at the Centre for Ion Beam Applications, National University of Singapore. MCF-7 breast cancer cells (American Type Culture Collection [ATCC]) were seeded on to silicon nitride windows, backed by a Hamamatsu pin diode acting as a particle detector. A reasonable contrast was obtained using 1 MeV protons and excellent contrast obtained using 1 MeV alpha parti…
Characterisation of beam focus quality in biomedical nuclear microscopy: A Fourier optics approach
Abstract The central peak widths of the 2D-autocorrelation function have been investigated as a Figure of Merit (FoM) of focus quality in nuclear microscopy using a quadrupole triplet lens system. The beam focus could be reliably characterised for direct scanning transmission ion microscopy (direct-STIM) images obtained with fluences as small as 5.8 × 10 9 ions cm - 2 which colocalisation tests showed did not introduce significant beam-induced changes in the cells.
Proton beam written hydrogen silsesquioxane (HSQ) nanostructures for Nickel electroplating
Abstract Hydrogen silsesquioxane (HSQ) behaves as a negative resist under MeV proton beam exposure. HSQ is a high-resolution resist suitable for production of tall ( 1.5 μ m ) high-aspect-ratio nanostructures with dimensions down to 22 nm. High-aspect-ratio HSQ structures are required in many applications, e.g. nanofluidics, biomedical research, etc. Since P-beam writing is a direct and hence slow process, it is beneficiary to fabricate a reverse image of the patterns in a metallic stamp, e.g. by Ni electroplating. The Ni stamp can then be used to produce multiple copies of the same pattern. In this study we investigate the possibility to produce Ni stamps from p-beam written HSQ samples. H…
Angular spreading measurements using MeV ion microscopes
Abstract The sharpness of MeV ion microscope images is governed by small-angle scattering and associated lateral spreading of the ion beam in the sample. We have investigated measurement of the half-angle of the angular spreading distribution by characterising the image blurring in direct-Scanning Transmission Ion Microscopy (direct-STIM). In these tests Mylar™ foils of 0.5–6 μm were used to induce angular spreading. Images were taken of an electron microscope grid using 2 MeV protons with, and without, the foils in the beam path. The blurring was measured by fitting the width of a circular Gaussian point spread function to the images with and without the foil in position. The results show …
Ion-induced fluorescence imaging of endosomes
Abstract Imaging laboratories at Jyvaskyla and Singapore are collaborating on the development of fluorescence imaging of cytoplasmic endosomes using a combination of proton induced fluorescence (PIF) with direct Scanning Transmission Ion Microscopy (direct-STIM) for sub-cellular structural imaging. A549 lung carcinoma cells were cultivated and stained for epidermal growth factor receptor (EGFR) and receptor α2β1 integrin. In this paper, we demonstrate that cells can be imaged at sub-150 nm resolution using the PIF technique. In addition, the same target cell was imaged at 50 and 25 nm resolution by using proton and He-STIM, respectively. The combination of both techniques offer a powerful t…
Angular and lateral spreading of ion beams in biomedical nuclear microscopy
Abstract Nuclear scattering from target atoms gives rise to a spatial broadening of energetic ion beams penetrating matter. The spatial broadening of the ion beam presents an ultimate limit to the resolving power that can be achieved in nuclear microscopy methods. The pressing of the attainable resolution limit in biomedical nuclear microscopy to dimensions approaching 10 nm, or so, implies the fundamental limitation from ion-target scattering will become increasingly significant. This effect has been investigated by a combined analytical and numerical computational approach to determine the extent and how single and multiple scattering processes limit the resolution for analysis with 2 MeV…