Ion Microbeam Irradiation Research Articles

The dual ion beam microprobe

A new experimental setup for dual ion microbeam irradiation and analysis of materials has been designed and commissioned at the Ruđer Bošković Institute (RBI). Ion beams in the MeV energy range are provided by two tandem accelerators, a 1.0 MV Tandetron and a 6.0 MV EN tandem Van de Graaff. The dual microprobe end station is primarily dedicated for the performance of experiments where one ion microbeam is used for the irradiation, while the other ion microbeam is used for the analysis or ‘probing’ the in-situ changes induced during the target irradiation. Likewise, both microbeams can be also used independently, for experiments involving either ion beam characterization or ion beam irradiation techniques. The ion beam from the small tandem accelerator is focused with a magnetic quadrupole triplet, which is more suitable for the high spatial resolution work with light ions. The ion beam from the larger accelerator is focused by an electrostatic quadruplet that enables easier focusing of heavy ions. Details of the experimental setup, its capabilities in terms of ion beam irradiation and analysis techniques, as well as the first applications, are presented.

High turnover and rescue effect of XRCC1 in response to heavy charged particle radiation

The DNA damage response is a highly orchestrated process. The involvement of the DNA damage response factors in DNA damage response depends on their biochemical reactions with each other and with chromatin. Using online live-cell imaging combined with heavy ion microbeam irradiation, we studied the response of the scaffold protein X-ray repair cross-complementary protein 1 (XRCC1) at the localized DNA damage in charged particle irradiated HT1080 cells expressing XRCC1-tagged RFP. The results showed that XRCC1 was recruited to the DNA damage with ultrafast kinetics in a poly ADP-ribose polymerase-dependent manner. The consecutive reaction model well explained the response of XRCC1 at ion hits, and we found that the XRCC1 recruitment was faster and dissociation was slower in the G2 phase than those in the G1 phase. The fractionated irradiation of the same cells resulted in accelerated dissociation at the previous damage sites, and the dissociated XRCC1 immediately recycled with a higher recruitment efficiency. Our data revealed XRCC1’s new rescue mechanism and its high turnover in DNA damage response, which benefits our understanding of the biochemical mechanism in DNA damage response.

Focused Ion Microbeam Irradiation Induces Clustering of DNA Double-Strand Breaks in Heterochromatin Visualized by Nanoscale-Resolution Electron Microscopy.

Background: Charged-particle radiotherapy is an emerging treatment modality for radioresistant tumors. The enhanced effectiveness of high-energy particles (such as heavy ions) has been related to the spatial clustering of DNA lesions due to highly localized energy deposition. Here, DNA damage patterns induced by single and multiple carbon ions were analyzed in the nuclear chromatin environment by different high-resolution microscopy approaches. Material and Methods: Using the heavy-ion microbeam SNAKE, fibroblast monolayers were irradiated with defined numbers of carbon ions (1/10/100 ions per pulse, ipp) focused to micrometer-sized stripes or spots. Radiation-induced lesions were visualized as DNA damage foci (γH2AX, 53BP1) by conventional fluorescence and stimulated emission depletion (STED) microscopy. At micro- and nanoscale level, DNA double-strand breaks (DSBs) were visualized within their chromatin context by labeling the Ku heterodimer. Single and clustered pKu70-labeled DSBs were quantified in euchromatic and heterochromatic regions at 0.1 h, 5 h and 24 h post-IR by transmission electron microscopy (TEM). Results: Increasing numbers of carbon ions per beam spot enhanced spatial clustering of DNA lesions and increased damage complexity with two or more DSBs in close proximity. This effect was detectable in euchromatin, but was much more pronounced in heterochromatin. Analyzing the dynamics of damage processing, our findings indicate that euchromatic DSBs were processed efficiently and repaired in a timely manner. In heterochromatin, by contrast, the number of clustered DSBs continuously increased further over the first hours following IR exposure, indicating the challenging task for the cell to process highly clustered DSBs appropriately. Conclusion: Increasing numbers of carbon ions applied to sub-nuclear chromatin regions enhanced the spatial clustering of DSBs and increased damage complexity, this being more pronounced in heterochromatic regions. Inefficient processing of clustered DSBs may explain the enhanced therapeutic efficacy of particle-based radiotherapy in cancer treatment.

Microcapacitors on graphene oxide and synthetic polymers prepared by microbeam lithography

Carbon-ion microbeam writing was employed for the mask-less production of microscale capacitors in insulating graphene oxide (GO), polyimide (PI) and poly(methyl-methacrylate) (PMMA) foils. The substrates were irradiated by a 5 MeV C beams with micrometer-scale resolution to create conductive strips. The morphology and quality of the created microstructures and compositional changes in the host matrix under the ion microbeam irradiation were studied using scanning electron microscopy and energy-dispersive X-ray spectroscopy. The changes in the structure and elemental composition of the irradiated areas were characterised by Raman micro-spectroscopy, X-ray photoelectron spectroscopy, Rutherford backscattering spectroscopy and elastic recoil detection analysis. The microcapacitors with the highest capacitance (in the order of pF) were those prepared on the GO surface. On the other hand, in PI and PMMA, the same carbon-ion irradiation does not induce such a significant enhancement of electric properties and the capacity of the resulting capacitor-like structures is substantially lower.

Micro ion beam used to optimize the quality of microstructures based on polydimethylsiloxane

Ion micro beam was employed to fabricate microstructures based on poly(dimethylsiloxane) (PDMS). Carbon ions with energy of 6.0 MeV were used. The ion treatment induces modifications in PDMS surface and on the optical properties as a function of the ion current and fluence. PDMS foils were produced from commercially available two component addition materials. The selective modifications of the polymer were studied for applications in micro-fluids, micro-optics and micro-electronics. The comparison between the unirradiated and irradiated PDMS was investigated in morphology and compactness using the atomic force microscopy. The wetting ability of the PDMS surface was also analyzed as a function of the ion microbeam irradiation. The compositional and optical changes in PDMS were monitored by Rutherford backscattering spectrometry, elastic recoil detection analysis and optical spectroscopy measurements.

In situ irradiation and IBIC probing using pulsed and dual ion microbeams

We present two new experimental arrangements designed to study time dependent processes during the irradiation of semiconductor detectors using ion microbeams. The first one is based on an upgraded pulsed beam irradiation setup of the RBI heavy ion microprobe. Controlled time-sequences of sample irradiation and intermediate IBIC probing is used to study dynamics of degradation of charge transport in detectors at different time scales. The second experimental arrangement is a dual ion microbeam irradiation setup that can accept simultaneously the ion beams from the two accelerators. In that case the first ion microbeam may be used as a damaging beam while the second one could be used as a probing beam. Probing techniques that have been used to study induced changes in crystalline materials are based on low microbeam currents and include STIM channeling, IBIC and IL.

From Energy Deposition of Ionizing Radiation to Cell Damage Signaling: Benchmarking Simulations by Measured Yields of Initial DNA Damage after Ion Microbeam Irradiation.

Advances in accelerator technology, which have enabled conforming radiotherapy with charged hadronic species, have brought benefits as well as potential new risks to patients. To better understand the effects of ionizing radiation on tumor and surrounding tissue, it is important to investigate and quantify the relationship between energy deposition at the nanometric scale and the initial biological events. Monte Carlo track structure simulation codes provide a powerful tool for investigating this relationship; however, their success and reliability are dependent on their improvement and development accordingly to the dedicated biological data to which they are challenged. For this aim, a microbeam facility that allows for fluence control, down to one ion per cell nucleus, was used to evaluate relative frequencies of DNA damage after interaction between the incoming ion and DNA according to radiation quality. Primary human cells were exposed to alpha particles of three different energies with respective linear energy transfers (LETs) of approximately 36, 85 or 170 keV·µm-1 at the cells' center position, or to protons (19 keV·µm-1). Statistical evaluation of nuclear foci formation (53BP1/γ-H2AX), observed using immunofluorescence and related to a particle traversal, was undertaken in a large population of cell nuclei. The biological results were adjusted to consider the factors that drive the experimental uncertainties, then challenged with results using Geant4-DNA code modeling of the ionizing particle interactions on a virtual phantom of the cell nucleus with the same mean geometry and DNA density as the cells used in our experiments. Both results showed an increase of relative frequencies of foci (or simulated DNA damage) in cell nuclei as a function of increasing LET of the traversing particles, reaching a quasi-plateau when the LET exceeded 80-90 keV·µm-1. For the LET of an alpha particle ranging from 80-90 to 170 keV·µm-1, 10-30% of the particle hits did not lead to DNA damage inducing 53BP1 or γ-H2AX foci formation.

Development of laser target sight-on system based on multiple transmission through a tapered glass capillary for ion microbeam irradiation

SynopsisAn estimation method of micro-meter sized laser profile created by the laser beam transmission through a tapered glass capillary is developed for the target sight-on system of ion microbeam irradiation to living cells. In our system, both the laser and the ions are designed to be transmitted through the same capillary so that the laser spot is surely on the ion track. By selecting the excitation light of a fluorescent protein in cell nucleus, the fluorescent volume can be confirmed as the targeted area.

Nitric oxide-mediated bystander signal transduction induced by heavy-ion microbeam irradiation

In general, a radiation-induced bystander response is known to be a cellular response induced in non-irradiated cells after receiving bystander signaling factors released from directly irradiated cells within a cell population. Bystander responses induced by high-linear energy transfer (LET) heavy ions at low fluence are an important health problem for astronauts in space. Bystander responses are mediated via physical cell–cell contact, such as gap-junction intercellular communication (GJIC) and/or diffusive factors released into the medium in cell culture conditions. Nitric oxide (NO) is a well-known major initiator/mediator of intercellular signaling within culture medium during bystander responses. In this study, we investigated the NO-mediated bystander signal transduction induced by high-LET argon (Ar)-ion microbeam irradiation of normal human fibroblasts. Foci formation by DNA double-strand break repair proteins was induced in non-irradiated cells, which were co-cultured with those irradiated by high-LET Ar-ion microbeams in the same culture plate. Foci formation was suppressed significantly by pretreatment with an NO scavenger. Furthermore, NO-mediated reproductive cell death was also induced in bystander cells. Phosphorylation of NF-κB and Akt were induced during NO-mediated bystander signaling in the irradiated and bystander cells. However, the activation of these proteins depended on the incubation time after irradiation. The accumulation of cyclooxygenase-2 (COX-2), a downstream target of NO and NF-κB, was observed in the bystander cells 6 h after irradiation but not in the directly irradiated cells. Our findings suggest that Akt- and NF-κB-dependent signaling pathways involving COX-2 play important roles in NO-mediated high-LET heavy-ion-induced bystander responses. In addition, COX-2 may be used as a molecular marker of high-LET heavy-ion-induced bystander cells to distinguish them from directly irradiated cells, although this may depend on the time after irradiation.

Radioprotection of targeted and bystander cells by methylproamine.

IntroductionRadioprotective agents are of interest for application in radiotherapy for cancer and in public health medicine in the context of accidental radiation exposure. Methylproamine is the lead compound of a class of radioprotectors which act as DNA binding anti-oxidants, enabling the repair of transient radiation-induced oxidative DNA lesions. This study tested methylproamine for the radioprotection of both directly targeted and bystander cells.MethodsT98G glioma cells were treated with 15 μM methylproamine and exposed to 137Cs γ-ray/X-ray irradiation and He2+ microbeam irradiation. Radioprotection of directly targeted cells and bystander cells was measured by clonogenic survival or γH2AX assay.ResultsRadioprotection of directly targeted T98G cells by methylproamine was observed for 137Cs γ-rays and X-rays but not for He2+ charged particle irradiation. The effect of methylproamine on the bystander cell population was tested for both X-ray irradiation and He2+ ion microbeam irradiation. The X-ray bystander experiments were carried out by medium transfer from irradiated to non-irradiated cultures and three experimental designs were tested. Radioprotection was only observed when recipient cells were pretreated with the drug prior to exposure to the conditioned medium. In microbeam bystander experiments targeted and nontargeted cells were co-cultured with continuous methylproamine treatment during irradiation and postradiation incubation; radioprotection of bystander cells was observed.Discussion and conclusionMethylproamine protected targeted cells from DNA damage caused by γ-ray or X-ray radiation but not He2+ ion radiation. Protection of bystander cells was independent of the type of radiation which the donor population received.

Magnetic patterning of FeRh thin films by energetic light ion microbeam irradiation

Magnetic patterning of FeRh thin films of various shapes and dimensions by 2 MeV H and He ion microbeam irradiation was examined by X-ray magnetic circular dichroism photoemission electron microscopy (XMCD-PEEM) at the photon energy of the Fe L3 absorption edge. The XMCD-PEEM images clearly show that the various magnetic patterns whose sizes are about 1–2 µm, can be successfully produced by controlling the scan condition and the size of the ion beam probe. Furthermore, the quantitative analysis of the XMCD-PEEM images reveals that the localized magnetization in the micrometer-sized magnetic patterns significantly depends on the irradiation ion fluence. In addition, the annealing condition is also found to be one of the predominant factors that determine the magnetic state even for the micrometer-sized magnetic patterns.

Continuous observation of polarization effects in thin SC-CVD diamond detector designed for heavy ion microbeam measurement

Continuous irradiation effects on a thin-film diamond detector were investigated for the utilization of these films as a detector for heavy ion microbeams. Temporal signal degradation in the energy spectrum was frequently observed during the focused heavy ion microbeam irradiation. To measure the temporal response to the each ion incidents, focused heavy ion microbeam with different beam fluence rates were irradiated to a single crystal (SC)-CVD diamond film detector with thickness of 50μm. The responses to each ion were continuously observed and characterized by ion beam-induced charge (IBIC) measurement system. Heavy ions with short penetration path in diamond generate the large difference in mean path of electrons and holes, which is inverted by changing bias polarity. Signal degradation condition was relied on the bias polarity under the irradiation of heavy ions with short penetration length in the diamond. The continuous observation of IBIC signals revealed that temporal degradation in pulse height of signals, so called polarization effects, seems to be mainly caused by the hole trapping in this diamond crystal.

MFM and PEEM observation of micrometre-sized magnetic dot arrays fabricated by ion-microbeam irradiation in FeRh thin films

FeRh thin films were irradiated with a 10 MeV iodine ion microbeam to produce micrometre-scale ferromagnetic microstructures by modifying the local magnetic character from antiferromagnetism to ferromagnetism using ion-microbeam irradiation. Two-dimensional magnetic dot arrays of dimensions ∼2 µm × 4 µm as well as 10 µm × 10 µm were successfully produced on the FeRh surface, which was confirmed by magnetic force microscopy (MFM). The results of photoemission electron microscopy (PEEM) combined with X-ray magnetic circular dichroism reveal that the easy axis of the magnetization of the ion-beam-irradiated ferromagnetism in the FeRh thin films lies in the film planes along the <001> direction of the MgO substrates.

High energy heavy ion microbeam irradiation facility at IMP

A high energy heavy ion microbeam irradiation facility was designed and installed in the Institute of Modern Physics (IMP) of Chinese Academy of Sciences (CAS). It can deliver well-focused ions into the targets at exact locations. With the advantage of allowing vertical irradiation as well as focusing ions from Carbon to Uranium with high energies in a broad energy range (7MeV/u to 100MeV/u for Carbon ions), this setup is able to deliver a preset number of ions into pre-selected target positions. Material specimens in vacuum and living cells in air can also be irradiated by this facility. Detailed description of this microbeam facility as well as its beam optics is presented in this paper. Some preliminary test results are also given.

Ion microbeam irradiation for radiobiology and radical chemistry: status and prospect

Ion microbeams are commonly used to study local irradiation effects in living cells, as it has been established that ion beam irradiations can lead to deleterious changes in cells that are not struck directly by the microbeam. Such changes, which take place over distances long compared to the size of the irradiation spot and for times long compared to the time of irradiation, are collectively termed radiation-induced bystander effect or RIBE. Free-radical chemistry is frequently invoked to explain the RIBE but no unified model is available at present. Ion microbeams when coupled with advanced methods for observing free radicals are the tools of choice for investigating the chemistry and biological processes governing RIBE.

Study of the aberrations of the IMP heavy ion microbeam irradiation system

A high energy heavy ion microbeam irradiation system is constructed at the Institute of Modern Physics (IMP) of the Chinese Academy of Sciences (CAS). A quadrupole focusing system, in combination with a series of slits, has been designed here. The IMP microbeam system is described in detail. The intrinsic and parasitic aberrations associated with the magnets are simulated. The ion beam optics of this microbeam system is investigated systematically. Then the optimized initial beam parameters are given for high spatial resolution and high hitting rates.

Design of the IMP microbeam irradiation system for 100 MeV/u heavy ions

A state-of-the-art high energy heavy ion microbeam irradiation system is constructed at the Institute of Modern Physics of the Chinese Academy of Sciences. This microbeam system operates in both full current intensity mode and single ion mode. It delivers a predefined number of ions to pre-selected targets for research in biology and material science. The characteristic of this microbeam system is high energy and vertical irradiation. A quadrupole focusing system, in combination with a series of slits, has been designed to optimize the spatial resolution. A symmetrically achromatic system leads the beam downwards and serves simultaneously as an energy analyzer. A high gradient quadrupole triplet finally focuses a C6+ ion beam to 1 μm in the vacuum chamber within the energy range from 10 MeV/u to 100 MeV/u. In this paper, the IMP microbeam system is described in detail. A systematic investigation of the ion beam optics of this microbeam system is presented together with the associated aberrations. Comparison is made between the IMP microbeam system and the other existing systems to further discuss the performance of this microbeam. Then the optimized initial beam parameters are given for high resolution and high hitting efficiency. At last, the experiment platform is briefly introduced.

Modification of structural and optical properties of silica glass induced by ion microbeam

Structural and optical properties of silica glass induced by ion microbeam irradiation were studied using micro- photoluminescence (μ-PL) spectroscopy and atomic force microscopy (AFM). Ion microbeam irradiation was performed using microbeam lines of 3-MV single-ended or tandem accelerators with various ion species including H +, He +, B 3+, C 4+, N 4+, O 4+, and Si 5+ at energies of 1.7 to 18 MeV. The beam was focused to about 1 μm and was scanned over the surface of high-purity silica glass with fluences of 10 13 to 10 18 ions/cm 2. The μ-PL spectrum in microbeam-irradiated silica shows two peaks at 540 and 650 nm. The mapping of the two PL bands reveals the distribution of defects induced along the track of ions. The compaction was observed in the form of groove at the surface of silica glass by AFM. The depth of the surface groove increases with increasing ion fluence and saturates at about several hundreds to 900 nm, depending on irradiated ion species. The mechanisms of structural and optical modifications of silica glass are can be understood in terms of energy loss due to electronic stopping and nuclear stopping powers. Refractive index changes with an order of 10 – 4 to 10 − 2 were estimated by a Lorentz–Lorenz relationship from the compaction. Technological implications of these results are also to be discussed.

Heavy Ion Microbeam Irradiation Induces Ultrastructural Changes in Isolated Single Fibers of Skeletal Muscle

The effects of heavy ion microbeams on muscle fibers isolated from mouse skeletal muscles were examined by electron microscopy. The plasma membranes of heavy ion beam-irradiated areas of muscle fibers showed irregular protrusions and invaginations. In the cytoplasm, an irregular distribution of microfilaments was found near the plasma membrane. Sarcoplasmic reticula in the irradiated regions showed a distended appearance with flocculent material within the lumen. These changes were seen as early as 2 min after irradiation, and persisted until as late as 22 min after irradiation. Many autophagic vacuoles could be seen at 7 min after irradiation. At 22 min, the vacuoles became more prominent and showed more variety. These observations suggest that heavy ion beam irradiation causes disruption of the cellular architecture and the autophagy is involved in removal of this disruption.

Biological effects of heavy ion microbeam irradiation to single tobacco BY-2 cells

Biological effects of heavy ion microbeam irradiation to single tobacco BY-2 cells