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Microbeam Radiation Biology and Its State-of-the-Art Technology
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Microbeam Radiation Biology and Its State-of-the-Art Technology

A special issue of Biology (ISSN 2079-7737). This special issue belongs to the section "Biotechnology".

Deadline for manuscript submissions: closed (30 September 2023) | Viewed by 25767

Special Issue Editors

Dr. Michiyo Suzuki

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Guest Editor
Quantum-Applied Biotechnology Project, Department of Quantum-Applied Biosciences, Takasaki Institute for Advanced Quantum Science, Foundational Quantum Technology Research Directorate, National Institutes for Quantum Science and Technology (QST), Takasaki 370-1292, Gunma, Japan
Interests: heavy-ion microbeam; neuroethology; C. elegans; targeted-irradiation technology; anesthesia-free irradiation method
Dr. Teruaki Konishi

E-Mail Website
Guest Editor
Single Cell Radiation Biology Group, Institute for Quantum Life Science, National Institutes for Quantum and Radiological Science and Technology, 4-9-1 Anagawa, Inage 263-8555, Chiba, Japan
Interests: radiation biology; single cell biology; microbeam; heavy ions; radiation induced bystander effect; cytoplasmic damage response; DNA damage and repair
Special Issues, Collections and Topics in MDPI journals
Dr. Tomoo Funayama

E-Mail Website
Guest Editor
Quantum-Applied Biotechnology Project, Department of Quantum-Applied Biosciences, Takasaki Institute for Advanced Quantum Science, Foundational Quantum Technology Research Directorate, National Institutes for Quantum Science and Technology (QST), Takasaki 370-1292, Gunma, Japan
Interests: heavy-ion microbeam; single cell irradiation; radiation induced bystander effect; dosimetry; targeted irradiation technology

Special Issue Information

Dear Colleagues,

Microbeam irradiation technology has great potential as a powerful tool for the analysis of cell- or tissue-specific radiation effects, as well as for radiotherapy. This Special Issue focuses mainly on microbeam irradiation studies using cultured cells and model animals to clarify the effects of radiation at the molecular (DNA), cell, tissue, or individual level. Specifically, we aim to compare and systematically discuss various radiation effects such as the bystander effect, abscopal effect, and individual-level effects of microbeam irradiation. We also welcome reports regarding the latest progress in the development of microbeam irradiation technology for various radiations, such as heavy ions, protons, and X-ray photons. It evaluates the potential value of microbeams for both biological research and radiation therapy, and contributes to facilitating discussions on radiobiology with microbeam state-of-the-art technology.

Dr. Michiyo Suzuki
Dr. Teruaki Konishi
Dr. Tomoo Funayama
Guest Editors

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Keywords

  • microbeam
  • heavy-ion beam
  • proton beam
  • X-ray photons
  • bystander effect
  • abscopal effect
  • targeted effect
  • non-targeted effect
  • irradiation device development
  • radiotherapy
  • radiosurgery

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Published Papers (12 papers)

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Research

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11 pages, 2233 KiB  
Article
Primary and Secondary Bystander Effects of Proton Microbeam Irradiation on Human Lung Cancer Cells under Hypoxic Conditions
by Narongchai Autsavapromporn, Alisa Kobayashi, Cuihua Liu, Aphidet Duangya, Masakazu Oikawa, Tengku Ahbrizal Tengku Ahmad and Teruaki Konishi
Biology 2023, 12(12), 1485; https://doi.org/10.3390/biology12121485 - 3 Dec 2023
Cited by 1 | Viewed by 1971
Abstract
Tumor hypoxia is the most common feature of radioresistance to the radiotherapy (RT) of lung cancer and results in poor clinical outcomes. High-linear energy transfer (LET) radiation is a novel RT technique to overcome this problem. However, a limited number of studies have [...] Read more.
Tumor hypoxia is the most common feature of radioresistance to the radiotherapy (RT) of lung cancer and results in poor clinical outcomes. High-linear energy transfer (LET) radiation is a novel RT technique to overcome this problem. However, a limited number of studies have been elucidated on the underlying mechanism(s) of RIBE and RISBE in cancer cells exposed to high-LET radiation under hypoxia. Here, we developed a new method to investigate the RIBE and RISBE under hypoxia using the SPICE-QST proton microbeams and a layered tissue co-culture system. Normal lung fibroblast (WI-38) and lung cancer (A549) cells were exposed in the range of 06 Gy of proton microbeams, wherein only ~0.04–0.15% of the cells were traversed by protons. Subsequently, primary bystander A549 cells were co-cultured with secondary bystander A549 cells in the presence or absence of a GJIC and NO inhibitor using co-culture systems. Studies show that there are differences in RIBE in A549 and WI-38 primary bystander cells under normoxia and hypoxia. Interestingly, treatment with a GJIC inhibitor showed an increase in the toxicity of primary bystander WI-38 cells but a decrease in A549 cells under hypoxia. Our results also show the induction of RISBE in secondary bystander A549 cells under hypoxia, where GJIC and NO inhibitors reduced the stressful effects on secondary bystander A549 cells. Together, these preliminary results, for the first time, represented the involvement of intercellular communications through GJIC in propagation of RIBE and RISBE in hypoxic cancer cells. Full article
(This article belongs to the Special Issue Microbeam Radiation Biology and Its State-of-the-Art Technology)
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14 pages, 2690 KiB  
Article
The COX-2/PGE2 Response Pathway Upregulates Radioresistance in A549 Human Lung Cancer Cells through Radiation-Induced Bystander Signaling
by Alisa Kobayashi, Yota Hiroyama, Taisei Mamiya, Masakazu Oikawa and Teruaki Konishi
Biology 2023, 12(11), 1368; https://doi.org/10.3390/biology12111368 - 25 Oct 2023
Viewed by 2176
Abstract
This study aimed to determine the mechanism underlying the modulation of radiosensitivity in cancer cells by the radiation-induced bystander effect (RIBE). We hypothesized that the RIBE mediates cyclooxygenase-2 (COX-2) and its metabolite prostaglandin E2 (PGE2) in elevating radioresistance in unirradiated cells. In this [...] Read more.
This study aimed to determine the mechanism underlying the modulation of radiosensitivity in cancer cells by the radiation-induced bystander effect (RIBE). We hypothesized that the RIBE mediates cyclooxygenase-2 (COX-2) and its metabolite prostaglandin E2 (PGE2) in elevating radioresistance in unirradiated cells. In this study, we used the SPICE-QST microbeam irradiation system to target 0.07–0.7% cells by 3.4-MeV proton microbeam in the cell culture sample, such that most cells in the dish became bystander cells. Twenty-four hours after irradiation, we observed COX-2 protein upregulation in microbeam-irradiated cells compared to that of controls. Additionally, 0.29% of the microbeam-irradiated cells exhibited increased cell survival and a reduced micronucleus rate against X-ray irradiation compared to that of non-microbeam irradiated cells. The radioresistance response was diminished in both cell groups with the hemichannel inhibitor and in COX-2-knockout cells under cell-to-cell contact and sparsely distributed conditions. The results indicate that the RIBE upregulates the cell radioresistance through COX-2/PGE2 intercellular responses, thereby contributing to issues, such as the risk of cancer recurrence. Full article
(This article belongs to the Special Issue Microbeam Radiation Biology and Its State-of-the-Art Technology)
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19 pages, 2117 KiB  
Article
Targeted Central Nervous System Irradiation with Proton Microbeam Induces Mitochondrial Changes in Caenorhabditis elegans
by Ahmad Sleiman, Kévin Lalanne, François Vianna, Yann Perrot, Myriam Richaud, Tanima SenGupta, Mikaël Cardot-Martin, Pascal Pedini, Christophe Picard, Hilde Nilsen, Simon Galas and Christelle Adam-Guillermin
Biology 2023, 12(6), 839; https://doi.org/10.3390/biology12060839 - 9 Jun 2023
Cited by 4 | Viewed by 2610
Abstract
Fifty percent of all patients with cancer worldwide require radiotherapy. In the case of brain tumors, despite the improvement in the precision of radiation delivery with proton therapy, studies have shown structural and functional changes in the brains of treated patients with protons. [...] Read more.
Fifty percent of all patients with cancer worldwide require radiotherapy. In the case of brain tumors, despite the improvement in the precision of radiation delivery with proton therapy, studies have shown structural and functional changes in the brains of treated patients with protons. The molecular pathways involved in generating these effects are not completely understood. In this context, we analyzed the impact of proton exposure in the central nervous system area of Caenorhabditis elegans with a focus on mitochondrial function, which is potentially implicated in the occurrence of radiation-induced damage. To achieve this objective, the nematode C. elegans were micro-irradiated with 220 Gy of protons (4 MeV) in the nerve ring (head region) using the proton microbeam, MIRCOM. Our results show that protons induce mitochondrial dysfunction, characterized by an immediate dose-dependent loss of the mitochondrial membrane potential (ΔΨm) associated with oxidative stress 24 h after irradiation, which is itself characterized by the induction of the antioxidant proteins in the targeted region, observed using SOD-1::GFP and SOD-3::GFP strains. Moreover, we demonstrated a two-fold increase in the mtDNA copy number in the targeted region 24 h after irradiation. In addition, using the GFP::LGG-1 strain, an induction of autophagy in the irradiated region was observed 6 h following the irradiation, which is associated with the up-regulation of the gene expression of pink-1 (PTEN-induced kinase) and pdr-1 (C. elegans parkin homolog). Furthermore, our data showed that micro-irradiation of the nerve ring region did not impact the whole-body oxygen consumption 24 h following the irradiation. These results indicate a global mitochondrial dysfunction in the irradiated region following proton exposure. This provides a better understanding of the molecular pathways involved in radiation-induced side effects and may help in finding new therapies. Full article
(This article belongs to the Special Issue Microbeam Radiation Biology and Its State-of-the-Art Technology)
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11 pages, 1895 KiB  
Article
Observation of Histone H2AX Phosphorylation by Radiation-Induced Bystander Response Using Titanium Characteristic X-ray Microbeam
by Masanori Tomita, Masaya Torigata, Tadayuki Ohchi and Atsushi Ito
Biology 2023, 12(5), 734; https://doi.org/10.3390/biology12050734 - 18 May 2023
Cited by 1 | Viewed by 1637
Abstract
Radiation-induced bystander response (RIBR) is a response induced in non-irradiated cells that receive bystander signals from directly irradiated cells. X-ray microbeams are useful tools for elucidating the mechanisms underlying RIBR. However, previous X-ray microbeams used low-energy soft X-rays with higher biological effects, such [...] Read more.
Radiation-induced bystander response (RIBR) is a response induced in non-irradiated cells that receive bystander signals from directly irradiated cells. X-ray microbeams are useful tools for elucidating the mechanisms underlying RIBR. However, previous X-ray microbeams used low-energy soft X-rays with higher biological effects, such as aluminum characteristic X-rays, and the difference from conventional X-rays and γ-rays has often been discussed. The microbeam X-ray cell irradiation system at the Central Research Institute of Electric Power Industry has been upgraded to generate higher energy titanium characteristic X-rays (TiK X-rays), which have a longer penetration distance sufficient to irradiate 3D cultured tissues. Using this system, we irradiated the nuclei of HeLa cells with high precision and found that the pan-nuclear induction of phosphorylated histone H2AX on serine 139 (γ-H2AX) in the non-irradiated cells increased 180 and 360 min after irradiation. We established a new method to quantitatively evaluate bystander cells, using the fluorescence intensity of γ-H2AX as an indicator. The percentage of bystander cells increased significantly to 23.2% ± 3.2% and 29.3% ± 3.5% at 180 and 360 min after irradiation, respectively. Our irradiation system and the obtained results may be useful for studies of cell competition as well as non-targeted effects. Full article
(This article belongs to the Special Issue Microbeam Radiation Biology and Its State-of-the-Art Technology)
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12 pages, 1960 KiB  
Article
Stimulation of Nuclear Factor (Erythroid-Derived 2)-like 2 Signaling by Nucleus Targeted Irradiation with Proton Microbeam
by Jun Wang, Masakazu Oikawa and Teruaki Konishi
Biology 2023, 12(3), 419; https://doi.org/10.3390/biology12030419 - 9 Mar 2023
Cited by 3 | Viewed by 1801
Abstract
Nuclear factor (erythroid-derived 2)-like 2 (NRF2), well-known as a master antioxidative response regulator in mammalian cells, is considered as a potential target for radiation protection and cancer therapy sensitization. We examined the response of NRF2 signaling in normal human lung fibroblast WI-38 cells [...] Read more.
Nuclear factor (erythroid-derived 2)-like 2 (NRF2), well-known as a master antioxidative response regulator in mammalian cells, is considered as a potential target for radiation protection and cancer therapy sensitization. We examined the response of NRF2 signaling in normal human lung fibroblast WI-38 cells to nucleus targeted irradiation by 3.4 MeV proton microbeam. Nucleus targeted irradiation stimulated the nucleus accumulation of NRF2 and the expression of its target gene, heme oxygenase 1 (HO-1). The nucleus accumulation of NRF2 increased from 3 h to 12 h post 500 proton irradiation. In the 500 protons range, higher number of protons resulted in increased NRF2 nucleus accumulation. Activating NRF2 with tert-butylhydroquinone reduced DNA double-strand break (DSB) formation in nucleus targeted irradiation by 15%. Moreover, ATM phosphorylation was found in nucleus targeted irradiation. Inhibiting ATM with ku55933 prevented NRF2 nucleus accumulation. Furthermore, nucleus targeted irradiation activated ERK 1/2, and ROS-ERK 1/2 signaling regulated NRF2 nucleus accumulation. Taken together, NRF2 signaling was activated by nucleus targeted irradiation and mitigated DNA DSB. The discovery of ATM and ERK 1/2 as upstream regulators of NRF2 signaling in nucleus targeted cells revealed new information regarding radiation protection. Full article
(This article belongs to the Special Issue Microbeam Radiation Biology and Its State-of-the-Art Technology)
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14 pages, 2040 KiB  
Article
Differential Recruitment of DNA Repair Proteins KU70/80 and RAD51 upon Microbeam Irradiation with α-Particles
by Laure Bobyk, François Vianna, Juan S. Martinez, Gaëtan Gruel, Marc Benderitter and Céline Baldeyron
Biology 2022, 11(11), 1652; https://doi.org/10.3390/biology11111652 - 11 Nov 2022
Cited by 3 | Viewed by 2216
Abstract
In addition to representing a significant part of the natural background radiation exposure, α-particles are thought to be a powerful tool for targeted radiotherapy treatments. Understanding the molecular mechanisms of recognition, signaling, and repair of α-particle-induced DNA damage is not only important in [...] Read more.
In addition to representing a significant part of the natural background radiation exposure, α-particles are thought to be a powerful tool for targeted radiotherapy treatments. Understanding the molecular mechanisms of recognition, signaling, and repair of α-particle-induced DNA damage is not only important in assessing the risk associated with human exposure, but can also potentially help in identifying ways of improving the efficacy of radiation treatment. α-particles (He2+ ions), as well as other types of ionizing radiation, and can cause a wide variety of DNA lesions, including DNA double-strand breaks (DSBs). In mammalian cells, DNA DSBs can be repaired by two major pathways: non-homologous end-joining (NHEJ) and homologous recombination (HR). Here, we investigated their dynamics in mouse NIH-3T3 cells through the recruitment of key proteins, such as the KU heterodimer for NHEJ and RAD51 for HR upon localized α-particle irradiation. To deliver α-particles, we used the MIRCOM microbeam, which allows targeting of subnuclear structures with submicron accuracy. Using mouse NIH-3T3 cells, we found that the KU heterodimer is recruited much earlier at DNA damage sites marked by H2AX phosphorylation than RAD51. We also observed that the difference in the response of the KU complex and RAD51 is not only in terms of time, but also in function of the chromatin nature. The use of a microbeam such as MIRCOM, represents a powerful tool to study more precisely the cellular response to ionizing irradiation in a spatiotemporal fashion at the molecular level. Full article
(This article belongs to the Special Issue Microbeam Radiation Biology and Its State-of-the-Art Technology)
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Review

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13 pages, 1089 KiB  
Review
Advances in Targeted Microbeam Irradiation Methods for Live Caenorhabditis elegans
by Michiyo Suzuki
Biology 2024, 13(11), 864; https://doi.org/10.3390/biology13110864 - 24 Oct 2024
Viewed by 702
Abstract
Charged-particle microbeam irradiation devices, which can convert heavy-ion or proton beams into microbeams and irradiate individual animal cells and tissues, have been developed and used for bioirradiation in Japan, the United States, China, and France. Microbeam irradiation technology has been used to analyze [...] Read more.
Charged-particle microbeam irradiation devices, which can convert heavy-ion or proton beams into microbeams and irradiate individual animal cells and tissues, have been developed and used for bioirradiation in Japan, the United States, China, and France. Microbeam irradiation technology has been used to analyze the effects of irradiation on mammalian cancer cells, especially bystander effects. In 2006, individual-level microbeam irradiation of the nematode Caenorhabditis elegans was first realized using JAEA-Takasaki’s (now QST-TIAQS’s) TIARA collimated microbeam irradiation device. As of 2023, microbeam irradiation of C. elegans has been achieved at five sites worldwide (one in Japan, one in the United States, one in China, and two in France). This paper summarizes the global progress in the field of microbeam biology using C. elegans, while focusing on issues unique to microbeam irradiation of live C. elegans, such as the method of immobilizing C. elegans for microbeam experiments. Full article
(This article belongs to the Special Issue Microbeam Radiation Biology and Its State-of-the-Art Technology)
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29 pages, 1901 KiB  
Review
Radiation-Induced Rescue Effect: Insights from Microbeam Experiments
by Kwan Ngok Yu
Biology 2022, 11(11), 1548; https://doi.org/10.3390/biology11111548 - 23 Oct 2022
Cited by 2 | Viewed by 1963
Abstract
The present paper reviews a non-targeted effect in radiobiology known as the Radiation-Induced Rescue Effect (RIRE) and insights gained from previous microbeam experiments on RIRE. RIRE describes the mitigation of radiobiological effects in targeted irradiated cells after they receive feedback signals from co-cultured [...] Read more.
The present paper reviews a non-targeted effect in radiobiology known as the Radiation-Induced Rescue Effect (RIRE) and insights gained from previous microbeam experiments on RIRE. RIRE describes the mitigation of radiobiological effects in targeted irradiated cells after they receive feedback signals from co-cultured non-irradiated bystander cells, or from the medium previously conditioning those co-cultured non-irradiated bystander cells. RIRE has established or has the potential of establishing relationships with other non-traditional new developments in the fields of radiobiology, including Radiation-Induced Bystander Effect (RIBE), Radiation-Induced Field Size Effect (RIFSE) and ultra-high dose rate (FLASH) effect, which are explained. The paper first introduces RIRE, summarizes previous findings, and surveys the mechanisms proposed for observations. Unique opportunities offered by microbeam irradiations for RIRE research and some previous microbeam studies on RIRE are then described. Some thoughts on future priorities and directions of research on RIRE exploiting unique features of microbeam radiations are presented in the last section. Full article
(This article belongs to the Special Issue Microbeam Radiation Biology and Its State-of-the-Art Technology)
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12 pages, 1141 KiB  
Review
Radiation-Induced Bystander Effect and Cytoplasmic Irradiation Studies with Microbeams
by Ziqi Zhang, Kui Li and Mei Hong
Biology 2022, 11(7), 945; https://doi.org/10.3390/biology11070945 - 21 Jun 2022
Cited by 6 | Viewed by 3039
Abstract
Although direct damage to nuclear DNA is considered as the major contributing event that leads to radiation-induced effects, accumulating evidence in the past two decades has shown that non-target events, in which cells are not directly irradiated but receive signals from the irradiated [...] Read more.
Although direct damage to nuclear DNA is considered as the major contributing event that leads to radiation-induced effects, accumulating evidence in the past two decades has shown that non-target events, in which cells are not directly irradiated but receive signals from the irradiated cells, or cells irradiated at extranuclear targets, may also contribute to the biological consequences of exposure to ionizing radiation. With a beam diameter at the micrometer or sub-micrometer level, microbeams can precisely deliver radiation, without damaging the surrounding area, or deposit the radiation energy at specific sub-cellular locations within a cell. Such unique features cannot be achieved by other kinds of radiation settings, hence making a microbeam irradiator useful in studies of a radiation-induced bystander effect (RIBE) and cytoplasmic irradiation. Here, studies on RIBE and different responses to cytoplasmic irradiation using microbeams are summarized. Possible mechanisms related to the bystander effect, which include gap-junction intercellular communications and soluble signal molecules as well as factors involved in cytoplasmic irradiation-induced events, are also discussed. Full article
(This article belongs to the Special Issue Microbeam Radiation Biology and Its State-of-the-Art Technology)
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Other

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15 pages, 9583 KiB  
Technical Note
A Method to Locally Irradiate Specific Organ in Model Organisms Using a Focused Heavy-Ion Microbeam
by Tomoo Funayama, Michiyo Suzuki, Nobumasa Miyawaki and Hirotsugu Kashiwagi
Biology 2023, 12(12), 1524; https://doi.org/10.3390/biology12121524 - 14 Dec 2023
Cited by 1 | Viewed by 1832
Abstract
The functions of organisms are performed by various tissues composed of different cell types. Localized irradiation with heavy-ion microbeams, which inactivate only a portion of the constituent cells without destroying the physical intercellular connections of the tissue, is a practical approach for elucidating [...] Read more.
The functions of organisms are performed by various tissues composed of different cell types. Localized irradiation with heavy-ion microbeams, which inactivate only a portion of the constituent cells without destroying the physical intercellular connections of the tissue, is a practical approach for elucidating tissue functions. However, conventional collimated microbeams are limited in the shape of the area that can be irradiated. Therefore, using a focused heavy-ion microbeam that generates a highly precise beam spot, we developed a technology to uniformly irradiate specific tissues of an organism with a defined dose, which conventional methods cannot achieve. The performance of the developed paint irradiation technology was evaluated. By irradiating the CR-39 ion track detector, we confirmed that the new method, in which each ion hit position is placed uniformly in the irradiated area, makes it possible to uniformly paint the area at a specified dose. The targeted irradiation of the pharynx and gonads of living Caenorhabditis elegans demonstrated that the irradiated ions were distributed in the same shape as the targeted tissue observed under a microscope. This technology will elucidate biological mechanisms that are difficult to analyze with conventional collimated microbeam irradiation. Full article
(This article belongs to the Special Issue Microbeam Radiation Biology and Its State-of-the-Art Technology)
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19 pages, 4134 KiB  
Technical Note
Proton Microbeam Targeted Irradiation of the Gonad Primordium Region Induces Developmental Alterations Associated with Heat Shock Responses and Cuticle Defense in Caenorhabditis elegans
by Pierre Beaudier, Guillaume Devès, Laurent Plawinski, Denis Dupuy, Philippe Barberet and Hervé Seznec
Biology 2023, 12(11), 1372; https://doi.org/10.3390/biology12111372 - 27 Oct 2023
Cited by 1 | Viewed by 1570
Abstract
We describe a methodology to manipulate Caenorhabditis elegans (C. elegans) and irradiate the stem progenitor gonad region using three MeV protons at a specific developmental stage (L1). The consequences of the targeted irradiation were first investigated by considering the organogenesis of [...] Read more.
We describe a methodology to manipulate Caenorhabditis elegans (C. elegans) and irradiate the stem progenitor gonad region using three MeV protons at a specific developmental stage (L1). The consequences of the targeted irradiation were first investigated by considering the organogenesis of the vulva and gonad, two well-defined and characterized developmental systems in C. elegans. In addition, we adapted high-throughput analysis protocols, using cell-sorting assays (COPAS) and whole transcriptome analysis, to the limited number of worms (>300) imposed by the selective irradiation approach. Here, the presented status report validated protocols to (i) deliver a controlled dose in specific regions of the worms; (ii) immobilize synchronized worm populations (>300); (iii) specifically target dedicated cells; (iv) study the radiation-induced developmental alterations and gene induction involved in cellular stress (heat shock protein) and cuticle injury responses that were found. Full article
(This article belongs to the Special Issue Microbeam Radiation Biology and Its State-of-the-Art Technology)
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15 pages, 3354 KiB  
Technical Note
Recruitment Kinetics of XRCC1 and RNF8 Following MeV Proton and α-Particle Micro-Irradiation
by Giovanna Muggiolu, Eva Torfeh, Marina Simon, Guillaume Devès, Hervé Seznec and Philippe Barberet
Biology 2023, 12(7), 921; https://doi.org/10.3390/biology12070921 - 27 Jun 2023
Cited by 1 | Viewed by 1342
Abstract
Time-lapse fluorescence imaging coupled to micro-irradiation devices provides information on the kinetics of DNA repair protein accumulation, from a few seconds to several minutes after irradiation. Charged-particle microbeams are valuable tools for such studies since they provide a way to selectively irradiate micrometric [...] Read more.
Time-lapse fluorescence imaging coupled to micro-irradiation devices provides information on the kinetics of DNA repair protein accumulation, from a few seconds to several minutes after irradiation. Charged-particle microbeams are valuable tools for such studies since they provide a way to selectively irradiate micrometric areas within a cell nucleus, control the dose and the micro-dosimetric quantities by means of advanced detection systems and Monte Carlo simulations and monitor the early cell response by means of beamline microscopy. We used the charged-particle microbeam installed at the AIFIRA facility to perform micro-irradiation experiments and measure the recruitment kinetics of two proteins involved in DNA signaling and repair pathways following exposure to protons and α-particles. We developed and validated image acquisition and processing methods to enable a systematic study of the recruitment kinetics of GFP-XRCC1 and GFP-RNF8. We show that XRCC1 is recruited to DNA damage sites a few seconds after irradiation as a function of the total deposited energy and quite independently of the particle LET. RNF8 is recruited to DNA damage sites a few minutes after irradiation and its recruitment kinetics depends on the particle LET. Full article
(This article belongs to the Special Issue Microbeam Radiation Biology and Its State-of-the-Art Technology)
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