Abstract
Past efforts in radiobiology, radio-biophysics, epidemiology and clinical research strongly contributed to the current understanding of ionizing radiation effects on biological materials like cells and tissues. It is well accepted that the most dangerous, radiation induced damages of DNA in the cell nucleus are double strand breaks, as their false rearrangements cause dysfunction and tumor cell proliferation. Therefore, cells have developed highly efficient and adapted ways to repair lesions of the DNA double strand. To better understand the mechanisms behind DNA strand repair, a variety of fluorescence microscopy based approaches are routinely used to study radiation responses at the organ, tissue and cellular level. Meanwhile, novel super-resolution fluorescence microscopy techniques have rapidly evolved and become powerful tools to study biological structures and bio-molecular (re-)arrangements at the nano-scale. In fact, recent investigations have increasingly demonstrated how super-resolution microscopy can be applied to the analysis of radiation damage induced chromatin arrangements and DNA repair protein recruitment in order to elucidate how spatial organization of damage sites and repair proteins contribute to the control of repair processes. In this chapter, we would like to start with some fundamental aspects of ionizing radiation, their impact on biological materials, and some standard radiobiology assays. We conclude by introducing the concept behind super-resolution radiobiology using single molecule localization microscopy (SMLM) and present promising results from recent studies that show an organized architecture of damage sites and their environment. Persistent homologies of repair clusters indicate a correlation between repair cluster topology and repair pathway at a given damage locus. This overview over recent investigations may motivate radiobiologists to consider chromatin architecture and spatial repair protein organization for the understanding of DNA repair processes.
Highlights
Past efforts in epidemiological and clinical research strongly contributed to the current understanding of ionizing radiation effects on human organs, tissues, and cells [1, 2]
The desoxyribonuleic acids (DNAs) damage response (DDR) against double-strand breaks (DSBs) is subject to intensive radiobiological investigation and fluorescence microscopy of in situ DSB repair proteins serves as state of the art biological dosimetry
We have addressed scientists, researchers, and clinicians working in interdisciplinary fields, which are searching for a brief introduction to current radiobiology, its fundamental principles and methodologies
Summary
Past efforts in epidemiological (nuclear power industry, atomic bomb explosions, nuclear reactor accidents, etc.) and clinical Methods based on stochastic reversible photobleaching [9–15] of single molecules called Single Molecule Localization Microscopy (SMLM) [16] reach effective resolutions down to 10 nm and have become popular among modern super-resolution imaging techniques as their realization is highly practical and straightforward using established specimen preparation methods of standard fluorescence microcopy [17]. As such resolutions allow the detection of single molecules, such as nucleosomes [18], proteins [19, 20], receptors and junction proteins [21, 22], or even single chromatin loops [23] etc., super-resolution microscopy opens new avenues for the research of radiation induced damaging and repair processes [5, 24]. The successful application of localization microscopy in radiation biology research is demonstrated along examples of current works
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