Patrono C, Monteiro Gil O, Giesen U, Langner F, Pinto M, Rabus H, Testa A
Technical Unit of Radiation Biology and Human Health, ENEA Casaccia, Rome, Italy
Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.
Radiat Prot Dosimetry. 2015 Sep;166(1-4):197-9. doi: 10.1093/rpd/ncv160. Epub 2015 Apr 15.
The aim of the 'BioQuaRT' (Biologically weighted Quantities in RadioTherapy) project is to develop measurement techniques for characterising charged particle track structure on different length scales, and to correlate at the cellular level the track structure properties with the biological effects of radiation. This multi-scale approach will allow characterisation of the radiation qualities used in radiotherapy and the related biological effects. Charged-particle microbeam facilities were chosen as the platforms for all radiobiology experiments in the 'BioQuaRT' project, because they allow targeting single cells (or compartments of a cell) with a predefined number of ionising particles and correlating the cell-by-cell induced damage with type and energy of the radiation and with the number of ions per cell. Within this project, a novel in situ protocol was developed for the analysis of the misrepaired and/or unrepaired chromosome damage induced by charged-particle irradiations at the Physikalisch-Technische Bundesanstalt (PTB) ion microbeam facility. Among the cytogenetic biomarkers to detect and estimate radiation-induced DNA damage in radiobiology, chromosomal aberrations and micronuclei were chosen. The characteristics of the PTB irradiation system required the design of a special in situ assay: specific irradiation dishes with a base made from a biofoil 25-µm thick and only 3000-4000 cells seeded and irradiated per dish. This method was developed on Chinese hamster ovary (CHO) cells, one of the most commonly used cell lines in radiobiology in vitro experiments. The present protocol allows the simultaneous scoring of chromosome aberrations and micronuclei on the same irradiated dish. Thanks to its versatility, this method could also be extended to other radiobiological applications besides the single-ion microbeam irradiations.
“生物放疗定量研究”（BioQuaRT，Biologically weighted Quantities in RadioTherapy）项目的目标是开发用于表征不同长度尺度上带电粒子径迹结构的测量技术，并在细胞水平上使径迹结构特性与辐射的生物学效应相关联。这种多尺度方法将能够表征放射治疗中使用的辐射质量及其相关生物学效应。在“生物放疗定量研究”项目中，带电粒子微束设施被选作所有放射生物学实验的平台，因为它们能够用预先确定数量的电离粒子靶向单个细胞（或细胞的特定区域），并将逐个细胞诱导的损伤与辐射类型、能量以及每个细胞的离子数量相关联。在该项目中，开发了一种新的原位实验方案，用于分析在德国物理技术研究院（PTB）离子微束设施中带电粒子辐照诱导的错误修复和/或未修复的染色体损伤。在放射生物学中用于检测和估计辐射诱导的DNA损伤的细胞遗传学生物标志物中，选择了染色体畸变和微核。PTB辐照系统的特性要求设计一种特殊的原位检测方法：使用由25微米厚的生物箔制成底部的特定辐照培养皿，每个培养皿仅接种3000 - 4000个细胞并进行辐照。该方法是在中国仓鼠卵巢（CHO）细胞上开发的，CHO细胞是放射生物学体外实验中最常用的细胞系之一。本实验方案允许在同一个辐照培养皿上同时对染色体畸变和微核进行评分。由于其通用性，除了单离子微束辐照外，该方法还可扩展到其他放射生物学应用。
