Characterization and implementation of a miniature X-ray system for live cell microscopy

IF 1.5 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis Pub Date : 2022-01-01 DOI:10.1016/j.mrfmmm.2021.111772

Surendra Prajapati , Maëlle Locatelli , Caleb Sawyer , Julia Holmes , Keith Bonin , Paul Black , Pierre-Alexandre Vidi

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引用次数: 1

Abstract

The study of radiation effects on biological tissues is a diverse field of research with direct applications to improve human health, in particular in the contexts of radiation therapy and space exploration. Understanding the DNA damage response following radiation exposure, which is a key determinant for mutagenesis, requires reproducible methods for delivering known doses of ionizing radiation (IR) in a controlled environment. Multiple IR sources, including research X-ray and gamma-ray irradiators are routinely used in basic and translational research with cell and animal models. These systems are however not ideal when a high temporal resolution is needed, for example to study early DNA damage responses with live cell microscopy. Here, we characterize the dose rate and beam properties of a commercial, miniature, affordable, and versatile X-ray source (Mini-X). We describe how to use Mini-X on the stage of a fluorescence microscope to deliver high IR dose rates (up to 29 Gy/min) or lower dose rates (≤ 0.1 Gy/min) in live cell imaging experiments. This article provides a blueprint for radiation biology applications with high temporal resolution, with a step-by-step guide to implement a miniature X-ray system on an imaging platform, and the information needed to characterize the system.

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用于活细胞显微镜的微型x射线系统的特性和实现

辐射对生物组织的影响研究是一个多样化的研究领域，可直接应用于改善人类健康，特别是在放射治疗和空间探索方面。了解辐射暴露后的DNA损伤反应，这是诱变的关键决定因素，需要在受控环境中提供已知剂量的电离辐射(IR)的可重复方法。多种红外光源，包括研究x射线和伽马射线辐照体，通常用于细胞和动物模型的基础和转化研究。然而，当需要高时间分辨率时，这些系统并不理想，例如用活细胞显微镜研究早期DNA损伤反应。在这里，我们描述了一种商业的、微型的、负担得起的、多功能的x射线源(Mini-X)的剂量率和光束特性。我们描述了如何在荧光显微镜台上使用Mini-X在活细胞成像实验中提供高IR剂量率(高达29 Gy/min)或更低剂量率(≤0.1 Gy/min)。本文提供了高时间分辨率辐射生物学应用的蓝图，逐步指导在成像平台上实现微型x射线系统，以及表征该系统所需的信息。

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来源期刊

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis 生物-毒理学

CiteScore

4.90

自引率

0.00%

发文量

审稿时长

51 days

期刊介绍： Mutation Research (MR) provides a platform for publishing all aspects of DNA mutations and epimutations, from basic evolutionary aspects to translational applications in genetic and epigenetic diagnostics and therapy. Mutations are defined as all possible alterations in DNA sequence and sequence organization, from point mutations to genome structural variation, chromosomal aberrations and aneuploidy. Epimutations are defined as alterations in the epigenome, i.e., changes in DNA methylation, histone modification and small regulatory RNAs. MR publishes articles in the following areas: Of special interest are basic mechanisms through which DNA damage and mutations impact development and differentiation, stem cell biology and cell fate in general, including various forms of cell death and cellular senescence. The study of genome instability in human molecular epidemiology and in relation to complex phenotypes, such as human disease, is considered a growing area of importance. Mechanisms of (epi)mutation induction, for example, during DNA repair, replication or recombination; novel methods of (epi)mutation detection, with a focus on ultra-high-throughput sequencing. Landscape of somatic mutations and epimutations in cancer and aging. Role of de novo mutations in human disease and aging; mutations in population genomics. Interactions between mutations and epimutations. The role of epimutations in chromatin structure and function. Mitochondrial DNA mutations and their consequences in terms of human disease and aging. Novel ways to generate mutations and epimutations in cell lines and animal models.