A standardized protocol for sample preparation for scanning electron microscopy to visualize extrachromosomal DNA.

IF 2.2 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS BioTechniques Pub Date : 2024-01-01 Epub Date: 2024-06-19 DOI:10.1080/07366205.2024.2346042
Jillann A Madren, Jingting Chen, William Dennis, Christina Ford, Kristen K White, Elizabeth Brunk
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Abstract

Extrachromosomal DNA (ecDNA) are circular DNA structures associated with cancer and drug resistance. One specific type, double minute (DM) chromosomes, has been studied since the 1960s using imaging techniques like cytogenetics and fluorescence microscopy. Specialized techniques such as atomic force microscopy (AFM) and scanning electron microscopy (SEM) offer micro to nano-scale visualization, but current sample preparation methods may not optimally preserve ecDNA structure. Our study introduces a systematic protocol using SEM for high-resolution ecDNA visualization. We have optimized the end-to-end procedure, providing a standardized approach to explore the circular architecture of ecDNA and address the urgent need for better understanding in cancer research.

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用于扫描电子显微镜观察染色体外 DNA 的样本制备标准化方案。
染色体外 DNA(ecDNA)是与癌症和耐药性有关的环状 DNA 结构。自 20 世纪 60 年代以来,人们一直在利用细胞遗传学和荧光显微镜等成像技术研究一种特殊类型的双微粒(DM)染色体。原子力显微镜(AFM)和扫描电子显微镜(SEM)等专业技术可提供微米到纳米尺度的可视化,但目前的样品制备方法可能无法最佳地保留 ecDNA 结构。我们的研究介绍了一种利用扫描电子显微镜进行高分辨率 ecDNA 可视化的系统方案。我们优化了端到端的程序,提供了一种标准化的方法来探索 ecDNA 的环状结构,满足了癌症研究中更好地了解其结构的迫切需要。
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来源期刊
BioTechniques
BioTechniques 工程技术-生化研究方法
CiteScore
4.40
自引率
0.00%
发文量
68
审稿时长
3.3 months
期刊介绍: BioTechniques is a peer-reviewed, open-access journal dedicated to publishing original laboratory methods, related technical and software tools, and methods-oriented review articles that are of broad interest to professional life scientists, as well as to scientists from other disciplines (e.g., chemistry, physics, computer science, plant and agricultural science and climate science) interested in life science applications for their technologies. Since 1983, BioTechniques has been a leading peer-reviewed journal for methods-related research. The journal considers: Reports describing innovative new methods, platforms and software, substantive modifications to existing methods, or innovative applications of existing methods, techniques & tools to new models or scientific questions Descriptions of technical tools that facilitate the design or performance of experiments or data analysis, such as software and simple laboratory devices Surveys of technical approaches related to broad fields of research Reviews discussing advancements in techniques and methods related to broad fields of research Letters to the Editor and Expert Opinions highlighting interesting observations or cautionary tales concerning experimental design, methodology or analysis.
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