Microscopy Services Laboratory, University of North Carolina at Chapel Hill, Chapel Hill, NC 27516, USA.
Department of Biochemistry & Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27516, USA.
Biotechniques. 2024;76(7):311-321. doi: 10.1080/07366205.2024.2346042. Epub 2024 Jun 19.
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.
染色体外 DNA（ecDNA）是与癌症和耐药性相关的环状 DNA 结构。自 20 世纪 60 年代以来，人们一直使用细胞遗传学和荧光显微镜等成像技术来研究一种特殊类型的双微体（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到纳米级的可视化，但目前的样品制备方法可能无法最佳保留 ecDNA 结构。我们的研究引入了一种使用 SEM 进行高分辨率 ecDNA 可视化的系统方案。我们已经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，并满足癌症研究中对更好理解的迫切需求。
