Williams Anthony, Chung Jaebum, Yang Changhuei, Cote Richard J
Department of Surgery, University of Chicago Medical Center, Chicago, IL, USA.
Department of Electrical Engineering, California Institute of Technology, Pasadena, CA, USA.
Methods Mol Biol. 2017;1634:107-117. doi: 10.1007/978-1-4939-7144-2_8.
Examining the hematogenous compartment for evidence of metastasis has increased significantly within the oncology research community in recent years, due to the development of technologies aimed at the enrichment of circulating tumor cells (CTCs), the subpopulation of primary tumor cells that gain access to the circulatory system and are responsible for colonization at distant sites. In contrast to other technologies, filtration-based CTC enrichment, which exploits differences in size between larger tumor cells and surrounding smaller, non-tumor blood cells, has the potential to improve CTC characterization through isolation of tumor cell populations with greater molecular heterogeneity. However, microscopic analysis of uneven filtration surfaces containing CTCs is laborious, time-consuming, and inconsistent, preventing widespread use of filtration-based enrichment technologies. Here, integrated with a microfiltration-based CTC and rare cell enrichment device we have previously described, we present a protocol for Fourier Ptychographic Microscopy (FPM), a method that, unlike many automated imaging platforms, produces high-speed, high-resolution images that can be digitally refocused, allowing users to observe objects of interest present on multiple focal planes within the same image frame. The development of a cost-effective and high-throughput CTC analysis system for filtration-based enrichment technologies could have profound clinical implications for improved CTC detection and analysis.
近年来，由于旨在富集循环肿瘤细胞（CTC）的技术的发展，肿瘤研究界对通过血行途径检查转移证据的关注度显著增加。循环肿瘤细胞是原发肿瘤细胞的亚群，它们进入循环系统并在远处部位定植。与其他技术不同，基于过滤的CTC富集技术利用较大肿瘤细胞与周围较小非肿瘤血细胞之间的大小差异，有潜力通过分离具有更大分子异质性的肿瘤细胞群体来改善CTC的表征。然而，对含有CTC的不均匀过滤表面进行显微镜分析既费力、耗时又不一致，这阻碍了基于过滤的富集技术的广泛应用。在此，结合我们之前描述的基于微滤的CTC和稀有细胞富集装置，我们提出了一种傅里叶叠层显微镜（FPM）方案。与许多自动成像平台不同，该方法能生成可数字重聚焦的高速、高分辨率图像，使用户能够在同一图像帧内观察存在于多个焦平面上的感兴趣物体。开发一种具有成本效益且高通量的基于过滤富集技术的CTC分析系统，可能对改进CTC检测和分析具有深远的临床意义。
