CAS Key Laboratory of Standardization and Measurement for Nanotechnology, CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, CAS Center for Excellence in Nanoscience, National Center for Nanoscience and Technology of China, Beijing 100190, China.
School of Information and Communication Engineering, North University of China, Taiyuan 030051, China.
Theranostics. 2017 Oct 17;7(19):4710-4721. doi: 10.7150/thno.20440. eCollection 2017.
Enumerating circulating tumor cells (CTCs) has been demonstrably useful in cancer treatment. Although there are several approaches that have proved effective in isolating CTC-like cells, the crucial identification of CTCs continues to rely on the manual interpretation of immunofluorescence images of all cells that have been isolated. This procedure is time consuming and more importantly, CTC identification relies on subjective criteria that may differ between examiners. In this study, we describe the design, testing, and verification of a microfluidic platform that provides accurate and automated CTC enumeration using a common objective criterion. The platform consists of a multi-functional microfluidic chip and a unique image processing algorithm. The microfluidic chip integrates blood filtering, cell isolation, and single cell positioning to ensure minimal cell loss, efficient cell isolation, and fixed arraying of single cells to facilitate downstream image processing. By taking advantage of the microfluidic chip design to reduce calculation loads and eliminate measurement errors, our specially designed algorithm has the capability of rapidly interpreting hundreds of images to provide accurate CTC counts. Following intensive optimization of the microfluidic chip, the image processing algorithm, and their collaboration, we verified the complete platform by enumerating CTCs from six clinical blood samples of patients with breast cancer. Compared to tube-based CTC isolation and manual CTC identification, our platform had better accuracy and reduced the time needed from sample loading to result review by 50%. This automated CTC enumeration platform demonstrates not only a sound strategy in integrating a specially designed multi-functional microfluidic chip with a unique image processing algorithm for robust, accurate, and "hands-free" CTC enumeration, but may also lead to its use as a novel diagnostic device used in clinics and laboratories as readily as a routine blood test.
循环肿瘤细胞 (CTC) 的计数在癌症治疗中已被证明是非常有用的。虽然有几种方法已被证明可以有效地分离出类 CTC 细胞，但 CTC 的关键鉴定仍然依赖于对所有已分离的细胞的免疫荧光图像进行人工解释。这个过程非常耗时，更重要的是，CTC 的鉴定依赖于可能因检查人员而异的主观标准。在这项研究中，我们描述了一种微流控平台的设计、测试和验证，该平台使用通用的客观标准提供准确和自动化的 CTC 计数。该平台由多功能微流控芯片和独特的图像处理算法组成。微流控芯片集成了血液过滤、细胞分离和单细胞定位，以确保最小的细胞损失、高效的细胞分离和单细胞的固定排列，从而便于下游图像处理。通过利用微流控芯片设计来降低计算负荷和消除测量误差，我们专门设计的算法具有快速解释数百张图像以提供准确 CTC 计数的能力。在对微流控芯片、图像处理算法及其协作进行了深入优化之后，我们通过对来自六名乳腺癌患者的六个临床血液样本进行 CTC 计数来验证了整个平台。与基于管的 CTC 分离和手动 CTC 鉴定相比，我们的平台具有更高的准确性，并且将从样本加载到结果审查所需的时间缩短了 50%。这个自动化 CTC 计数平台不仅展示了一种明智的策略，即将专门设计的多功能微流控芯片与独特的图像处理算法相结合，用于稳健、准确和“无需人工干预”的 CTC 计数，而且还可能使其成为一种新的诊断设备，像常规血液测试一样在临床和实验室中得到广泛应用。
Zotero 插件
