基于微流控芯片、上转换发光纳米标记和多光镊分选循环肿瘤细胞

The detection of circulating tumor cells (CTCs) in blood, which is a major task for “liquid biopsy”, is of great importance for cancer diagnosis and prognosis. However, due to the heterogeneity of the cancer cells, how to differentiate subpopulations that may have varying phenotypes while a bulk population of CTCs is isolated, still remains a major challenge. Aiming for this we put forward a novel microfluidic chip and up-converting nano-labling based technique “luminescence activated cell soring (FACS) using optical tweezers”, and a novel method of multiple-channel sorting of CTC based on the detection of two-color luminescence and optical trapping array is proposed. Our aim is to solve the scientific issue “simultaneous sorting and phenotype differentiation in microfluidic chip”, and finally develop a new technology for cancer liqid biopsy. For the proof-of-principle study, two-color upconversion luminescence nanoprobes are to be prepared for dual-labelling of the tumor markers epithelial cell adhesion molecule (EpCAM) and human epidermal growth factor receptor 2 (HER2) on breast cells of three different phenotypes, and CTC capturing and sorting will be realized through accurately controlling of the laser intensity and movement of the multiple optical tweezers, and moreover, three different phenotypes of breast cancers can be simultaneously differentiated. With this approach, CTC counting, rapid detection and sorting/classification can be attained on condition that the viability of the CTCs is kept.

以血液中循环肿瘤细胞（CTC）为主要检测指标的“液体活检”对于癌症的诊断和预后研究有重要价值，但由于癌细胞具有异质性，如何在高效捕获CTC的同时实现其分子分型，是新一代CTC检测技术面临的巨大挑战。本项目针对上述挑战，提出基于微流控芯片和上转换纳米标记构建一种“发光激活细胞光镊分选”技术，并提出一种双色发光检测结合阵列多光阱的多通道快速分选CTC的新方法，通过解决“在微流控通道中同时实现CTC分选和分型”这一关键科学问题，为癌症的“液体活检”发展一种新技术。拟以三种不同分型的乳腺癌细胞为研究对象，通过制备双色上转换发光纳米探针，对其表面的上皮细胞粘附分子(EpCAM)和人表皮生长因子受体-2 (HER2)进行双重标记，通过精密控制多光阱的激光强度和快速位移，实现癌细胞的分选，并同时实现三种不同乳腺癌细胞的分型。本方法在保持CTC活性的前提下，同时实现CTC的计数、快速检测和分选/分型。

以血液中循环肿瘤细胞（CTC）为主要检测指标的“液体活检”对于癌症的诊断和预后研究有重要价值，但由于癌细胞具有异质性，如何在高效捕获CTC的同时实现其分子分型，是新一代CTC检测技术面临的巨大挑战。本项目针对上述挑战，提出基于微流控芯片和上转换纳米标记构建一种“发光激活细胞光镊分选”技术，并提出一种双色发光检测结合阵列多光阱的多通道快速分选CTC的新方法，通过解决“在微流控通道中同时实现CTC分选和分型”这一关键科学问题，为癌症的“液体活检”发展一种新技术。拟以三种不同分型的乳腺癌细胞为研究对象，通过制备双色上转换发光纳米探针，对其表面的上皮细胞粘附分子(EpCAM)和人表皮生长因子受体-2 (HER2)进行双重标记，通过精密控制多光阱的激光强度和快速位移，实现癌细胞的分选，并同时实现三种不同乳腺癌细胞的分型。本方法在保持CTC活性的前提下，同时实现CTC的计数、快速检测和分选/分型。