一个支持多维度生物力学与生物化学表征的自动化微型流式细胞仪的开发和对于癌细胞分选的应用

Over the past years, many microfluidic devices have been developed for quantitative measurements of both mechanical and biochemical cell characteristics. In cancer research, it has been well proven that metastatic cancer cells share some common mechanical and biochemical characteristics, which can potentially be considered as diagnostic biomarkers. Metastasis involves migration of cancer cells from sites to sties through tiny tissue gaps or vessel walls in a patient’s body; therefore cell-cell and cell-matrix interactions are important criteria in such process. However, the effective identification of the metastasis using the cancer patient’s blood is yet challenging, mainly due to the very limited number of metastatic cells existing in the blood (<10 cells/ml) The measurement sensitivity is yet to be improved largely for clinical applications...Here, we propose to develop a novel surface-functioned elasticity microcytometer associated with the automation platform to simultaneously quantify for the physical properties and multiple surface protein expressions of every cell (out of 10^6 cells) in a bio-sample in order to precisely categorize small subsets of cells (may down to a group of <10 cells) based on selected ranges of cell properties, e.g. cell viscoelasticity, and epithelial cadherin adhesion molecule (EpCAM) expression. Technically, we will design the device with multiple stages of funnel-shaped confining microchannels based on multilayer soft lithography. Under well-defined physical configurations (e.g. microstructure dimensions and flow conditions), the dynamics of cell deformations under compression of the confining channel side-walls can reveal the cell viscoelasticity quantities. We will derive a first-principle model for the first to effectively convert transient deformations of encapsulated suspending cells to the important cell parameters. Further applying multiple stages of the confining channels functionalized with different surface protein antibodies, this integrated strategy can recognize multiple cell surface protein expressions as biomarkers for identification of the metastatic potential. Importantly, the system automation is essential to achieve high-throughput cell characteristics quantification of a large cell number (~10^6 cells) within a manageable time frame. ..Collectively, considering both the mechanical and biomechanical characteristics can achieve a more specific identification of highly metastatic cells. The automated elasticity microcytometer can be applied as an effective diagnostic tool for metastatic potential of cancer cells. We anticipate this measurement method can examine cell samples for the existence of cancer cells from specific primary tumor sites with higher metastatic potentials and may achieve an effective metastasis diagnosis of clinical samples from cancer patients.

近年，转移性癌细胞所共有一些生物力学与生物化学特征被视为潜在的癌症诊断标记。然而，病人血液中癌细胞的密度极少（小于每毫升十个），以血液检测癌细胞依然具有挑战性。对于临床应用，其检测的精度与特异性更亟待提高。.在此，我们提出一种高通量自动化微流体细胞仪。这可以同时定量检测细胞的力学性质（如细胞黏弹性）以及表面蛋白的表达如上皮细胞粘附分子（EpCAM），并进一步区分不同的细胞群体。该细胞仪有一个通过多层软光刻方法制作的漏斗形微通道。细胞在通过微通道时的动态形变可以揭示细胞的粘弹性性质。然后，让细胞通过不同层次的微通道，每个层次的通道表面用不同的细胞表面蛋白抗体功能化；这种集成化的结构可以识别多种可用来判断癌细胞转移性的细胞表面蛋白。.总之，通过对细胞力学特性和生化特性的综合考量，该细胞仪能够更加精确地诊断转移性的癌细胞。我们预计这个自动化的平台可作为一个检测病人血液样品中转移癌细胞的有效工具。

近年，转移性癌细胞所共有一些生物力学与生物化学特征被视为潜在的癌症诊断标记。然而，病人血液中癌细胞的密度极少（小于每毫升十个），以血液检测癌细胞依然具有挑战性。对于临床应用，其检测的精度与特异性更亟待提高。.在此，我们提出一种高通量自动化微流体细胞仪。这可以同时定量检测细胞的力学性质（如细胞黏弹性）以及表面蛋白的表达如上皮细胞粘附分子（EpCAM），并进一步区分不同的细胞群体。该细胞仪有一个通过多层软光刻方法制作的漏斗形微通道。细胞在通过微通道时的动态形变可以揭示细胞的粘弹性性质。然后，让细胞通过不同层次的微通道，每个层次的通道表面用不同的细胞表面蛋白抗体功能化；这种集成化的结构可以识别多种可用来判断癌细胞转移性的细胞表面蛋白。.总之，通过对细胞力学特性和生化特性的综合考量，该细胞仪能够更加精确地诊断转移性的癌细胞。我们预计这个自动化的平台可作为一个检测病人血液样品中转移癌细胞的有效工具。