SBIR Fast Track: Development of a High-throughput Magnetic Cytometer for Single Cell Sorting

项目摘要 这个小型企业创新研究项目旨在开发一种高通量、易于使用的细胞分选仪 MagiCyte™以单细胞分辨率快速精确分选完整和活的稀有细胞。 从异质细胞群中分选单细胞对于生命科学应用是必不可少的，例如 单克隆抗体生产和细胞系开发。近年来，随着精密度的快速增长， 在医学上，对于稀有细胞的单细胞分选技术也有强烈的需求，例如 循环肿瘤细胞（CTC）、循环胎儿细胞、干细胞和抗原特异性T细胞或B细胞。然而，在这方面， 以单细胞分辨率收集这样的稀有细胞以满足单细胞分析的要求是不可行的 用于常规的单细胞分选技术。显微操作和连续稀释是耗时的， 低通量，而荧光激活细胞分选（FACS）遭受显著的细胞损失， 在分离稀有细胞方面效率低下。此外，许多这样的技术需要细胞固定和/或荧光标记。 染色，这降低了细胞活力，从而阻止了许多下游分子分析，以及 和细胞培养。因此，存在对稀有细胞的高通量单细胞分选的未解决的需求。 我们的目标是开发一种基于芯片的高通量流式细胞仪MagiCyte™，以快速分离、鉴定和分选 来自异质细胞群的单个完整和活的靶细胞。MagiCyte的核心组件是一个 半导体芯片MagiChip™用于单细胞识别和单细胞分选。在这种快速- 跟踪项目，我们将首先开发控制软件并演示自动化单细胞分选; 第二，我们将开发下一代MagiChip，构建MagiCyte beta产品并在 临床样本。

Project Summary This Small Business Innovation Research project aims to develop a high-throughput easy-to-use cell sorter MagiCyte™ to rapidly and precisely sort intact and viable rare cells at single-cell resolution. Single-cell sorting from a heterogeneous cell population has been essential to life science applications such as monoclonal antibody production and cell line development. Recently, with the rapid growth of precision medicine, there has also been a strong demand for single-cell sorting technologies for rare cells, such as circulating tumor cells (CTCs), circulating fetal cells, stem cells and antigen-specific T- or B-cells. However, collecting such rare cells at single-cell resolution to meet the requirements of single-cell analysis is not feasible for conventional single-cell sorting techniques. Micromanipulation and serial dilution are time consuming and low-throughput, while fluorescent-activated cell sorting (FACS) suffers from remarkable cell loss and is inefficient in isolating rare cells. In addition, many such techniques require cell fixation and/or fluorescent staining, which decreases cell viability and thus prohibits many downstream molecular analyses, as well as and cell culture. As such, there is an unaddressed need for high throughput single-cell sorting of rare cells. We aim to develop a high-throughput chip-based cytometer MagiCyte™ to rapidly isolate, identify and sort single intact and viable target cells from heterogeneous cell population. The core component of MagiCyte is a semiconductor chip MagiChip™ which is used for single cell identification and single cell sorting. In this fast- track project, we will first develop the control software and demonstrate automated single-cell sorting; In Phase II, we will develop the next-generation MagiChip, build the MagiCyte beta product and validate the system on clinical samples with our collaborators.