Microscopy-based cell separation, selective propagation, and selective single cell sequencing

基于显微镜的细胞分离、选择性增殖和选择性单细胞测序

• 批准号: RGPIN-2020-05412
• 负责人: Ma, Hongshen
• 金额: $ 2.84万
• 依托单位: University of British Columbia
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=717628
• 关键词: Microscopy; based; cell; separation; selective

The analysis of cells, or cytometry, is fundamentally important to many aspects of biological research. Flow cytometry has been the dominant cytometry technology since the 1960s because it is simple, high-throughput, and can be coupled with fluorescence activated cell sorting to separate specific cell populations. However, flow cytometry also has a number of limitations including (1) shear stress on the cell sample; (2) significant cell loss during processing; (3) difficulty in analyzing non-spherical cells, delicate cells, and cell clusters; (4) inability to assess sub-cellular structure and morphology. Many of these limitations could be addressed using fluorescence microscopy, but microscopy has the critical limitation of not being able to separate target cells, which in turn excludes many subsequent analyses, such as propagation to form clonal populations and single cell sequencing. Current technologies for isolating cells identified by microscopy involve physically transferring single cells using a micromanipulator, but this approach is difficult, low-throughput, and prone to contamination. The long-term vision of this research program is to develop microscopy-based technologies for cell separation, selective propagation, and selective single cell sequencing. Leveraging our recent work in this area, as well as advances in wide-field microscopy and image-processing, we believe our work will ultimately enable microscopy to compete with flow cytometry as a standard tool for cytometry, cell separation, and single cell analysis. This discovery grant will train HQPs at graduate, undergraduate, and post-doctoral levels. Trainees involved in the proposed research program will receive world-class training in an equitable, diverse, and inclusive environment that is dynamic and multi-disciplinary. Specific areas of research and training include optics, microscopy, instrumentation, image-processing, machine learning, polymer materials, cell biology, molecular biology, and genomics. Trainees will be able to leverage the high-level skills, accomplishments, and engineering acumen developed through their research projects into fantastic careers in industry and academia.

细胞分析或细胞计数对生物学研究的许多方面都至关重要。自20世纪60年代以来，流式细胞术一直是占主导地位的细胞术技术，因为它简单、高通量，并且可以与荧光激活细胞分选相结合以分离特定的细胞群。然而，流式细胞术也具有许多限制，包括（1）细胞样品上的剪切应力;（2）处理期间的显著细胞损失;（3）难以分析非球形细胞、脆弱细胞和细胞簇;（4）不能评估亚细胞结构和形态。这些限制中的许多可以使用荧光显微镜来解决，但是显微镜具有不能分离靶细胞的关键限制，这反过来又排除了许多后续分析，例如繁殖以形成克隆群体和单细胞测序。目前用于分离通过显微镜鉴定的细胞的技术涉及使用显微操作器物理地转移单个细胞，但是这种方法是困难的、低通量的并且易于污染。 该研究计划的长期愿景是开发基于显微镜的细胞分离，选择性繁殖和选择性单细胞测序技术。利用我们最近在这一领域的工作，以及在宽视场显微镜和图像处理的进步，我们相信我们的工作最终将使显微镜与流式细胞术竞争，作为细胞计数，细胞分离和单细胞分析的标准工具。 这项发现补助金将在研究生，本科和博士后水平上培训HQP。参与拟议研究计划的学员将在一个公平，多元化和包容性的环境中接受世界一流的培训，该环境是动态和多学科的。研究和培训的具体领域包括光学，显微镜，仪器，图像处理，机器学习，聚合物材料，细胞生物学，分子生物学和基因组学。学员将能够利用通过研究项目开发的高水平技能、成就和工程头脑，在工业和学术界获得出色的职业生涯。