Microfluidic Sample Handling for the Spectroscopic Integration (SPIN) Program

用于光谱积分 (SPIN) 计划的微流体样品处理

• 批准号: RGPIN-2020-05371
• 负责人: Foulds, Ian
• 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: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=740536
• 关键词: Microfluidic; Sample; Handling; Spectroscopic; Integration

Long Term Vision To develop a microfluidic platform to integrate distinct single cell spectroscopic technologies for the study of heterogeneous cell populations with single cell granularity across an unprecedented spectral range. The Full SPectrum INterrogation Project (Full SPIN) is an initiative to incorporate multiple spectral modalities in order to gain a complete picture of dynamics within heterogeneous cell populations with single cell granularity. Full SPIN take disaggregated cell samples, image the cells, and then characterize single cell transmission spectra using four complementary technologies: Radio Frequency spectroscopy, Terahertz spectroscopy, Raman spectroscopy, and (cavity-reflection-enhanced) Visible spectroscopy. This system will allow for the cell-indexed correlation of the spectra at a single cell level so that their contribution to the bulk response can be inferred and the heterogeneous nature of the population can be better understood. The proposed work spans the fabrication of microfluidic modules with optimized materials for each spectroscopic technology, the cell handling needed to index a library of cells so that the spectra can be correlated at the single cell level across all the interrogation wavelengths, and the demonstration of the platform's utility by studying the cell population's response to stimulus. The work supported in this proposal will involve 2 PhD, 1 MASc and 2 undergraduate students per year for the duration of the funding. The HQP will be focused on the development and demonstration of the various modules that make up the Full SPIN platform. Human samples will not be used in the work proposed, which will focus on technology development and use standard cell lines to demonstrate the functionality and utility of the platform. The modular platform consists of several components: an indexing chip to generate a library of single cells stored in a water-in-oil emulsion, an index cartridge that will store the library and maintain cell viability, and a set of interrogation chips customized to each of the spectral interrogation methods. The library of cells will be transferred in the index cartridge to each interrogation chip in turn and a second index cartridge will collect the library again at the output of the interrogation chip and then transfer the library to the next interrogation module in turn. The final milestone for the proposed work is processing an indexed cell library of >1000 cells, imaging the cells, and acquiring correlated spectra in all of the spectral windows, both before and after a stimulus event. The resulting data will then be used to enable the statistical modelling of the individual cellular contribution population's response to the stimuli. The applications of this technology platform are broad, answering basic questions about heterogeneous cellular response.

开发一个微流控平台，整合不同的单细胞光谱技术，在前所未有的光谱范围内研究具有单细胞粒度的异质细胞群。全光谱询问项目（Full SPIN）是一项整合多光谱模式的倡议，目的是获得单细胞粒度的异质细胞群体内动态的完整图像。全自旋采集分解的细胞样品，对细胞成像，然后使用四种互补技术表征单个细胞的透射光谱：射频光谱、太赫兹光谱、拉曼光谱和（腔反射增强）可见光谱。该系统将允许在单个细胞水平上的光谱的细胞索引相关性，以便可以推断它们对总体响应的贡献，并且可以更好地理解种群的异质性。提出的工作包括为每种光谱技术使用优化材料制作微流体模块，对细胞库进行索引所需的细胞处理，以便在单个细胞水平上对所有的探测波长进行光谱关联，以及通过研究细胞群体对刺激的反应来展示平台的实用性。在资助期内，每年将有2名博士、1名硕士和2名本科生参与。HQP将侧重于开发和演示构成全自旋平台的各种模块。拟议的工作将不使用人类样本，其重点是技术开发和使用标准细胞系来展示平台的功能和效用。模块化平台由几个组件组成：一个索引芯片，用于生成存储在油包水乳液中的单个细胞库，一个索引盒，用于存储库并保持细胞活力，以及一套针对每种光谱询问方法定制的询问芯片。细胞库将在索引盒中依次转移到每个审讯芯片，第二个索引盒将在审讯芯片的输出处再次收集细胞库，然后依次将细胞库转移到下一个审讯模块。这项工作的最后一个里程碑是处理一个包含bb101000个细胞的索引细胞库，对细胞进行成像，并在刺激事件前后的所有光谱窗口中获取相关光谱。得到的数据将用于建立个体细胞贡献群体对刺激反应的统计模型。该技术平台的应用是广泛的，回答了有关异质细胞反应的基本问题。