Microfluidic Sample Handling for the Spectroscopic Integration (SPIN) Program

Long Term Vision开发一个微流体平台，集成不同的单细胞光谱技术，用于研究具有前所未有光谱范围的单细胞粒度的异质细胞群体。 全谱查询项目(Full Spin)是一项将多种光谱模式结合在一起的倡议，以获得单细胞粒度的不同细胞群体内动态的完整图景。Full Spin采集分解的细胞样本，对细胞成像，然后使用四种互补技术来表征单细胞的传输光谱：射频光谱、太赫兹光谱、拉曼光谱和(腔反射增强的)可见光光谱。这一系统将允许在单个细胞一级对光谱进行细胞索引相关，从而可以推断它们对总体响应的贡献，并可以更好地了解群体的异质性。这项拟议的工作包括为每种光谱技术制造具有优化材料的微流控模块，为细胞库编制索引以使光谱可以在单个细胞水平上跨所有询问波长进行关联所需的细胞处理，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。 这项提案支持的工作在资助期间每年将涉及2名博士、1名硕士和2名本科生。HQP将专注于组成全旋转平台的各种模块的开发和演示。拟议的工作将不使用人体样本，该工作将侧重于技术开发，并使用标准细胞系来展示平台的功能和用途。 该模块化平台由几个部件组成：用于生成存储在油包水乳剂中的单细胞库的索引芯片、将存储该库并保持细胞活力的索引盒、以及为每种光谱询问方法定制的一组询问芯片。单元库将在索引盒中被依次转移到每个询问芯片，并且第二索引盒将在询问芯片的输出处再次收集该库，然后将该库依次转移到下一个询问模块。 这项拟议工作的最后一个里程碑是处理1000个细胞的索引细胞库，对这些细胞成像，并在所有光谱窗口中获取相关光谱，包括在刺激事件之前和之后。然后，所得到的数据将被用于对单个细胞贡献群体对刺激的反应进行统计建模。这个技术平台的应用范围很广，回答了关于异质细胞响应的基本问题。

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.