Microfluidic Sample Handling for the Spectroscopic Integration (SPIN) Program

长期愿景开发一个微流体平台，集成不同的单细胞光谱技术，用于研究跨前所未有的光谱范围的单细胞粒度的异质细胞群。 全光谱询问项目 (Full SPIN) 是一项整合多种光谱模式的举措，旨在获得单细胞粒度的异质细胞群动态的完整图像。 Full SPIN 采集分解的细胞样本，对细胞进行成像，然后使用四种互补技术表征单细胞透射光谱：射频光谱、太赫兹光谱、拉曼光谱和（腔体反射增强）可见光谱。该系统将允许在单细胞水平上对光谱进行细胞索引相关性，以便可以推断它们对整体响应的贡献，并可以更好地理解群体的异质性。拟议的工作涵盖为每种光谱技术使用优化材料制造微流体模块、索引细胞库所需的细胞处理，以便光谱可以在所有询问波长的单细胞水平上相互关联，以及通过研究细胞群对刺激的反应来演示该平台的实用性。 该提案支持的工作将在资助期间每年涉及 2 名博士生、1 名硕士生和 2 名本科生。 HQP 将专注于构成 Full SPIN 平台的各种模块的开发和演示。 拟议的工作将不会使用人体样本，该工作将专注于技术开发并使用标准细胞系来展示该平台的功能和实用性。 该模块化平台由多个组件组成：一个索引芯片，用于生成存储在油包水乳液中的单细胞文库；一个索引盒，用于存储文库并维持细胞活力；以及一组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询问芯片，并且第二个索引盒将在询问芯片的输出处再次收集库，然后将库依次转移到下一个询问模块。 拟议工作的最后一个里程碑是处理超过 1000 个细胞的索引细胞库，对细胞进行成像，并在刺激事件之前和之后获取所有光谱窗口中的相关光谱。 然后，所得数据将用于对单个细胞贡献群体对刺激的反应进行统计建模。 该技术平台的应用广泛，可以回答有关异质细胞反应的基本问题。

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.