SBIR Phase I: High Throughput Cell Sorting with Coherent Raman Scattering (CRS )

SBIR 第一阶段：利用相干拉曼散射 (CRS) 进行高通量细胞分选

• 批准号: 1248414
• 负责人: Christian Freudiger
• 金额: $ 15万
• 依托单位: Invenio
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1248414&HistoricalAwards=false
• 关键词: SBIR; Phase; Throughput; Cell; Sorting

This Small Business Innovation Research (SBIR) Phase I project proposes to develop a flow cytometer for sorting cells based on chemical phenotyping, which (1) provides quantitative analysis of specific molecular species, (2) has single cell sensitivity, (3) is high throughput, (4) label-free, and (5) non-destructive to living organisms. The proposed system uses coherent Raman scattering (CRS), a laser spectroscopy based on analysis of the vibrations of chemical bonds. Compared to conventional Raman scattering, the CRS signal is coherently amplified, which should achieve sorting speeds up to 10,000 cells/s, which is 10,000x faster than what can currently be achieved with spontaneous Raman. Reaching this goal requires development of (1) a spectrally multiplexed CRS system based on a narrowband pump laser and a broadband probe laser, and (2) a high-speed, high-sensitivity multichannel detection system to simultaneously probe multiple spectral components of the broadband pump laser.The broader impact/commercial potential of this project derives from the variety of potential applications of this novel phenotyping technology in basic and applied R&D. One example is in the field of products from biological organisms (e.g., algae or yeast) including fuels and specialty chemicals, where the organism synthesizes the target chemical using energy either harvested from sunlight or from a low-value feedstock, such as glucose. Genetic modification is used to improve yield and specificity for the target chemical species. The process of mutation and selection can be repeated until an optimized outcome is obtained. High-throughput chemical phenotyping is required to sample a large number of genetic mutations and speed up the directed evolution process. It would be advantageous to perform such biochemical analysis non-destructively, so that the fitness of candidates could be tested in growth and stress assays after selection. In addition to biofuel production, there are numerous applications in basic research, as well as medical applications in both diagnostics and treatment.

本小型企业创新研究（SBIR）一期项目提出开发一种基于化学表型的细胞分选流式细胞仪，该流式细胞仪（1）提供特定分子种类的定量分析，（2）具有单细胞灵敏度，（3）高通量，（4）无标记，（5）对生物体无破坏性。所提出的系统使用相干拉曼散射（CRS），一种基于化学键振动分析的激光光谱学。与传统的拉曼散射相比，CRS信号被相干放大，这将实现高达10，000个细胞/秒的分选速度，这比目前利用自发拉曼可以实现的速度快10，000倍。达到该目标需要开发（1）基于窄带泵浦激光器和宽带探测激光器的光谱复用CRS系统，以及（2）高速，高灵敏度多通道检测系统，可同时探测宽带泵浦激光器的多个光谱分量。该项目的商业潜力来自于这种新型表型分析技术在基础和应用研发中的各种潜在应用&。一个例子是来自生物有机体的产品领域（例如，藻类或酵母），包括燃料和专用化学品，其中生物体使用从阳光或从低价值原料（如葡萄糖）收获的能量合成目标化学品。遗传修饰用于提高目标化学物质的产量和特异性。可以重复突变和选择的过程，直到获得优化的结果。需要高通量化学表型分析来对大量基因突变进行采样并加速定向进化过程。有利的是非破坏性地进行这种生物化学分析，使得候选物的适合性可以在选择后在生长和胁迫测定中进行测试。除了生物燃料生产之外，在基础研究以及诊断和治疗方面的医学应用中也有许多应用。