PFI:AIR - TT: Multifocal Laser Tweezers Raman Spectroscopy for Parallel Spectral Analysis of Biological Cells

这个PFI：加速创新研究（AIR）技术翻译（TT）项目专注于翻译单细胞拉曼光谱，以填补细胞术市场对新技术的需求，这些技术不需要使用外源性标签来分析活细胞的生物化学。多焦点激光镊子拉曼光谱（M-LTRS）技术很重要，因为它提供了通过直接询问其内在生物化学来评估细胞组成、结构和功能来分析单个活细胞的新能力。这种独特的能力有可能克服现有细胞术方法的局限性，这将导致生物学和生物医学发现的新进展，最终将对医疗保健产生重大影响。该项目将产生一个概念验证m - lts系统，该系统能够分析溶液中的二维（2-D）光学捕获细胞阵列。这种m - lts方法具有以下独特的特点：多达100个细胞的平行拉曼光谱分析，活细胞的无创和无标记化学分析，以及细胞动力学和对环境刺激的反应的长期监测。这些特性提供了更高的分析效率、独特的分析能力和性能，无论是与研究界的LTRS仪器的现状相比，还是与目前市场上领先的基于荧光细胞术的竞争方法相比。由于拉曼散射过程本身的微弱信号，单细胞拉曼光谱具有较低的分析通量。一次只能分析少数细胞的能力是目前拉曼光谱在许多生物应用中的主要限制，这些应用通常需要分析大量细胞以确定系统的异质性。如何通过同时检测许多单个细胞的拉曼光谱来进一步提高通量，目前存在技术差距。随着技术从研究发现向商业应用的转变，该项目解决了这一差距。本项目采用的策略是开发一种多焦点LTRS系统，该系统能够光学捕获二维细胞阵列并同时询问其光谱。将开发一种新的检测方案，该方案涉及不同激光焦点组合的快速快门，允许单个相机探测器检测不同叠加的拉曼光谱模式。将开发一种反卷积算法，当应用于叠加光谱时，将能够从每个细胞中并行检索单个拉曼光谱，而不会产生光谱串扰。在这个概念验证项目的结论中，预计这种新的检测方案将使单细胞拉曼光谱的吞吐量提高100倍。此外，参与该项目的人员，包括本科生和研究生，将通过加州大学戴维斯分校的既定项目（如创业学院）接受创业培训，该项目侧重于将科学创新商业化，并通过每周与外部合作伙伴的互动。该项目邀请Desatoya有限责任公司在从研究发现到商业现实的技术转化工作的商业化和科学方面提供必要的指导。

This PFI: Accelerating Innovation Research (AIR) Technology Translation (TT) project focuses on translating single cell Raman spectroscopy to fill the need for new techniques in the cytometry market that do not require the use of exogenous labels for analyzing the biochemistry of living cells. The multifocal laser tweezers Raman spectroscopy (M-LTRS) technology is important because it offers new capabilities for analyzing single living cells by directly interrogating their intrinsic biochemistry to assess cellular composition, structure, and function. Such unique capabilities have the potential to overcome limitations of existing cytometry methods, which will lead to new advancements in biological and biomedical discoveries that ultimately will have a significant impact on healthcare. The project will result in a proof-of-concept M-LTRS system that is capable of analyzing a two-dimensional (2-D) array of optically trapped cells in solution. This M-LTRS approach has the following unique features: parallel Raman spectral analysis of up to one hundred cells, non-invasive and label-free chemical analysis of living cells, and long-term monitoring of cell dynamics and response to environmental stimuli. These features provide improved analytical efficiency, unique analytical capabilities, and improved performance when compared to the current state of LTRS instruments in the research community or when compared to the leading competing fluorescence based cytometry methods currently in this market space. Single cell Raman spectroscopy has a low analytical throughput due to the intrinsically weak signals of a Raman scattering process. The ability to analyze only a few cells at a time is currently a major limitation of Raman spectroscopy for many biological applications, which often require the analysis of a large number of cells in order to determine the heterogeneity of the system. A technology gap currently exists on how to further improve the throughput by detecting the Raman spectra of many individual cells simultaneously. This project addresses this gap as the technology translates from research discovery toward commercial application. The strategy applied in this project is to develop a multifocal LTRS system that is capable of optically trapping a 2-D array of cells and to simultaneously interrogate their spectra. A novel detection scheme will be developed that involves the rapid shuttering of different combinations of the laser foci, allowing for different superimposed Raman spectral patterns to be detected by a single camera detector. A deconvolution algorithm will be developed that, when applied to the superimposed spectra, will enable the parallel retrieval of the individual Raman spectra from each cell with no spectral crosstalk. At the conclusion of this proof-of-concept project, demonstration of a 100-fold improvement in the throughput of single cell Raman spectroscopy with this new detection scheme is expected. In addition, personnel involved in this project, including undergraduate and graduate students, will receive training in entrepreneurship through established programs at UC Davis (e.g. Entrepreneurship Academy) that focus on commercializing science innovations as well as through weekly interactions with the external collaborative partner.The project engages Desatoya, LLC to provide essential guidance in both the commercialization and scientific aspects in this technology translation effort from research discovery toward commercial reality.