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

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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.