Collaborative Research: Fast Spectrally-Encoded Photoacoustic Microscopy for Multi-Parameter Bioenergetic Characterization of Heterogeneous Cancer Cells

与正常细胞相比，癌细胞具有独特的生物能量(能量产生)功能，与细胞新陈代谢和致癌(致癌)特性有关，细胞之间存在显著差异。因此，在单细胞水平上的生物能量学表征对于揭示不同的癌细胞能量产生途径是必要的。这种认识不仅对癌症的基础研究，而且对准确的癌症诊断和预后，以及个性化抗癌治疗的发展都是重要的。在该项目中，将开发一种新的高速高分辨率光谱成像技术和高密度微孔阵列。如果成功，该项目将为实现研究癌细胞代谢和异质性的重要方法奠定坚实的基础。将为研究生和本科生以及7-12年级的教师和学生提供学习和培训机会。技术翻译和产品开发将通过与生物仪器公司的合作进行探索。这项研究的目标是实现大量癌细胞的单细胞耗氧率(OCR)和胞外酸化率(ECAR)表征。OCR和ECAR与细胞内氧化磷酸化和糖酵解这两条主要代谢途径密切相关，因此可以作为其生物能量表型的主要指标。到目前为止，已经开发了电学或荧光方法来表征少数细胞的OCR。然而，很难将这些方法放大以用于大细胞群体的多参数表征。为了实现这一目标，将开发一种快速光谱编码光声显微镜，以在单一测量平台上实现高通量和同时进行OCR和ECAR表征。具体地说，将采用阿达玛变换和数字微镜器件的光开关来实现激发波长的快速调谐。一种新的水浸式微型扫描镜将被开发出来，以提供宽视场光学和声学共扫描。将这两种新技术结合起来，将允许在可编程或可针对特定目标重新配置的多个激发波长进行高速光声数据采集。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Compared to their normal counterparts, cancer cells have distinctive bioenergetic (energy generating) functions related to cell metabolism and tumorigenic (tumor causing) properties with significant cell-to-cell variations. Thus, bioenergetic characterization at the single-cell level is necessary to reveal the differing cancer cell energy producing pathways. Such understanding is important for not only fundamental cancer research but also for accurate cancer diagnosis and prognosis and for the development of personalized anti-cancer therapies. In this project, a new high-speed and high-resolution spectroscopic imaging technology and a high-density microwell array will be developed. If successful, this project will lay a solid foundation to achieve a significant method for studying cancer cell metabolism and heterogeneity. Learning and training opportunities will be provided for graduate and undergraduate students as well as Grade 7-12 teachers and students. Technical translation and product development will be explored through teaming with bioinstrumentation companies.The goal of this research is to achieve single-cell oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) characterization for a large population of cancer cells. OCR and ECAR are closely related to the two main metabolic pathways in cells, oxidative phosphorylation and glycolysis, and therefore serve as primary indicators of their bioenergetic phenotypes. To date, electrical or fluorescence methods have been developed to characterize the OCR of a small number of cells. However, it is difficult to scale these methods up for multi-parameter characterization of a large cell population. To accomplish this goal, a fast spectrally-encoded photoacoustic microscopy will be developed to enable high-throughput and simultaneous OCR andECAR characterization on a single measurement platform. Specifically, the Hadamard transformation and optical switching with digital micromirror device will be employed to enable fast tuning of the excitation wavelength. A new water-immersible micro scanning mirror will be developed to provide wide-field optical and acoustic co-scanning. Combining these two new techniques will allow high-speed photoacoustic data acquisition at multiple excitation wavelengths that are programmable or reconfigurable for a specific target.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.