Brillouin Microscope for Biomedical Research

这一建议的动机是需要评估微观粘弹性，这已经出现， 是许多疾病，如癌症、动脉粥样硬化、镰状细胞病等的有力生物标志物，但 也被认为是许多生物过程的驱动力，如致癌作用， 血管生成，形态发生等。再生医学的新型生物材料的出现也要求 以更好地理解生物力学细胞水平的相互作用。 在过去，对组织的弹性性质的评估大多限于使用以下技术的大规模成像： 以及涉及使用光学或磁共振成像的纳米级接触评估 镊子或原子力显微镜仪器，这为我们更好地了解 细胞和组织的粘弹性及其对生物医学研究的重要性。与此同时， 现在意识到，在能够非侵入性评估的仪器方面存在巨大的技术差距， 在微观尺度上具有足够高的空间分辨率、高灵敏度和高的粘弹性能。 速度最近，成功地开发了光学相干弹性成像以评估 15-100μ l的组织。理想的情况是，这样一项文书应与现有文书完全兼容， 使用荧光和拉曼显微镜系统的仪器，以提供额外的能力， 那些.布里渊显微镜正在成为一个强大的工具，非侵入性生物医学成像。开发它 成为生物医学研究和潜在临床应用的强大工具被认为是 这一提案的总体目标。 两个战略将通过这一赠款申请。第一种方法是依靠自发 布里渊显微镜，其使用更简单，并且通过相对较小的修改，可以实现为 一种用于大型生物医学团体的现有商业荧光或拉曼显微镜的选择。 第二种策略是利用非线性布里渊光谱学和显微镜来提高 信号和数据采集速率提高了惊人的5个数量级。这种方法利用超短 脉冲激发，并与多光子荧光显微镜，第二和第三谐波完全兼容 显微镜和相干反斯托克斯拉曼显微镜。非线性布里渊的另一个好处 显微镜是改进的切片能力。总体战略是设计、构建和表征 这两个显微镜并行，因为每一个都为一组特定的应用提供了独特的优势， 并展示其用于生物相关系统的成像能力，以成像细胞生长， 发育响应于局部粘弹性环境和图像开发斑马鱼胚胎在 受精后72小时内。

This proposal is motivated by the need to assess microscopic viscoelastic properties, which have emerged as a powerful biomarker for a number of diseases, such as cancer, atherosclerosis, sickle cell disease, etc., but also have been identified as a driving force for many biological processes, such as carcinogenesis, angiogenesis, morphogenesis, etc. The emergence of novel biomaterials for regenerative medicine also calls for a better understanding of biomechanical cellular-level interactions. In the past, assessment of elastic properties of tissues was mostly limited to large-scale imaging using ultrasound and magnetic resonant imaging and to nanoscopic contact assessment using either optical tweezers or atomic force microscopy instruments, which paved the way to our better understanding of viscoelastic properties of cells and tissues and their importance for biomedical research. In the same time, it is now realized that there is a substantial technology gap in instrumentation capable of assessing non-invasively viscoelastic properties on a microscopic scale with high enough spatial resolution, high sensitivity and high speed. Recently, optical coherence elastography was successfully developed to assess elastic properties of tissues on the scale of 15-100 𝜇𝑚. Ideally, such an instrument should be fully compatible with existing instrumentation using fluorescence and Raman microscopy systems to provide an additional capability to those. Brillouin microscopy is emerging as a powerful tool for non-invasive biomedical imaging. Developing it into a powerful instrument for biomedical research and, potentially, clinical applications is considered to be the overarching goal of this proposal. Two strategies will be pursued through this grant application. The first approach is relying on spontaneous Brillouin microscopy, which is simpler in use, and, with relatively minor modifications, can be implemented as an option in already existing commercial fluorescent or Raman microscopes for a large biomedical community. The second strategy is to utilize nonlinear Brillouin spectroscopy and microscopy to boost the efficiency of the signal and data acquisition rate by astonishing 5 orders of magnitude. This methodology utilizes ultrashort pulse excitation and is fully compatible with multiphoton fluorescence microscopy, second- and third-harmonic microscopies and coherent anti-Stokes Raman microscopy. An additional benefit of nonlinear Brillouin microscopy is improved sectioning capabilities. The overall strategy is to design, construct and characterize both microscopes in parallel, since each of those offers distinct advantages for a particular set of applications, and to demonstrate their imaging capabilities for biologically relevant systems to image cells growth and development in response to a local viscoelastic environment and image developing zebrafish embryo during the first 72 hours post fertilization.