Brillouin Microscope for Biomedical Research

用于生物医学研究的布里渊显微镜

• 批准号: 10015304
• 负责人: Vladislav V. Yakovlev
• 金额: $ 28.9万
• 依托单位: TEXAS ENGINEERING EXPERIMENT STATION
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-10 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10015304
• 关键词: Applications Grants; Atherosclerosis; Atomic Force Microscopy; Biocompatible Materials; Biological; Biological Markers; Biological Process; Biomechanics; Biomedical Research; Cells; Communities; Development; Device or Instrument Development; Disease; Embryo; Environment; Feedback; Fertilization; Fluorescence; Goals; Growth and Development function; Hour; Image; Imaging Device; Magnetism; Malignant Neoplasms; Measurement; Methodology; Microscope; Microscopic; Microscopy; Minor; Modality; Modification; Morphogenesis; Multiphoton Fluorescence Microscopy; Noise; Optics; Performance; Physiologic pulse; Property; Regenerative Medicine; Research; Resolution; Sampling; Sickle Cell Anemia; Signal Transduction; Spectrum Analysis; Speed; System; Technology; Time; Tissues; Ultrasonography; Validation; Zebrafish; angiogenesis; base; bioimaging; carcinogenesis; cellular imaging; clinical application; data acquisition; design and construction; driving force; elastography; imaging capabilities; imaging modality; improved; instrument; instrumentation; laser tweezer; microscopic imaging; novel; optical imaging; response; technology development; technology research and development; temporal measurement; tool; viscoelasticity

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.

该提案是出于评估微观粘弹性特性的需要的动机，这些特性已出现为 用于多种疾病的强大生物标志物，例如癌症，动脉粥样硬化，镰状细胞疾病等，但 也已被确定为许多生物过程的驱动力，例如癌变， 血管生成，形态生成等。新型生物材料的再生医学出现也称为 为了更好地理解生物力学细胞水平相互作用。 过去，对组织的弹性特性的评估主要限于大规模成像 超声和磁谐振成像以及使用两种光学的纳米镜接触评估 镊子或原子力显微镜仪器，这为我们更好地理解铺平了道路 细胞和组织的粘弹性及其对生物医学研究的重要性。同时，是 现在意识到，能够无侵入性评估的仪器中存在巨大的技术差距 具有足够高的空间分辨率，高灵敏度和高的显微镜尺度上的粘弹性特性 速度。最近，成功开发了光学连贯性弹性图，以评估 尺度上的组织15-100 𝜇𝑚。理想情况下，这样的工具应与现有的 使用荧光和拉曼显微镜系统的仪器提供了额外的能力 那些。布里鲁因显微镜已成为非侵入性生物医学成像的强大工具。开发它 进入生物医学研究强大的工具，并可能被认为是 该提议的总体目标。 通过此赠款申请将采取两种策略。第一种方法是依靠赞助 Brillouin显微镜使用更简单，并且具有相对较小的修改，可以将其实现为 大型生物医学界已经存在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鲁因光谱和显微镜来提高效率 信号和数据采集速率令人惊讶的5个数量级。这种方法利用超平 脉冲兴奋，与多光子荧光显微镜，第二和第三谐波完全兼容 显微镜和连贯的抗烟拉曼显微镜。非线性布里鲁因的另一个好处 显微镜是改进的分段功能。总体策略是设计，构建和表征 这两个显微镜都并行，因为每个显微镜都为特定应用程序集提供了不同的优势，所以 并展示其对生物学相关系统的成像功能，以形象细胞的生长和 响应局部粘弹性环境和图像在期间开发斑马鱼胚胎的图像 受精后的前72小时。