Sensing local nano-environment with coherent Raman microspectroscopy

使用相干拉曼显微光谱检测局部纳米环境

• 批准号: 10218816
• 负责人: Vladislav V. Yakovlev
• 金额: $ 18.08万
• 依托单位: TEXAS ENGINEERING EXPERIMENT STATION
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10218816
• 关键词: Affect; Biochemical Reaction; Biological; Biological Process; Cell physiology; Cells; Chemicals; Concentration measurement; Detection; Environment; Goals; Heterogeneity; Image; In Vitro; Label; Life; Lipids; Measures; Methods; Microscope; Microscopic; Microscopy; Modality; Molecular; Monitor; Phase; Photobleaching; Physiologic pulse; Raman Spectrum Analysis; Relaxation; Research; Research Proposals; Resolution; Sensitivity and Specificity; Signal Transduction; Speed; Structure; System; Testing; Time; Work; contrast imaging; detection sensitivity; experimental study; imaging modality; improved; in vivo; live cell imaging; microscopic imaging; molecular imaging; multiplexed imaging; nanoenvironment; novel; novel strategies; optical imaging; programs; prototype; response; single molecule; spectroscopic imaging; temporal measurement; tool; vibration

ABSTRACT This exploratory research proposal which aims to investigate a novel approach for interrogating local nano- environments within a living cell by improving the detection sensitivity of coherent Raman microspectroscopy as applied to live-cell imaging. The proposed new approach overcomes the existing shortcomings of coherent Raman spectroscopy. It provides an exquisite tool of microscopic chemically specific imaging with greatly improved sensitivity of detection and exceptional sensitivity to the local nano-environment. The central hypothesis is that vibrational coherence excited by two broadband pulses is sensitive to the local environment and can be precisely measured by interrogating this coherence using a third pulse at a variable time delay. By choosing a molecular vibration with a relatively narrow spectral line that corresponds to a long relaxation time, and measuring time-delayed response, background signal can be substantially reduced at long delay times of the probe pulse. This adjustment makes low-concentration measurements possible. The overall research goal of the proposed program is to validate this hypothesis by demonstrating sensitivity of coherent Raman microspectroscopy to a local nano-environment and by improving the detection sensitivity of Raman spectroscopy. The specific aims of the proposed activities are focused on constructing a prototype of a microscope and exploring its capabilities using both in vitro and in vivo systems. The proposed research program is expected to deliver a novel and label-free optical imaging modality with enhanced sensitivity and specificity that is suitable for live-cell imaging.

抽象的 这项探索性研究计划旨在研究一种询问局部纳米的新方法 通过提高相干拉曼显微光谱的检测灵敏度来提高活细胞内的环境 应用于活细胞成像。所提出的新方法克服了相干的现有缺点 拉曼光谱。它提供了一种精巧的显微化学特异性成像工具，具有极大的 提高了检测灵敏度以及对局部纳米环境的卓越灵敏度。中央 假设是由两个宽带脉冲激发的振动相干性对当地环境敏感 并且可以通过使用可变时间延迟的第三个脉冲询问这种相干性来精确测量。经过 选择具有相对较窄谱线的分子振动，对应于较长的弛豫时间， 并测量延时响应，在较长的延迟时间下可以大大减少背景信号 探测脉冲。这种调整使得低浓度测量成为可能。研究总体目标 拟议程序的目的是通过证明相干拉曼的灵敏度来验证这一假设 对局部纳米环境进行显微光谱分析并提高拉曼的检测灵敏度 光谱学。拟议活动的具体目标集中于构建一个原型 显微镜并利用体外和体内系统探索其功能。拟议的研究 该计划预计将提供一种新颖的、无标记的光学成像方式，具有增强的灵敏度和 适合活细胞成像的特异性。