High-resolution spectroscopic imaging with infrared nonlinear optical (IR-NLO) microscopy

项目摘要 一个真正的无标记成像技术，振动显微镜提供细胞和组织的地图 具有非常高的化学对比度，因为它直接探测的基本振动模式 样品振动成像方法包括红外吸收显微光谱法和共焦显微光谱法。 拉曼显微镜，已成功商业化的方法（不断增长的5亿） 美元市场），现在是分析和生物实验室中常见的调查工具。 在过去的四十年里，这些技术在生物学领域产生了重大影响。 和生物医学，提供健康和患病组织的空间分辨评估， 分子视角该建议旨在显著提高振动的能力， 显微镜我们提出了一种新的成像方法，融合了IR的理想特性 具有相干非线性光学（NLO）特性的吸收显微镜 分子的激发。这种新颖的红外非线性光学技术提高了红外吸收的空间分辨率 显微镜的10倍，同时提供更高的灵敏度指纹分子振动相对于 拉曼显微镜方法。 我们的团队由相干拉曼散射显微镜和红外 显微光谱成像我们的创新使快速获取红外吸收图像成为可能 指纹振动模式的分辨率为0.5微米或更好。初步数据 表明IR-NLO方法可以成功地应用于快速激光扫描显微镜， 允许组织样本的方便的振动成像。在我们的建议中，我们开发，测试， 改进新的IR-NLO技术。该技术的验证是通过广泛的 生物医学成像研究和比较与最先进的红外显微镜今天可用。 拟议的计划解决了红外光谱显微镜的一个主要挑战，即 提高成像分辨率。这一新功能意义重大，因为更高的分辨率 组织中亚微米细胞内和细胞外结构的识别， 在红外成像中仍然不可见。因此，高分辨率成像特性 提高了该技术的诊断能力。通过设定新的分辨率标准， 指纹振动成像，IR-NLO技术很可能在组织中产生重大影响 成像，并且可以使其能够用于病理学的研究和临床领域。

Project Summary A genuine label-free imaging technology, vibrational microscopy provides maps of cells and tissues with exceptionally high chemical contrast as it directly probes the fundamental vibrational modes of samples. Vibrational imaging approaches include IR-absorption micro-spectroscopy and confocal Raman microscopy, methods that have been successfully commercialized (a growing 500 million dollar market) and are now common tools of inquiry found in analytical and biological laboratories. Over the past four decades, these techniques have had a measurable impact in the fields of biology and biomedicine, offering a spatially resolved assessment of healthy and diseased tissues from a molecular perspective. This proposal aims to significantly improve the capabilities of vibrational microscopy. We propose a new imaging approach that merges the desirable properties of IR absorption microscopy with some of the unique properties of coherent, nonlinear optical (NLO) excitation of molecules. This novel IR-NLO technique improves the spatial resolution of IR absorption microscopy by tenfold, while offering higher sensitivity to fingerprint molecular vibrations relative to Raman-based microscopy methods. Our team is comprised of experts in coherent Raman scattering microscopy and IR microspectroscopic imaging. Our innovation makes it possible to rapidly acquire IR absorption images of fingerprint vibrational modes with a resolution of 0.5 micrometer or better. The preliminary data shows that the IR-NLO approach can be successfully adopted in a rapid laser-scanning microscope, allowing convenient vibrational imaging of tissue specimens. In our proposal we develop, test, and improve the new IR-NLO technology. The validation of the technology is achieved through extensive biomedical imaging studies and comparison with the state of the art IR microscopy available today. The proposed program tackles a major challenge in IR spectroscopic microscopy, namely the improvement of imaging resolution. This new capability is significant, as the higher resolution enables the identification of sub-micrometer intra- and extra-cellular structures in the tissue, which hitherto have remained invisible in IR-imaging. The high-resolution imaging property thus dramatically improves the diagnostic capabilities of the technique. By setting a new resolution standard for fingerprint vibrational imaging, the IR-NLO technology is likely to have a significant impact in tissue imaging and can enable its use in both research and clinical domains for pathology.