Super-multiplex optical imaging: development of novel spectroscopy and probes to illuminate complex biomedicine

超级多重光学成像：开发新型光谱学和探针来阐明复杂的生物医学

• 批准号: 10622905
• 负责人: Wei Min
• 金额: $ 88.19万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10622905
• 关键词: 3-Dimensional; Address; Architecture; Biological; Brain Mapping; Brain imaging; Cells; Color; Complex; Confocal Microscopy; Data; Development; Electronics; Fluorescence; Fluorescence Microscopy; Image; Imaging technology; Length; Microscopy; Monitor; Nature; Optics; Organ; Organelles; Pathology; Phenotype; Proteins; Raman Spectrum Analysis; Resolution; Sampling; Specimen; Spectrum Analysis; System; Techniques; Technology; Time; Tissues; accurate diagnosis; biological systems; cell type; complex biological systems; design; disease diagnosis; image archival system; imaging probe; in vivo; innovation; light microscopy; multiplexed imaging; new technology; novel; optical imaging; protein complex; single molecule; spatiotemporal; technology platform; temporal measurement; two-dimensional; two-photon; ultra high resolution

Biological systems are inherently heterogeneous in space, dynamic in time, and complex in nature. A grand challenge is how to study the vast number of interacting players at every relevant length scale, ranging from protein network in protein complexes, to interacting organelles within cells, to various cell types within tissues, and to synergistic tissues within functional organs. Hence, the ability to simultaneously monitor a large number of interacting species inside biological systems with sufficient spatial-temporal resolution is indispensable for characterization and understanding of the underlying complexity. However, there is currently no suitable technology that can meet this grand challenge. The prevalent “omics” technologies do not have the required spatial-temporal resolution, especially for three-dimensional samples or living specimen. Optical microscopy can only image a few (2~5) different targets at once, limited by the fundamental “color barrier” of fluorescence. To break the color barrier of light microscopy and to bridge the gap between “omics” and imaging, here we propose a radically new technology platform. Novel vibrational spectroscopy including electronic-pre-resonance stimulated Raman scattering (epr-SRS) and stimulated Raman excited fluorescence (SREF) will be exploited, to achieve the most sensitive Raman imaging to date. Our preliminary data have proved single-molecule sensitivity. We will further develop the technique by exploring the two-dimensional excitation spectroscopy to reach ~100 colors, designing and synthesizing a library of imaging probes, opening up super-resolution super- multiplex imaging. The imaging technology will then be implemented in several broad-impact applications including super-multiplex tissue pathology, mapping brain-wide architecture complexity, and super-multiparameter deep phenotyping of living cells. Innovations in optical microscopy have changed the way many biological problems are addressed. Just like confocal microscopy is the work-horse in biomedical labs and two-photon fluorescence microscopy has transformed in vivo brain imaging, we envision our newly proposed super-multiplex spectroscopy and microscopy will break the current technical bottleneck, revolutionize multicolor optical imaging, and become a new standard for system-wide study of complex systems in general.

生物系统在空间上本质上是异质的，时间动态的，并且在 自然。一个巨大的挑战是如何研究每个相关的大量互动玩家 长度尺度，范围从蛋白质复合物中的蛋白质网络到在相互作用的细胞器中 细胞，组织中各种细胞类型，以及功能器官中的协同组织。因此， 简单地监测生物学内大量相互作用物种的能力 具有足够时空分辨率的足够时空分辨率的系统对于表征和 了解潜在的复杂性。 但是，目前尚无适合这一巨大挑战的合适技术。 普遍的“ OMICS”技术没有所需的时空分辨率，尤其是 用于三维样品或活标本。光学显微镜只能成像一些（2〜5） 一次不同的目标受到荧光的基本“色屏障”的限制。 打破光学显微镜的颜色屏障，并弥合“幻象”和 成像，在这里，我们提出了一个激进的新技术平台。新型振动光谱 包括电子抗性刺激拉曼散射（EPR-SR）并刺激 将探索拉曼激发荧光（SREF），以达到最敏感的拉曼 迄今为止的成像。我们的初步数据已证明是单分子灵敏度。我们将进一步 通过探索二维兴奋光谱来开发该技术，达到〜100 颜色，设计和综合成像问题库，打开超分辨率超级 - 多重成像。然后，成像技术将在几个宽大影响中实施 应用包括超级肌肉组织病理学，绘制脑部整体结构 复杂性和活细胞的超级型深度表型。 光学显微镜的创新改变了许多生物学问题的方式 解决。就像共聚焦显微镜是生物医学实验室和两光子的工作马 荧光显微镜已在体内脑成像中转化，我们设想了我们新提出的 超级肌光谱和显微镜将破坏当前的技术瓶颈， 革新多色光学成像，并成为全系统研究的新标准 一般而言，复杂的系统。