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.

生物系统本质上在空间上是异质的，在时间上是动态的，在本质上是复杂的。 一个巨大的挑战是如何研究大量玩家在各个相关领域的互动。 长度尺度，范围从蛋白质复合物中相互作用的蛋白质网络到内部的细胞器 细胞、组织内的各种细胞类型以及功能器官内的协同组织。 同时监测生物体内大量相互作用物种的能力 具有足够时空分辨率的系统对于表征和分析是必不可少的。 了解潜在的复杂性。 然而，目前还没有合适的技术可以应对这一巨大挑战。 流行的“组学”技术不具备所需的时空分辨率，尤其是 对于三维样品或活体标本，光学显微镜只能对少数（2~5）个样品进行成像。 一次不同的目标，受到荧光基本“色垒”的限制。 打破光学显微镜的颜色障碍，弥合“组学”和“组学”之间的差距 成像，在这里我们提出了一个全新的振动光谱技术平台。 包括电子预共振受激拉曼散射 (epr-SRS) 和受激 将利用拉曼激发荧光（SREF）来实现最灵敏的拉曼 迄今为止，我们的初步数据已经证明了单分子敏感性。 通过探索二维激发光谱来开发该技术以达到~100 颜色，设计并合成成像探针库，开启超分辨率超- 该成像技术随后将在几个具有广泛影响的领域得到应用。 应用包括超级多重组织病理学、绘制全脑架构 复杂性和活细胞的超多参数深度表型分析。 光学显微镜的创新改变了许多生物问题的解决方式 就像共焦显微镜是生物医学实验室和双光子的主力一样。 荧光显微镜已经改变了体内脑成像，我们设想我们新提出的 超多重光谱和显微镜将打破当前的技术瓶颈， 彻底改变多色光学成像，并成为全系统研究的新标准 一般复杂系统。