【摘要】 光学显微镜的技术革新将极大地改变生物系统的研究方式。虽然荧光显微成像是目前细胞成像的首选方法,但在"组学"时代,荧光探针分子量过大且通常不能应用于5种颜色以上的标记和成像,其应用受到极大的限制。因此提出了两种化学成像策略。首先,设计了一种适于探测生物小分子动力学的活细胞生物正交化学成像平台。该方案将新兴的受激拉曼散射显微镜与微小的拉曼探针(例如炔烃、腈和包括²H和¹³C的稳定同位素)结合,并应用在众多生物医学研究中,如脂肪酸代谢和毒性、葡萄糖摄取和代谢、药物传输、脑内蛋白质合成、DNA复制、蛋白质降解、RNA合成和肿瘤代谢等。其次,发明了一种超多路复用光学成像技术。开发了电子预共振受激拉曼散射（EPR-SRS）显微镜,实现了优良的振动选择性,具有高的通用性和灵敏度。化学上,创建了由独特的新型染料组成的振动调色板,结合共轭和同位素编辑的三键,在拉曼频谱寂默窗口具有良好和单一的拉曼峰。目前可标记24种不同的颜色,并具有进一步扩展的巨大潜力。使用这种方法,监测了培养神经元和脑组织中的DNA及蛋白质的代谢。这种超复用光学成像方法有望促进复杂生物系统中相互作用关系的解开,并且可以在光子学和生物技术中找到更广泛的应用。更多还原

【Abstract】 Innovations in light microscopy have tremendously revolutionized the way researchers study biological systems.Although fluorescence microscopy is currently the method of choice for cellular imaging,it faces fundamental limitations such as the bulky fluorescent tags and limited multiplexing ability in the era of"omics". In this paper,two chemical imaging strategies are presented respectively. First,a live-cell bio-orthogonal chemical imaging platform suited for probing the dynamics of small bio-molecules,which can not be effectively labeled by bulky fluorophores is devised. This scheme couples the emerging stimulated Raman scattering microscopy with tiny and Raman-active vibrational probes(e. g.,alkynes,nitriles and stable isotopes including 2H and 13C). Exciting biomedical applications such as imaging fatty acid metabolism related to lipotoxicity,glucose uptake and metabolism,drug trafficking,protein synthesis in brain,DNA replication,protein degradation,RNA synthesis and tumor metabolism will be presented. Second,a super-multiplex optical imaging technique is invented. Electronic pre-resonance stimulated Raman scattering(epr-SRS)microscopy is developed,achieving exquisite vibrational selectivity with high versatility and sensitivity. Chemically,a unique vibrational palette is created,consisting of novel dyes bearing conjugated and isotopically-edited triple bonds,each displaying a single epr-SRS peak in the cell-silent spectra window. Up to 24 resolvable colors are currently achieved with great potential for further expansion. Using this approach,we monitored DNA and protein metabolism in neuronal co-cultures and brain tissues. This super-multiplex optical imaging approach promises to facilitate untangling the intricate interactions in complex biological systems,and find broad applications in photonics and biotechnology in general.更多还原