Imaging the Invisible with No Labels: A Major Opportunity in Biology and Medicine

在没有标签的情况下对不可见的物体进行成像：生物学和医学的一个重大机遇

• 批准号: 8137677
• 负责人: XIAOLIANG SUNNEY XIE
• 金额: $ 77.23万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8137677
• 关键词: Adoption; Area; Biochemistry; Biology; Biomedical Research; Brain Neoplasms; Cells; Chemicals; Cholesterol; Communities; DNA; Development; Drug Kinetics; Folate; Frequencies; Glucose; Image; Insulin Resistance; Knowledge; Label; Laboratories; Lead; Life; Lipids; Lipolysis; Malignant Neoplasms; Maps; Medicine; Messenger RNA; Microscopy; Molecular; Monitor; Operative Surgical Procedures; Optics; Organism; Outcome; Pharmaceutical Preparations; Proteins; Specificity; Spectrum Analysis; Techniques; Technology; Time; Tissues; Uncertainty; Unsaturated Fats; base; cancer diagnosis; chemical property; drug distribution; improved; interest; lipid biosynthesis; lipid metabolism; optical imaging; public health relevance; research study; saturated fat; small molecule; tumor

DESCRIPTION (provided by applicant): Small molecules such as metabolites and drugs are crucial to the biochemistry of living organisms because of their large diversity and the sheer number present. Unlike their large molecule counter parts such proteins, DNA and mRNA, most of these species are invisible in a living cell or organism because of technical difficulties. Tagging them with bulky fluorescent labels perturbs their function. Label-free optical imaging based on Raman scattering is highly desirable because it offers contrast based on the intrinsic chemical properties of molecules via molecular vibrational spectroscopy, but has been limited to low sensitivity and long acquisition times. Our group has recently developed stimulated Raman scattering (SRS) microscopy, which offers an unprecedented combination of high sensitivity, rapid image acquisition, chemical specificity and noninvasiveness. The development of SRS microscopy has generated a great of interest from the biomedical research community and studies of lipid metabolism and drug distributions in tissue are already underway in our laboratory. We propose to continue the pioneering development in order to probe lipid metabolism, including lipogenesis, lipolysis and insulin resistance. We will image not only different types of lipids, such as saturated and unsaturated lipids and cholesterol, but also key metabolites, such as glucose, ATP, ADP, and small molecule drugs as well, all in living cells and tissue without the use of perturbative labels. These new experiments will no doubt allow new observations that will enrich our knowledge of cellular biochemistry. We will also develop all-optical cancer markers and use SRS to bring about a new paradigm for intra-operative imaging of brain tumors, improving the surgical outcome by shortening surgery and allowing more precise definition of tumor margins. The widespread adoption of SRS and the discovery of new applications will revolutionize many areas of biomedical research, including lipid metabolism, pharmacokinetics and cancer diagnosis. PUBLIC HEALTH RELEVANCE: The ability to visualize small molecules such as metabolites and drugs in living cells and organisms without labels will revolutionize biomedical research, particularly lipid metabolism, pharmacokinetics and cancer diagnosis. We propose to extend the recently developed stimulated Raman scattering microscopy technique to offer rapid, sensitive and noninvasive imaging based on intrinsic vibrational frequencies of molecules, in order to selectively map the distributions of lipids, protein, glucose, cholesterol, ADP, ATP, folate and small molecule drugs, among other species. This technology will make an unprecedented transformation of our ability to monitor biochemistry in real time in living systems, and lead to a potential breakthrough in intra-operative imaging for rapid and precise tumor identification.

描述（由申请人提供）：小分子如代谢物和药物对生物体的生物化学至关重要，因为它们具有很大的多样性和存在的绝对数量。与它们的大分子对应物如蛋白质、DNA和mRNA不同，由于技术困难，这些物种中的大多数在活细胞或生物体中是不可见的。用大体积的荧光标记物标记它们会干扰它们的功能。基于拉曼散射的无标记光学成像是高度期望的，因为其经由分子振动光谱提供基于分子的固有化学性质的对比度，但是受限于低灵敏度和长采集时间。我们的小组最近开发了受激拉曼散射（SRS）显微镜，它提供了一个前所未有的高灵敏度，快速图像采集，化学特异性和非侵入性的组合。SRS显微镜的发展已经产生了极大的兴趣，从生物医学研究界和组织中的脂质代谢和药物分布的研究已经在我们的实验室进行。我们建议继续开拓性的发展，以探测脂质代谢，包括脂肪生成，脂解和胰岛素抵抗。我们不仅将成像不同类型的脂质，如饱和和不饱和脂质和胆固醇，而且还将成像关键代谢物，如葡萄糖，ATP，ADP和小分子药物，所有这些都在活细胞和组织中，而无需使用微扰标记。这些新的实验无疑将允许新的观察，这将丰富我们的细胞生物化学知识。我们还将开发全光学癌症标记物，并使用SRS为脑肿瘤的术中成像带来新的范例，通过缩短手术时间和更精确地定义肿瘤边缘来改善手术结果。SRS的广泛采用和新应用的发现将彻底改变生物医学研究的许多领域，包括脂质代谢、药代动力学和癌症诊断。 公共卫生相关性：可视化小分子的能力，如代谢物和药物在活细胞和生物体中没有标签，将彻底改变生物医学研究，特别是脂质代谢，药代动力学和癌症诊断。我们建议扩展最近开发的受激拉曼散射显微镜技术，以提供快速，灵敏和非侵入性成像的基础上的固有振动频率的分子，以选择性地映射分布的脂质，蛋白质，葡萄糖，胆固醇，ADP，ATP，叶酸和小分子药物，以及其他物种。这项技术将使我们在生命系统中真实的实时监测生物化学的能力发生前所未有的转变，并导致术中成像的潜在突破，以快速准确地识别肿瘤。