Small Animal Tomography System for Green Fluorescent Protein Imaging

用于绿色荧光蛋白成像的小动物断层扫描系统

• 批准号: 8137808
• 负责人: ANDREAS H HIELSCHER
• 金额: $ 33.61万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-20 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8137808
• 关键词: Accounting; Affect; Algorithms; Animal Model; Animals; Area; Autopsy; Behavior; Biology; Bioluminescence; Brain; Cell Line; Cells; Characteristics; Chimeric Proteins; Code; Data; Data Set; Development; Devices; Diffusion; Disease; Disease Progression; Equation; Equipment; Fluorescence; Fluorescent Probes; Frequencies; Gene Proteins; Goals; Green Fluorescent Proteins; Growth; Herpesvirus 1; Histocompatibility Testing; Image; Imaging Techniques; In Vitro; Knowledge; Lead; Light; Liver; Location; Lung; Magnetic Resonance Imaging; Malignant Neoplasms; Measurement; Measures; Methods; Microscopic; Modeling; Molds; Molecular; Mus; Neoplasm Metastasis; Noise; Optical Methods; Optical Tomography; Optics; Organ; Organelles; Pathway interactions; Performance; Phase; Positioning Attribute; Positron-Emission Tomography; Process; Property; Proteins; Radiolabeled; Reading; Reporter; Reporter Genes; Research; Research Personnel; Resolution; Scheme; Signal Transduction; Signaling Molecule; Simulate; Solutions; Source; Spatial Distribution; Staging; Surface; System; Techniques; Testing; Thymidine Kinase; Time; Tissues; Work; Xenograft procedure; absorption; anticancer research; base; design; detector; experience; fluorescence imaging; fluorophore; human disease; image reconstruction; improved; in vivo; innovation; insight; instrumentation; interest; light scattering; mouse model; novel; optical imaging; protein transport; radiotracer; reconstruction; single molecule; theories; tomography; treatment effect; treatment response; tumor; tumor progression; tumorigenesis

DESCRIPTION (provided by applicant): The overall goal of this proposal is to develop image reconstruction schemes and instrumentation for tomographic imaging of green fluorescent proteins (GFP) in small animals. Small animal imaging has gained considerable importance in recent years as more and more animal models for human diseases have become available. Imaging of disease progression and effects of treatments in animals has many scientific and economical advantages, as sacrificing animals at various disease stages and performing necropsy and histopathological studies can be sharply reduced. Optical techniques have proven to be very valuable when applied to small animal imaging because of an abundance of optical markers that can target and visualize various disease related processes on the cellular and molecular level. However, to date most optical imaging studies have only explored whole animal surface imaging without tomographic reconstruction. Only a few groups including our team have presented first tomographic results that reveal three-dimensional fluorescent probe distribution in small animals. In these cases the image reconstructions were exclusively based on the diffusion model of light propagation in tissue, which is an approximation to the more generally applicable transport model. This has limited the application of such codes to studies involving fluorophores that emit in the near-infrared, where tissue absorption is smaller than in the visible spectrum. Furthermore, only steady-state instrumentation or frequency-domain devices with modulation frequencies smaller than 150 MHz were employed. While promising, these systems still suffer from cross-talk between absorption and scattering effects, limited spatial resolution, and limited accuracy in fluorescence absorption and lifetime determination. It is highly desirable to extend optical tomographic methods to shorter wavelengths, which would provide a means to image green fluorescence proteins that emit light in the visible spectrum. Using GFP and its derivatives, it is now possible to visualize nearly any protein of interest in any cell, tissue, or species. Researchers working at all levels of biology, such as single-molecule dynamics, protein trafficking within cells, organelle dynamics, and cell and tissue behaviors during development, cancer progression, and other diseases, are nowadays making use of GFP mostly in vitro studies. The availability of an optical tomography system for in vivo GFP imaging would have a significant impact on this large area of research. Themain hypothesis of this proposal is that limitations related to existing instrumentation for optical tomography can be overcome by an imaging system that uses a frequency-domain light propagation model that is based on the equation of radiative transfer (also called "transport equation"). Combined with instrumentation that allows for modulation frequencies higher than the currently available 150MHz, this will lead to better spatial resolution and reduced cross-talk between scattering and absorption effects. Furthermore, using a multi-wavelength and multi-frequency imaging system offers unique opportunities to reduce undesirable background signal due to autofluorescence. The overall goal of this proposal is to develop novel optical methods for volumetric imaging of small animals that are used in research to better understand and treat numerous diseases. For example, the imaging system could help visualize tumor location and growth. If successfully implemented the new imaging system will provide new insights into how various diseases, such as cancer, spread and affect the body.

描述（由申请人提供）：该提案的总体目标是开发图像重建方案和仪器，用于小动物中绿色荧光蛋白（GFP）的层析成像。近年来，随着越来越多的人类疾病动物模型已获得，小动物成像已变得非常重要。疾病进展的成像和在动物中的治疗作用具有许多科学和经济的优势，因为在各种疾病阶段牺牲动物并进行了死金和组织病理学研究可以大大降低。光学技术已被证明是非常有价值的，因为大量的光学标记物可以靶向和可视化各种疾病与细胞和分子水平的相关过程。然而，迄今为止，大多数光学成像研究仅探索了整个动物表面成像，而无需重构。只有包括我们团队在内的少数小组提出了首个层析成果结果，这些结果揭示了小动物中的三维荧光探针分布。在这些情况下，图像重建仅基于组织中光传播的扩散模型，这是对更普遍适用的运输模型的近似。这将此类代码的应用限制在涉及在近红外发出的荧光团的研究，在近红外发出的荧光团比可见的光谱小。此外，仅采用了小于150 MHz的调制频率的稳态仪器或频域设备。尽管有希望，但这些系统仍然遭受吸收和散射效应，有限的空间分辨率以及荧光吸收和寿命确定的精度有限的跨越。非常需要将光学层析成像方法扩展到较短的波长，这将提供一种图像图像绿色荧光蛋白的手段，该绿色荧光蛋白在可见光光谱中发出光。使用GFP及其衍生物，现在可以在任何细胞，组织或物种中可视化几乎所有感兴趣的蛋白质。如今，在发育，癌症进展和其他疾病期间，细胞内的单分子动力学，细胞动力学以及细胞和组织行为等生物学的研究人员如今主要是在使用GFP，主要是在体外研究。用于体内GFP成像的光学断层扫描系统的可用性将对这一大型研究产生重大影响。他们对该提案的假设是，与现有仪器相关的光学层析成像相关的局限性可以通过使用频域光传播模型的成像系统来克服，该模型基于辐射传递方程（也称为“传输方程”）。结合仪器的调制频率高于当前可用的150MHz，这将导致更好的空间分辨率，并减少散射和吸收效应之间的串扰。此外，使用多波长和多频成像系统提供了独特的机会，可以减少由于自动荧光而导致的不良背景信号。该提案的总体目标是开发新型的光学方法，用于对小动物的体积成像进行体积成像，这些动物被用于更好地理解和治疗多种疾病。例如，成像系统可以帮助可视化肿瘤的位置和生长。如果成功实施新的成像系统将提供有关各种疾病（例如癌症）如何传播和影响身体的新见解。

项目成果

Non-contact small animal fluorescence imaging system for simultaneous multi-directional angular-dependent data acquisition.

非接触式小动物荧光成像系统，用于同时多方向角度相关数据采集。

• DOI: 10.1364/boe.5.002301
• 发表时间: 2014
• 期刊: Biomedical optics express
• 影响因子: 3.4
• 作者: Lee,JongHwan;Kim,HyunKeol;Chandhanayingyong,Chandhanarat;Lee,FrancisYoung-In;Hielscher,AndreasH
• 通讯作者: Hielscher,AndreasH