Optical Coherence Tomography for Embryology

用于胚胎学的光学相干断层扫描

基本信息

批准号：
BB/E002870/1
负责人：
Adrian Podoleanu
金额：
$ 72万
依托单位：
University of Kent
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2006
资助国家：
英国
起止时间：
2006 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FE002870%2F1
关键词：
Optical Coherence Tomography Embryology

项目摘要

The proposed research programme aims to develop novel high-resolution imaging tools for Embryology based on optical coherence tomography (OCT), a non-invasive imaging technology that, compared to confocal microscopy, provides enhanced depth resolution and penetration, especially when the sample is several millimetres away from the microscope objective. The activity will be carried jointly, by the Applied Optics Group (AOG) of the School of Physical Sciences and the Cell and Developmental Biology group of the Biosciences Department at the University of Kent at Canterbury. The two teams will amalgamate expertise in complementary areas and pool resources to form an interdisciplinary team to investigate the potential of OCT based measurement and imaging platforms on well-characterised as well as on novel animal models for embryology and cell imaging. The research will focus on: 1. New approaches for label-less analysis and 4D imaging of cells and embryos; 2. Development of a combined simultaneous or sequential OCT/fluorescence system for allowing direct comparison of OCT with fluorescence imaging and generating a platform capable of 4-D imaging GFP-expressing and fluorescently labelled tissues and embryos; 3. Development of contrast enhancement procedures, mainly based on protein-tagging. The work will benefit from prior expertise of the AOG in developing several innovative aspects of the OCT technology for in-vivo imaging of the eye and for in-vitro imaging of several types of tissue. The activity will initially use fully functional OCT systems within the AOG implemented over the last 5 years of active research in the field of OCT. Development of novel non-invasive imaging systems is aimed at providing the much higher resolution required for imaging cells and embryos. Therefore, at the start, the research will embark on microscopy related improvements such as developing specialised high resolution probe heads to respond to the needs of biological imaging. We will test this specialised OCT microscope system by imaging the morphogenetic process of dorsal closure in live wild type and mutant Drosophila embryos, analysing for the first time cell and tissue movements at the dorsal and ventral surfaces in a single Z-series. Further, the research activity will develop a combined OCT/fluorescence system to address a double target: (i) fluorescence imaging simultaneously or sequentially with OCT, allowing direct comparison of OCT with fluorescence imaging and generating a platform capable of 4-D imaging GFP-expressing and fluorescently labelled tissues and embryos, (ii) development of contrast enhancement procedures for OCT imaging, allowing for protein-tagging and deep imaging of cellular structure. A versatile platform will be devised to accommodate several combinations of fluorescence and OCT bands, the exact configuration depending upon whether the fluorescence band is close or superposes to the OCT bandwidth. Accommodating different bands for OCT operation, which requires single mode fibre delivery is an expensive exercise. Therefore, we will devise a configuration that will allow, with minimal changes, adaptation to the widest variety of fluorescence/OCT band pairs. We will then embark on evaluating Phytochrome A as an in vivo contrast agent for OCT imaging. Novel nonfluorescent and nonbioluminescent molecular imaging probes have been proposed recently, such as pump probe OCT, pump suppression OCT, ground state recovery OCT that will initiate new directions in coherent optical molecular imaging. Probes and techniques designed for coherent molecular imaging are likely to improve the detection and diagnostic capabilities of OCT. Therefore it is timely to consider such avenues. We aim to further improve our OCT imaging capability for 4-D cellular imaging at depth in studies of embryonic development by genetically engineering Drosophila strains to express photoactive Phytochrome A cytoplasmically and as a protein tag.

拟议的研究计划旨在开发基于光学相干断层扫描（OCT）的新型高分辨率成像工具，该工具是一种非侵入性成像技术，与共聚焦显微镜相比，该技术提供了增强的深度分辨率和渗透率，尤其是当样本是几个时毫米远离显微镜目标。这项活动将由肯特大学肯特大学的生物科学系的物理科学学院以及坎特伯雷大学生物科学系的细胞和发育生物学小组共同进行。这两个团队将在互补领域和集合资源中融合专业知识，以组建一个跨学科的团队，以研究基于OCT的测量和成像平台的潜力，并在新颖的动物模型和细胞成像的新型动物模型上进行了研究。该研究将重点介绍：1。无标记分析的新方法和细胞和胚胎的4D成像； 2。开发同时或顺序的OCT/荧光系统，用于将OCT与荧光成像进行直接比较，并生成能够表达GFP的4-D成像和荧光标记的组织和胚胎的平台； 3。发展对比度增强程序，主要基于蛋白质标记。这项工作将受益于AOG的先前专业知识，以开发OCT技术的几个创新方面，用于对眼睛的体内成像和几种类型的组织的体外成像。该活动最初将在OCT领域的过去5年中实施的AOG中使用功能齐全的OCT系统。新型非侵入成像系统的开发旨在提供成像细胞和胚胎所需的更高的分辨率。因此，一开始，研究将开始与显微镜相关的改进，例如开发专门的高分辨率探针头以响应生物成像的需求。我们将通过对活野生型和突变的果蝇胚胎中的背闭合的形态发生过程进行成像，从而分析单个Z系的背侧和腹侧表面的首次时间细胞和组织运动。此外，研究活动将开发一个联合的OCT/荧光系统，以解决双重目标：（i）荧光成像与OCT同时或顺序成像，从而可以将OCT与荧光成像进行直接比较，并生成能够使用4D成像GFP-的平台表达和荧光标记的组织和胚胎，（ii）开发对比度成像的对比度增强程序，从而允许对细胞结构进行蛋白质标记和深层成像。将设计一个多功能平台以适应荧光和OCT频段的几种组合，确切的配置取决于荧光频带是接近还是超级置于OCT带宽。容纳不同的频段进行OCT操作，这需要单模式纤维交付是一项昂贵的练习。因此，我们将设计一种配置，该配置将允许最小的变化适应最宽的荧光/OCT带对。然后，我们将开始评估植物色素A作为OCT成像的体内对比剂。最近已经提出了新型的非荧光和非发光分子成像探针，例如泵探针OCT，泵抑制OCT，基态回收OCT，它将启动连贯的光学分子成像中的新方向。专为相干分子成像设计的探针和技术可能会改善OCT的检测和诊断能力。因此，考虑此类途径是及时的。我们旨在进一步提高我们的OCT成像能力，以通过遗传工程果蝇菌株对胚胎发育的深度进行4-D细胞成像，以表达光活性植物色素A细胞质量和蛋白质标签。