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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）的新型高分辨率胚胎学成像工具，这是一种非侵入性成像技术，与共聚焦显微镜相比，它提供了更高的深度分辨率和穿透性，特别是当样品距离显微镜物镜几毫米时。该活动将由物理科学学院的应用光学小组（AOG）和坎特伯雷肯特大学生物科学系的细胞与发育生物学小组联合进行。这两个团队将整合互补领域的专业知识，并汇集资源，组成一个跨学科团队，研究基于OCT的测量和成像平台在胚胎学和细胞成像的良好特征以及新型动物模型上的潜力。研究将集中在：1。细胞和胚胎无标记分析和4D成像的新方法2. 开发联合同步或顺序OCT/荧光系统，用于直接比较OCT与荧光成像，并生成能够对表达gfp和荧光标记的组织和胚胎进行4-D成像的平台；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在开发OCT技术的几个创新方面的专业知识，用于眼睛的体内成像和几种组织的体外成像。该活动最初将在过去5年在OCT领域积极研究的AOG中使用功能齐全的OCT系统。新型非侵入性成像系统的开发旨在提供成像细胞和胚胎所需的更高分辨率。因此，在一开始，研究将着手于显微镜相关的改进，如开发专门的高分辨率探头，以满足生物成像的需求。我们将通过在活的野生型和突变型果蝇胚胎中成像背部闭合的形态发生过程来测试这种专门的OCT显微镜系统，首次分析单个z系列中背部和腹部表面的细胞和组织运动。此外，该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)与OCT同时或顺序进行荧光成像，允许直接比较OCT和荧光成像，并生成一个能够对表达gfp和荧光标记的组织和胚胎进行4-D成像的平台；（ii）开发OCT成像的对比度增强程序，允许蛋白质标记和细胞结构的深度成像。将设计一个多功能平台，以适应荧光和OCT波段的几种组合，确切的配置取决于荧光波段是否接近或叠加到OCT带宽。适应不同波段的OCT操作需要单模光纤传输，这是一项昂贵的工作。因此，我们将设计一种结构，允许以最小的变化适应最广泛的荧光/OCT波段对。然后，我们将着手评估光敏色素A作为OCT成像的体内造影剂。近年来，新型的非荧光和非生物发光分子成像探针被提出，如泵浦探针OCT、泵浦抑制OCT、基态恢复OCT等，将为相干光学分子成像开辟新的方向。为相干分子成像设计的探针和技术可能会提高oct的检测和诊断能力，因此考虑这些途径是及时的。我们的目标是进一步提高我们的OCT成像能力，深入研究胚胎发育的4-D细胞成像，通过基因工程果蝇菌株表达光活性光敏色素A和作为蛋白质标签。