Optical Coherence Tomography for Embryology

用于胚胎学的光学相干断层扫描

• 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

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）的新型胚胎学高分辨率成像工具，这是一种非侵入性成像技术，与共焦显微镜相比，可提供增强的深度分辨率和穿透力，特别是当样本为多个时距显微镜物镜几毫米。该活动将由坎特伯雷肯特大学物理科学学院应用光学组（AOG）和生物科学系细胞与发育生物学组联合开展。两个团队将合并互补领域的专业知识并汇集资源，组建一个跨学科团队，研究基于 OCT 的测量和成像平台在特征良好的以及胚胎学和细胞成像的新型动物模型上的潜力。该研究将重点关注： 1. 细胞和胚胎无标记分析和 4D 成像的新方法； 2. 开发同时或连续OCT/荧光组合系统，允许直接比较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4D成像的平台； 3. 开发对比度增强程序，主要基于蛋白质标记。这项工作将受益于 AOG 先前在开发用于眼睛体内成像和多种类型组织体外成像的 OCT 技术的几个创新方面的专业知识。该活动最初将使用 AOG 内功能齐全的 OCT 系统，这些系统是过去 5 年在 OCT 领域的积极研究中实施的。新型非侵入性成像系统的开发旨在提供细胞和胚胎成像所需的更高的分辨率。因此，一开始，该研究将着手显微镜相关的改进，例如开发专门的高分辨率探头来满足生物成像的需求。我们将通过对活体野生型和突变果蝇胚胎中背侧闭合的形态发生过程进行成像来测试这种专门的 OCT 显微镜系统，并首次分析单个 Z 系列中背侧和腹侧表面的细胞和组织运动。此外，该研究活动将开发一种组合的 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 与荧光成像，并生成能够 4-D 成像 GFP- 的平台。表达和荧光标记的组织和胚胎，(ii) 开发用于 OCT 成像的对比度增强程序，允许对细胞结构进行蛋白质标记和深度成像。将设计一个多功能平台来适应荧光和 OCT 波段的多种组合，具体配置取决于荧光波段是否接近或叠加于 OCT 带宽。适应 OCT 操作的不同频段需要单模光纤传输，这是一项昂贵的工作。因此，我们将设计一种配置，以最小的变化即可适应最广泛的荧光/OCT 波段对。然后我们将开始评估光敏色素 A 作为 OCT 成像的体内造影剂。最近提出了新型非荧光和非生物发光分子成像探针，例如泵浦探针OCT、泵浦抑制OCT、基态恢复OCT，这将开创相干光学分子成像的新方向。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，现在是考虑这些途径的时候了。我们的目标是通过对果蝇菌株进行基因改造，使其在细胞质中表达光活性光敏色素 A 并作为蛋白质标签，进一步提高我们在胚胎发育研究中进行 4-D 细胞成像的 OCT 成像能力。

项目成果

Quantitative assessment of rat bone regeneration using complex master-slave optical coherence tomography

使用复杂主从光学相干断层扫描定量评估大鼠骨再生

• DOI: http://dx.10.21037/qims.2019.05.03
• 发表时间: 2019
• 期刊: Quantitative Imaging in Medicine and Surgery
• 影响因子: 2.8
• 作者: Luca R

Multiple-depth en face optical coherence tomography using active recirculation loops

使用主动再循环环路的多深度正面光学相干断层扫描

• DOI: http://dx.10.1364/ol.35.002296
• 发表时间: 2010
• 期刊: Optics Letters
• 影响因子: 3.6
• 作者: Neagu L

Dual optical coherence tomography/fluorescence microscopy for monitoring of Drosophila melanogaster larval heart.

双光学相干断层扫描/荧光显微镜监测果蝇幼虫心脏。

• DOI: http://dx.10.1002/jbio.200910021
• 发表时间: 2009
• 期刊: Journal of biophotonics
• 作者: Bradu A

Fiber Optics, From Sensing to Non Invasive High Resolution Medical Imaging

光纤，从传感到非侵入性高分辨率医学成像

• DOI: http://dx.10.1109/jlt.2009.2032787
• 发表时间: 2010
• 期刊: Journal of Lightwave Technology
• 影响因子: 4.7
• 作者: Podoleanu A

Sequential en-face optical coherence tomography imaging and monitoring of Drosophila Melanogaster larval heart

果蝇幼虫心脏的顺序正面光学相干断层扫描成像和监测

• DOI: http://dx.10.1117/12.808159
• 发表时间: 2009
• 作者: Bradu A