Structural and Molecular Phenotyping of Embryonic Development through Multi-Modal Optical Imaging

通过多模态光学成像进行胚胎发育的结构和分子表型

• 批准号: 10378053
• 负责人: David Mayerich
• 金额: $ 76万
• 依托单位: UNIVERSITY OF HOUSTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10378053
• 关键词: 3-Dimensional; Address; Adopted; Affect; Algorithms; Atlases; Benchmarking; Biological; Biological Assay; Blood Circulation; Blood Vessels; Blood flow; Cardiovascular system; Cell physiology; Cells; Central Artery; Chorion; Clinical; Complex; Computer software; Data; Data Collection; Data Set; Data Storage and Retrieval; Defect; Development; Developmental Biology; Developmental Process; Disease; Embryo; Embryonic Development; Erythroblasts; Event; Exhibits; Fetal Development; Fetal Growth Retardation; Fetus; Fluorescence; Goals; Growth; Health; Human; Hybrids; Image; Imaging Techniques; In Vitro; Infertility; Label; Lead; Ligands; Light; Methods; Microscopy; Modality; Modeling; Modification; Molecular; Morphogenesis; Mothers; Multi-modal optical imaging; Multimodal Imaging; Mus; Nature; Newborn Infant; Optical Coherence Tomography; Optics; Organ; Phenotype; Placenta; Pre-Eclampsia; Process; Proteins; Public Health; Reporter; Research; Research Personnel; Resolution; Risk; Rotation; Sampling; Scanning; Series; Signal Transduction; Software Tools; Specificity; Spontaneous abortion; Structure; Structure of umbilical artery; System; Techniques; Testing; Three-Dimensional Imaging; Time; Tissue imaging; Tissues; Transgenic Organisms; Translating; Umbilical vein; Venous; Work; allantois; base; computerized data processing; design; endothelial stem cell; experimental study; fetal; fetus circulation; hemodynamics; imaging modality; imaging platform; imaging system; in vivo; instrument; instrumentation; lens; loss of function; microscopic imaging; molecular phenotype; mouse model; multimodal data; multimodality; notch protein; novel; prevent; reconstruction; temporal measurement; terabyte; tool

PROJECT SUMMARY The ability to correlate between large-scale developmental milestones and micro-scale cellular and protein- specific changes is a significant unmet need in the study of developmental biology. The overall objective of this work is to develop a multi-modality imaging platform that can provide time resolved three-dimensional images of tissue development, with high temporal and spatial resolutions at a molecular level. Current studies rely on multiple imaging modalities to collect information on critical stages of vertebrate embryogenesis, and most offer only static snapshots of a single developmental stage. Live imaging with optical coherence tomography (OCT) provides high temporal resolution with contrast between tissue structures, allowing researchers to identify and test the mechanisms underlying developmental processes. Three- dimensional fluorescence approaches such as confocal and light sheet microscopy (LSM) provide increased resolution and molecular specificity which can be used to observe cellular mechanisms, such as the presence of erythroblasts indicating active blood flow, that are inaccessible to lower-resolution techniques. This will be accomplished by designing and developing a novel microscopic imaging system that provides spatially and temporally aligned OCT and light sheet microscopy images. Simultaneous images will be collected through OCT scanning and fluorescent light sheet excitation of the same sample plane. Fluorescence emission will be imaged through a second objective, while the OCT signal will be collected through the same lens in reflection mode. Software will be designed to synchronize data collection with an integrated high-precision rotational stage. A novel software toolkit will be developed to analyze this rich multi-modal data. Novel reconstruction methods will be designed to fuse both modalities, while addressing the sparse and multiplex nature of the LSM images and high frame rate of OCT. Finally, we'll use this tool to test the central hypothesis that a combined LSM+OCT imaging system can reveal the precise structural and molecular events required to form a circulatory loop between the embryo and maternal chorio-allantoic placenta. Successful accomplishment of the proposed work will generate a novel, integrated imaging platform, including instrumentation and analytical software, which could be widely adopted by developmental biologists to bridge the gap between large-scale developing phenotypes and the underlying molecular and cellular processes. We will benchmark this accomplishment by identifying currently unknown critical milestones in murine embryonic development. Specifically, LSM+OCT will be used to define the precise series of events necessary to form the umbilical artery (UA) and umbilical vein (UV). This research will clarify the sequence of events, including cellular, molecular, and global phenotypic changes, that lead to the establishment of an embryonic circulatory system between the mother and developing fetus, a critical prerequisite for embryonic survival.

项目总结 在大规模发育里程碑和微观细胞和蛋白质之间建立联系的能力-- 在发育生物学的研究中，特定的变化是一个重要的未得到满足的需求。这样做的总体目标是 工作是开发一个多通道成像平台，可以提供时间分辨率的三维图像 在分子水平上具有很高的时间和空间分辨率。 目前的研究依赖于多种成像方式来收集脊椎动物关键阶段的信息 胚胎发育，大多数只提供单个发育阶段的静态快照。使用光学技术进行实时成像 相干断层扫描(OCT)提供具有组织结构之间的对比度的高时间分辨率， 使研究人员能够识别和测试潜在的发育过程的机制。三个- 空间荧光方法，如共聚焦和光片显微镜(LSM)，提供了 分辨率和分子特异性，可用于观察细胞机制，如存在 表明血液流动活跃的红细胞，这是较低分辨率技术无法获得的。 这将通过设计和开发一种新型的显微成像系统来实现，该系统提供 在空间和时间上对齐的OCT和光片显微镜图像。将同时收集图像 通过OCT扫描和同一样品平面的荧光光片激发。荧光发射 将通过第二个物镜成像，而OCT信号将通过相同的透镜收集 反射模式。将设计软件，使数据收集与集成的高精度同步 旋转阶段。将开发一种新的软件工具包来分析这些丰富的多模式数据。小说 重建方法将被设计为融合这两种模式，同时解决稀疏和多路传输 LSM图像的性质和OCT的高帧频。最后，我们将使用该工具来检验中心假设 LSM+OCT组合成像系统可以揭示所需的精确结构和分子事件 在胚胎和母体绒毛尿囊膜胎盘之间形成循环循环。成功 拟议工作的完成将产生一个新颖的综合成像平台，包括 仪器和分析软件，可被发育生物学家广泛采用 大规模发育表型与潜在的分子和细胞过程之间的差距。我们 将通过确定小鼠胚胎中目前未知的关键里程碑来确定这一成就的基准 发展。具体地说，LSM+OCT将用于定义形成 脐动脉(UA)和脐静脉(UV)。这项研究将澄清事件的顺序，包括细胞， 导致胚胎循环系统建立的分子和全球表型变化 在母亲和发育中的胎儿之间，是胚胎存活的关键先决条件。