In utero mouse embryo phenotyping with high-frequency ultrasound

项目总结/摘要 这项计划的目标是通过分割选择的器官来对早期至中期妊娠的小鼠胚胎进行表型分析 在子宫内使用高频超声（HFU）采集的3D数据集中使用系统。国际小鼠Phe- 包括NIH敲除（KO）小鼠表型分型计划（KOMP 2）在内的非分型联盟（IMPC）将 在未来十年内产生20，000种小鼠品系，包括许多重要的人类结构出生模型 缺陷和先天性疾病。开发表型分析方法，提供有效的管道 对KO小鼠品系中胚胎生长缺陷的分析是这一努力的高度优先事项。子宫内3D 成像方法，能够在一定范围内对各种器官系统进行体积和纵向分析。 早期至中期妊娠期小鼠胚胎，将提供额外的贝内和关键的额外体内数据 目前不可用。商业HFU系统在许多研究中心广泛使用，这主要归功于 美国国立卫生研究院资助的小动物成像研究计划和共享仪器计划。HFU是 因此，这是一种提供可定量分析的子宫内3D图像数据的优秀候选模式 并存档以支持KOMP 2/IMPC胚胎致死表型分析管道和未来的表型分析工作。 我们建议开发和验证子宫内3D HFU图像采集协议和图像处理方法， ODS允许对胚胎发育进行非侵入性的纵向研究，特别是检测和 KO表型的表征。将在子宫内从分期的小鼠胚胎中收集体积HFU数据 在E9.5至15.5之间，以建立正常发育的数据库。算法将被开发， 分割3D区域并提取量化胚胎阶段的参数， 正常和KO胚胎。我们将使用定制的环形阵列系统和VisualSonics Vevo 2100.高分辨率环形阵列数据将用于初步开发图像处理算法， 则算法将适用于Vevo 2100数据。我们将比较得出的定量参数 从两台扫描仪获得的分割结果，以确保Vevo 2100能够提供 等同的突变体检测和定量。将使用野生型和En 1和Gli 2进行初始测试 有已知缺陷的突变体最后，将采集和处理协议应用于3D Vevo 来自5-10个在各种器官系统中具有胚胎缺陷的KOMP 2 KO小鼠品系的2100个数据，以验证 用于检测和表征这些突变胚胎中的表型的HFU方法。

Project Summary/Abstract The goal of this proposal is to phenotype early- to mid-gestational mouse embryos by segmenting select organ systems in 3D data sets acquired in utero with high-frequency ultrasound (HFU). The International Mouse Phe- notyping Consortium (IMPC), which includes the NIH Knockout (KO) Mouse Phenotyping Program (KOMP2), will generate 20,000 mouse strains in the next decade, including many important models of human structural birth defects and congenital diseases. The development of phenotyping methods that provide for efficient pipeline analyses of defects in embryonic growth in the KO mouse strains is a high priority for this effort. An in utero 3D imaging approach, enabling volumetric and longitudinal analyses of a variety of organ systems over a range of early- to mid-gestational stage mouse embryos, would provide added benefit and critical additional in vivo data not currently available. Commercial HFU systems are widely available in many research centers largely thanks to the NIH-funded Small Animal Imaging Research Programs and Shared Instrumentation Programs. HFU is therefore an excellent candidate modality to provide in utero 3D image data that can be quantitatively analyzed and archived to support the KOMP2/IMPC embryonic lethal phenotyping pipeline and future phenotyping efforts. We propose to develop and validate in utero 3D HFU image-acquisition protocols and image-processing meth- ods that permit noninvasive, longitudinal studies of embryonic development and, in particular, the detection and characterization of KO phenotypes. Volumetric HFU data will be collected in utero from mouse embryos staged between E9.5 to 15.5 in order to establish a database of normal development. Algorithms will be developed to segment 3D regions and extract parameters that quantify embryonic stage and identify regional changes between normal and KO embryos. We will acquire data with a custom, annular-array system and with a VisualSonics Vevo 2100. The fine-resolution annular-array data will be used to initially develop the image-processing algorithms and then the algorithms will be adapted for Vevo 2100 data. We will compare the quantitative parameters derived from the segmentation results obtained from the two scanners to ensure that the Vevo 2100 is able to provide equivalent mutant detection and quantification. Initial testing will be undertaken using wild-type and En1 and Gli2 mutants that have known defects. Finally, the acquisition and processing protocols will be applied to 3D Vevo 2100 data from 5-10 KOMP2 KO mouse lines with embryonic defects in a variety of organ systems to validate the HFU methods for detecting and characterizing phenotypes in these mutant embryos.