Virtual histology of transgenic mouse embryos for high-throughput phenotyping.
Virtual histology of transgenic mouse embryos for high-throughput phenotyping.
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DOI：
10.1371/journal.pgen.0020061
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发表时间：
2006-04
期刊：
影响因子：
4.5
通讯作者：
Keller C
中科院分区：
文献类型：
--
作者：
Johnson JT;Hansen MS;Wu I;Healy LJ;Johnson CR;Jones GM;Capecchi MR;Keller C
一项大胆的新研究试图破坏小鼠基因组中的每一个基因，这需要系统的、跨学科的方法来分析小鼠胚胎中的模式缺陷。我们提出了一种新的、快速的、廉价的方法来获得高分辨率的虚拟组织学用于小鼠胚胎的表型评估。使用四氧化锇对组织进行差异染色，然后使用体积x射线计算机断层扫描对整个胚胎进行成像，扫描时间分别为2 h和12 h，获得27 μm和8 μm的等长分辨率，使用妊娠中期的E9.5-E12.5胚胎。由这种方法产生的数据集立即适用于最先进的器官模式分析计算方法。这种评估胚胎解剖的技术在分辨率、时间和费用上都有了显著的提高，可以对由基因工程突变和化学诱导的胚胎毒性引起的发育模式缺陷进行定量、三维分析。由于成像技术、仪器和软件的进步，发育生物学正在进入数字时代。这些进步与发育生物学的发现相结合，为我们提供了前所未有的见解，了解我们使用的产品、我们生活的环境和我们的基因组成是如何影响人类发展的。工业社会越来越关注妇女及其未出生胎儿暴露于药品和普通家用化学品的问题。此外，通过分离特定基因，现在可以了解出生缺陷的遗传原因，这些基因可以在胚胎中有效地破坏，随后观察出生缺陷。这类胚胎毒性研究通常涉及使用小鼠胚胎。然而，过去对小鼠胚胎的评估涉及昂贵和繁琐的外部检查和在显微镜下观察的薄切片。因此，发育生物学家热切地期待着工具的出现，这些工具将使他们能够使用快速而廉价的技术，对胚胎的复杂和动态组织进行常规评估。在这篇文章中，作者介绍了一种快速、高质量和廉价的技术，用于使用x射线计算机断层扫描对小鼠胚胎进行三维可视化，非常适合制药、工业和学术实验室的研究人员。
A bold new effort to disrupt every gene in the mouse genome necessitates systematic, interdisciplinary approaches to analyzing patterning defects in the mouse embryo. We present a novel, rapid, and inexpensive method for obtaining high-resolution virtual histology for phenotypic assessment of mouse embryos. Using osmium tetroxide to differentially stain tissues followed by volumetric X-ray computed tomography to image whole embryos, isometric resolutions of 27 μm or 8 μm were achieved with scan times of 2 h or 12 h, respectively, using mid-gestation E9.5–E12.5 embryos. The datasets generated by this method are immediately amenable to state-of-the-art computational methods of organ patterning analysis. This technique to assess embryo anatomy represents a significant improvement in resolution, time, and expense for the quantitative, three-dimensional analysis of developmental patterning defects attributed to genetically engineered mutations and chemically induced embryotoxicity. Developmental biology is entering the digital age, thanks to advancements in imaging technologies, instrumentation, and software. These advancements are converging with discoveries in developmental biology to deliver unprecedented insight into how human development is impacted by the products we use, the environment we live in, and our genetic composition. Industrial societies are becoming increasingly concerned with the exposure of women and their unborn fetuses to pharmaceuticals and commonplace household chemicals. In addition, understanding genetic causes of birth defects is now possible through the isolation of specific genes, which can be efficiently disrupted in embryos, and subsequently observed for birth defects. Such studies of embryotoxicity typically involve the use of mouse embryos. However, evaluation of mouse embryos in the past has involved expensive and cumbersome external inspection and thin sectioning for view under the microscope. As such, developmental biologists have eagerly anticipated the advent of tools that would allow them to routinely assess the complex and dynamic organization of embryos using techniques that are fast and inexpensive. In this article, the authors introduce a rapid, high-quality, and inexpensive technique for the three-dimensional visualization of mouse embryos using X-ray computed tomography that is ideally suited for researchers in pharmaceutical, industrial, and academic laboratories.
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DOI：
10.1073/pnas.95.13.7485
发表时间：
1998-06-23
影响因子：
11.1
作者：
Kasarskis, A;Manova, K;Anderson, KV
通讯作者：
Anderson, KV
影响因子：
5.7
作者：
Jacobs, RE;Ahrens, ET;Laidlaw, D
通讯作者：
Laidlaw, D
影响因子：
30.8
作者：
Weninger, WJ;Mohun, T
通讯作者：
Mohun, T
影响因子：
5.7
作者：
Zhang, JY;Miller, MI;Mori, S
通讯作者：
Mori, S
影响因子：
2.4
作者：
Schneider, JE;Bamforth, SD;Bhattacharya, S
通讯作者：
Bhattacharya, S