Johnson John T, Hansen Mark S, Wu Isabel, Healy Lindsey J, Johnson Christopher R, Jones Greg M, Capecchi Mario R, Keller Charles
Scientific Computing and Imaging Institute, University of Utah, Salt Lake City, Utah, United States of America.
PLoS Genet. 2006 Apr;2(4):e61. doi: 10.1371/journal.pgen.0020061. Epub 2006 Apr 28.
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 mum or 8 mum 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.
一项旨在破坏小鼠基因组中每个基因的大胆新举措需要系统的、跨学科的方法来分析小鼠胚胎中的模式缺陷。我们提出了一种新颖、快速且廉价的方法，用于获得高分辨率虚拟组织学，以对小鼠胚胎进行表型评估。使用四氧化锇对组织进行差异染色，然后通过容积X射线计算机断层扫描对整个胚胎进行成像，使用妊娠中期E9.5 - E12.5胚胎时，分别在2小时或12小时的扫描时间内实现了27微米或8微米的等距分辨率。通过这种方法生成的数据集可立即用于器官模式分析的最新计算方法。这种评估胚胎解剖结构的技术在分辨率、时间和费用方面有显著改进，可用于对由基因工程突变和化学诱导的胚胎毒性导致的发育模式缺陷进行定量三维分析。
