An improved method for three-dimensional reconstruction of protein expression patterns in intact mouse and chicken embryos and organs
An improved method for three-dimensional reconstruction of protein expression patterns in intact mouse and chicken embryos and organs
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DOI：
10.1369/jhc.7a7226.2007
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发表时间：
2007-09-01
影响因子：
3.2
通讯作者：
Hecksher-Sorensen, Jacob
中科院分区：
文献类型：
--
作者：
Ahnfelt-Ronne, Jonas;Jorgensen, Mette C.;Hecksher-Sorensen, Jacob
我们已经开发了一套完整的免疫荧光方案，可以同时检测小鼠和鸡胚胎中的多达三种蛋白质。与Murray的清洗剂(BABB)和安装在共焦显微镜上的具有长工作范围和高数值孔径的显微镜物镜相结合，可以在深处获得细胞分辨率，从而有可能检查整个器官或胚胎的表达模式。光学共焦段的三维投影可以用允许绕任何轴旋转的计算机软件来计算。该方案是健壮的，我们发现大多数作用于组织切片的抗体也适用于该方案。本手稿包含http://www.jhc.org.上的在线补充材料请在线访问这篇文章以查看这些材料。
We have developed a whole mount immunofluorescence protocol for the simultaneous detection of up to three proteins in mouse and chicken embryos. Combined with Murray's clearing reagent (BABB) and microscope objectives with long working ranges and high numerical apertures mounted on a confocal microscope, cellular resolution can be obtained in depths offering the possibility of examining expression patterns in entire organs or embryos. Three-dimensional projections of the optical confocal sections can be computed with computer software allowing rotation around any axis. The protocol is robust and we find that most antibodies working on tissue sections also work with this protocol. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials.