Department of Pathology, Stanford University School of Medicine, Stanford, California, USA.
Dev Dyn. 2012 Aug;241(8):1301-9. doi: 10.1002/dvdy.23818. Epub 2012 Jun 23.
Polarization of tissue is achieved by asymmetric distribution of proteins and organelles within individual cells. However, existing quantitative assays to measure this asymmetry in an automated and unbiased manner suffer from significant limitations.
Here, we report a new way to assess protein and organelle localization in tissue based on correlative fluorescence analysis. As a proof of principle, we successfully characterized planar cell polarity dependent asymmetry in developing Drosophila melanogaster tissues on the single cell level using fluorescence cross-correlation.
Systematic modulation of signal strength and distribution show that fluorescence cross-correlation reliably detects asymmetry over a broad parameter space. The novel method described here produces robust, rapid, and unbiased measurement of biometrical properties of cell components in live tissue that is readily applicable in other model systems.
组织的极化是通过单个细胞内蛋白质和细胞器的不对称分布来实现的。然而,现有的定量检测方法在自动化和无偏倚的方式下存在显著的局限性。
在这里,我们报告了一种基于荧光相关分析来评估组织中蛋白质和细胞器定位的新方法。作为原理验证,我们成功地使用荧光互相关在单细胞水平上对发育中的果蝇组织中平面细胞极性依赖的不对称性进行了特征描述。
系统地调节信号强度和分布表明,荧光互相关可以在广泛的参数空间中可靠地检测不对称性。这里描述的新方法能够快速、稳健、无偏地测量活组织中细胞成分的生物计量特性,在其他模型系统中也很容易应用。