Raykova Doroteya, Koos Björn, Asplund Anna, Gelléri Márton, Ivarsson Ylva, Danielson U Helena, Söderberg Ola
Department of Pharmaceutical Biosciences, Pharmaceutical Cell Biology, Biomedical Center, Box 594, Uppsala University, SE-751 08 Uppsala, Sweden.
Department of Systemic Cell Biology, Max Planck Institute of Molecular Physiology, Otto-Hahn-Straße 11, 44227 Dortmund, Germany.
Proteomes. 2016 Nov 29;4(4):36. doi: 10.3390/proteomes4040036.
The invention of the microscope has been fundamental for the understanding of tissue architecture and subcellular structures. With the advancement of higher magnification microscopes came the development of various molecular biology tools such as Förster resonance energy transfer (FRET) and proximity ligation assay ( PLA) to monitor protein interactions. Microscopy has become a commonly used method for the investigation of molecular events within the cell, for the identification of key players in signaling networks, and the activation of these pathways. Multiple approaches are available for functional analyses in single cells. They provide information not only on the localization of proteins at a given time point, but also on their expression levels and activity states, allowing us to pinpoint hallmarks of different cellular identities within tissues in health and disease. Clever solutions to increase the sensitivity of molecular tools, the possibilities for multiplexing, as well as image resolution have recently been introduced; however, these methods have their pros and cons. Therefore, one needs to carefully consider the biological question of interest along with the nature of the sample before choosing the most suitable method or combination of methods. Herein, we review a few of the most exciting microscopy-based molecular techniques for proteomic analysis and cover the benefits as well as the disadvantages of their use.
显微镜的发明对于理解组织结构和亚细胞结构至关重要。随着更高放大倍数显微镜的发展，诸如荧光共振能量转移（FRET）和邻近连接分析（PLA）等各种分子生物学工具得以开发，用于监测蛋白质相互作用。显微镜检查已成为研究细胞内分子事件、识别信号网络中的关键参与者以及激活这些途径的常用方法。单细胞功能分析有多种方法可用。它们不仅能提供蛋白质在给定时间点的定位信息，还能提供其表达水平和活性状态信息，使我们能够在健康和疾病状态下的组织中确定不同细胞身份的特征。最近已经引入了提高分子工具灵敏度、多重分析可能性以及图像分辨率的巧妙解决方案；然而，这些方法各有优缺点。因此，在选择最合适的方法或方法组合之前，需要仔细考虑感兴趣的生物学问题以及样本的性质。在此，我们综述了一些最令人兴奋的基于显微镜的蛋白质组学分析分子技术，并阐述了其使用的优点和缺点。
