Salipalli Sandeep, Singh Prafull Kumar, Borlak Jürgen
Centre for Pharmacology and Toxicology, Hannover Medical School, Carl-Neuberg-Str, 1, 30625 Hannover, Germany.
BMC Cell Biol. 2014 Jul 8;15:26. doi: 10.1186/1471-2121-15-26.
Live cell imaging enables the study of dynamic processes of living cells in real time by use of suitable reporter proteins and the staining of specific cellular structures and/or organelles. With the availability of advanced optical devices and improved cell culture protocols it has become a rapidly growing research methodology. The success of this technique relies mainly on the selection of suitable reporter proteins, construction of recombinant plasmids possessing cell type specific promoters as well as reliable methods of gene transfer. This review aims to provide an overview of the recent developments in the field of marker proteins (bioluminescence and fluorescent) and methodologies (fluorescent resonance energy transfer, fluorescent recovery after photobleaching and proximity ligation assay) employed as to achieve an improved imaging of biological processes in hepatoma cells. Moreover, different expression systems of marker proteins and the modes of gene transfer are discussed with emphasis on the study of lipid droplet formation in hepatocytes as an example.
活细胞成像通过使用合适的报告蛋白以及对特定细胞结构和/或细胞器进行染色,能够实时研究活细胞的动态过程。随着先进光学设备的出现和细胞培养方案的改进,它已成为一种迅速发展的研究方法。这项技术的成功主要依赖于合适报告蛋白的选择、具有细胞类型特异性启动子的重组质粒的构建以及可靠的基因转移方法。本综述旨在概述标记蛋白(生物发光和荧光)领域的最新进展以及用于改善肝癌细胞生物过程成像的方法(荧光共振能量转移、光漂白后荧光恢复和邻近连接分析)。此外,还讨论了标记蛋白的不同表达系统和基因转移模式,并以肝细胞中脂滴形成的研究为例进行了重点阐述。