Application of GFP:Time-Lapse Multi-Wavelength Fluorescence Imaging of Living Mammalian Cells

Abstract

The jellyfish green fluorescent protein(GFP) is a powerful molecular tool to fluorescently label specific proteins by fusing the GFP−coding gene to the gene of interest; the dynamic behavior of these fusion proteins can be examined in living cells. In order to examine the spatial and temporal coordination of the GFP fusion protein with other intracellular structures, we use a time−lapse multiple−wavelength fluorescence microscope system that is capable of recording simultaneously multiple cellular components in the living state. For example, using the system, we have observed dynamic behavior of chromosomes and several GFP fusion proteins−such as lamin B receptor−GFP, cyclin B1−GFP and CENP−B−GFP−in living human cells during mitosis. This cytological technology is also applicable for visualization of mitotic and meiotic events in yeast cells. Thus, live observation of GFP fusion proteins is useful for understanding the full relevance of the temporal and spatial relationships between multiple cellular components.