T. Haraguchi et al., Application of GFP: Time-lapse multi-wavelength fluorescence imaging of living mammalian cells, ACT HIST CY, 33(3), 2000, pp. 169-175
The jellyfish green fluorescent protein (GFP) is a powerful molecular tool
to fluorescently label specific proteins by fusing the GFP-coding gene to t
he gene of interest; the dynamic behavior of these fusion proteins can be e
xamined in living cells. In order to examine the spatial and temporal coord
ination of the GFP fusion protein with other intracellular structures, we u
se a time-lapse multiple-wavelength fluorescence microscope system that is
capable of recording simultaneously mutiple cellular components in the livi
ng 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 c
ytological technology is also applicable for visualization of mitotic and m
eiotic 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 relationships components.