Tracking the subcellular protein localization process by combining the time-gated method and photoconvertible fluorescence protein

Abstract

Chlorophyll within chloroplast emits high-intensity autofluorescence, which interferes with the observation of exogenous fluorescent substance such as fluorescent protein in the plant cell. We previously reported that clear fluorescence observation can be done by using the time-gated method that eliminates the autofluorescence. Recently, we succeeded in tracking the subcellular localization process of the blue-light photoreceptor phototropin (Phot) to the chloroplast periphery in the liverwort Marchantia polymorpha by a combination of the confocal laser microscopy for the time-gated method and a photoconvertible fluorescent protein. Here, as an example of the time-gated fluorescence imaging, we introduce a study about intracellular localization and movement of Phot in M. polymorpha.