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BBB seminar: Akihiko Nakano

State-of-the-art live cell imaging at high-speed and super-resolution - A dream to see real vesicular trafficking has come true

Akihiko Nakano
Live Cell Super-Resolution Imaging Research Team, RIKEN Center for Advanced Photonics, Wako, and Department of Biological Sciences, Graduate School of Science, University of Tokyo, Japan

Live imaging has proved powerful to solve many problems in cell biology, especially in the field of membrane trafficking. We have made several important observations such as the demonstration of Golgi cisternal maturation (Ref. 1), visualization of cargo transport from the ER exit site to the cis-Golgi by the hug-and-kiss action (Ref. 2), and from an early zone to a late zone in a maturing Golgi cisterna (Ref. 3) by pulse-chase imaging using the super-resolution confocal live imaging microscopic (SCLIM) method we have developed. Spatiotemporal resolution is critical in these studies, because the dynamics of vesicles often holds the clue to be able to understand underlying mechanisms. We have recently built up a next-generation SCLIM, SCLIM2. This employs an ultrahigh-sensitivity, high-speed and multicolor detection system and enables us to perform 3-color 4D imaging at the resolution of 70-100 nm in space and 20 volumes/sec in time by single-photon counting and a new precise deconvolution algorithm we have invented. I will present movies taken by SCLIM2, which show amazingly dynamic behaviors of Golgi cisternae, clathrin-coated buds and vesicles, as well as cargo, in yeast, plant and animal cells.

References:

1) Nakano et al. Live imaging of yeast Golgi cisternal maturation. Nature 441:1007, 2006

2) Nakano et al. Contact of cis-Golgi with ER exit sites executes cargo capture and delivery from the ER. Nat. Commun. 5:3653, 2014

3) Nakano et al. Visualization of secretory cargo transport within the Golgi apparatus. J. Cell Biol. 218:1602, 2019
 

Chairperson: Jaakko Saraste, Department of Biomedicine