Time-lapse or live-cell fluorescence microscopy was performed using a Zeiss Axiovert 200 M microscope (Plan Apochromat 1006, 1.4NA objective) with an Ultra-View RS-3 spinning disk confocal system (PerkinElmer Inc., Shelton, CT, USA) which was equipped with a CSU21 confocal optical scanner, 12-bit digital cooled Hamamatsu Orca-ER camera (OPELCO, Sterling, VA, USA) and a 491 nm 100 mW and a 561 nm 50 mW laser illumination under the control of MetaMorph Premier Software (Universal Imaging, New York, NY, USA) [39 (link),40 (link)]. Briefly, z-stacks comprised of 0.5 µm-spaced sections that were captured. GFP excitation were performed at 491 nm (Emission 525/40 nm).
Bright-field microscopy was carried out by an Olympus IX71 microscope (Olympus, Tokyo, Japan) equipped with a Plan APO 100X/1.45 objective. Images were captured by Photometrics CoolSNAP HQ camera (Tucson, AZ, USA) and processed with MetaVue (Universal Imaging, Downingtown, PA, USA), Adobe Illustrator (Adobe Inc., San Jose, CA, USA), and ImageJ (LOCI, University of Wisconsin, Madison, WI, USA).
Bright-field microscopy was carried out by an Olympus IX71 microscope (Olympus, Tokyo, Japan) equipped with a Plan APO 100X/1.45 objective. Images were captured by Photometrics CoolSNAP HQ camera (Tucson, AZ, USA) and processed with MetaVue (Universal Imaging, Downingtown, PA, USA), Adobe Illustrator (Adobe Inc., San Jose, CA, USA), and ImageJ (LOCI, University of Wisconsin, Madison, WI, USA).
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