Fluorescence microscopy was performed using a CSU-X spinning-disk confocal microscopy system (Intelligent Imaging Innovations) or a DeltaVision Elite system (GE Healthcare Life system). The CSU-X spinning-disk confocal microscopy system was equipped with a DMI 6000B microscope (Leica), 100×/1.45-NA objective, and a QuantEM electron-multiplying charge-coupled device camera (Photometrics). Imaging was done at room temperature in YNB medium using GFP and mCherry channels with different exposure times according to the fluorescence intensity of each protein. Images were analyzed and processed with SlideBook 6.0 software (Intelligent Imaging Innovations).
The DeltaVision Elite system is equipped with an Olympus IX-71 inverted microscope, DV Elite complementary metal-oxide semiconductor camera, a 100×/1.4-NA oil objective, and a DV Light SSI 7 Color illumination system with Live Cell Speed Option with DV Elite filter sets. Imaging was done at room temperature in YNB medium using GFP and mCherry channels with different exposure times according to the fluorescence intensity of each protein. Image acquisition and deconvolution (conservative setting; five cycles) were performed using DeltaVision software softWoRx 6.5.2 (Applied Precision).
The DeltaVision Elite system is equipped with an Olympus IX-71 inverted microscope, DV Elite complementary metal-oxide semiconductor camera, a 100×/1.4-NA oil objective, and a DV Light SSI 7 Color illumination system with Live Cell Speed Option with DV Elite filter sets. Imaging was done at room temperature in YNB medium using GFP and mCherry channels with different exposure times according to the fluorescence intensity of each protein. Image acquisition and deconvolution (conservative setting; five cycles) were performed using DeltaVision software softWoRx 6.5.2 (Applied Precision).