X100 na 1

Fluorescence microscopy was performed using an Olympus IX81 microscope equipped with a x100/NA 1.40 (Olympus) objective and Orca-AG cooled CCD camera (Hamamatsu, JAPAN), using Metamorph software (Universal Imaging). Simultaneous imaging of red and green fluorescence was performed using an Olympus IX81 microscope, described above, and an image splitter (Dual-View; Optical Insights) that divided the red and green components of the images with a 565-nm dichroic mirror and passed the red component through a 630/50 nm filter and the green component through a 530/30 nm filter. These split signals were taken simultaneously with one CCD camera, described above.

Fluorescence microscopy was performed using an Olympus IX83 microscope equipped with a x100/NA 1.40 (Olympus) objective and Orca-R2 cooled CCD camera (Hamamatsu), using Metamorph software (Universal Imaging). FM4-64 staining was performed as described previously [32 (link)]. The fluorescence intensities were analyzed by using the program ImageJ V1.44.