The microscope was an inverted Nikon Eclipse
TE200. The band pass filters used for the red and green channels were
605/90 and 525/50, respectively. The excitation wavelength was set at
800 nm for all experiments. The anesthetized rat was placed on the microscope
stage with the exposed kidney in direct contact with the cover
glass (see Animal preparation for imaging). A 60× Nikon Plan Apo water
immersion objective (NA 1.2) was used for imaging. All images were
collected at a constant pixel dwell time, which yielded a 1.1-s frame
time for the frame size of 512×512. All images for a given study were
collected at approximately the same focal plane at a depth of approximately
10–20 μm.