For intravital microscopy, only STZ-diabetic mice were used. First, the mice were anesthetized with isoflurane (0.8 l/min, 2.5%, Baxter Deutschland GmbH), and an abdominal body window was implanted for repeated kidney imaging as previously described (Schiessl et al., 2019 (link)).
The next day, mice were anesthetized with isoflurane (0.8 l/min, 1.5%), intubated, and catheterized into the lateral tail vein. To maintain body temperature during intravital microscopy, the mice were kept on a heating plate.
Image acquisition was performed using an upright Leica SP8 multiphoton laser scanning microscope with a 40x/1.1 NA water immersion objective at the Core Facility Cellular Imaging at the Technical University Dresden. Multiphoton imaging was performed with 860 nm laser excitation to visualize Angiospark680® (Perkin&Elmer, 30 µl), Hoechst 33,342 (Thermo Fischer, 50 µl of 2 mg/ml stock), and Lucifer Yellow (LY; Sigma-Aldrich).
The next day, mice were anesthetized with isoflurane (0.8 l/min, 1.5%), intubated, and catheterized into the lateral tail vein. To maintain body temperature during intravital microscopy, the mice were kept on a heating plate.
Image acquisition was performed using an upright Leica SP8 multiphoton laser scanning microscope with a 40x/1.1 NA water immersion objective at the Core Facility Cellular Imaging at the Technical University Dresden. Multiphoton imaging was performed with 860 nm laser excitation to visualize Angiospark680® (Perkin&Elmer, 30 µl), Hoechst 33,342 (Thermo Fischer, 50 µl of 2 mg/ml stock), and Lucifer Yellow (LY; Sigma-Aldrich).