Journal: Cell Reports

Article Title: Pancreatic Islet Blood Flow Dynamics in Primates

doi: 10.1016/j.celrep.2017.07.039

Figure Lengend Snippet: High-Resolution In Vivo Longitudinal Imaging of Islet Re-vascularization in the ACE of Monkeys (A) Overview of the in vivo imaging rig setup based on the Leica SP8 microscope that can accommodate monkeys under the objective holder (for a full description and dimensions, see ). 1, manual mechanical jack setup on top of a high-precision scanning stage with a heating mat to maintain the animal’s core temperature at 37°C. 2, M250 Leica stereomicroscope setup on a boom stand that swings on/off the top of the scanning stage. 3, high-speed Hamamatsu electron multiplier charge-coupled device (EMCCD) camera used for wide-field microscopy. 4, confocal and two-photon scanning head. 5, custom-built head holder attached to the scanning stage and used to accommodate and stabilize the monkey’s head comfortably. Below the scanning head, the light protection box can be seen, with the integrated setup (green tubing) for ventilated anesthesia used during imaging sessions. (B) Close-up view of the inside of the light protection box, showing the microscope body with the transmitted light removed and ample space under the objective holder. The anesthesia tubes can be seen entering the box in the back. (C) Close-up top view of the scanning stage fitted with the head (1) and eye (2) holders. (D) Close-up view of the complete setup under the objective holder. In the top left corner, the enclosure for 2 external non-descanned detectors can be seen. (E) Longitudinal confocal imaging of the re-vascularization process of islets engrafted in the eye of monkeys on days 20 and 30 post-transplantation. Vessels (green) were stained in vivo with FITC-labeled 150-kDa dextran. The iris is shown in gray. Both vessels and the iris were imaged by fluorescence emitted after excitation by a 920-nm laser beam. The yellow dotted-line circle demarks the limits of the engrafted islet. (F) Quantification of the islet re-vascularization process and vascular density over time from two separate animals (B116 and C282). (G) Measurements of the vascular diameter of engrafted islets over time. Data are shown as SEM, acquired from n = 3 animals with n = 3 islets per animal, and all data points are shown. Scale bar, 100 μm (E).

Article Snippet: 3, high-speed Hamamatsu electron multiplier charge-coupled device (EMCCD) camera used for wide-field microscopy.

Techniques: In Vivo, Imaging, In Vivo Imaging, Microscopy, Transplantation Assay, Staining, Labeling, Fluorescence