Immunofluorescent (IF) staining for insulin was performed on human pancreatic sections adjacent to those used for in vitro autoradiography binding studies. Briefly, sections were stained using Insulin A SC-7839 (Santa Cruz, Dallas, TX, USA; goat-polyclonal 1:1000). The sections were then incubated with secondary antibody Alexa fluor 488 (Invitrogen, Carlsbad, CA, USA; donkey anti-goat; dilution 1:100). Tile scan images were acquired with a Zeiss LSM780 confocal microscope.
Immunohistochemical (IHC) staining for insulin on NHP pancreatic sections was performed by primary antibody A0564 (Agilent, Santa Clara, CA, USA). The stains were developed by the Envision DAB system K4010 using an antirabbit secondary antibody. All sections were counterstained using Mayer’s Hematoxylin. The slides were captured digitally in bright-field mode at × 10/0.45 magnification, using an Axio Imager 2 microscope (Carl Zeiss Microscopy GmbH, Germany) mounted with an Axiocam MRc camera. Images were analyzed in Zen Blue 2.0 (Carl Zeiss Microscopy GmbH, Germany).
Immunohistochemical (IHC) staining for insulin on NHP pancreatic sections was performed by primary antibody A0564 (Agilent, Santa Clara, CA, USA). The stains were developed by the Envision DAB system K4010 using an antirabbit secondary antibody. All sections were counterstained using Mayer’s Hematoxylin. The slides were captured digitally in bright-field mode at × 10/0.45 magnification, using an Axio Imager 2 microscope (Carl Zeiss Microscopy GmbH, Germany) mounted with an Axiocam MRc camera. Images were analyzed in Zen Blue 2.0 (Carl Zeiss Microscopy GmbH, Germany).