Jsm 1400 transmission electron microscope

Amniotic fluid samples were centrifuged at 2,300 × g for 5 min at room temperature and the supernatant was discarded. Electron microscopy fixative [2.5% glutaraldehyde in 0.1 M phosphate buffer, pH 7.4 (Cat# 16537–05, Electron Microscopy Science; Hatfield, PA, USA)] was carefully added to the cell pellet. Following fixation for 2 h at 4°C, the cell pellet was washed with 1X electron microscopy wash buffer [Sorensen’s phosphate buffer 0.2 M, pH 7.4 (Cat# 11601–10, Electron Microscopy Science)] and gently resuspended in 1 mL of the same buffer. Cell pellets were transported to the Microscopy & Image Analysis Laboratory at the University of Michigan. Images were obtained using an AMRAY 1910 Field Emission Scanning Electron Microscope (SEMTechSolutions; North Billerica, MA, USA) and a JSM-1400 Transmission Electron Microscope (JEOL USA, Inc.; Peabody, MA, USA).

Murine FAC-sorted β cells were placed in EM Fix solution containing 4% paraformaldehyde, 2.5% glutaraldehyde, and 0.02% picric acid, in 0.1M sodium cacodylate buffer. After fixation overnight at 4°C, cells were post-fixed for 1h in 1% OsO₄ −1.5% K-ferricyanide. Then, cells were dehydrated, infiltrated with Embed 812 epoxy resin, and transferred to resin models at 50°C for 36 hours. Samples were viewed using a JEOL JSM1400 transmission electron microscope operating at 100 KV (JEOL USA, Peabody, MA) and images were captured on a Veleta 2K x 2K CCD camera (EMSIS, GbhM, Miuenster, Germany).