Rapid Automated Preparation for Serial Block Face Scanning Electron Microscopy

Abstract

By providing automated acquisition of large volume electron microscope image stacks, serial block-face electron microscopy (SBEM) creates exciting new possibilities for life science discovery. While many repetitive aspects of microscopy are now computer-controlled, sample preparation remains a hurdle. SBEM sample preparation uses chemical fixation, heavy metal staining, and resin embedding protocols based on those developed for TEM, plus additional toxic and reactive steps. While TEM preparation typically requires 1-2 days, nearly week-long SBEM protocols are used to provide the heavy metal staining and conductivity required for high-contrast back-scattered SEM imaging at 1-3 KeV.