Ultrathin sectioning and staining artifacts in transmission electron microscopy specimen preparation

Abstract

Ultrathin sectioning followed by staining of sections are the final two steps in the preparation procedure for transmission electron microscope (TEM) specimens. While each requires an understanding of the scientific principles involved, these steps, perhaps more than any others, also require meticulous attention to detail and, for ultrathin sectioning, considerable skill and practice. Even with a properly fixed and infiltrated specimen in a well-polymerized block, there are a number of factors which can contribute to artifacts in ultrathin sections. Most can be corrected or at least minimized assuming that the ultramicrotome environment is well-situated; i.e., free of vibration, excessive heat, humidity, and air currents.