Helium Ion Microscopy (HIM) for the imaging of biological samples at sub-nanometer resolution

Sci Rep. 2013 Dec 17:3:3514. doi: 10.1038/srep03514.


Scanning Electron Microscopy (SEM) has long been the standard in imaging the sub-micrometer surface ultrastructure of both hard and soft materials. In the case of biological samples, it has provided great insights into their physical architecture. However, three of the fundamental challenges in the SEM imaging of soft materials are that of limited imaging resolution at high magnification, charging caused by the insulating properties of most biological samples and the loss of subtle surface features by heavy metal coating. These challenges have recently been overcome with the development of the Helium Ion Microscope (HIM), which boasts advances in charge reduction, minimized sample damage, high surface contrast without the need for metal coating, increased depth of field, and 5 angstrom imaging resolution. We demonstrate the advantages of HIM for imaging biological surfaces as well as compare and contrast the effects of sample preparation techniques and their consequences on sub-nanometer ultrastructure.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arabidopsis / ultrastructure
  • Bacteria / ultrastructure
  • HeLa Cells / ultrastructure
  • Helium*
  • Humans
  • Ions*
  • Microscopy / methods*
  • Microscopy, Electron, Scanning / methods
  • Nematoda / ultrastructure


  • Ions
  • Helium