Abstract
Scanning helium-ion microscopy (HIM) is an imaging technique with sub-nanometre resolution and is a powerful tool to resolve some of the tiniest structures in biology. In many aspects, the HIM resembles a field-emission scanning electron microscope (FE-SEM), but the use of helium ions rather than electrons provides several advantages, including higher surface sensitivity, larger depth of field, and a straightforward charge-compensating electron flood gun, which enables imaging of non-conductive samples, rendering HIM a promising high-resolution imaging technique for biological samples. Starting with studies focused on medical research, the last decade has seen some particularly spectacular high-resolution images in studies focused on plants, microbiology, virology, and geomicrobiology. However, HIM is not just an imaging technique. The ability to use the instrument for milling biological objects as small as viruses offers unique opportunities which are not possible with more conventional focused ion beams, such as gallium. Several pioneering technical developments, such as methods to couple secondary ion mass spectrometry (SIMS) or ionoluminescence with the HIM, also offer the possibility for new and exciting research on biological materials. In this review, we present a comprehensive overview of almost all currently published literature which has demonstrated the application of HIM for imaging of biological specimens. We also discuss some technical features of this unique type of instrument and highlight some of the new advances which will likely become more widely used in the years to come.
Highlights
Since its commercialisation in 2006 [1,2,3,4,5], the helium-ion microscope (HIM) has become a well-established tool for nanoscale imaging and nanoscale fabrication in physics and materials science
We present a comprehensive overview of almost all currently published literature which has demonstrated the application of helium-ion microscopy (HIM) for imaging of biological specimens
It is as flexible and straightforward to use as a scanning electron microscope (SEM) but with a five times larger depth of field [1] and a lateral resolution of about 0.5 nm [4], which is between high-end field-emission SEMs (FE-SEM) and transmission electron microscopes (TEM)
Summary
Since its commercialisation in 2006 [1,2,3,4,5], the helium-ion microscope (HIM) has become a well-established tool for nanoscale imaging and nanoscale fabrication in physics and materials science. Scanning helium-ion microscopy (HIM) is an imaging technique with sub-nanometre resolution and is a powerful tool to resolve some of the tiniest structures in biology. The HIM resembles a field-emission scanning electron microscope (FESEM), but the use of helium ions rather than electrons provides several advantages, including higher surface sensitivity, larger depth of field, and a straightforward charge-compensating electron flood gun, which enables imaging of non-conductive samples, rendering HIM a promising high-resolution imaging technique for biological samples.
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