Larry Scipioni

High Resolution Helium Ion Scanning Microscopy of the Rat Kidney

Helium ion scanning microscopy is a novel imaging technology with the potential to provide sub-nanometer resolution images of uncoated biological tissues. So far, however, it has been used mainly in materials science applications. Here, we took advantage of helium ion microscopy to explore the epithelium of the rat kidney with unsurpassed image quality and detail. In addition, we evaluated different tissue preparation methods for their ability to preserve tissue architecture. We found that high contrast, high resolution imaging of the renal tubule surface is possible with a relatively simple processing procedure that consists of transcardial perfusion with aldehyde fixatives, vibratome tissue sectioning, tissue dehydration with graded methanol solutions and careful critical point drying. Coupled with the helium ion system, fine details such as membrane texture and membranous nanoprojections on the glomerular podocytes were visualized, and pores within the filtration slit diaphragm could be seen in much greater detail than in previous scanning EM studies. In the collecting duct, the extensive and striking apical microplicae of the intercalated cells were imaged without the shrunken or distorted appearance that is typical with conventional sample processing and scanning electron microscopy. Membrane depressions visible on principal cells suggest possible endo- or exocytotic events, and central cilia on these cells were imaged with remarkable preservation and clarity. We also demonstrate the use of colloidal gold probes for highlighting specific cell-surface proteins and find that 15 nm gold labels are practical and easily distinguishable, indicating that external labels of various sizes can be used to detect multiple targets in the same tissue. We conclude that this technology represents a technical breakthrough in imaging the topographical ultrastructure of animal tissues. Its use in future studies should allow the study of fine cellular details and provide significant advances in our understanding of cell surface structures and membrane organization.

Understanding imaging modes in the helium ion microscope

Recent investigations are gaining us a better understanding of the nature of the beam-sample interactions in the helium ion microscope and what they mean for the image information provided. In secondary electron (SE) imaging, for example, the surface sensitivity is attributed to the low SE-II fraction. Voltage contrast imaging shows the ability to see both buried structures and to probe the conductance to ground of surface contacts. It is found, however, that the prominence of these two types of contrast varies oppositely with beam energy, yielding information about the nature of the interactions that gives rise to them. Transmission ion imaging can yield information about material density, atomic number, grain structure, and electronic structure. It is possible to capture the top-side SE signal, bright field signal, and dark field signal from a given sample simultaneously. The detection of diffraction contrast is under investigation.

Fabrication and initial characterization of ultrahigh aspect ratio vias in gold using the helium ion microscope

Toward the end goal of creating transducers with nanometer scale sensing features, the helium ion microscope (HIM) has been employed to create and characterize high aspect ratio features in gold films. The HIM has a spot size less than 1 nm, uses a chemically inert noble gas (He), which does not deposit/implant any species that may contaminate the material being patterned, and is able to rapidly generate arrays of vias in Au. Hence, the HIM is an ideal tool to generate these ultrahigh aspect ratio features. The authors characterize the vias, also using HIM, by measurements of feature size, lateral milling resolution, sidewall angle, and fabrication speed. Two novel methods were employed to enable the characterization due to the very small size of the features. A significant reduction in via width is achieved, as compared with traditional focused ion beam milling.

Analysis and metrology with a focused helium ion beama)

The newly introduced ORION™ helium ion microscope has been used for high resolution imaging and nanofabrication. More recently, an energy sensitive detector has been developed that permits the measurement of the energy spectrum of the backscattered helium ions. The spectra can be analyzed directly or compared with the simulated spectra from hypothetical models of the specimen. The technique can provide information about the elemental composition of the specimen or structural information (for example, layer thickness) of the specimen.

Scanning Helium Ion Microscopy

Abstract Recent developments in the area of gas field ion sources, coupled with knowledge gained from field ion microscopy, have made the realization of very high brightness ion sources a reality. In particular, an ion source using helium has been produced with a brightness equal to (or even exceeding) that of a cold field emission electron source. The other optical properties of this source (energy spread and angular intensity) are also very favorable, enabling the development of high resolution ion microscopes. One such development is a scanning helium ion microscope (HIM) based on this source technology. In many ways, the HIM has similarities to the scanning electron microscope in terms of construction and use. However the use of helium ions over electrons offers some unique advantages, including smaller focused probe size, beam/sample interactions, and sample charge control. This article describes the principles of operation for the helium ion source. Consideration is then given to a scanning column design, highlighting the small focused probe size that is to be expected. A description of the helium ion beam/sample interaction is presented followed by a review of the signals generated at the sample that can then be used for imaging or analytical purposes. Finally, a series of example applications are presented that highlight the unique capabilities of scanning helium ion microscopy.

Ionoluminescence in the helium ion microscope

Ionoluminescence (IL) is the emission of light from a material due to excitation by an ion beam. In this work, a helium ion microscope (HIM) has been used in conjunction with a luminescence detection system to characterize IL from materials in an analogous way to how cathodoluminescence (CL) is characterized in a scanning electron microscope (SEM). A survey of the helium ion beam induced IL characteristics, including images and spectra, of a variety of materials known to exhibit CL in an SEM is presented. Direct band-gap semiconductors that luminesce strongly in the SEM are found not do so in the HIM, possibly due to defect-related nonradiative pathways created by the ion beam. Other materials do, however, exhibit IL, including a cerium-doped garnet sample, quantum dots, and rare-earth doped LaPO4 nanocrystals. These emissions are a result of transitions between f electron states or transitions across size dependent band gaps. In all these samples, IL is found to decay with exposure to the beam, fitting well to double exponential functions. In an exploration of the potential of this technique for biological tagging applications, imaging with the IL emitted by rare-earth doped LaPO4 nanocrystals, simultaneously with secondary electron imaging, is demonstrated at a range of magnifications.

Helium Ion Secondary Electron Mode Microscopy For Interconnect Material Imaging

The recently developed helium ion microscope (HIM) is now capable of 0.35 nm secondary electron (SE) mode image resolution. When low-k dielectrics or copper interconnects in ultra large scale integrated circuits (ULSI) interconnect structures were imaged in this mode, it was found that unique pattern dimension and fidelity information at sub-nanometer resolution was available for the first time. This paper will discuss the helium ion microscope architecture and the SE imaging techniques that make the HIM observation method of particular value to the low-k dielectric and dual damascene copper interconnect technologies.

Creating nanohole arrays with the helium ion microscope

Helium Ion Microscopy has been established as a powerful imaging technique offering unique contrast and high resolution surface information. More recently, the helium ion beam has been used for nanostructuring applications similar to a gallium focused ion beam. A key difference between helium and gallium induced sputtering is the less intense damage cascade which lends this technique to precise and controlled milling of different materials enabling applications. The helium ion beam has been used for drilling 5nm holes in a 100nm gold foil (20:1 aspect ratio) while the gallium beam sputtered holes of a similar aspect ratio seem to be limited to a 50nm hole size. This paper explores the drilling of nanopores in gold films and other materials and offers an explanation for the observed differences in results between helium and gallium ions.

Structure and properties of polymer core-shell systems: Helium ion microscopy and electrical conductivity studies

Peculiarities of the structural organization and electrical properties of two core-shell polymer systems under different fabrication protocols have been studied with a combination of helium ion microscopy (HIM) and current-voltage characterization. The systems under study included a submicrometer core of a ferroelectric polymer polyvinylidene fluoride and a shell of intrinsically conducting or semiconducting polymer polyaniline (PANI) or poly(3-methylthiophene) (P3MT). Application of HIM allowed identification of the polymer components, visualization of the electrically conductive percolation network of PANI or P3MT, and its variation due to thermal annealing and/or interaction with the environment. HIM is proved to be a powerful tool for characterization of not only the morphology but also of the charge distribution and conductivity properties on the nanoscale. The specific contrast formation in HIM imaging is due to differences in local electrical conductivity of the components. The authors have demonst...

Material analysis with a helium ion microscope

The helium ion microscope, a new imaging technology, is being applied also to sample modification. The application opportunity exists due to the extreme high resolution and the ability to gather analytical data as well as images. Possible applications include inspection, elemental analysis, and dopant concentration measurements.