This PDF file contains the front matter associated with SPIE Proceedings Volume 9636, including the Title Page, Copyright information, Table of Contents, Introduction (if any), and Conference Committee listing.

This is the fourth part of a series of tutorial papers discussing various causes of measurement uncertainty in scanned particle beam instruments, and some of the solutions researched and developed at NIST and other research institutions. Scanned particle beam instruments, especially the scanning electron microscope (SEM), have gone through tremendous evolution to become indispensable tools for many and diverse scientific and industrial applications. These improvements have significantly enhanced their performance and made them far easier to operate. But, the ease of operation has also fostered operator complacency. In addition, the user-friendliness has reduced the apparent need for extensive operator training. Unfortunately, this has led to the idea that the SEM is just another expensive "digital camera" or another peripheral device connected to a computer and that all of the problems in obtaining good quality images and data have been solved. Hence, one using these instruments may be lulled into thinking that all of the potential pitfalls have been fully eliminated and believing that, everything one sees on the micrograph is always correct. But, as described in this and the earlier papers, this may not be the case. Care must always be taken when reliable quantitative data are being sought. The first paper in this series discussed some of the issues related to signal generation in the SEM, including instrument calibration, electron beam-sample interactions and the need for physics-based modeling to understand the actual image formation mechanisms to properly interpret SEM images. The second paper has discussed another major issue confronting the microscopist: specimen contamination and methods to eliminate it. The third paper discussed mechanical vibration and stage drift and some useful solutions to mitigate the problems caused by them, and here, in this the fourth contribution, the issues related to specimen "charging" and its mitigation are discussed relative to dimensional metrology.

Serial sectioning using the FIB and subsequent imaging of the same FIB-exposed surface by both FIB microscopy and scanning electron microscopy (SEM) in a DualBeam has proven especially useful to study the three-dimensional (3D) morphology of complex engineered materials systems. The technique was first introduced as an automated process in 2004 and since then has established itself as one of the primary applications for FIB and DualBeams. While state-of-the-art systems can produce datasets with a z-axis slice thickness of 3-5 nm, FIB nanotomography remains a destructive technique and is limited in resolution by the z-axis slice thickness. Electron tomography is another technique used to visualize 3D structures within a transmission electron microscope used in TEM or STEM mode. Using a thin sample focused on a region of interest, the electron beam passes through the specimen incrementally tilting around the center of the region of interest as images are acquired sequentially on a camera (TEM) or a Detector (STEM). The resulting images are reconstructed into a 3D volume using a variety of algorithms including Weighted Back Projection (WBP), or Serial Iterative Reconstruction Technique (SIRT). Low energy STEM in SEM is a routine analysis in SEMs and DualBeam FIB-SEM instrumentation for morphological characterization and ultra high-resolution imaging. With a DualBeam or SEM configured with a solid state silicon diode STEM detector and a stage with adequate tilt freedom, it is possible to acquire a sufficient number of images for 3D reconstruction using STEM tomography in SEMs and DualBeam instruments.

A thin section sample of gadolinium nanoparticles ranging in size up to 50 nm mounted on an aluminum substrate was prepared using in-situ lift-out (INLO) by FIB. The sample was thinned using 30 kV Ga⁺ FIB to approximately 125 nm. Using an in-situ stage with 360 degree continuous tilt, the thin section was imaged every 1 degree with 30 keV SEM and the STEM detector through approximately 125 degrees of tilt. The data set was then reconstructed into a 3D rendering using FEI's tomography reconstruction software, Inspect3D Express, and visualized using FEI's Avizo image and data analysis software. The technique and results compared with conventional TEM and STEM tomography using a 200 keV FEI Talos TEM will be discussed.

Preparation of lamellae from bulk to grid for Cs-corrected Transmission Electron Microscope (TEM) observation has mostly become routine work on the latest FIB-SEM systems, with standardized techniques that often are left to automation for the initial steps. The finalization of lamellae however, has mostly become, non-routine, non-repeatable and often driven by user experience level in most cases to produce high quality damage-less cross section. Materials processing of the latest technologies, with ever-shrinking Nano-sized structures pose challenges to modern FIB-SEM systems. This can often lead to specialized techniques and hyper-specific functions for producing ultra-thin high quality lamellae that often are lab specific, preventing practical use of such techniques across multiple materials and applications. Several factors that should be incorporated in processing fine structured materials successfully include how the use of electron and ion scan conditions can affect a thin section during ion milling, the type of ion species applied for material processing during the finalization of lamellae with gallium ions or of a smaller ion species type such as Ar/Xe, sample orientation of the lamella during the thinning process which is linked to ion beam incident angle as a direct relationship in the creation of waterfall effects or curtain effects, and how software can be employed to aid in the reduction of these artifacts with reproducible results regardless of FIB-SEM experience for site-specific lift outs. A traditional TEM preparation was performed of a fine structure specimen in pursuit of a process technique to produce a high quality TEM lamella which would address all of the factors mentioned. These new capabilities have been refined and improved upon during the FIB-SEM design and development stages with an end result of a new approach that yields an improvement in quality by the reduction of common ion milling artifacts such as curtain effects, amorphous material, and better pin pointing of the area of interest while reducing overall processing time for the TEM sample preparation process and enhancing repeatability through ease of use via software controls. The development of these new technologies, incorporating a third Ar/Xe ion beam column in conjunction with the electron and gallium ion beam column, a 7-axis stage for enhanced sample orientation with tilt functions in two axes and automated swing control along with a host of additional functions which address the factors aforementioned such as electron and ion scan techniques and curtain effect removal by the use of hardware and software components that are key to reduce typical FIB related artifacts, all of which are called “ACE [Anti Curtaining Effect] Technologies” are explained. The overall developments of these technologies are to address a significant point that productivity, throughput and repeatability are comprised by synergy between the user, application, software and hardware within a FIB-SEM system. The latest Hitachi FIB-SEM platform offers these innovations for reliability, repeatability and high quality lamella preparation for Cs-corrected (S)TEMs.

The industrial market for processing large-scale films has seen dramatic changes since the 1980s and has almost completely been replaced by lasers and digital processes. A commonly used technology for engraving screens, print and embossing forms in the printing industry, well known since then, is the use of RF-excited CO₂ lasers with a beam power up to about 1 kW, modulated in accordance to the pattern to be engraved. Future needs for high-security printing (banknotes, security papers, passports, etc.) will require laser engraving of at least half a million or even more structured elements with a depth from some μm up to 500 μm. Industry now wants photorealistic pictures in packaging design, which requires a similar performance. To ensure ’trusted pulses’ from the digital process to the print result the use of correlative microscopy (CLSM and SEM) is demonstrated as a complete chain for a correlative print process in this paper.

Simultaneous two-color subtraction microscopy using mode multiplexing is realized experimentally. The samples are irradiated with single laser diode at wavelength of 445 nm. Then the beam split laser spots generate separate solid and donut spatial modes and are multiplexed with modulators for simultaneous excitation. The produced fluorescence signals are back collected and further divided into two color bands with dichroic mirrors. Then they are detected with two photomultipliers and demultiplexed in four lock-in amplifiers. Four fluorescence images are recorded in every scan and resolution enhanced images are obtained in two color channels after applying the subtraction strategy. With this method, imaging results of microspheres stained with organic dyes and mesenteric lymph nodes of a mouse labeled with quantum dots (Q525/650) are realized. Improvement of 20% ~ 30% in resolving power of the two color channels compared with confocal microscopy is achieved in with corresponding subtraction factor of about 0.3.

There have been tremendous interests about the fabrication of the Au plasmonic nanopore due to its capability of the nanosize optical biosensor. We have investigated the influence of low energy electron beam irradiation on an Au nanomembrane during Au nanopore formation. In this report, the influence of electron beam irradiation on the Au nanopore formation will be reported. The nanopores on the 200 nm thick Au membrane were initially fabricated using focused ion beam (FIB) and high energy electron beam techniques such as transmission electron microscopy (TEM), and field emission scanning electron microscopy (FESEM). During high energy electron beam by using TEM, either a "shrinking" or a "opening" phenomenon is reported dependent on the ratio of thickness to aperture diameter. However, for a FESEM electron beam irradiation, a shrinking phenomenon is always observed. In this report, the nanopore formation during FESEM electron beam irradiation will be reported depending upon energy absorption and thermal diffusivity.

Scanning electron microscopy combined with microchemical analysis has evolved into one of the most widely used instruments in forensic science today. In particular, the environmental scanning electron microscope (SEM) in conjunction with energy dispersive spectroscopy (EDS), has created unique opportunities in forensic science in regard to the examination of trace evidence; i.e. the examination of evidence without altering the evidence with conductive coatings, thereby enabling criminalists to solve cases that were previously considered unsolvable. Two cold cases were solved at URI using a JEOL 5900 LV SEM in conjunction with EDS. A cold case murder and a cold missing person case will be presented from the viewpoint of the microscopist and will include sample preparation, as well as image and chemical analysis of the trace evidence using electron microscopy and optical microscopy.

For forensic investigation in the food industry, scanning electron microscopy (SEM) in conjunction with energy dispersive X-ray spectrometry (EDS) is a powerful, often non-destructive, instrumental analysis tool to provide information about:

• Identification of inorganic (and some organic) materials found as foreign contaminants in food products returned by consumers or detected during quality control inspections in the production facilities

• Identification of wear particles found in production lines

• Distribution of materials within a matrix

• Corrosion and failure analysis of production equipment

The identification of materials by SEM/EDS is accomplished through a combination of morphology by SEM imaging and the elemental composition of the material by EDS. Typically, the EDS analysis provides a qualitative spectrum showing the elements present in the sample. Further analysis can be done to quantify the detected elements in order to further refine the material identification. Metal alloys can often be differentiated even within the same family such as 300 Series stainless steels. Glass types can be identified by the elemental composition where the detected elements are quantified as the oxides of each element. In this way, for example, common window glass is distinguishable from insulation glass used in many ovens. Wear particles or fragments from breakage can find their way into food products. SEM/EDS analysis of the materials is an important resource to utilize when trying to determine the original source. Suspected source materials can then be sampled for comparative analysis. EDS X-ray mapping is another tool that is available to provide information about the distribution of materials within a matrix. For example, the distribution of inorganic ingredients in a dried food helps to provide information about the grind and blend of the materials.

During the reconstruction of a shooting incident, several analytical techniques are at the disposal of forensic scientists. The Netherlands Forensic Institute is exploring the opportunities in which mXRF can be used to obtain information in these reconstructions. In this paper the possibilities of using the 2D-mXRF instrument as a screening tool are explored. Primarily the focus is on using mXRF to obtain information regarding the elemental composition of gunshot residue from cartridge cases. Secondly, the possibilities of using the 2D-mXRF instrument to, quickly and easily, set up databases for gunshot residue analysis are investigated.

When placed at a high school in one of the nation’s regions often cited in the press for negative metrics, the Hitachi TM3030 has proved to be a device that engenders a broad range of educational cooperative efforts. In this article, the author describes how the scanning electron microscope was used to connect learners from elementary school to university. Students from across the high school curriculum spectrum, including artists, poets, musicians and videographers used the device for advanced explorations. Unanticipated connections were made between at-risk and underserved groups with science learning using state of the art tabletop SEM technology. Teachers experienced new ways to motivate their students, and model curricula was developed that is now being used in educator training.

The National Nanotechnology Infrastructure Network (NNIN)¹is an integrated partnership of 14 universities across the US funded by NSF to support nanoscale researchers. The NNIN education office is located at the Institute of Electronics and Nanotechnology at the Georgia Institute of Technology. At Georgia Tech we offer programs that integrate the facility and its resources to educate the public about nanotechnology. One event that has proved highly successful involves using microscopes in our characterization suite to educate a diverse audience about a variety of imaging instruments. As part of the annual Atlanta Science Festival (ATLSF)² we provided an event entitled: “What’s all the Buzz about Nanotechnology?” which was open to the public and advertised through a variety of methods by the ATLSF. During the event, we provided hands-on demos, cleanroom tours, and activities with three of our microscopes in our recently opened Imaging and Characterization Facility: 1. Keyence VHX-600 Digital Microscope; 2. Hitachi SU823 FE-SEM; and 3. Hitachi TM 3000. During the two hour event we had approximately 150 visitors including many families with school-aged children. Visitors were invited to bring a sample for scanning with the TM-3000. This paper will discuss how to do such an event, lessons learned, and visitor survey results.

One of the goals of the National Nanotechnology Infrastructure Network (NNIN) is workforce development. If K-12 students are to be ready for the jobs of the 21st century they must be able to work with tools such as microscopes. This study was conducted to determine if the NNIN lesson Taking a Closer Look at Objects helped grades K-5 students connect what they were learning about light and its uses, to how microscopes work. The results of this study indicate that the lesson not only helped students improve their knowledge of how light is used but also helped them become more knowledgeable about how microscopes work. Students learning how light is used to form an image in an optical microscope, lays the foundation for them to learn how other microscopes such as the Scanning Electron Microscope and Atomic Force Microscope are used to form images. This knowledge is important in supporting the continued research and development in the field of nanoscale science and engineering.

We present a continuous tip monitoring method during atomic force microscopy imaging based on the use of higher harmonics, which are generated in the repulsive regime as a result of the nonlinear interactions between the cantilever tip and the surface under study. We have applied this method to commercial rectangular microfabricated silicon cantilevers with force constants in the 45 N/m range and fundamental frequencies in the 300-400 kHz range and with tip radii below 10 nm. We have focused in the resonance of the 2^nd flexural mode and the 6^th harmonic using polystyrene surfaces. The simultaneous acquisition of topographic and higher harmonic images allows a continuous control of the state of the tip. The experimental results have been rationalized with computer simulations taking into account both the cantilever dynamics and the tip-surface interactions.

Critical dimension atomic force microscopes (CD-AFMs) use flared tips and two-dimensional sensing and control of the tip-sample interaction to enable scanning of features with near-vertical or even reentrant sidewalls. Sidewall sensing in CD-AFM usually involves lateral dither of the tip, which was the case in the first two generations of instruments. Current, third generation instruments also utilize a control algorithm and fast response piezo actuator to position the tip in a manner that resembles touch-triggering of coordinate measuring machine (CMM) probes. All methods of tip position control, however, induce an effective tip width that may deviate from the actual geometrical tip width. The National Institute of Standards and Technology (NIST) has been investigating the dependence of effective tip width on the dither settings and lateral stiffness of the tip, as well as the possibility of material effects due to sample composition. We have concluded that these effects will not generally result in a residual bias, provided that the tip calibration and sample measurement are performed under the same conditions. To further validate our prior conclusions about the dependence of effective tip width on lateral stiffness, we recently performed experiments using a very large non-CD tip with an etched plateau of approximately 2 μm width. The effective lateral stiffness of these tips is at least 20 times greater than typical CD-AFM tips, and these results supported our prior conclusions about the expected behavior for larger tips. The bottom-line importance of these latest observations is that we can now reasonably conclude that a dither slope of 3 nm/V is the baseline response due to the induced motion of the cantilever base.

Three-coordinate laser interferometer was designed to enable metrological measurements using conventional scanning probe microscopes. This article presents the results of the errors sources investigation in three-coordinate laser interferometer, describes the features of the optical scheme and data processing system that ensure sub-nanometer accuracy of the measurements.

We report the initial results of a recent bilateral comparison of linewidth or critical dimension (CD) calibrations on photomask line features between two national metrology institutes (NMIs): the National Institute of Standards and Technology (NIST) in the United States and the Physikalisch-Technische Bundesanstalt (PTB) in Germany. For the comparison, a chrome on glass (CoG) photomask was used which has a layout of line features down to 100 nm nominal size. Different measurement methods were used at both institutes. These included: critical dimension atomic force microscopy (CD-AFM), CD scanning electron microscopy (CD-SEM) and ultraviolet (UV) transmission optical microscopy. The measurands are CD at 50 % height of the features as well as sidewall angle and line width roughness (LWR) of the features. On the isolated opaque features, we found agreement of the CD measurements at the 3 nm to 5 nm level on most features – usually within the combined expanded uncertainties of the measurements.

X-ray spectra suffer significantly degraded spatial resolution when measured in the variable-pressure scanning electron microscope (VPSEM, chamber pressure 1 Pa to 2500 Pa) as compared to highvacuum SEM (operating pressure < 10 mPa). Depending on the gas path length, electrons that are scattered hundreds of micrometers outside the focused beam can contribute 90% or more of the measured spectrum. Monte Carlo electron trajectory simulation, available in NIST DTSA-II, models the gas scattering and simulates mixed composition targets, e.g., particle on substrate. The impact of gas scattering at the major (C > 0.1 mass fraction), minor (0.01 ≤ C ≤ 0.1), and trace (C < 0.01) constituent levels can be estimated. NIST DTSA-II for Java-platforms is available free at: http://www.cstl.nist.gov/div837/837.02/epq/dtsa2/index.html).