Three-dimensional atom probe tomography (APT) is successfully used to analyze the near-apex regions of an atomic force microscope (AFM) tip. Atom scale material structure and chemistry from APT analysis for standard silicon AFM tips and silicon AFM tips coated with a thin film of Cu is presented. Comparison of the thin film data with that observed using transmission electron microscopy indicates that APT can be reliably used to investigate the material structure and chemistry of the apex of an AFM tip at near atomic scales.

Multispectral analysis combined with the Periodic Acid-Schiff method was used to investigate cytological features of insoluble polysaccharides and changes in total insoluble polysaccharide content (TPC) during pollen development in rice, including four cytoplasmic male sterility lines (MSLs) and their corresponding fertility-maintaining lines (FMLs). The multispectral curves of the relative transmittance value (RTV) and the images of developing pollen cells were obtained across a range of successive wavelengths (400–720 nm). A minimum RTV was found near 550 nm indicating an absorption peak of the TPC. Thus, the TPC was measured using the RTV of 550 nm. In the four FMLs, the minimum TPC of developing pollen cells occurred at the late microspore stage, while the maximum TPC occurred at the mature pollen grain stage. The TPC levels of pollen cells were significantly higher in the four FMLs than in their corresponding MSLs during and after pollen abortion. Notably, a steep decrease of multispectral curves at 420 nm appeared before the occurrence of abortion, implying a marker associated with pollen abortion in rice. Our results will be helpful for exploring the changes in TPC during pollen ontogenesis in rice and provide a novel method for the study of bio-macromolecules.

Extended abstract of a paper presented at Microscopy and Microanalysis 2010 in Portland, Oregon, USA, August 1 – August 5, 2010.

Scanning transmission electron microscope (STEM) images of three-dimensional (3D) samples were simulated. The samples consisted of a micrometer(s)-thick substrate and gold nanoparticles at various vertical positions. The atomic number (Z) contrast as obtained via the annular dark-field detector was generated. The simulations were carried out using the Monte Carlo method in the CASINO software (freeware). The software was adapted to include the STEM imaging modality, including the noise characteristics of the electron source, the conical shape of the beam, and 3D scanning. Simulated STEM images of nanoparticles on a carbon substrate revealed the influence of the electron dose on the visibility of the nanoparticles. The 3D datasets obtained by simulating focal series showed the effect of beam broadening on the spatial resolution and on the signal-to-noise ratio. Monte Carlo simulations of STEM imaging of nanoparticles on a thick water layer were compared with experimental data by programming the exact sample geometry. The simulated image corresponded to the experimental image, and the signal-to-noise levels were similar. The Monte Carlo simulation strategy described here can be used to calculate STEM images of objects of an arbitrary geometry and amorphous sample composition. This information can then be used, for example, to optimize the microscope settings for imaging sessions where a low electron dose is crucial for the design of equipment, or for the analysis of the composition of a certain specimen.

We describe a feedback mechanism in the gas cascade amplification process used in magnetic immersion lens environmental scanning electron microcopy (ESEM). Feedback dominates gas gain under the conditions typically used for ultra-high-resolution ESEM and gives rise to novel dependencies of the imaging signal and noise on microscope operating parameters. It is ascribed tentatively to the generation of free electrons upon de-excitation of metastable species in the gas cascade. The results have implications for optimization of ESEM systems for applications such as critical dimension metrology and real-time imaging of nanostructure growth by gas mediated electron beam induced deposition.

Extended abstract of a paper presented at Microscopy and Microanalysis 2010 in Portland, Oregon, USA, August 1 – August 5, 2010.

Local magnification effects and trajectory overlaps related to the presence of a second phase (clusters) are key problems and still open issues in the assessment of quantitative composition data in three-dimensional atom probe tomography (APT) particularly for tiny solute-enriched clusters. A model based on the distribution of distance of first nearest neighbor atoms has been developed to exhibit the variations in the apparent atomic density in reconstructed volumes and to correct compositions that are biased by local magnification effects. This model was applied to both simulated APT reconstructions and real experimental data and shows an excellent agreement with the expected composition of clusters.

Extended abstract of a paper presented at Microscopy and Microanalysis 2010 in Portland, Oregon, USA, August 1 – August 5, 2010.

Following is a list of microscopy-related meetings and courses. The editors would greatly appreciate input to this list via the electronic submission form found in the MSA World-Wide Web page at www.msa.microscopy.com. We will gladly add hypertext links to the notice on the web and insert a listing of the meeting in the next issue of the Journal. Send comments and questions to Nan Yao, nyao@Princeton.edu.

Despite the many demonstrated applications of factor analysis (FA) in analyzing hyperspectral materials images, FA does have inherent mathematical limitations, preventing it from solving certain materials characterization problems. A notable limitation of FA is its parsimony restriction, referring to the fact that in FA the number of components cannot exceed the chemical rank of a dataset. Clustering is a promising alternative to FA for the phase classification of hyperspectral materials images. In contrast with FA, the phases extracted by clustering do not have to be parsimonious. Clustering has an added advantage in its insensitivity to spectral collinearity that can result in phase mixing using FA. For representative energy dispersive X-ray spectroscopy materials images, namely a solder bump dataset and a braze interface dataset, clustering generates phase classification results that are superior to those obtained using representative FA-based methods. For the solder bump dataset, clustering identifies a Cu-Sn intermetallic phase that cannot be isolated using FA alone due to the parsimony restriction. For the braze interface sample that has collinearity among the phase spectra, the clustering results do not exhibit the physically unrealistic phase mixing obtained by multivariate curve resolution, a commonly utilized FA algorithm.

Extended abstract of a paper presented at Microscopy and Microanalysis 2010 in Portland, Oregon, USA, August 1 – August 5, 2010.

Following is a list of microscopy-related meetings and courses. The editors would greatly appreciate input to this list via the electronic submission form found in the MSA World-Wide Web page at www.msa.microscopy.com. We will gladly add hypertext links to the notice on the web and insert a listing of the meeting in the next issue of the Journal. Send comments and questions to Nan Yao, nyao@Princeton.edu.