Microscope and fiber-optic spectrophotometry of transmittance and backscattering both showed moss leaves to be capable of casting strong shadows, with a single leaf blocking approximately 90% of incident light from a point source. In leaves with only one layer of cells, the transmittance through the cytoplasm of single cells was similar to that for whole leaves. Analysis of cell wall birefringence by polarized-light interferometry indicated that cell walls might normally scatter rather than transmit light. Spectra transmitted through, or backscattered from, the upper green layers of moss were dominated by selective absorbance from chlorophyll, but there was also evidence of wavelength-dependent scattering, as detected in the lower layers of brown, dead moss. Specular reflectance from moss leaves was detected by polarimetry and may have contributed to the relatively high macroscopic transmittance of stationary moss in water. Shadowing by moss leaves was confirmed by dynamic measurements of mosses in turbulent water without bubbles. Flicker patterns from leaves were superimposed on the underwater flicker pattern created at the air-water interface, thus flecks of light were reduced in intensity, increased in frequency, and decreased in duration. This was detected with both point source and diffuse illumination of samples.

The transition from 10-fold to 5-fold symmetry was observed during the analysis of nanodiffraction patterns of a gold decahedral multiple twinned nanoparticle of 15 nm in diameter. The analysis shows that as the convergence of the beam is increased, the rotational symmetry of the diffraction pattern shifts from 10- to 5-fold. The 10-fold symmetry predicted by Friedel's law is lost by the asymmetric shift of the diffraction spots, an effect that becomes more noticeable as the electron beam convergence increases. Dynamical and kinematical diffraction calculations indicate this decrease in symmetry is the result of a double refraction effect coupled with the variation of the dynamical diffraction conditions arising from a varying electron beam convergence.

The objective of this study was to evaluate the influence of a cola soft drink (CSD) and coffee on the microhardness and surface roughness of composite resin. Fifty cavities were prepared on the vestibular surface of bovine incisors and restored with nanoparticulate resin. The teeth were divided into five groups (n = 10): group A (control), immersion in artificial saliva (AS) for 14 days; group B, immersion in coffee for 15 min (3×/day) for 7 days followed by immersion in AS for another 7 days; group C, immersion in CSD for 15 min (3×/day) for 7 days followed by immersion in AS for another 7 days; group D, immersion in AS for 7 days, immersion in coffee for 15 min (3×/day) for 7 days; group E, immersion in AS for 7 days, immersion in CSD for 15 min (3×/day) for 7 days. After the immersion periods the specimens were analyzed for their microhardness and surface roughness. The data were subjected to analysis of variance (ANOVA) followed by t-test with 5% significance. Group A presented the highest average microhardness and lowest surface roughness, so it was possible to conclude that the consumption of CSD and coffee alters the microhardness and surface roughness of new restorations.

Energy dispersive X-ray microanalysis of micron-sized inclusions in steel is of considerable industrial importance. Measured spectra and Monte Carlo simulations show a significant effect of the steel matrix on analysis of CaO-Al2O3-MgO inclusions: the steel matrix filters the softer (Al and Mg) characteristic X-rays, increasing the relative height of the Ca peak. Bulk matrix correction methods would not result in correct inclusion compositions, but operating at a lower acceleration voltage shifts the effect to smaller inclusion sizes.

Light microscopy images suffer from poor contrast due to light absorption and scattering by the media. The resulting decay in contrast varies exponentially across the image along the incident light path. Classical space invariant deconvolution approaches, while very effective in deblurring, are not designed for the restoration of uneven illumination in microscopy images. In this article, we present a modified radiative transfer theory approach to solve the contrast degradation problem of light sheet microscopy (LSM) images. We confirmed the effectiveness of our approach through simulation as well as real LSM images.

Closed cell polymer foam skin thickness can be assessed by taking backscatter electron (BSE) images in a scanning electron microscope (SEM) at a series of accelerating voltages. Under a given set of experimental conditions, the electron beam mostly passes through thin polymer skin cell walls. That cell appears dark compared to adjacent thicker-skinned cells. Higher accelerating voltages lead to a thicker skin being penetrated. Monte Carlo modeling of beam-sample interactions indicates that at 5 keV, skin less than ∼0.5 μm in thickness will appear dark, whereas imaging at 30 keV allows skin thicknesses up to ∼4 μm to be identified. The distribution of skin thickness can be assessed over square millimeters of foam surface in this manner. Qualitative comparisons of the skin thicknesses of samples can be made with a simple visual inspection of the images. A semiquantitative comparison is possible by applying image analysis. The proposed method is applied to two example foams. Characterizing foam skin thickness by this method is possible using any SEM that is capable of collecting useful BSE images over a range of accelerating voltages. Imaging in low vacuum, where an electrically conductive metal coating is not required, leads to more sensitivity in skin thickness characterization.

Extended abstract of a paper presented at Microscopy and Microanalysis 2011 in Nashville, Tennessee, USA, August 7–August 11, 2011.

In scattering specimens, multiphoton excitation and nondescanned detection improve imaging depth by a factor of 2 or more over confocal microscopy; however, imaging depth is still limited by scattering. We applied the concept of clearing to deep tissue imaging of highly scattering specimens. Clearing is a remarkably effective approach to improving image quality at depth using either confocal or multiphoton microscopy. Tissue clearing appears to eliminate the need for multiphoton excitation for deep tissue imaging.

Persistence of gunshot residue (GSR) simultaneously collected from hands, face and hair, and clothing of the shooting person was examined. Samples were collected from five shooters in nine time intervals after a single shoot with a Luger 9 mm pistol, in the range of 0–4 h and examined with scanning electron microscopy and energy dispersive X-ray spectrometry. Numbers of particles, frequencies of occurrence of certain compositions of particles, and their sizes in function of the time intervals were inspected. The greatest numbers of particles were observed in samples collected from hands right after shooting, but they decrease quickly with time. In samples collected from the face smaller initial numbers of particles were found, but they lasted at a similar level longer. The estimated half-life times of particles were less than 1 h for samples taken from the hands, over 1 h for clothing and about 2–3 h for the face. In samples collected at longer intervals after shooting, there were particles present of small sizes and irregular shapes. The results demonstrate that including evidence collected from the suspect's face and hair may increase the probability of detection of GSR in cases when the suspect has not been apprehended immediately after the investigated incident.

Herein, we characterized lead pipe scale removed from a drinking water distribution system using argon ion beam etching and a variety of solids analysis approaches. Specifically, pipe scale cross sections and solids were analyzed using scanning electron microscopy, energy dispersive X-ray analysis, X-ray diffraction, and acid digestion followed by inductively coupled plasma mass spectrometry analyses. The pipe scale consisted of at least five layers that contained Pb(II) and Pb(IV) minerals, and magnesium, aluminum, manganese, iron, and silicon solids. The outer layer was enriched with crystalline amorphous manganese and iron, giving it a dark orange to red color. The middle layers consisted of hydrocerussite and plattnerite, and the bottom layer consisted primarily of litharge. Over the litharge layer, hydrocerussite crystals were grown vertically away from the pipe wall, which included formations of plattnerite. Significant amounts of trace contaminant vanadium, likely in the form of vanadinite, and copper accumulated in the scale as well.

Extended abstract of a paper presented at Microscopy and Microanalysis 2011 in Nashville, Tennessee, USA, August 7–August 11, 2011.

Ligament healing of a grade III injury (i.e., a complete tear) involves a multifaceted chain of events that forms a neoligament, which is more scar-like in character than the native tissue. The remodeling process may last months or even years with the injured ligament never fully recovering pre-injury mechanical properties. With tissue engineering and regenerative medicine, understanding the normal healing process in ligament and quantifying it provide a basis to create and assess innovative treatments. Ligament fibroblasts produce a number of extracellular matrix (ECM) components, including collagen types I and III, decorin and fibromodulin. Using a combination of advanced histology, molecular biology, and nonlinear optical imaging approaches, the early ECM events during ligament healing have been better characterized and defined. First, the dynamic changes in ECM factors after injury are shown. Second, the factors associated with creeping substitution are identified. Finally, a method to quantify collagen organization is developed and used. Each ECM factor described herein as well as the temporal quantification of fiber organization helps elucidate the complexity of ligament healing.

To date, there are limited studies on loggerhead turtle (Caretta caretta) eggshell ultrastructure and its elemental composition. Eggs were collected from turtle nests immediately after oviposition and post hatching. Three eggshell layers were recognized. The outer calcareous layer consists of loose nodular units of different shapes and sizes with loose attachment between the units, resulting in numerous spaces and openings. Each unit consists of CaCO3 crystals in aragonite (99%) and calcite (1%). The middle layer has several strata with numerous openings connecting the calcareous and the inner shell membrane. Crystallites of the middle layer are a mix of amorphous material with aragonite (62%) and calcite (38%). The inner shell membrane has numerous reticular fibers mixed predominantly with halite (NaCl) and small amounts of sylvite. Thermogravimetry analysis of the calcareous showed a low exothermic peak at 425°C, which corresponds to a transitional phase from aragonite to calcite. A high endothermic peak at 814°C corresponds to decomposition of calcite CaCO3 to CaO and CO2. Electron diffraction confirmed the presence of NaCl halite crystal. A significant difference was found in the percentage of elements and crystal configurations in the three layers. This study has value in assessing the emergence success in this endangered species.

Focused ion beam (FIB) induced damage in nanocrystalline Al thin films has been characterized using advanced transmission electron microscopy techniques. Electron tomography was used to analyze the three-dimensional distribution of point defect clusters induced by FIB milling, as well as their interaction with preexisting dislocations generated by internal stresses in the Al films. The atomic structure of interstitial Frank loops induced by irradiation, as well as the core structure of Frank dislocations, has been resolved with aberration-corrected high-resolution annular dark-field scanning TEM. The combination of both techniques constitutes a powerful tool for the study of the intrinsic structural properties of point defect clusters as well as the interaction of these defects with preexisting or deformation dislocations in irradiated bulk or nanostructured materials.

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.

A confocal scanning laser holography (CSLH) microscope that uniquely combines the concepts of confocal microscopy with holography has been validated for making nonintrusive, full three-dimensional (3D) intensity and phase measurements of objects from a single viewpoint of observation without loss of object information. The phase measurements have been used to determine the 3D refractive indices of a point source heated silicone oil. The refractive indices are converted to 3D temperature measurements, which are useful for heat transfer studies. An important feature of CSLH is its nonintrusive 3D scanning method, which enables its application to the study of Marangoni convection in microgravity with minimal operational vibrations affecting the motion of fluid in the specimen.