The parasitic dinoflagellate Haplozoon is found in the intestine of marine polychaetes. It is composed of a chain of cells that hang from the intestinal wall into the lumen, and releases daughter cells from the posterior end of the chain which leave the host and reinfect other polychaetes. Few studies exist in the recent literature regarding Haplozoon and it has not been reported from the Gulf Coast of the United States. This study reports the genus Haplozoon from the maldanid polychaete Axiothella mucosa in St. Andrew Bay, Florida, and examines the structure of the organism by light and electron microscopy.

Axiothella mucosa was collected in St. Andrew Bay, Florida and maintained in seawater at the Troy State University campus. Haplozoon spp. was prepared for whole mounts by smearing minced setigers of the worms onto a slide with a drop of seawater.

Nanometer-resolution surface imaging by secondary electrons (SE) is now routinely obtainable in field-emission SEM or STEM microscopes. The resolution in SE images obtained in a modern field-emission (FE) SEM is still limited by the electron probe size although the localization of the SE generation processes sets the ultimate resolution limit that will be achievable in SE images. The combination of a FE gun with improved probe forming lenses and detection systems makes it possible to image nanometer-scale surface features at low voltages. Because of the reduced electron beam range and the new SE detection system, image contrast formation in low voltage FE-SEMs is quite different to that in conventional SEMs. In a FE-SEM, SEs are detected through the combination of a strong magnetic field of the lens and a weak electrostatic extraction field of the detector. The detection efficiency varies with the energy of the emitted SEs.

The aim of this study was to compare surface structures of laser-irradiated dental hard tissues using confocal (CFM), atomic force (AFM), and scanning electron microscopy (SEM). The general potential of the AFM in analyzing laser-irradiated surfaces was determined in this context. Specimens of human enamel and dentin were irradiated using an 8.6 W Nd:YVO4 laser with a pulse duration of 8 ps, λCenter=1,064 nm, and a pulse repetition rate of 500 kHz. Surface topology of irradiated areas (1 mm2) was investigated using AFM, CFM, and SEM. Surface roughness Rz was measured only with the AFM and the CFM. For non-irradiated enamel and dentin surfaces, roughnesses for CFM and AFM are in the nanometer range. However, major differences in roughness were determined for laser-prepared surfaces. For enamel, Rz(CFM)=2.33 μm is much higher compared with Rz(AFM)=0.09 μm; in the case of dentin, Rz(CFM)=5.35 μm is also much higher compared with Rz(AFM)=0.093 μm. Information regarding structural properties of surfaces needs real dimensions, particularly for use in dentistry. In this respect, AFM technology provides no additional results that lead to a significant improvement.

Extended abstract of a paper presented at Microscopy and Microanalysis 2006 in Chicago, Illinois, USA, July 30 – August 3, 2005

Energy Dispersive Spectrometry (EDS) is an ubiquitous method of elemental analysis for SEM, TEM, and STEM applications. The elements of interest are generally quantified without standards using theoretical calculations or by using standards that are high purity specimens of the elements measured. However, EDS is often used to determine a small percentage of an element in a matrix. The accuracy and limit of detection of these low concentration measurements has not been established. An earlier report proved the concept that a cross section high dose BF2 implanted specimen could provide a standard for EDS measurement of F. This study extends this quantification approach to transition elements of importance to the semiconductor industry.

The Fe and Co standards were created by high dose ion implantation. For ions implanted into silicon, a dose of lxl016 atoms/cm2 results in a peak concentration of approximately lxl021 atoms/cm3 or 2% atomic. The exact concentration can be determined using methods such as Rutherford Backscattering Spectrometry (RBS) and Secondary Ion Mass Spectrometry (SIMS).

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

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Rechargeable Lithium ion batteries are widely used as portable power source in communication and computer technology, prospective uses include medical implantable devices and electric vehicles. The safety and cycle life of Li ion batteries is improved over that of batteries containing metallic lithium anodes because the insertion of Li between the crystal layers of both electrodes was proved to be safer than the electroplating of Li onto a metallic Lithium anode. in Li-ion batteries, the charge transport is governed by the oscillation of Li ions between anode and cathode. They are sometimes called “rocking-chair“ batteries. The most common materials for these batteries are lithiated carbons for anodes, and transition metal oxides (LixCoO2) as cathodes.

LixCoO2 has an ordered rhombohedral Rm structure consisting of alternating layers of Co-O-Li-O-Co. The capacity and energy density of the batteries is limited by the amount of Li that can be stored in the anode and cathode materials.

Experimental conditions and setting up of an electron microprobe is directly linked to the assumption or the knowledge that can make the operator on the results of the measurements. Consequently, there is an increasing interest in modelling the X-ray spectrum in order to predict accurately and rapidly the response of the microprobe when a sample of approximately known composition is submitted to specific excitation conditions. Even if the spectra simulation is a current tool proposed in EDS, it always remains a challenge to create a complete absolute WDS spectrum. Indeed, the definition of the efficiency function was a limit to the calculation of absolute intensities.

In this work, we propose a new generation software for EPMA which uses the absolute simulation spectra to assist the operator in the choices of experimental conditions to achieve accurate measurements in a short time as well as to create new functionalities.

Extended abstract of a paper presented at Microscopy and Microanalysis 2006 in Chicago, Illinois, USA, July 30 – August 3, 2005

Light microscopy is a powerful tool that allows for many types of samples to be examined in a rapid, easy, and nondestructive manner. Subsequent image analysis, however, is compromised by distortion of signal by instrument optics. Deconvolution of images prior to analysis allows for the recovery of lost information by procedures that utilize either a theoretically or experimentally calculated point spread function (PSF). Using a laser scanning confocal microscope (LSCM), we have imaged whole impact tracks of comet particles captured in silica aerogel, a low density, porous SiO2 solid, by the NASA Stardust mission. In order to understand the dynamical interactions between the particles and the aerogel, precise grain location and track volume measurement are required. We report a method for measuring an experimental PSF suitable for three-dimensional deconvolution of imaged particles in aerogel. Using fluorescent beads manufactured into Stardust flight-grade aerogel, we have applied a deconvolution technique standard in the biological sciences to confocal images of whole Stardust tracks. The incorporation of an experimentally measured PSF allows for better quantitative measurements of the size and location of single grains in aerogel and more accurate measurements of track morphology.

This is a report of the adaptation of microwave processing in the preparation of liver biopsies for transmission electron microscopy (TEM) to examine ultrastructural damage of mitochondria in the setting of metabolic stress. Hemorrhagic shock was induced in pigs via 35% total blood volume bleed and a 90-min period of shock followed by resuscitation. Hepatic biopsies were collected before shock and after resuscitation. Following collection, biopsies were processed for TEM by a rapid method involving microwave irradiation (Giberson, 2001). Samples pre- and postshock of each of two animals were viewed and scored using the mitochondrial ultrastructure scoring system (Crouser et al., 2002), a system used to quantify the severity of ultrastructural damage during shock. Results showed evidence of increased ultrastructural damage in the postshock samples, which scored 4.00 and 3.42, versus their preshock controls, which scored 1.18 and 1.27. The results of this analysis were similar to those obtained in another model of shock (Crouser et al., 2002). However, the amount of time used to process the samples was significantly shortened with methods involving microwave irradiation.

Extended abstract of a paper presented at Microscopy and Microanalysis 2006 in Chicago, Illinois, USA, July 30 – August 3, 2006

This report summarizes our experiences gained with deconvolving 3-d images of various types of biological samples - animal and plant cells; live and fixed. We compared raw, unmodified images with images processed with standard filtering methods and images improved by using deconvolution. The blind decovolution software package from AutoQuant, Inc. was used. All images were acquired using a BioRad MRC 1024 confocal microscope (attached to a Nikon Diaphot microscope), using fluorescence or reflected light. For thin (up to 25 μm) flat specimens a 60x PlanApo NA1.4 lens was used while for thick (25-200 μm) water containing samples a 40x Fluor NA1.15 water immersion lens was used.

In an attempt to test the usefulness and validity of deconvolution for various types of biological samples we adopted the following approaches:

1. collect images of structures that are known to be both fluorescent and reflective - compare the results of deconvolution of fluorescence images with reflected light images that are not deconvolved.

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

Fluorescein and the 1.4 nm Nanogold® gold cluster label may be incorporated into a single Fab’ immunoprobe by separate cross-linking reactions, to give a probe which labels antigenic sites in a single step for correlative fluorescence and electron microscope visualization. These probes show high labeling density, labeling a pre-mRNA splicing factor in the HeLa cell nucleus; Microtubules were also densely labeled using fluorescence, other optical modalities, and electron microscopy; in a parallel experiment, a 5 nm colloidal gold probe gave only occasional labeling. We now describe Fab’ and streptavidin probes containing both Nanogold® and the fluorescent cyanine dye, Cy3.

F(ab’)2 Goat anti-Mouse IgG and F(ab’)2 goat anti-rabbit IgG fragments were reductively cleaved to Fab’ fragments using dithiothreitol (DTT) or mercaptoethylamine hydrochloride (MEA), which selectively reduce the F(ab’)2 hinge disulfide bonds, with 5 mm EDTA to prevent reoxidation. Fab’ fragments were isolated by gel filtration (coarse gel: GH25, Amicon) then labeled with Monomaleimido- Nanogold® which reacts site-specifically with thiols. Streptavidin was labeled using Mono- Sulfo-NHS-Nanogold® at pH 7.5. Nanogold® conjugates were isolated by gel filtration (Superose-12 column, Pharmacia), then reacted with excess Cy3 monofunctional NHS ester (labeling kit, Amersham Life Sciences) at pH 7.5; dual-labeled conjugates were isolated by gel filtration (Superose-12).

Extended abstract of a paper presented at Microscopy and Microanalysis 2006 in Chicago, Illinois, USA, July 30 – August 3, 2005

Electron tomography is the most widely applicable method for obtaining three-dimensional information by electron microscopy. It is, in fact, the only method suitable for investigating pleomorphic structures, such as many supramolecular assemblies, organelles and cells. With the recent development of automated low-dose data-acquisition schemes, it is now possible to study molecules and cells embedded in vitreous ice. This opens up new horizons for investigating the functional organization of cellular components with minimal perturbation of the cellular context.

In the present study we used automated electron tomography in conjunction with cryomicroscopy to reveal the internal organization and ultrastructure of mitochondria. The whole isolated cell organelles from Neurospora crassa were quick-frozen and examined in vitrified ice. Single-axis tilt series were recorded with a Philips CM 120 Biotwin under low-dose conditions; the estimated total exposure was 10,000 e−nm−2. The tilt increment was 1° and the tilt series ranged from −65° to +65°. In order to enhance the contrast of the rather thick samples zero-loss energy filtering was employed. The images were recorded with a Ik × Ik CCD camera. The experimental setup has been described by Koster et al2 Three-dimensional reconstructions were performed by weighted backprojection. Prior to the three-dimensional visualization of 3-D data sets a denoising technique based on nonlinear anisotropic diffusion was applied (Figs. 2, 3).