NanoSIMS imaging of Bacillus spores sectioned by focused ion beam.

Preparation and sectioning of bacterial spores by focused ion beam and subsequent high resolution secondary ion mass spectrometry analytical imaging is demonstrated. Scanning transmission electron microscopy mode imaging in a scanning electron microscope is used to show that the internal structure of the bacterial spore can be preserved during focused ion beam sectioning and can be imaged without contrast staining. Ion images of the sections show that the internal elemental distributions of the sectioned spores are preserved. A rapid focused ion beam top-sectioning method is demonstrated to yield comparable ion images without the need for sample trenching and section lift-out. The lift-out and thinning method enable correlated transmission electron microscopy and high resolution secondary ion mass spectrometry analyses. The top-cutting method is preferable if only secondary ion mass spectrometry analyses are performed because this method is faster and yields more sample material for analysis; depth of useful sample material is approximately 300 nm for top-cut sections versus approximately 100 nm for electron-transparent sections.

Investigation of cryoprotectant toxicity to porcine embryos.

The objective of this study was to examine the potential toxicity of sucrose (Experiment 1) and of various cryoprotectants (Experiment 2) to porcine preimplantation embryos. In Experiment 1, 65 embryos, ranging from compact morulae to hatched blastocysts, were allocated within donor female across 5 concentrations of sucrose (0, 0.25, 0.50, 1.0, 2.0 M) to determine the highest concentration that would not inhibit subsequent embryo development. After a 48-h post-treatment culture period, the embryos were stained and cell nuclei were counted. The concentration of sucrose affected embryo development (P < 0.001) and embryo quality (P < 0.001). Embryos placed into 2.0 M sucrose exhibited poorer development and quality than embryos at the lower 4 concentrations, which were not different from one another. In Experiment 2, 182 embryos of the same developmental stages as in Experiment 1 were collected from 16 donors. Embryos were allotted within donor female to 2 of the 5 concentrations (10, 20, 30, 40, or 50%) of each of 3 cryoprotectants (ethylene glycol, propylene glycol, glycerol). After a 30-sec exposure to a cryoprotectant, the embryos were cultured and stained as in Experiment 1. As the concentration of an individual cryoprotectant increased beyond 30%, embryo development decreased. Embryos exposed to glycerol or propylene glycol exhibited poorer development than did embryos placed into ethylene glycol, especially at concentrations of 40% or higher.

Systems for research and development in medical ultrasound imaging.

The system requirements of engineers conducting research and development into new ultrasonic methods for medical applications are reported. The design criteria for these systems are explained together with a review of what is currently available.

Identification of Desulfobacterales as primary hydrogenotrophs in a complex microbial mat community.

Hypersaline microbial mats have been shown to produce significant quantities of H2 under dark, anoxic conditions via cyanobacterial fermentation. This flux of a widely accessible microbial substrate has potential to significantly influence the ecology of the mat, and any consumption will affect the net efflux of H2 that might otherwise be captured as a resource. Here, we focus on H2 consumption in a microbial mat from Elkhorn Slough, California, USA, for which H2 production has been previously characterized. Active biologic H2 consumption in this mat is indicated by a significant time-dependent decrease in added H2 compared with a killed control. Inhibition of sulfate reduction, as indicated by a decrease in hydrogen sulfide production relative to controls, resulted in a significant increase in H2 efflux, suggesting that sulfate-reducing bacteria (SRB) are important hydrogenotrophs. Low methane efflux under these same conditions indicated that methanogens are likely not important hydrogenotrophs. Analyses of genes and transcripts that encode for rRNA or dissimilatory sulfite reductase, using both PCR-dependent and PCR-independent metatranscriptomic sequencing methods, demonstrated that Desulfobacterales are the dominant, active SRB in the upper, H2-producing layer of the mat (0-2 mm). This hypothesis was further supported by the identification of transcripts encoding hydrogenases derived from Desulfobacterales capable of H2 oxidation. Analysis of molecular data provided no evidence for the activity of hydrogenotrophic methanogens. The combined biogeochemical and molecular data strongly indicate that SRB belonging to the Desulfobacterales are the quantitatively important hydrogenotrophs in the Elkhorn Slough mat.

Constraints on the formation age of cometary material from the NASA Stardust mission.

We measured the 26Al-26Mg isotope systematics of a approximately 5-micrometer refractory particle, Coki, returned from comet 81P/Wild 2 in order to relate the time scales of formation of cometary inclusions to their meteoritic counterparts. The data show no evidence of radiogenic 26Mg and define an upper limit to the abundance of 26Al at the time of particle formation: 26Al/27Al < 1 x 10(-5). The absence of 26Al indicates that Coki formed >1.7 million years after the oldest solids in the solar system, calcium- and aluminum-rich inclusions (CAIs). The data suggest that high-temperature inner solar system material formed, was subsequently transferred to the Kuiper Belt, and was incorporated into comets several million years after CAI formation.