Anther development of maize (Zea mays) and longstamen rice (Oryza longistaminata) revealed by cryo-SEM, with foci on locular dehydration and pollen arrangement.

Key message: Pollen maturation in Poaceae. Another development has been extensively examined by various imaging tools, including transmission electron microscopy, scanning electron microscopy, and light microscopy, but none is capable of identifying liquid water. Cryo-scanning electron microscopy with high-pressure rapid freeze fixation is excellent in preserving structures at cellular level and differentiating gas- versus liquid-filled space, but rarely used in anther study. We applied this technique to examine anther development of Poaceae because of its economic importance and unusual peripheral arrangement of pollen. Maize and longstamen rice were focused on. Here, we report for the first time that anthers of Poaceae lose the locular free liquid during late-microspore to early pollen stages; the majority of pollen grains arranged in a tight peripheral whorl develops normally and reaches maturity in the gas-filled loculus. Occasionally, pollen grains are found situated in the locular cavity, but they remain immature or become shrunk at anthesis. At pollen stage, microchannels and cytoplasmic strands are densely distributed in the entire pollen exine and intine, respectively, suggesting that nutrients are transported into the pollen from the entire surface. We propose that in Poaceae, the specialized peripheral arrangement of pollen grains is crucial for pollen maturation in the gas-filled loculus, which enables pollen achieving large surface contact area with the tapetum and neighboring grains to maintain sufficient nutrient flow. This report also shows that the single aperture of pollen in Poaceae usually faces the tapetum, but other orientation is also common; pollen grains with different aperture orientations show no morphological differences.

An engineered microenvironment for multidimensional microscopy of live cells.

Multidimensional imaging (MD) of live cells is gaining importance in biomedical research as the commercial availability of confocal, nonlinear optical microscopes, environmental chambers, and specific fluorescence probes grows. One crucial aspect of the MD live cell imaging involves the proper immobilization of cells, which refers to the rapid and sufficient immobilization of cells on the microscope stage, neither disrupting the cellular structure and functions nor affecting the optical properties of the cells and the environments. Conventional cell immobilization methods glue the anchoring cells to coated surfaces, but such methods require centrifugation or extended incubation and are not suitable for cells in suspension. Most of the current three-dimensional (3-D) gels either exhibit unsatisfactory optical properties or have adverse effects on cell functions in culture. Recently, an engineered 3-D microcapsule has been developed that involves the complex coacervation of a positively charged collagen and a negatively charged polymer of 2-hydroxyethyl methacrylate--methacrylic acid--methyl methacrylate (HEMA-MMA-MAA). Hence, confocal imaging of live cells in this engineered 3-D microenvironment was investigated for its optical properties and cellular function compatibility. We report here that this microenvironment facilitates efficient cell immobilization, exhibits good optical properties, and can preserve cellular structures and functions, which will be useful in MD imaging of live cells for various applications.

The morphology and stability of resin-modified glass-ionomer adhesive at the dentin/resin-based composite interface.

PURPOSE: To evaluate the interface with tooth structure, mode of failure and stability of Fuji Bond LC (FBLC) resin-modified glass-ionomer bonding system. MATERIALS AND METHODS: The study was in three main parts. The first part with interfacial characteristics of FBLC. The micropermeability of the FBLC/dentin interface of restored cavities in extracted teeth, after fluorescent dye was introduced into the pulp chamber, was assessed by confocal microscopy. In other teeth, confocal microscopy was also used to examine trhe failure of FBLC bonded to flat dentin surfaces, on shear loading. In the second part, cervical cavities restored with FBLC (with and without light-curing) and a resin-based composite were examined for stability of the interface over the first 15 minutes. In the last part of the study, a new fluorescent dye was mixed with FBLC for restoration of cavities and subsequent examination using 2-photon imaging techniques. RESULTS: FBLC adapted well to tooth structure, had an interface with dentin that was permeable to fluid and displayed the absorption layer. Shear loading at the dentin interface resulted in complex failure with areas of cohesive failure where the absorption layer was present. Light-curing of FBLC seemed to produce a stable restorative while "co-curing" resulted in an unstable situation, with the appearance of an emmisable exudate from the FBLC. Advanced fluorescent imaging of the material itself seemed to suggest that there was a movement of aluminum ions in a manner similar to other glass-ionomer cements. The presence of HEMA in the matrix of the cement associated with the absorption layer was also demonstrated.

Fast Algorithm for X-ray Cone-beam Microtomography.

Cone-beam X-ray microtomography attracts increasing attention due to its applications in biomedical sciences, material engineering, and industrial nondestructive evaluation. Rapid volumetric image reconstruction is highly desirable in all these areas for prompt visualization and analysis of complex structures of interest. In this article, we reformulate a generalized Feldkamp cone-beam image reconstruction algorithm, utilize curved voxels and mapping tables, improve the reconstruction efficiency by an order of magnitude relative to a direct implementation of the standard algorithm, and demonstrate the feasibility with numerical simulation and experiments using a prototype cone-beam X-ray microtomographic system. Our fast algorithm reconstructs a 256-voxel cube from 100 projections within 2 min on an Intel Pentium II(R) 233 MHz personal computer, produces satisfactory image quality, and can be further accelerated using special hardware and/or parallel processing techniques.