Investigation of aspect ratio of hole drilling from micro to nanoscale via focused ion beam fine milling

Holes with different sizes from microscale to nanoscale were directly fabricated by focused ion beam (FIB) milling in this paper. Maximum aspect ratio of the fabricated holes can be 5:1 for the hole with large size with pure FIB milling, 10:1 for gas assistant etching, and 1:1 for the hole with size below 100 nm. A phenomenon of volume swell at the boundary of the hole was observed. The reason maybe due to the dose dependence of the effective sputter yield in low intensity Gaussian beam tail regions and redeposition. Different materials were used to investigate variation of the aspect ratio. The results show that for some special material, such as Ni-Be, the corresponding aspect ratio can reach 13.8:1 with Cl₂ assistant etching, but only 0.09:1 for Si(100) with single scan of the FIB.

Inverse Metabolic Engineering of Synechocystis PCC 6803 for Improved Growth Rate and Poly-3-hydroxybutyrate Production

Synechocystis PCC 6803 is a photosynthetic bacterium that has the potential to make bioproducts from carbon dioxide and light. Biochemical production from photosynthetic organisms is attractive because it replaces the typical bioprocessing steps of crop growth, milling, and fermentation, with a one-step photosynthetic process. However, low yields and slow growth rates limit the economic potential of such endeavors. Rational metabolic engineering methods are hindered by limited cellular knowledge and inadequate models of Synechocystis. Instead, inverse metabolic engineering, a scheme based on combinatorial gene searches which does not require detailed cellular models, but can exploit sequence data and existing molecular biological techniques, was used to find genes that (1) improve the production of the biopolymer poly-3-hydroxybutyrate (PHB) and (2) increase the growth rate. A fluorescence activated cell sorting assay was developed to screen for high PHB producing clones. Separately, serial sub-culturing was used to select clones that improve growth rate. Novel gene knock-outs were identified that increase PHB production and others that increase the specific growth rate. These improvements make this system more attractive for industrial use and demonstrate the power of inverse metabolic engineering to identify novel phenotype-associated genes in poorly understood systems.