Visually-guided protein crystal manipulation using micromachined silicon tools

We present a system for protein crystal micro-manipulation with focus on automated crystal mounting for the purposes of X-ray data collection. The system features a set of newly designed micropositioner end-effectors we call microshovels which address some limitations of the traditional cryogenic loops. We have used micro-electrical mechanical system (MEMS) techniques to design and manufacture various shapes and quantities of microshovels. Visual feedback from a camera mounted on the microscope is used to control the micropositioner as it lowers a microshovel into the liquid containing the crystals and approaches a selected crystal for pickup. We present experimental results that illustrate the applicability of our approach

Visually-guided protein crystal manipulation using micromachined silicon tools

We present a system for protein crystal micro-manipulation with focus on automated crystal mounting for the purposes of X-ray data collection. The system features a set of newly designed micropositioner end-effectors we call microshovels which address some limitations of the traditional cryogenic loops. We have used micro-electrical mechanical system (MEMS) techniques to design and manufacture various shapes and quantities of microshovels. Visual feedback from a camera mounted on the microscope is used to control the micropositioner as it lowers a microshovel into the liquid containing the crystals and approaches a selected crystal for pickup. We present experimental results that illustrate the applicability of our approach

A Microrobotic System For Protein Streak Seeding

We present a microrobotic system for protein crystal micromanipulation tasks. The focus in this report is on a task called streak seeding, which is used by crystallographers to entice certain protein crystals to grow. Our system features a set of custom designed micropositioner end-effectors we call microshovels to replace traditional tools used by crystallographers for this task. We have used micro-electrical mechanical system (MEMS) techniques to design and manufacture various shapes and quantities of microshovels. Visual feedback from a camera mounted on the microscope is used to control the micropositioner as it lowers a microshovel into the liquid containing the crystals for poking and streaking. We present experimental results that illustrate the applicability of our approach

Automated streak-seeding with micromachined silicon tools

This report presents a new approach to streak-seeding based on custom-designed silicon microtools. Experimental data show that the microtools produce similar results to the commonly used boar bristles. One advantage to using silicon is that it is rigid and can easily serve as an accurately calibrated end-effector on a micro-robotic system. Additionally, the fabrication technology allows the production of microtools of various shapes and sizes. A working prototype of an automatic streak-seeding system based on these microtools was built and successfully applied for protein crystallization

Howard walnut trees can be brought into bearing without annual pruning

In traditionally managed Howard walnut orchards, trees are pruned annually during the orchard development phase, an expensive operation in terms of labor and prunings disposal costs. Our observations and some prior research by others had suggested that pruning may not be necessary in walnut. In a trial of pruned and unpruned hedgerow trees over 8 years, beginning a year after planting, we documented canopy growth, tree height, yield and nut quality characteristics and also the effects of fruit removal. Pruning altered canopy shape but did not lead to increases in canopy development, yield or nut quality. Although fruit removal stimulated more vegetative growth in both the pruned and unpruned treatments, fruit removal did not result in an increase in midday canopy photosynthetically active radiation interception or cumulative yield when fruit removal was stopped after year 4. After 8 years, there were no significant differences in tree height, nut quality or cumulative yield among any of the treatments, which suggests that not pruning young Howard orchards could provide a net benefit to growers

A Possible Role for Metallic Ions in the Carbohydrate Cluster Recognition Displayed by a Lewis Y Specific Antibody

BACKGROUND:Lewis Y (Le(y)) is a blood group-related carbohydrate that is expressed at high surface densities on the majority of epithelial carcinomas and is a promising target for antibody-based immunotherapy. A humanized Le(y)-specific antibody (hu3S193) has shown encouraging safety, pharmacokinetic and tumor-targeting properties in recently completed Phase I clinical trials. METHODOLOGY/PRINCIPAL FINDINGS:We report the three-dimensional structures for both the free (unliganded) and bound (Le(y) tetrasaccharide) hu3S193 Fab from the same crystal grown in the presence of divalent zinc ions. There is no evidence of significant conformational changes occurring in either the Le(y) carbohydrate antigen or the hu3S193 binding site, which suggests a rigid fit binding mechanism. In the crystal, the hu3S193 Fab molecules are coordinated at their protein-protein interface by two zinc ions and in solution aggregation of Fab can be initiated by zinc, but not magnesium ions. Dynamic light scattering revealed that zinc ions could initiate a sharp transition from hu3S193 Fab monomers to large multimeric aggregates in solution. CONCLUSIONS/SIGNIFICANCE:Zinc ions can mediate interactions between hu3S193 Fab in crystals and in solution. Whether metallic ion mediated aggregation of antibody occurs in vivo is not known, but the present results suggest that similar clustering mechanisms could occur when hu3S193 binds to Le(y) on cells, particularly given the high surface densities of antigen on the target tumor cells