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1.
Heliyon ; 10(1): e23202, 2024 Jan 15.
Article in English | MEDLINE | ID: mdl-38169844

ABSTRACT

Laser-powder bed fusion additive manufacturing (LPBF-AM) of metals is rapidly becoming one of the most important materials processing pathways for next-generation metallic parts and components in a number of important applications. However, the large parametric space that characterizes laser-based LPBF-AM makes it challenging to understand what are the variables controlling the microstructural and mechanical property outcomes. Sensitivity studies based on direct LPBF-AM processing are costly and lengthy to conduct, and are subjected to the specifications and variability of each printer. Here we develop a fast-throughput numerical approach that simulates the LPBF-AM process using a cellular automaton model of dynamic solidification and grain growth. This is accompanied by a polycrystal plasticity model that captures grain boundary strengthening due to complex grain geometry and furnishes the stress-strain curves of the resulting microstructures. Our approach connects the processing stage with the mechanical testing stage, thus capturing the effect of processing variables such as the laser power, laser spot size, scan speed, and hatch width on the yield strength and tangent moduli of the processed materials. When applied to pure Cu and stainless 316L steel, we find that laser power and scan speed have the strongest influence on grain size in each material, respectively.

2.
Mol Cell ; 69(2): 321-333.e3, 2018 01 18.
Article in English | MEDLINE | ID: mdl-29351850

ABSTRACT

We have developed a highly parallel strategy, systematic gene-to-phenotype arrays (SGPAs), to comprehensively map the genetic landscape driving molecular phenotypes of interest. By this approach, a complete yeast genetic mutant array is crossed with fluorescent reporters and imaged on membranes at high density and contrast. Importantly, SGPA enables quantification of phenotypes that are not readily detectable in ordinary genetic analysis of cell fitness. We benchmark SGPA by examining two fundamental biological phenotypes: first, we explore glucose repression, in which SGPA identifies a requirement for the Mediator complex and a role for the CDK8/kinase module in regulating transcription. Second, we examine selective protein quality control, in which SGPA identifies most known quality control factors along with U34 tRNA modification, which acts independently of proteasomal degradation to limit misfolded protein production. Integration of SGPA with other fluorescent readouts will enable genetic dissection of a wide range of biological pathways and conditions.


Subject(s)
High-Throughput Nucleotide Sequencing/methods , High-Throughput Screening Assays/methods , Cyclin-Dependent Kinase 8/genetics , Gene Regulatory Networks , Genotype , Mediator Complex/genetics , Oligonucleotide Array Sequence Analysis , Phenotype , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae Proteins/genetics
3.
Appl Environ Microbiol ; 81(16): 5639-49, 2015 Aug 15.
Article in English | MEDLINE | ID: mdl-26070672

ABSTRACT

Agar, a seaweed extract, has been the standard support matrix for microbial experiments for over a century. Recent developments in high-throughput genetic screens have created a need to reevaluate the suitability of agar for use as colony support, as modern robotic printing systems now routinely spot thousands of colonies within the area of a single microtiter plate. Identifying optimal biophysical, biochemical, and biological properties of the gel support matrix in these extreme experimental conditions is instrumental to achieving the best possible reproducibility and sensitivity. Here we systematically evaluate a range of gelling agents by using the yeast Saccharomyces cerevisiae as a model microbe. We find that carrageenan and Phytagel have superior optical clarity and reduced autofluorescence, crucial for high-resolution imaging and fluorescent reporter screens. Nutrient choice and use of refined Noble agar or pure agarose reduce the effective dose of numerous selective drugs by >50%, potentially enabling large cost savings in genetic screens. Using thousands of mutant yeast strains to compare colony growth between substrates, we found no evidence of significant growth or nutrient biases between gel substrates, indicating that researchers could freely pick and choose the optimal gel for their respective application and experimental condition.


Subject(s)
Agar , Culture Media/chemistry , Gels , Microbiological Techniques/methods , Chemical Phenomena , High-Throughput Screening Assays , Saccharomyces cerevisiae/growth & development
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