Search NASA⌕ Search

Engineering topics

Lin, Wei

Publications and source records attributed to Lin, Wei.

13 records
All sourcesNASA NTRSDOE OSTI
DOE OSTI2023

Light-sheet autofluorescence lifetime imaging with a single-photon avalanche diode array

Significance: Fluorescence lifetime imaging microscopy (FLIM) of the metabolic co-enzyme nicotinamide adenine dinucleotide (phosphate) [NAD(P)H] is a popular method to monitor single-cell metabolism within unperturbed, living 3D systems. However, FLIM of NAD(P)H has not been performed in a light-sheet geometry, which is advantageous for rapid imaging of cells within live 3D samples. Aim: We aim to design, validate, and demonstrate a proof-of-concept light-sheet system for NAD(P)H FLIM. Approach: A single-photon avalanche diode camera was integrated into a light sheet microscope to achieve optical sectioning and limit out-of-focus contributions for NAD(P)H FLIM of single cells. Results: An NAD(P)H light-sheet FLIM system was built and validated with fluores cence lifetime standards and with time-course imaging of metabolic perturbations in pancreas cancer cells with 10 s integration times. NAD(P)H light-sheet FLIM in vivo was demonstrated with live neutrophil imaging in a larval zebrafish tail wound also with 10 s integration times. Finally, the theoretical and practical imaging speeds for NAD(P)H FLIM were compared across laser scanning and light-sheet geometries, indicating a 30× to 6× acquisition speed advantage for the light sheet compared to the laser scanning geometry. Conclusions: FLIM of NAD(P)H is feasible in a light-sheet geometry and is attrac tive for 3D live cell imaging applications, such as monitoring immune cell metabolism and migration within an organism.

47 OTHER INSTRUMENTATION↗
DOE OSTI2021

Functional Characterization and Structural Insights Into Stereoselectivity of Pulegone Reductase in Menthol Biosynthesis

Monoterpenoids are the main components of plant essential oils and the active components of some traditional Chinese medicinal herbs like Mentha haplocalyx Briq ., Nepeta tenuifolia Briq ., Perilla frutescens (L.) Britt and Pogostemin cablin (Blanco) Benth. Pulegone reductase is the key enzyme in the biosynthesis of menthol and is required for the stereoselective reduction of the Δ 2,8 double bond of pulegone to produce the major intermediate menthone, thus determining the stereochemistry of menthol. However, the structural basis and mechanism underlying the stereoselectivity of pulegone reductase remain poorly understood. In this study, we characterized a novel (-)-pulegone reductase from Nepeta tenuifolia ( Nt PR), which can catalyze (-)-pulegone to (+)-menthone and (-)-isomenthone through our RNA-seq, bioinformatic analysis in combination with in vitro enzyme activity assay, and determined the structure of (+)-pulegone reductase from ( Mp PR) by using X-ray crystallography, molecular modeling and docking, site-directed mutagenesis, molecular dynamics simulations, and biochemical analysis. We identified and validated the critical residues in the crystal structure of Mp PR involved in the binding of the substrate pulegone. We also further identified that residues Leu56, Val282, and Val284 determine the stereoselectivity of the substrate pulegone, and mainly contributes to the product stereoselectivity. This work not only provides a starting point for the understanding of stereoselectivity of pulegone reductases, but also offers a basis for the engineering of menthone/menthol biosynthetic enzymes to achieve high-titer, industrial-scale production of enantiomerically pure products.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗
DOE OSTI2021

Submonolayer Is Enough: Switching Reaction Channels on Pt/SiO 2 by Atomic Layer Deposition

The reaction mechanism of CO 2 with H 2 is studied on platinum nanoparticles supported on fumed silica. It is found that platinum nanoparticle size, reaction temperature, and metal oxide promoters play important roles in determining the reaction rate and the mechanism of forming surface carbonyl species. Metal oxide promoters consist of sub-monolayer titanium oxide or aluminum oxide overcoated onto the catalysts by atomic layer deposition (ALD). Furthermore, these alter the CO formation rate, influence the adsorption and desorption behavior, and switch the surface reaction channel from Eley-Rideal to Langmuir-Hinshelwood mechanism due to an enhancement of CO 2 affinity to the metal-metal oxide interface. At the temperatures relevant for catalytic turnover, ALD overcoating significantly increases catalytic activity in CO 2 hydrogenation to CH 4 and CO, while the identity of the oxide overcoat helps control product selectivity.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2021

Investigating Single-Molecule Fluorescence Spectral Heterogeneity of Rhodamines Using High-Throughput Single-Molecule Spectroscopy

We experimentally investigated several intra-molecular coordinate and environmental changes as potential causes of single-molecule fluorescence spectral heterogeneities (smFSH). Here, we developed a high-throughput single-molecule spectroscopy technique to analyze more than 5,000 single-molecule emission spectra from each of nine commonly used fluorophores with different structural rigidities deposited on substrates with different polarities. We observed an unexpected high smFSH from structurally rigid Rhodamine B compared with a structurally flexible Cyanine dye - Alexa Fluor 647. Based on experimentally measured smFSH, we ruled out the system’s noise uncertainty, single-molecule spectral diffusion, and environmental polarity as the primary causes of the high smFSH. We found that the rotational flexibility of N, N-dialkylated groups contributed to the smFSH. With the high smFSH observed in structurally more rigid model fluorophores, we speculated that other intra-molecular coordinate and environmental changes might also contribute to the high smFSH in Rhodamines.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2019

Distributed formation control with open-loop Nash strategy

In this paper, we investigate game theoretical multiagent formation control problem. The main challenge of this problem is how to enable local execution of game strategies, e.g., Nash equilibrium, which generally requires global information in the presence of communication topology among agents. Toward this, we first derive open-loop Nash equilibrium for agents minimizing formation error based performance indices and proceed to introduce a distributed estimation scheme to facilitate local implementation of the derived Nash strategies. In addition, a shrinking horizon control technique is integrated into the proposed scheme to handle unexpected state changes. Finally, the proposed scheme is fully distributed and ensures an ϵ-Nash equilibrium. An illustrative examples is presented to verify the effectiveness.

42 ENGINEERING↗
DOE OSTI2019

Structural basis of ECF-σ-factor-dependent transcription initiation

Extracytoplasmic (ECF) σ factors, the largest class of alternative σ factors, are related to primary σ factors, but have simpler structures, comprising only two of six conserved functional modules in primary σ factors: region 2 (σR2) and region 4 (σR4). Here, we report crystal structures of transcription initiation complexes containing Mycobacterium tuberculosis RNA polymerase (RNAP), M. tuberculosis ECF σ factor σL, and promoter DNA. The structures show that σR2 and σR4 of the ECF σ factor occupy the same sites on RNAP as in primary σ factors, show that the connector between σR2 and σR4 of the ECF σ factor–although shorter and unrelated in sequence–follows the same path through RNAP as in primary σ factors, and show that the ECF σ factor uses the same strategy to bind and unwind promoter DNA as primary σ factors. The results define protein-protein and protein-DNA interactions involved in ECF-σ-factor-dependent transcription initiation.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Pore structure development during hydration of tricalcium silicate by X-ray nano-imaging in three dimensions

We report that tricalcium silicate (C 3 S) is the most important component among the four main clinker phases of Portland cement. Pure C 3 S is widely used as a simplified model system of cement in various researches. However, the spatial structure development of cement, even pure C 3 S during hydration at the nano-scale has been rarely directly reported. In this work, X-ray nano-computed tomography (X-ray Nano-CT), a non-destructive X-ray analytical method, was used to study the hydration of a pure C 3 S sample with a water/C 3 S mass ratio of 0.5. The three-dimensional (3D) structure of the hydrating C 3 S specimen was investigated to see the internal pore structure evolution within the hardened C 3 S paste. Investigation of the 3D structural development of the C 3 S specimen at different hydration times was performed to monitor the changes of pore shapes and sizes/volumes. Lastly, it is found that volumes of the sealed pores generally grow larger, and the open pores become smaller while the volume and the external morphology of the whole hardened C 3 S paste remains almost the same during hydration.

36 MATERIALS SCIENCE↗
DOE OSTI2018

Coherent diffraction study of calcite crystallization during the hydration of tricalcium silicate

The aim of this work is using Bragg coherent X-ray diffraction imaging (BCDI) to study the calcite crystallization during carbonation of hydrated tricalcium silicate (C 3 S). Portland cement is a very complex synthesized product whose 50–70% mass is composed of C 3 S, which is the most important phase to produce calcium silicate hydrates and calcium hydroxide. Hence, its hydration contributes greatly to the hydration of cement and later to the carbonation of cement products when it reacts with CO 2 , often from the air, to form calcium carbonates. BCDI has emerged in the last decade as a promising high-resolution lens-less imaging approach for characterization of various samples. It has made significant progress with the development of X-ray sources and phase-retrieval algorithms. BCDI allows for imaging the whole three-dimensional structure of micro- and sub-micro- crystalline materials and can show the strain distribution at the nanometer spatial resolution. Finally, results show that calcite crystallization follows a through-solution reaction and the growth model of the calcite crystal can be explained by using “phase domain” theory. In conclusion, during carbonation, calcite crystals grow by increasing the number of phase domains within them while the domain size remains at about 200–300 nm.

36 MATERIALS SCIENCE↗
NASA NTRS2013

Developing IVHM Requirements for Aerospace Systems

The term Integrated Vehicle Health Management (IVHM) describes a set of capabilities that enable sustainable and safe operation of components and subsystems within aerospace platforms. However, very little guidance exists for the systems engineering aspects of design with IVHM in mind. It is probably because of this that designers have to use knowledge picked up exclusively by experience rather than by established process. This motivated a group of leading IVHM practitioners within the aerospace industry under the aegis of SAE's HM-1 technical committee to author a document that hopes to give working engineers and program managers clear guidance on all the elements of IVHM that they need to consider before designing a system. This proposed recommended practice (ARP6883 [1]) will describe all the steps of requirements generation and management as it applies to IVHM systems, and demonstrate these with a "real-world" example related to designing a landing gear system. The team hopes that this paper and presentation will help start a dialog with the larger aerospace community and that the feedback can be used to improve the ARP and subsequently the practice of IVHM from a systems engineering point-of-view.

IVHM↗
NASA NTRS2013

Towards Requirements in Systems Engineering for Aerospace IVHM Design

Health management (HM) technologies have been employed for safety critical system for decades, but a coherent systematic process to integrate HM into the system design is not yet clear. Consequently, in most cases, health management resorts to be an after-thought or 'band-aid' solution. Moreover, limited guidance exists for carrying out systems engineering (SE) on the subject of writing requirements for designs with integrated vehicle health management (IVHM). It is well accepted that requirements are key to developing a successful IVHM system right from the concept stage to development, verification, utilization, and support. However, writing requirements for systems with IVHM capability have unique challenges that require the designers to look beyond their own domains and consider the constraints and specifications of other interlinked systems. In this paper we look at various stages in the SE process and identify activities specific to IVHM design and development. More importantly, several relevant questions are posed that system engineers must address at various design and development stages. Addressing these questions should provide some guidance to systems engineers towards writing IVHM related requirements to ensure that appropriate IVHM functions are built into the system design.

Requirements↗
NASA NTRS2012

NASA Integrated Model Centric Architecture (NIMA) Model Use and Re-Use

This whitepaper accepts the goals, needs and objectives of NASA's Integrated Model-centric Architecture (NIMA); adds experience and expertise from the Constellation program as well as NASA's architecture development efforts; and provides suggested concepts, practices and norms that nurture and enable model use and re-use across programs, projects and other complex endeavors. Key components include the ability to effectively move relevant information through a large community, process patterns that support model reuse and the identification of the necessary meta-information (ex. history, credibility, and provenance) to safely use and re-use that information. In order to successfully Use and Re-Use Models and Simulations we must define and meet key organizational and structural needs: 1. We must understand and acknowledge all the roles and players involved from the initial need identification through to the final product, as well as how they change across the lifecycle. 2. We must create the necessary structural elements to store and share NIMA-enabled information throughout the Program or Project lifecycle. 3. We must create the necessary organizational processes to stand up and execute a NIMA-enabled Program or Project throughout its lifecycle. NASA must meet all three of these needs to successfully use and re-use models. The ability to Reuse Models a key component of NIMA and the capabilities inherent in NIMA are key to accomplishing NASA's space exploration goals. 11

Conroy, Mike↗