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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 307 records · Page 17
All sourcesNASA NTRSDOE OSTI
DOE OSTI2025

Conformal duality of the nonlinear Schrödinger equation: Theory and applications to parameter estimation

The nonlinear Schrödinger equation (NLSE) in one spatial dimension has stationary solutions similar to those of the linear Schrödinger equation (LSE) as well as more exotic solutions such as solitary waves and quantum droplets. Here, we present a newly discovered conformal duality which unifies the stationary and time-dependent traveling-wave solutions of the one-dimensional cubic-quintic NLSE, the cubic NLSE and LSE. Any two systems that are classified by the same single number called the cross ratio are related by this symmetry. Notably, the conformal duality can also be adapted in Newtonian mechanics and serves as a powerful tool for investigating physical systems that otherwise cannot be directly accessed in experiments. Further, we show that the conformal symmetry is a valuable resource to substantially improve NLSE parameter estimation from noisy empirical data by introducing an optimization afterburner. The new method therefore has far reaching practical applications for nonlinear physical systems. Published by the American Physical Society 2025

Reinhardt, David B. (ORCID:0009000409812838)↗
DOE OSTI2020

The ASD Scorpius Accelerator at the NNSS. Recruitment Slides for the University of Nevada, Reno [Slides]

The Scorpius Accelerator: Supports the maintenance of the US nuclear stockpile, helps the US not return to nuclear testing, is one of four diagnostics being installed 963 feet below ground, probes subcritical implosions of nuclear material, and supports the validation of LANL and LLNL computer codes. We are searching engineers and scientists with experienced in: Mechanical Engineering, Electrical Engineering, Pulsed Power, Plasma Physics, Controls, Diagnostics, and Fluids and pressure systems.

42 ENGINEERING↗
DOE OSTI2021

Effects of 18 O Exchange on Neutron Emission Rates of Aging 238 PuO 2 LWRHUs

The Lightweight Radioisotope Heater Unit (LWRHU) provides about 1 Watt of thermal power to critical electrical and mechanical systems in the extreme temperatures of space, maintaining the operating temperature of the spacecraft’s scientific and general operating equipment. Los Alamos National Laboratory (LANL) designed and tested the LWRHU in the 1970s through early 1980s. The first production campaign at LANL was in the early 1980s in support of the Galileo mission in which over 100 LWRHUs were manufactured. In the mid-1990s, almost 200 LWRHUs were manufactured in support of the Cassini mission. After the primary Cassini manufacturing campaign, the Department of Energy (DOE) approved the fabrication of more than 10 additional LWRHUs as spares in 1998. The LWRHU assembly contains the following components: a fuel pellet, a vented capsule, a pyrolytic graphite insulator, and a fine-weave pierced fabric (FWPF) graphite aeroshell. The fuel pellet is a hot-pressed 238 PuO 2 cylindrical pellet that is sintered to create a ceramic pellet. The pellet is encapsulated in a platinum-rhodium encapsulation. The welded fueled clad is placed inside a pyrolytic graphite thermal insulation to protect the fuel from the heat produced in reentry events. The exterior layer of the LWRHU assembly is the FWPF aeroshell. Before sintering, PuO 2 granules are heated in a furnace at temperatures greater than or equal to 700°C under an enriched 16 O atmosphere in order to reduce trace quantities of 18 O. Because 18 O undergoes an (α,n) reaction, 18 O needs to be removed as much as possible to meet radiation emission requirements set by the Jet Propulsion Laboratory (JPL) and the National Aeronautics and Space Administration (NASA), as well as other agencies. The relative abundances of the oxygen isotopes are indicated by neutron emission rate (NER) measurements. LWRHUs are vented with a sintered platinum frit vent (George 1986). During extraterrestrial operation, the vent in the cladding allows helium from the decay of plutonium to escape and relieves pressure from the system while preventing the solids from escaping (Rinehart 1996, Tate 1982 & 1985). This vent is protected during manufacture by a platinum-30 rhodium cover. The capsule vent is activated by milling a 0.025-inch diameter hole through the protective cover to a nominal depth of 0.015 inches, generally immediately before the LWRHU is loaded into the aeroshell (George 1986). Prior to long-term storage, LWRHUs are vented and thus potentially allow infiltration of 18 O from the ambient atmosphere. Although NER decreases with age due to the reduction in emitted alpha particles, it might be expected that NER reduction is mitigated by the exchange of 18 O. However, this is not necessarily the case and no trend was observed relating NER to age other than the natural decay of 238 Pu (Mulford 2021). The most probable and dominant factor for determining specific NER was manufacturing discrepancies between individual units. It is important to understand the lack of observed 16 O- 18 O exchange and to compare LWRHU pellets as much as possible.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗
DOE OSTI2024

Materials Assessment for FLiBe Fusion Blankets

This evaluation was prepared at the request of DOE-FES and expands on an earlier evaluation prepared by Wilson, Wiffen and Keiser in 2015 (attached as Appendices A and B). Additional useful information for materials in fission molten salt reactor systems that use FLiBe can be found in a prior NRC report [Busby 2019]. The purpose of this report is to review the considerations that will determine the fusion blanket structural materials that will be suitable for use in a FLiBe breeder-coolant fusion power plant (FPP). Identifying a fusion-relevant structural material that is sufficiently compatible with FLiBe is a key issue to be addressed. The constraints and trade-offs made in structural materials selection include the usual power generation requirements such as mechanical properties, fabricability and durability, which assumes reasonable compatibility with the working fluid. For an FPP, additional constraints include neutron economy (breeding ratio), resistance to irradiation damage and low activation (i.e. waste disposal) characteristics.

36 MATERIALS SCIENCE↗
DOE OSTI2024

Multi-Physics Topology Optimization for Dual Flow Heat Exchangers

Current heat exchanger (HX) designs are subjected to strict constraints pertaining to their manufacturability through conventional lengthy and expensive processes consisting of subtractive, forming and brazing processes. Therefore, existing HXs are typically restricted to the classical and rather simple geometries, such as plate-fin or shell-tube geometries, with lower efficiency and larger size. To increase HX efficiencies, novel designs are needed. Recent advances in topology optimization (TO) and additive manufacturing (AM) offer significant flexibility in design and fabrication of complex geometries, with potentially superior thermo-mechanical performance and high-power density. On the other hand, there is no commercially available software tool that addresses the combined coupling of structural, fluid dynamics, and thermal with multiple materials (hot and cold streams, solid, and void space) in TO of HXs. The goal of this project is to develop a multi-physics multi-phase TO approach to enhance the performance of HXs. The optimized design will be additively manufactured and tested in a physical environment to quantify its performance.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗
DOE OSTI2025

Cyber Conservative Operations

In an era of increasingly sophisticated and pervasive cyber threats, robust cyber resilience strategies are more critical than ever. This paper introduces the concept of Cyber Conservative Operations, a proactive approach designed to assist critical infrastructure owners and operators in managing risk and maintaining resilience in the face of imminent, yet not occurring, cyber events. By leveraging the principles of Cyber-Informed Engineering (CIE) and the energy sector’s practice of conservative operations, Cyber Conservative Operations offer a framework for planning and executing coordinated active defense and resilience-oriented actions ahead of cyber events. This approach aims to minimize the consequences of digitally-enabled hazards and ensure swift recovery from anticipated cyber threats. Cyber Conservative Operations enhance the ability of owners and operators to execute pre-planned defense and resilience actions, thereby reducing the impact of digitally-enabled hazards. These operations are crucial for addressing impacts that cannot be precisely quantified or forecasted and for preparing organizations for rapid recovery from imminent digital threats. The paper begins with a discussion of conservative operations as applied to the bulk power system (BPS), a current mechanism allowing BPS entities to enact defensive operating plans to mitigate impending grid unreliability. Building on this model, we present a concept for implementing cyber conservative operations at asset owner facilities. The paper concludes with two case studies of cyber conservative operations drawn from high-profile cyber events, illustrating the practical application and benefits of this proactive approach.

42 - ENGINEERING↗
DOE OSTI2021

Real-Time Nonlinear Model Predictive Controller for Multiple Degrees of Freedom Wave Energy Converters with Non-Ideal Power Take-Off

An increase in wave energy converter (WEC) efficiency requires not only consideration of the nonlinear effects in the WEC dynamics and the power take-off (PTO) mechanisms, but also more integrated treatment of the whole system, i.e., the buoy dynamics, the PTO system, and the control strategy. It results in an optimization formulation that has a nonquadratic and nonstandard cost functional. This article presents the application of real-time nonlinear model predictive controller (NMPC) to two degrees of freedom point absorber type WEC with highly nonlinear PTO characteristics. The nonlinear effects, such as the fluid viscous drag, are also included in the plant dynamics. The controller is implemented on a real-time target machine, and the WEC device is emulated in real-time using the WECSIM toolbox. The results for the successful performance of the design are presented for irregular waves under linear and nonlinear hydrodynamic conditions.

Haider, Ali S. (ORCID:0000000246513157)↗
DOE OSTI2020

The Gaia–Kepler Stellar Properties Catalog. II. Planet Radius Demographics as a Function of Stellar Mass and Age

Studies of exoplanet demographics require large samples and precise constraints on exoplanet host stars. Using the homogeneous Kepler stellar properties derived using the Gaia Data Release 2 by Berger et al., we recompute Kepler planet radii and incident fluxes and investigate their distributions with stellar mass and age. We measure the stellar mass dependence of the planet radius valley to be dlogR{sub p}/dlogM{sub ⋆} = 0.26{sub −0.16}{sup +0.21}, consistent with the slope predicted by a planet mass dependence on stellar mass (0.24–0.35) and core-powered mass loss (0.33). We also find the first evidence of a stellar age dependence of the planet populations straddling the radius valley. Specifically, we determine that the fraction of super-Earths (1–1.8 R{sub ⊕}) to sub-Neptunes (1.8–3.5 R{sub ⊕}) increases from 0.61 ± 0.09 at young ages (<1 Gyr) to 1.00 ± 0.10 at old ages (>1 Gyr), consistent with the prediction by core-powered mass loss that the mechanism shaping the radius valley operates over Gyr timescales. Additionally, we find a tentative decrease in the radii of relatively cool (F {sub p} < 150 F{sub ⊕}) sub-Neptunes over Gyr timescales, which suggests that these planets may possess H/He envelopes instead of higher mean molecular weight atmospheres. We confirm the existence of planets within the hot sub-Neptunian “desert” (2.2 R {sub ⊕} < R {sub p} < 3.8 R{sub ⊕}, F {sub p} > 650 F{sub ⊕}) and show that these planets are preferentially orbiting more evolved stars compared to other planets at similar incident fluxes. In addition, we identify candidates for cool (F {sub p} < 20 F{sub ⊕}) inflated Jupiters, present a revised list of habitable zone candidates, and find that the ages of single and multiple transiting planet systems are statistically indistinguishable.

79 ASTRONOMY AND ASTROPHYSICS↗
DOE OSTI2020

Mechanical failure of fresh nuclear grade iron–chromium–aluminum (FeCrAl) cladding under simulated hot zero power reactivity initiated accident conditions

The reactivity initiated accident (RIA) is a postulated accident in light water reactors initiated by a rapid increase in reactivity which causes an increase in fission rate and fuel temperature. One potential mode of fuel system failure during RIA is pellet cladding mechanical interaction (PCMI) due to the rapid thermal expansion of the fuel pellet. Simulated PCMI experiments were performed by rapidly pressurizing iron-chromium-aluminum (FeCrAl) cladding tube samples using a hydraulic modified burst test system, causing the specimens to burst under a biaxial loading path. Deformation and rupture of the specimens were tracked with a telecentric lens and high-speed camera system. Outer surface strains were calculated using digital image correlation (DIC) on speckle patterns painted on the specimens’ outer surfaces. Experiments were conducted at approximately 275 °C, representative of hot zero power RIA conditions. The failure hoop strain was DIC-calculated between approximately 1.8–3.4%, corresponding to quasi-static energy depositions of approximately 110–260 calories per gram UO 2 assuming initial pellet-cladding contact. These conditions are very similar to the proposed energy deposition limit of 150 calories per gram UO 2 for unirradiated zirconium-based cladding in US Nuclear Regulatory Commission (NRC) Draft Regulatory Guide 1327. A very small strain rate dependence was observed in the data, with the magnitude of the failure hoop strain decreasing slightly with increasing strain rate. This observed dependence may be relevant because the FeCrAl cladding strain rate will be approximately 20% higher than in zirconium-based cladding due to the harder neutron spectrum and resultant shorter neutron generation time.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗
DOE OSTI2025

Distinct Dynamics of Lithium Intercalation and Plating on Graphite Anode for Li‐Ion Batteries in eVTOL Applications

In the absence of viable high-energy-density battery alternatives, lithium-ion (Li-ion) batteries remain essential for enabling electric vertical take-off and landing (eVTOL) platforms in advanced air mobility. Unlike Li-ion batteries used in electric vehicles and portable electronics, eVTOL battery systems operate under distinct high-power demands, which necessitate an independent assessment of material degradation mechanisms. This study presents a case analysis of graphite anode evolution under high-power cycling conditions. The findings reveal lithium entrapment within graphite particles, potentially resulting from incomplete Li-ion de-intercalation during a high-rate discharge event that is characteristic of eVTOL take-off and landing. This phenomenon leads to a progressive reduction in graphite-specific capacity and, over time, promotes lithium metal plating on the anode. Notably, the Li-metal plating observed in this study differs from that associated with fast-charging conditions, as it is primarily governed by concentration polarization-induced overpotential in the latter case. In conclusion, these findings highlight the inherent challenges of utilizing graphite in high-power Li-ion battery applications and elucidate the unique degradation mechanisms that arise due to the sluggish reaction kinetics of Li-ion intercalation and de-intercalation within graphite.

Li plating↗
DOE OSTI2022

Large-scale frictionless jamming with power-law particle size distributions

Due to significant computational expense, discrete element method simulations of jammed packings of size-dispersed spheres with size ratios greater than 1:10 have remained elusive, limiting the correspondence between simulations and real-world granular materials with large size dispersity. Here, invoking a recently developed neighbor binning algorithm, we generate mechanically stable jammed packings of frictionless spheres with power-law size distributions containing up to nearly 4 000 000 particles with size ratios up to 1:100. By systematically varying the width and exponent of the underlying power laws, we analyze the role of particle size distributions on the structure of jammed packings. The densest packings are obtained for size distributions that balance the relative abundance of large-large and small-small particle contacts. Although the proportion of rattler particles and mean coordination number strongly depend on the size distribution, the mean coordination of nonrattler particles attains the frictionless isostatic value of six in all cases. The size distribution of nonrattler particles that participate in the load-bearing network exhibits no dependence on the width of the total particle size distribution beyond a critical particle size for low-magnitude exponent power laws. This signifies that only particles with sizes greater than the critical particle size contribute to the mechanical stability. However, for high-magnitude exponent power laws, all particle sizes participate in the mechanical stability of the packing.

72 PHYSICS OF ELEMENTARY PARTICLES AND FIELDS↗
DOE OSTI2025

Understanding the structural mechanics of ligated DNA crystals via molecular dynamics simulation

DNA self-assembly is a highly programmable method to construct arbitrary architectures based on sequence complementarity. Among various constructs, DNA crystals are macroscopic crystalline materials formed by assembling motifs via sticky end association. Due to their high structural integrity and size ranging from tens to hundreds of micrometers, DNA crystals offer unique opportunities to study the structural properties and deformation behaviors of DNA assemblies. For example, enzymatic ligation of sticky ends can selectively seal nicks resulting in more robust structures with enhanced mechanical properties. However, the research efforts have been mostly on experiments involving different motif designs, structural optimization, or new synthesis methods, while their mechanics are not yet fully understood. The complex properties of DNA crystals are difficult to study via experiments alone, and numerical simulation can complement and aid the experiments. The coarse-grained molecular dynamics (MD) simulation is a powerful tool that can probe the mechanics of DNA assemblies. Here, we investigate DNA crystals made of four different motif lengths with various ligation patterns (full ligation, major directions, connectors, and in-plane) using oxDNA, an open-source, coarse-grained MD platform. We found that several distinct deformation stages emerge in response to mechanical loading and that the number and the location of ligated nucleotides can significantly modulate structural behaviors. These findings should be useful for predicting crystal properties and thus improving the design.

DNA crystal↗
DOE OSTI2020

Elucidating nanoscale mechanical properties of diabetic human adipose tissue using atomic force microscopy

Abstract Obesity-related type 2 diabetes (DM) is a major public health concern. Adipose tissue metabolic dysfunction, including fibrosis, plays a central role in DM pathogenesis. Obesity is associated with changes in adipose tissue extracellular matrix (ECM), but the impact of these changes on adipose tissue mechanics and their role in metabolic disease is poorly defined. This study utilized atomic force microscopy (AFM) to quantify difference in elasticity between human DM and non-diabetic (NDM) visceral adipose tissue. The mean elastic modulus of DM adipose tissue was twice that of NDM adipose tissue (11.50 kPa vs. 4.48 kPa) to a 95% confidence level, with significant variability in elasticity of DM compared to NDM adipose tissue. Histologic and chemical measures of fibrosis revealed increased hydroxyproline content in DM adipose tissue, but no difference in Sirius Red staining between DM and NDM tissues. These findings support the hypothesis that fibrosis, evidenced by increased elastic modulus, is enhanced in DM adipose tissue, and suggest that measures of tissue mechanics may better resolve disease-specific differences in adipose tissue fibrosis compared with histologic measures. These data demonstrate the power of AFM nanoindentation to probe tissue mechanics, and delineate the impact of metabolic disease on the mechanical properties of adipose tissue.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

High Performance SOFCs with a Superior Stability for Reliable and Durable Power Systems

Next generation of fuel cells, electrolyzers, and batteries requires higher power, faster kinetics, and larger energy density, which necessitate the use of compositionally complex oxides to achieve multifunctionalities and activity. These compositionally complex oxides may change their phases and structures during an electrochemical process – a so-called “electrochemically driven phase transformation”. The origin for such a phase change has remained obscure. More importantly, there is a need to develop high performance solid oxide fuel cells with an enhanced stability. In this work, the La 0.6 Sr 0.4 Co 0.2 Fe 0.8 O 3 (LSCF) cathode surface is modified by infiltration of Pr 6 O 11 and the power density at 0.8V and 750 °C is improved by 21%. In addition, by replacing the traditional barrier layer Gd 0.2 Ce 0.8 O 1.9 with mixed conducting Pr 0.1 Gd 0.1 Ce 0.8 O 1.9 , the power density increases by 38%. The different mechanism of promotions was investigated by electrochemical impedance spectroscopy. The ohmic resistance is dramatically reduced by applying the PGCO interlayer, and the distribution of relaxation time was used to analyze the mechanism for which the polarization resistance was decreased attributing to the mixed conduction nature in PrO x . An increase of power density at 0.8 V of 0.358 W/cm 2 (71%) is achieved with the implementation of both surface modification and buffer layer engineering. An experimental study and a theoretical analysis were then carried out on phase evolution in praseodymium nickelates. Nickelate-based electrodes show up to 60× greater phase transformation during operation when compared to thermally annealed ones. Theoretical analysis suggests that the presence of a reduced oxygen partial pressure at the interface between the oxygen electrode and the electrolyte is the origin for the phase change in an oxygen electrode. Guided by the theory, an addition of the electronic conduction in the interface layer leads to the significant suppression of phase change, while improving cell performance and performance stability. When an oxygen electrode is under polarization, the oxygen partial pressure at the interface between the oxygen electrode and the electrolyte is lower than that of incoming oxidant. Under a high polarization, the environment at the aforementioned interface may lead to phase transformation of the oxygen electrode. The local oxygen partial pressure is determined by the transport properties at the interfaces. An addition of the electronic conduction in the interface layer, for instance using (Pr,Gd)-doped ceria to replace Gd-doped ceria, results in improved cell performance and performance stability, while the phase transformation is significantly suppressed. This work provides a fundamental understanding of the origin for phase transformation in oxygen electrodes during operation and use this knowledge to develop a high-performance electrode that exhibits improved performance stability.

30 DIRECT ENERGY CONVERSION↗
DOE OSTI2021

Waste heat recovery lube oil management

A waste heat recovery system comprising a thermal circuit. The thermal circuit includes a boiler and an expander fluidly coupled to the boiler. The thermal circuit further includes a power transfer system integrated to the expander. The power transfer system is configured to receive mechanical energy from the expander. The thermal circuit further includes an ejector fluidly coupled to the boiler and to the power transfer system. The ejector is configured to receive a motive flow of working fluid from the boiler. The ejector is further configured to receive a suction flow of working fluid from the power transfer system. The ejector is further configured to combine the motive flow of working fluid and the suction flow of working fluid.

Ernst, Timothy C.↗
DOE OSTI2020

Universal finite-time thermodynamics of many-body quantum machines from Kibble-Zurek scaling

We demonstrate the existence of universal features in the finite-time thermodynamics of quantum machines by considering a many-body quantum Otto cycle in which the working medium is driven across quantum critical points during the unitary strokes. Specifically, we consider a quantum engine powered by dissipative energizing and relaxing baths. We show that under very generic conditions, the output work is governed by the Kibble-Zurek mechanism; i.e., it exhibits a universal power-law scaling with the driving speed through the critical points. We also optimize the finite-time thermodynamics as a function of the driving speed. The maximum power and the corresponding efficiency take a universal form, and are reached for an optimal speed that is governed by the critical exponents. We exemplify our results by considering a transverse-field Ising spin chain as the working medium. For this model, we also show how the efficiency and power vary as the engine becomes critical.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗
DOE OSTI2023

Maritime Fuel Cell Generator Project: 2018 – 2023

This report summarizes activity in the Maritime Fuel Cell (MarFC) Generator Project from 2018 – 2023. FY 2018 saw the implementation of upgrades and repairs, making the unit more reliable and operator friendly. In FY2019 the team engaged the Scripps Institution of Oceanography (SIO) to use the MarFC to provide zero-emission shore power to the research vessel R/V Robert Gordon Sproul while in port at the Nimitz Marine Facility in San Diego, CA. In FY2020, the MarFC unit was shipped to San Diego, CA. A fueling contract with IGX was established to support MarFC operations at SIO, with renewable hydrogen provided by the California State University Los Angeles (CSULA) hydrogen station. The project team (Sandia, Cummins/Hydrogenics) provided training to SIO staff on the technical details of the MarFC, the safe use of hydrogen in general and the MarFC in particular. The first fueling of the unit at the SIO pier was successfully completed by IGX. The first powering of a vessel with fuel-cell shore power was conducted with the R/V Robert Gordon Sproul. While the mechanical systems (lights, AC, ventilation, hydraulics, pumps and cranes) were powered without incident, problems arose when powering the computer systems. Inspections carried out in FY2021 revealed the MarFC needed routine maintenance. Maintenance was performed and the unit was upgraded. The MarFC was turned on after the year pause, and initial test data on power levels and stability were collected. FY2022 was a year spent repairing, upgrading and testing the MarFC unit. Spikes in power and voltage were observed above 60 kW that could potentially extend below 60 kW with time. Such spikes could cause problems with the Sproul electrical systems. These age-related problems, the extended time for the Sproul spent in dry dock for scheduled upgrades, and the associated need to reschedule the vessel’s high-priority science missions made it no longer possible to deploy the unit at SIO. After due consideration, the decision was made by DOE, MARAD and the project team to cease the deployment, remove the MarFC from SIO, and not pursue further deployment activities. On December 2, 2022, the MarFC unit was removed from the Scripps Nimitz Marine Facility and shipped to Fridley, Michigan. The Cummins/Hydrogenics plan for the unit is to assess the condition of the MarFC subcomponents, and then use it as a training/learning system for technical employees new to hydrogen fuel-cell technology. After summarizing project activity from 2018 – 2023, this report provides a review of lessons learned. This report provides next steps in contemplating a follow-on project that would further advance the use of fuel-cell-based shore power in a marine setting. A comparison is made of the project results to the original objectives. This report ends with an accounting of presentations stemming from the project, and a list of references.

08 HYDROGEN↗
DOE OSTI2023

Characterization of mApple as a Red Fluorescent Protein for Cryogenic Single-Molecule Imaging with Turn-Off and Turn-On Active Control Mechanisms

Single-molecule superresolution microscopy is a powerful tool for the study of biological structures on size scales smaller than the optical diffraction limit. Imaging samples at cryogenic temperatures (77 K) reduces the quantum yield of photobleaching for many fluorescent labels, yielding localization precisions below 10 nm. Cryogenic imaging further enables correlation with cryogenic electron tomography. A key limitation in applying methods such as PALM and STORM to samples maintained at 77 K is the limited number of fluorophores known to undergo efficient turn-on and turn-off mechanisms necessary to control the sparsity of active emitters. Here we find that mApple, a red-emitting fluorescent protein, undergoes a novel turn-off mechanism in response to simultaneous illumination with two colors of light. This turn-off mechanism enables localization of many individual molecules in initially bright samples, but the final density of localizable emitters is limited by relatively inefficient turn-on (photoactivation). Bulk excitation and emission spectroscopy shows that mApple has access to two distinct emissive states as well as dark states accessible optically or through changes in pH. The bright and stable emission of mApple enables widefield collection of single-molecule emission spectra, which highlight the complex nature and environmental sensitivity of states observed in red fluorescent proteins.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗