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At least 1,081 records · Page 60

Data Analyst - A Device for Continuous Repeating Real-Time Assay of HPGe, CZT, or Scintillation Gamma Spectra - 18574

There are many situations where a continuous repeating series of quantitative gamma spectral assays is desirable. These include measurements of fluid going through a pipe, of items on a conveyor passing under a detector, a detector aimed at the ground on a moving platform, or a detector in free air as a plume of radioactivity drifts by. While Canberra and many others have built and deployed many of these devices, they have all been custom built for that particular application, and rather expensive. The Data Analyst [DA] is a general purpose device that is designed to fulfill many of these applications without modification, but can be fairly-easily modified if needed. The DA is a small [12 x 18 x 5 cm], low-power [3 W] box that accepts the input from external MCAs, controls the acquisition cycle, analyzes the results, and transmits them to the outside world when needed. The detector-MCAs supported are HPGe with Lynx MCA, NaI or CeBr or LaBr with Osprey MCA, and CZT or CsI with internal MCAs. The DA internal software used for analysis is the full standard Genie analysis software, which is very widely used and well accepted in the industry. This allows exported spectral files to be reviewed and confirmed by Experts, and reanalyzed if needed. The DA hardware is totally autonomous. As soon as power is applied, it starts the pre-programmed sequences of acquisition and analysis. Multiple analysis sequences [called Workflows] can be in operation simultaneously, e.g. with different count times and different nuclide libraries. Only the detector with MCA, and the DA are required in the field. A PC can be connected to the DA locally or remotely via WiFi, USB, or Ethernet. The PC is needed to setup the DA, and to readout the results. The DA also supports [with application-specific programming] external Inputs/Outputs either for recording with the analysis results or for use in the computation of the result, e.g. temperature, flow rate, pressure, ... ]. The DA also has an internal GPS receiver with external antenna. Applications of the DA that are discussed include stack gas effluent activity where a portion of the effluent stream was extracted to flow through a shield with a HPGe detector and Lynx MCA, and measurement of reactor coolant activity where a shielded CZT detector and MCA were aimed at various sections of reactor primary coolant during a maintenance outage. Potential other applications are also described. (author)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Large Volume Airborne Contamination Monitoring To Support Nuclear Processes' Deactivation and Decommissioning

Current D and D operations at Hanford have demonstrated a flaw in the current state of the art capability of defining airborne contamination boundaries - Airborne particulate emissions of Pu-239 from CM2H operations on the Hanford Pu Finishing Plant were detected well beyond areas controlled for airborne Pu, putting numerous workers at risk for radiological assimilations - Current air samplers and CAM systems were surveying insufficient volumes of air to accurately predict where respiratory protection was required - Hanford is located in a high radon environment, making air monitoring for alpha emitting actinides challenging in high alpha radon induced backgrounds. - Inexpensive HEPA based home and industrial air purifiers filter significantly higher volumes of air than commercially available continuous-air monitoring (CAM) systems. - Inexpensive models capable of filtered air volumes exceeding 1000 times that of a CAM. - Detector system can be built to detect x-rays generated from actinide decay. - Branching ratios of x-rays are 4 orders of magnitude more intense from Plutonium decay than its gamma emissions. - Detector system can be a low resolution systems, as x-ray region of interest is not terribly congested. - Detection systems evaluated will be designed based on slabs of NaI or pixelated NaI or CsI panels. - Simple graphical user interface software will be developed to operate the detection system. - Concept is to deploy some of these air-purifying systems in nonradiological areas around SRNL to confirm radon can be rejected with confidence. - Generate some filters contaminated with plutonium via electroplated Pu or lab generated simulated particles. - then deploy units in known airborne radiological environments to establish systems ability to accurately measure actinide based hot particles Lab analyses will follow up the system analyses to ensure hot particles were correctly identified. Analysis by Scintillation: - Application of scintillation media to the surface of the filters is being explored. - Evaluating slabs of ZnS(Ag), application of powdered ZnS(Cu) and spray on Perkin Elmer Enhance. - Scintillation events would then be digitized with a digital camera and quantified. - Comparing against sensitivity of a PMT or SiPD readout. - Currently evaluating a Thorlabs 8 Megapixel Monochrome Scientific CCD Camera, hermetically sealed cooled package with a wide angle lens. - Wide angle lens allows complete view of HEPA filter from 7 inches away. - Images taken of glow in the dark paint, as well as plutonium-induced fluorescence. - Plutonium was flamed mounted on a 1 inch diameter stainless steel planchet, covered with a layer of mylar and a section of scintillating ZnS obtained from Eljon. - Currently working on reducing signal-to-noise levels to boost sensitivity. - Dark box to hold contaminated or electroplated source covered filters under fabrication. Analysis by x-ray Spectroscopy: - Branching ratios for x-ray emissions from Plutonium isotopes are orders of magnitude more intense than gamma-ray emissions. - The nuclear databases are incomplete on the branching ratios of some of the isotopes. - Evaluating some custom built SrI2 x-ray spectrometers vs a conventional windowed NaI detector. - SrI2 detectors are carbon-composite-windowed 51 mm x 51 mm. - MCNP calculations to establish self absorbance of filter media on 17 keV x-ray. Radon Rejection: - Activated charcoal pre-filter acts as a Radon trap. - Adding a time lapse feature to camera to aid in radon rejection. SRS Plutonium Fuel Form D and D Operations: Currently deploying air sampler to D and D operation of Pu-238 facility at SRS to generate some field samples to analyze.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Reconstructing Solar Particle Event Spectra from Absorbed Dose Measurements

The space radiation environment is unlike anything encountered on Earth. It is dominated by heavy charged particles consisting of highly charged nuclei with atomic numbers ranging from 1 to 94 with a broad spectrum of kinetic energies. The three primary sources of radiation in space are trapped belts, galactic cosmic rays (GCRs), and solar particle events (SPEs). The trapped belts are only of concern near bodies with significant magnetic fields and GCR flux spectra are relatively well-predicted using models that account for solar modulation of the local interstellar spectrum. SPEs, which consist primarily of protons, are difficult to predict in terms of occurrence, duration, intensity, and energy spectrum. SPE intensity and energy spectrum are critical for understanding radiation exposure-related health effects for astronauts in space. The National Aeronautics and Space Administration (NASA) relies on data from the Geostationary Operational Environmental Satellite series to characterize intensities and energy spectra of SPEs near Earth. However, these data are not applicable for spacecraft in other regions of the solar system. To understand the radiation environment for astronauts embarking on travel beyond Earth orbit, NASA is developing the Hybrid Electronic Radiation Assessor (HERA), a distributed dosimetry system that employs a solid-state silicon sensor coupled with the Timepix readout chip. The pixelated nature of the detector permits detailed analyses of charged particle tracks which can then be used to characterize the fluence spectrum at the location of the detector. Despite the unprecedented level of detail regarding the radiation environment provided by these detectors, knowledge of the fluence spectrum at all locations within the vehicle, specifically inside of astronauts, is desired to fully characterize the associated risk of harmful health effects. Radiation transport of the external radiation environment, considering the shielding provided by the vehicle, other objects, and even astronauts, is performed to obtain this information. Thus, there is clearly a need to define the external radiation environment using measurements performed within the vehicle. The number of detectors that may be flown as a part of HERA is limited by logistics, including data, power, and mass restrictions. The current configuration for crewed flight consists of six detectors at various locations with in the Orion Multi-Purpose Crew Vehicle (MPCV). It follows that an external proton fluence spectrum can be characterized with at most six energy bins with traditional matrix inversion techniques. The purpose of the present study is to determine whether it is possible to reconstruct SPE spectra with greater resolution from absorbed dose measurements and knowledge of the response of individual detectors using linear programming.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Tunable 3D Nanoresonators for Gas-Sensing Applications

Here, the detection of gas species with high sensitivity is a significant task for fundamental sciences as well as for industrial applications. Similarly, the ongoing trend for device miniaturization brings new challenges for advanced fabrication including on–demand functionality tuning. Following this motivation, here the additive, direct–write fabrication of freestanding 3D nanoarchitectures is introduced, which can be brought into mechanical resonance via electric AC fields. Specifically, this study focuses on the 3D nanostructure synthesis, the subsequent determination of Young's modulus, and demonstrates a postgrowth procedure, which can precisely tune the material modulus. As–fabricated resonators reveal a Young's modulus of 9–13 GPa, which can be increased by a factor greater than 5. Next, the electric readout of the resonance behavior is demonstrated via electric current measurement as an essential element for the resonance sensor applications. Finally, the implications of gas–physisorption and gas–chemisorption on the resonance frequencies are studied, representing a proof–of–principle for sensing applications by the here presented approach.

47 OTHER INSTRUMENTATION↗

Chemiresistors for the Real‐Time Wireless Detection of Anions

Abstract Reported is an electrical transduction platform for real‐time wireless anion sensing using single‐walled carbon nanotubes (SWCNTs) noncovalently functionalized with squaramide‐based anion binding selectors. Systematically studied are anion‐binding properties and efficiency of the electrical transduction of the functionalized SWCNT composites using the squaramide‐based selectors with two similar electron‐withdrawing groups, 3,5‐bis(trifluoromethyl)benzyl ( 1 ) and 3,5‐bis(trifluoromethyl)phenyl ( 2 ), which induce hydrogen‐bonding interaction with anions and deprotonation of a squaramide N–H proton upon addition of acetate (AcO − ), respectively. Charge transduction occurs with AcO − as a result of charge transfer from the deprotonated selector 2 , whereas less sensitive transduction is observed with selector 1 via hydrogen‐bonding interaction. These results provide guidelines to efficiently transduce the chemical interaction between selectors and anions to create resistive transduction with functionalized SWCNTs. Electron‐withdrawing groups adjacent to the squaramide as well as proximate cationic pyridyl groups, enhance the anion binding affinity and also lower the selector's p K a . The chemiresistive sensor arrays are readily integrated with a wireless sensing module and demonstrated real‐time sensing of multiple anions with a smartphone readout.

Choi, Seon‐Jin↗

Emergent Ferroelectric Switching Behavior from Polar Vortex Lattice

We report topologically protected polar textures have provided a rich playground for the exploration of novel, emergent phenomena. Recent discoveries indicate that ferroelectric vortices and skyrmions not only host properties markedly different from traditional ferroelectrics, but also that these properties can be harnessed for unique memory devices. Using a combination of capacitor-based capacitance measurements and computational models, it is demonstrated that polar vortices in dielectric–ferroelectric–dielectric trilayers exhibit classical ferroelectric bi-stability together with the existence of low-field metastable polarization states. This behavior is directly tied to the in-plane vortex ordering, and it is shown that it can be used as a new method of non-destructive readout-out of the poled state.

36 MATERIALS SCIENCE↗

Automated Label‐Free Assay for Viral Detection and Inhibitor Screening via Biomembrane‐Functionalized Microelectrode Arrays

Most virus infection assays have indirect readout such as virus number following entry (e.g., PCR, cell lysis). While effective, these technologies are labor‐intensive, require specialized environments (e.g., sterile or RNA‐free), and detect later‐stage viral events like lysis or cell death, lacking sensitivity to early fusion events. To address these limitations, we present biologically relevant 2D membrane materials, host‐cell‐derived supported lipid bilayers (hcd‐SLBs), integrated with organic microelectrode arrays (OMEAs) for detection of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) fusion. By overexpressing angiotensin‐converting enzyme 2 (ACE2) receptors on the native membranes, the platform functions as a viral sensor capable of detecting virus pseudo particles (VPPs) through the late pathway. Additionally, hcd‐SLBs extracted from human lung epithelium expressing native ACE2 detect fusion events through the early pathway. The platform's utility as a drug‐screening tool is demonstrated by testing antibodies targeting either the ACE2 on the host membrane or the viral spike (S) proteins. To enhance the throughput, microfluidics are integrated for automation and OMEAs are incorporated within each channel, miniaturizing the testing units. This system supports high‐throughput data generation, automation, and scalability, providing an efficient platform for viral fusion detection that advances the study of pathogen‐host interactions and accelerates antiviral drug discovery.

Biology↗

Nanoscale Compositional and Strain Gradients Enable High‐Speed and Amplitude‐Resolved Pyroelectric Sensing

The frequency response of pyroelectric sensors is fundamentally governed by thermal time constant (τth, determined by thermal mass and thermal conductance) and electrical impedance arising from film capacitance and readout circuit. Conventional bulk LiTaO3 detectors are optimized for high responsivity at low modulation frequencies (0.1-10 Hz), possessing a large τth that thermally averages rapid temperature oscillations at elevated modulation frequencies, limiting fidelity in resolving dynamic varying thermal signals. Here, compositional and strain gradients are introduced into 100-nm-thick relaxor-ferroelectric films reducing τth to ≈2 µs and producing built-in potentials (≈1.45 V or 145 kV cm-1) that enhance the pyroelectric coefficient and suppress the dielectric constant. This enables complementary dual-mode operation by enhancing current-mode electrical responsivity and improving the voltage-mode figure of merit - advantageous for superior temperature resolution (ΔTmin ≈ 30 µK). The responsivity peak shifts to near 1 kHz (>2500-times higher than conventional bulk sensors), with measurable responsivity extending to a carrier frequency of 100 kHz and amplitude-resolved detection at modulation frequencies up to 15 kHz. These results establish nanoscale internal-field engineering can reshape electro-thermal trade-off in pyroelectric thin films toward zero-bias, high-thermal-sensitivity, and amplitude-resolved thermal sensing across a wide frequency bandwidth.

Lin, Ching‐Che↗

Molecularly Imprinted Polymer Sensor Empowered by Bound States in the Continuum for Selective Trace-Detection of TGF-beta

The integration of advanced materials and photonic nanostructures can lead to enhanced biodetection capabilities, crucial in clinical scenarios and point-of-care diagnostics, where simplified strategies are essential. Herein, a molecularly imprinted polymer (MIP) photonic nanostructure is demonstrated, which selectively binding to transforming growth factor-beta (TGF-β), in which the sensing transduction is enhanced by bound states in the continuum (BICs). The MIP operating as a synthetic antibody matrix and coupled with BIC resonance, enhances the optical response to TGF-β at imprinted sites, leading to an augmented detection capability, thoroughly evaluated through spectral shift and optical lever analogue readout. The validation underscores the MIP-BIC sensor capability to detect TGF-β in spiked saliva, achieving a limit of detection of 10 fM and a resolution of 0.5 pM at physiological concentrations, with a precision of two orders of magnitude above discrimination threshold in patients. The MIP tailored selectivity is highlighted by an imprinting factor of 52, showcasing the sensor resistance to interference from other analytes. The MIP-BIC sensor architecture streamlines the detection process eliminating the need for complex sandwich immunoassays and demonstrates the potential for high-precision quantification. This positions the system as a robust tool for biomarker detection, especially in real-world diagnostic scenarios.

77 NANOSCIENCE AND NANOTECHNOLOGY↗

Electrocatalytic Effects of Au Nanoclusters in its Electrochemiluminescence Enhanced by Coreactant Tertiary Amines

Abstract Atomically precise metal nanoclusters are increasingly exploited in electrocatalysis such as water electrolysis and CO 2 reduction. The catalytic effects are generally evaluated by conventional electrochemical methods or end‐product analysis. Herein, near infrared electrochemiluminescence (ECL) of the nanoclusters is introduced as a new signaling mechanism to complement classical voltammetric analysis for elucidating essential electrocatalytic parameters. Coreactant ECL from an aqueous soluble Au 22 nanocluster (NCs) enhanced by a common pH buffer materials 4‐(2‐hydroxyethyl)‐1‐piperazineethanesulfonic acid (HEPES) and by a piperazine drug hydroxyzine (HDZ) is studied under oxidative reduction pathways. Foot of the wave analysis on voltametric features and the kinetics profiles in potential‐step experiments are studied in different nanocluster and coreactant concentrations. Benchmark parameters such as rate constants are determined under EC (Electron‐transfer‐Chemical‐reaction) mechanism as feasibility validations. Because the luminescence properties of AuNCs are sensitive to the surface ligands or adsorbents, ECL is proposed as a new signal readout for operando studies providing insights for governing reaction mechanism and kinetics during catalytic reactions. Retrospectively, mechanistic understanding of the complex multi‐step reactions during ECL generation can provide guidance for the evaluation and optimization of ECL property for better quantitative applications.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Room‐Temperature Mid‐Infrared Detection Using Metasurface‐Absorber‐Integrated Phononic Crystal Oscillator

Mid-infrared (MIR) detectors find extensive applications in chemical sensing, spectroscopy, communications, biomedical diagnosis, and space exploration. Alternative to semiconductor MIR photodiodes and bolometers, mechanical-resonator-based MIR detectors show advantages in higher sensitivity and lower noise at room temperature, especially toward longer wavelength infrared. Here, uncooled room-temperature MIR detectors based on lithium niobate surface acoustic wave phononic crystal (PnC) resonators integrated with wavelength-and-polarization-selective metasurface absorber arrays are demonstrated. The detection is based on the resonant frequency shift induced by the local temperature change due to MIR absorptions. The PnC resonator is configured in an oscillating mode, enabling active readout and low-frequency noise. The 1-GHz oscillator-based MIR detector shows a relative frequency deviation of 5.24 × 10 −10 Hz −1/2 at an integration time of 50 µs, leading to an incident noise equivalent power of 197 pW Hz −1/2 when input 6-µm MIR light is modulated at 1.8 kHz, and a large dynamic range of 10 7 in incident MIR power. The device architecture is compatible with the scalable manufacturing process and can be readily extended to a broader spectral range by tailoring the absorbing wavelengths of metasurface absorbers.

Xi, Zichen [Virginia Polytechnic Inst. and State U↗

Improved reconstruction stability for chemical shift encoded hyperpolarized 13 C magnetic resonance spectroscopic imaging using k‐t spiral acquisitions

Purpose A multiecho, field of view (FOV)‐oversampled k‐t spiral acquisition and direct iterative decomposition of water and fat with echo asymmetry and least‐squares estimation reconstruction is demonstrated to improve the stability of hyperpolarized 13 C magnetic resonance spectroscopic imaging (MRSI) in the presence of signal ambiguities attributed to low‐SNR (signal‐to‐noise‐ratio) species, local uncertainties in metabolite peaks, and echo‐to‐echo signal inconsistencies. Theory k‐t spiral acquisitions redistribute readout points to be more densely spaced radially in k‐space by acquiring an FOV and matrix that are oversampled by η . These more densely spaced spiral turns constitute effective intraspiral echoes and can supplement conventional interspiral echoes to improve spectral separation and reduce spectral cross‐talk to better resolve 13 C‐labeled species for spectroscopic imaging. Methods Digital simulations and imaging phantom experiments were performed for a range of interspiral echo spacings and η using multiecho, k‐t spiral acquisitions. Image spectral cross‐talk artifacts were evaluated both qualitatively and quantitatively as the percent error in measured metabolite ratios. In vivo murine experiments evaluated the feasibility of multiecho, k‐t spiral [1‐ 13 C]pyruvate MRSI to reduce spectral cross‐talk for 3 scenarios of different expected reconstruction stability. Results Digital simulations and imaging phantom experiments both demonstrated reduced or comparable image spectral cross‐talk and percent errors in measured metabolite ratios with increasing η and better choices of echo spacings. In vivo images displayed markedly reduced spectral cross‐talk in lactate images acquired with η = 7 versus η = 1. Conclusion The precision of hyperpolarized 13 C metabolic imaging and quantification in the presence of low‐SNR species, local uncertainties in metabolite resonances, and echo‐to‐echo signal inconsistencies can be improved with the use of FOV‐oversampled k‐t spiral acquisitions.

Macdonald, Erin B.↗

Single‐Cell Nanodroplet Processing Proteomics Pipeline for Analysis of Human‐Derived Microglia

Single-cell omics tools provide unique insights into heterogeneous cell populations and their responses to stimuli. For example, single-cell RNA sequencing has identified several transcriptionally distinct populations of microglia, which are resident immune cells of the central nervous system (CNS) that are responsive to CNS injury, infection, and neurodegeneration. To date, single-cell studies of microglia have focused on RNA-sequencing or cytometry by time of flight (CyTOF), which provide indirect readouts of protein abundance or quantification of a limited number of targets. Herein, we present a workflow based on FACS-assisted isolation, cryopreservation, and nanodroplet-based processing for single-cell mass spectrometry proteomics analysis of the postmortem human brain cortex-derived microglia. From a single microglial cell, 1039 proteins could be identified on average. As a proof-of-principle, we applied single-cell proteomics for exploring the heterogeneity of brain microglia at the cellular level. This pilot proteomics data partially recapitulates the prior microglia subtypes. Specifically, we determined that mitochondrial proteins, in particular members of NADH dehydrogenase (Complex I), cytochrome b-c1 (Complex III), cytochrome c oxidase (Complex IV), F1-ATPase (Complex V), and Na+/K+-ATPase complex, drive variation across microglia. This pipeline offers the potential for identifying functionally and analytically relevant protein targets for microglia in Alzheimer's disease and other neurological disorders.

59 BASIC BIOLOGICAL SCIENCES↗

Engineering highly stable variants of Corynactis californica green fluorescent proteins

Fluorescent proteins (FPs) are versatile biomarkers that facilitate effective detection and tracking of macromolecules of interest in real time. Engineered FPs such as superfolder green fluorescent protein (sfGFP) and superfolder Cherry (sfCherry) have exceptional refolding capability capable of delivering fluorescent readout in harsh environments where most proteins lose their native functions. Our recent work on the development of a split FP from a species of strawberry anemone, Corynactis californica, delivered pairs of fragments with up to threefold faster complementation than split GFP. We present the biophysical, biochemical, and structural characteristics of five full-length variants derived from these split C. californica GFP (ccGFP). These ccGFP variants are more tolerant under chemical denaturation with up to 8 kcal/mol lower unfolding free energy than that of the sfGFP. It is likely that some of these ccGFP variants could be suitable as biomarkers under more adverse environments where sfGFP fails to survive. A structural analysis suggests explanations of the variations in stabilities among the ccGFP variants.

59 BASIC BIOLOGICAL SCIENCES↗

Anomalous Loss Reduction Below Two‐Level System Saturation in Aluminum Superconducting Resonators

Superconducting resonators are widely used in many applications such as qubit readout for quantum computing, and kinetic inductance detectors. These resonators are susceptible to numerous loss and noise mechanisms, especially the dissipation due to two-level systems (TLS) which become the dominant source of loss in the few-photon and low temperature regime. In this study, capacitively-coupled aluminum half-wavelength coplanar waveguide resonators are investigated. Surprisingly, the loss of the resonators is observed to decrease with a lowering temperature at low excitation powers and temperatures below the TLS saturation. This behavior is attributed to the reduction of the TLS resonant response bandwidth with decreasing temperature and power to below the detuning between the TLS and the resonant photon frequency in a discrete ensemble of TLS. When response bandwidths of TLS are smaller than their detunings from the resonance, the resonant response and thus the loss is reduced. At higher excitation powers, the loss follows a logarithmic power dependence, consistent with predictions from the generalized tunneling model (GTM). A model combining the discrete TLS ensemble with the GTM is proposed and matches the temperature and power dependence of the measured internal loss of the resonator with reasonable parameters.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗

Quantum Enhanced Probes of Magnetic Circular Dichroism

Magneto-optical microscopies, including optical measurements of magnetic circular dichroism, are increasingly ubiquitous tools for probing spin-orbit coupling, charge-carrier g-factors, and chiral excitations in matter, but the minimum detectable signal in classical magnetic circular dichroism measurements is fundamentally limited by the shot-noise limit of the optical readout field. Here, a two-mode squeezed light source is used to improve the minimum detectable signal in magnetic circular dichroism measurements by 3 decibels compared with state-of-the-art classical measurements, even with relatively lossy samples like terbium gallium garnet. Furthermore, these results provide a framework for new quantum-enhanced magneto-optical microscopies that are particularly critical for environmentally sensitive materials and for low temperature measurements where increased optical power can introduce unacceptable thermal perturbations.

42 ENGINEERING↗

Steric Modulation of Protein‐Mediated Nanoparticle Assembly: Controlling Cluster Size, Polydispersity, and FRET Responses by Rebalancing Short‐ and Long‐Range Interactions

Understanding and manipulating protein-nanoparticle interactions is of broad interest to fields ranging from nanomedicine to the biological fabrication of functional hierarchical materials. This study investigates how steric forces introduced by a pegylated derivative of superfolder green fluorescent protein (sfGFP) that is monofunctional for silica binding modulate the delicate interplay of long-range (electrostatic and van der Waals) and short-range (protein-mediated) interactions in pH-responsive silica nanoparticle (SiNP) assembly by bifunctional silica-binding sfGFP. Increasing the length of the PEG segment and pre-incubating SiNPs with increasing concentrations of pegylated proteins enables precise control over cluster size within the 800–1450 nm range with a sixfold decrease in polydispersity index to a remarkable 0.1 endpoint. Weakening short-range attractive interactions via mutagenesis extends this control to clusters in the 50–250 nm range and reveals that the Förster resonance energy transfer (FRET) efficiency of clusters scales linearly with cluster diameter below 230 nm but increases only by 15% as clusters grow to 1450 nm. Furthermore, these findings enable the development of a system that provides an optical readout to dynamic changes in solution conditions enacted by a combination of pH adjustment and ion charge screening.

77 NANOSCIENCE AND NANOTECHNOLOGY↗

Single-Cell Metabolomics with Rapid Determination of Chemical Formulas from Isotopic Fine Structures

Metabolomic measurements can provide functional readouts of cellular states and phenotypes. Here, we present a protocol for single-cell metabolomics that permits direct untargeted detection of a broad number of metabolites under ambient conditions, without the need for sample processing, and with high confidence in the discovery and identification of the molecular formulas for detected metabolites. This protocol describes combining fiber-based laser ablation electrospray ionization (f-LAESI) with a 21 Tesla Fourier transform ion cyclotron resonance mass spectrometer (21T-FTICR-MS) to obtain high confidence molecular formula information about detected metabolites. The f-LAESI source utilizes mid-infrared laser ablation through a sharp optical fiber tip, affording direct ambient analysis of cells without the need for sample processing. Using the 21T-FTICR-MS as a mass analyzer enabled measurement of the isotopic fine structure (IFS) for numerous metabolites simultaneously from single cells, and the IFSs were in turn computationally processed to rapidly determine the corresponding elemental compositions. This metabolomics technique complements other single cell omics measurement methods, helping to resolve complex molecular interactions that take place within cells unattainable from single cell transcriptomic and proteomics methods.

mass spectrometry, FTICR, FTMS, ultrahigh mass res↗