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At least 307 records · Page 17

Rapid discovery and evolution of nanosensors containing fluorogenic amino acids

Binding-activated optical sensors are powerful tools for imaging, diagnostics, and biomolecular sensing. However, biosensor discovery is slow and requires tedious steps in rational design, screening, and characterization. Here we report on a platform that streamlines biosensor discovery and unlocks directed nanosensor evolution through genetically encodable fluorogenic amino acids (FgAAs). Building on the classical knowledge-based semisynthetic approach, we engineer ~15 kDa nanosensors that recognize specific proteins, peptides, and small molecules with up to 100-fold fluorescence increases and subsecond kinetics, allowing real-time and wash-free target sensing and live-cell bioimaging. An optimized genetic code expansion chemistry with FgAAs further enables rapid (~3 h) ribosomal nanosensor discovery via the cell-free translation of hundreds of candidates in parallel and directed nanosensor evolution with improved variant-specific sensitivities (up to ~250-fold) for SARS-CoV-2 antigens. Altogether, this platform could accelerate the discovery of fluorogenic nanosensors and pave the way to modify proteins with other non-standard functionalities for diverse applications.

Biosensors↗

Pyrrole‐Imine Macrocycle: Self‐Organizing Cross‐Reactive Anion Receptor and Sensor

Self-organizing macrocyclic receptor-sensors for phosphorus oxyanions, phosphates, and phosphonates comprising imine moieties were prepared by condensation of dipyrrolylmethane dicarbaldehyde with diethylene triamine. The incorporation of flexible ethylene moieties endows the macrocycle with unprecedented flexibility and ability to accommodate numerous phosphorus oxyanions from orthophosphate to large anions such as ATP or phosphonate glyphosate. The anion binding was elucidated by NMR titrations, low-temperature NMR, and NOESY NMR. The incorporation of dansyl fluorophore enables sensing of anions using the fluorescence signal, whereas the changes in fluorescence intensity, width of the fluorescence band, and position of the maxima are analyte-specific and useful in recognition and identification of eleven different P-oxyanions in water. The affinity (K assoc ) for Na + salts was H 2 PO 4 − ≈ Methylphosphonate > H 2 P 2 O 7 2− > Phenylphosphonate- > Glyphosate 2− > AMP 2− > ADP 2− > ATP 2− . Interestingly, phosphonates, including methylphosphonate and glyphosate anions, were also found to display a strong affinity (K assoc ∼10 6 M −1 ) while halides, nitrate, carbonates, or hydrogen sulfate did not show a significant affinity. The determined fluorescence spectral parameters were used to classify the 12 analytes (11 anions and water) using Linear Discriminant Analysis (LDA). Quantification was performed using LDA and Support Vector Machine (SVM), and the phosphonate concentrations in unknown samples were determined with an error of 3.5% or lower.

anions↗

NASA Tech Briefs, April 2004

Topics covered include: Analysis of SSEM Sensor Data Using BEAM; Hairlike Percutaneous Photochemical Sensors; Video Guidance Sensors Using Remotely Activated Targets; Simulating Remote Sensing Systems; EHW Approach to Temperature Compensation of Electronics; Polymorphic Electronic Circuits; Micro-Tubular Fuel Cells; Whispering-Gallery-Mode Tunable Narrow-Band-Pass Filter; PVM Wrapper; Simulation of Hyperspectral Images; Algorithm for Controlling a Centrifugal Compressor; Hybrid Inflatable Pressure Vessel; Double-Acting, Locking Carabiners; Position Sensor Integral with a Linear Actuator; Improved Electromagnetic Brake; Flow Straightener for a Rotating-Drum Liquid Separator; Sensory-Feedback Exoskeletal Arm Controller; Active Suppression of Instabilities in Engine Combustors; Fabrication of Robust, Flat, Thinned, UV-Imaging CCDs; Chemical Thinning Process for Fabricating UV-Imaging CCDs; Pseudoslit Spectrometer; Waste-Heat-Driven Cooling Using Complex Compound Sorbents; Improved Refractometer for Measuring Temperatures of Drops; Semiconductor Lasers Containing Quantum Wells in Junctions; Phytoplankton-Fluorescence-Lifetime Vertical Profiler; Hexagonal Pixels and Indexing Scheme for Binary Images; Finding Minimum-Power Broadcast Trees for Wireless Networks; and Automation of Design Engineering Processes.

Source record↗

Compensation of Oxygen Transmittance Effects for Proximal Sensing Retrieval of Canopy-Leaving Sun-Induced Chlorophyll Fluorescence

Estimates of Sun-Induced vegetation chlorophyll Fluorescence (SIF) using remote sensing techniques are commonly determined by exploiting solar and/or telluric absorption features. When SIF is retrieved in the strong oxygen (O2) absorption features, atmospheric effects must always be compensated. Whereas correction of atmospheric effects is a standard airborne or satellite data processing step, there is no consensus regarding whether it is required for SIF proximal-sensing measurements nor what is the best strategy to be followed. Thus, by using simulated data, this work provides a comprehensive analysis about how atmospheric effects impact SIF estimations on proximal sensing, regarding: (1) the sensor height above the vegetated canopy; (2) the SIF retrieval technique used, e.g., Fraunhofer Line Discriminator (FLD) family or Spectral Fitting Methods (SFM); and (3) the instrument's spectral resolution. We demonstrate that for proximal-sensing scenarios compensating for atmospheric effects by simply introducing the O2 transmittance function into the FLD or SFM formulations improves SIF estimations. However, these simplistic corrections still lead to inaccurate SIF estimations due to the multiplication of spectrally convolved atmospheric transfer functions with absorption features. Consequently, a more rigorous oxygen compensation strategy is proposed and assessed by following a classic airborne atmospheric correction scheme adapted to proximal sensing. This approach allows compensating for the O2 absorption effects and, at the same time, convolving the high spectral resolution data according to the corresponding Instrumental Spectral Response Function (ISRF) through the use of an atmospheric radiative transfer model. Finally, due to the key role of O2 absorption on the evaluated proximal-sensing SIF retrieval strategies, its dependency on surface pressure (p) and air temperature (T) was also assessed. we combined simulated spectral data with p and T measurements obtained for a one-year period in the Hyytiala Forestry Field Station in Finland. Of importance hereby is that seasonal dynamics in terms of T and p, if not appropriately considered as part of the retrieval strategy, can result in erroneous SIF seasonal trends that mimic those of known dynamics for temperature-dependent physiological responses of vegetation.

"sun–induced chlorophyll fluorescence (SIF); pro↗

Noise Reduction for Solar-Induced Fluorescence Retrievals Using Machine Learning and Principal Component Analysis: Simulations and Applications to GOME-2 Satellite Retrievals

We use a spectral-based approach that employs principal component analysis along with a relatively shallow artificial neural network (NN) to substantially reduce noise and other artifacts in terrestrial chlorophyll solar-induced fluorescence (SIF) retrievals. SIF is a very small emission at red and far-red wavelengths that is difficult to measure and is highly sensitive to random errors and systematic artifacts. Our approach relies upon an assumption that a trained NN can effectively reconstruct the total SIF signal from a relatively small number of leading principal components of the satellite-observed far-red radiance spectra without using information from the trailing modes that contain most of the random errors. We test the approach with simulated reflectance spectra produced with a full atmospheric and surface radiative transfer model using different observing and geophysical parameters and various noise levels. Resulting noisy and noise-reduced retrieved SIF values are compared with true values to assess performance. We then apply our noise reduction approach to SIF derived from two different satellite spectrometers. For evaluation, since the truth in this case is unknown, we compare SIF retrievals from two independent sensors with each other. We also compare the noise-reduced SIF temporal variations with those from an independent gross primary product (GPP) product that should display similar variations. Results show that our noise reduction approach improves capture of SIF seasonal and inter-annual variability. Our approach should be applicable to many noisy data products derived from spectral measurements. Our methodology does not replace the original retrieval algorithms; rather the original noisy retrievals are needed as the target for the NN training process.

noise reduction, solar-induced fluorescence, SIF, ↗

Investigations of Arcjet Flow Thermochemistry

An experimental and numerical investigation of the thermochemical state of arcjet flows is currently being conducted at NASA Ames Research Center. The experimental approach relies on the use of laser- and emission-spectroscopic diagnostic techniques in three regions of the flow. A fiber optic sensor is used to record spectrally resolved emission signals from the electrode package region, where the flow is most likely to be in thermochemical equilibrium. A second emission diagnostic measurement is made in the shock layer formed over a blunt-body test article placed in the stream, and a CCD camera is used to simultaneously record spectral emission from several measurement locations along the stagnation streamline. Downstream of the nozzle exit, but upstream of the test article, Laser-Induced Fluorescence (LIF) of atomic nitrogen is used to assess the nonequilibrium distribution of flow enthalpy in the free stream. Results from the measurements are compared with predictions from a two-temperature, axisymmetric flow model that solves the nozzle and shock-layer flows.

Fletcher, Douglas G.↗

A theoretical/experimental program to develop active optical pollution sensors: Quantitative remote Raman lidar measurements of pollutants from stationary sources

Typical pollutant gas concentrations at the stack exits of stationary sources can be estimated to be about 500 ppm under the present emission standards. Raman lidar has a number of advantages which makes it a valuable tool for remote measurements of these stack emissions. Tests of the Langley Research Center Raman lidar at a calibration tank indicate that night measurements of SO2 concentrations and stack opacity are possible. Accuracies of 10 percent are shown to be achievable from a distance of 300 m within 30 min integration times for 500 ppm SO2 at the stack exits. All possible interferences were examined quantitatively (except for the fluorescence of aerosols in actual stack emissions) and found to have negligible effect on the measurements. An early test at an instrumented stack is strongly recommended.

Poultney, S. K.↗

Oceanographic applications of laser technology

Oceanographic activities with the Airborne Oceanographic Lidar (AOL) for the past several years have primarily been focussed on using active (laser induced pigment fluorescence) and concurrent passive ocean color spectra to improve existing ocean color algorithms for estimating primary production in the world's oceans. The most significant results were the development of a technique for selecting optimal passive wavelengths for recovering phytoplankton photopigment concentration and the application of this technique, termed active-passive correlation spectroscopy (APCS), to various forms of passive ocean color algorithms. Included in this activity is use of airborne laser and passive ocean color for development of advanced satellite ocean color sensors. Promising on-wavelength subsurface scattering layer measurements were recently obtained. A partial summary of these results are shown.

Hoge, F. E.↗

Electrochemical Switching of a Fluorescent Molecular Rotor Embedded within a Bistable Rotaxane

We report how the nanoconfined environment, introduced by the mechanical bonds within an electrochemically switchable bistable [2]rotaxane, controls the rotation of a fluorescent molecular rotor, namely, an 8-phenyl-substituted boron dipyrromethene (BODIPY). The electrochemical switching of the bistable [2]rotaxane induces changes in the ground-state coconformation and in the corresponding excited-state properties of the BODIPY rotor. In the starting redox state, when no external potential is applied, the cyclobis(paraquat-p-phenylene) (CBPQT4+) ring component encircles the tetrathiafulvalene (TTF) unit on the dumbbell component, leaving the BODIPY rotor unhindered and exhibiting low fluorescence. Upon oxidation of the TTF unit to a TTF2+ dication, the CBPQT4+ ring is forced toward the molecular rotor, leading to an increased energy barrier for the excited state to rotate the rotor into the state with a high nonradiative rate constant, resulting in an overall 3.4-fold fluorescence enhancement. On the other hand, when the solvent polarity is high enough to stabilize the excited charge-transfer state between the BODIPY rotor and the CBPQT4+ ring, movement of the ring toward the BODIPY rotor produces an unexpectedly strong fluorescence signal decrease as the result of photoinduced electron transfer from the BODIPY rotor to the CBPQT4+ ring. The nanoconfinement effect introduced by mechanical bonding can effectively lead to modulation of the physicochemical properties as observed in this bistable [2]rotaxane. On account of the straightforward synthetic strategy and the facile modulation of switchable electrochromic behavior, our approach could pave the way for the development of new stimuli-responsive materials based on mechanically interlocked molecules for future electro-optical applications, such as sensors, molecular memories, and molecular logic gates.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Membrane potential sensing: Material design and method development for single particle optical electrophysiology

We review the development of “single” nanoparticle-based inorganic and organic voltage sensors, which can eventually become a viable tool for “non-genetic optogenetics.” The voltage sensing is accomplished with optical imaging at the fast temporal response and high spatial resolutions in a large field of view. Inorganic voltage nanosensors utilize the Quantum Confined Stark Effect (QCSE) to sense local electric fields. Engineered nanoparticles achieve substantial single-particle voltage sensitivity (~2% Δλ spectral Stark shift up to ~30% ΔF/F per 160 mV) at room temperature due to enhanced charge separation. A dedicated home-built fluorescence microscope records spectrally resolved images to measure the QCSE induced spectral shift at the single-particle level. Biomaterial based surface ligands are designed and developed based on theoretical simulations. The hybrid nanobiomaterials satisfy anisotropic facet-selective coating, enabling effective compartmentalization beyond non-specific staining. Self-spiking- and patched-HEK293 cells and cortical neurons, when stained with hybrid nanobiomaterials, show clear photoluminescence intensity changes in response to membrane potential (MP) changes. Organic voltage nanosensors based on polystyrene beads and nanodisk technology utilize Fluorescence (Förster) Resonance Energy Transfer (FRET) to sense local electric fields. Voltage sensing FRET pairs achieve voltage sensitivity up to ~35% ΔF/F per 120 mV in cultures. Non-invasive MP recording from individual targeted sites (synapses and spines) with nanodisks has been realized. However, both of these QCSE- and FRET-based voltage nanosensors yet need to reach the milestone of recording individual action potentials from individual targeted sites.

59 BASIC BIOLOGICAL SCIENCES↗

Planetary exploration sensor systems

Most of the instruments that have been used in planetary exploration have been either spectrometers or imaging devices. Instruments of these types are being developed for the Galileo and VOIR (Venus Orbiting Imaging Radar) missions. Galileo will take advantage of new CCD (charge-coupled device) technology, and VOIR will use techniques of synthetic aperture radar developed for Seasat. For determining the macrostructure of mineral resources, the best approach is believed to involve acoustic imaging, essentially a seismic data processing technique. Determinations of microstructure would require a light microscope and an electron microscope. For determining the nature and form of volatiles, a differential scanning calorimeter could be used. To determine the mineral composition, an electron beam microprobe with X-ray fluorescence and spectroscopy would be used.

Saunders, R. S.↗

Planetary and satellite x ray spectroscopy: A new window on solid-body composition by remote sensing

The rings and most of the satellites of the outer planets orbit within the radiation belts of their parent bodies. This is an environment with intense fluxes of energetic electrons. As a result, these objects are strong emitters of X-rays. The characteristic X-ray lines from these bodies depend on atomic composition, but they are not sensitive to how the material is arranged in compounds or mixtures. X-ray fluorescence spectral analysis has demonstrated its unique value in the laboratory as a qualitative and quantitative analysis tool. This technique has yet to be fully exploited in a planetary instrument for remote sensing. The characteristic X-ray emissions provide atomic relative abundances. These results are complementary to the molecular composition information obtained from IR, visible, and UV emission spectra. The atomic relative abundances are crucial to understanding the formation and evolution of these bodies. They are also crucial to the proper interpretation of the molecular composition results from the other sensors. The intensities of the characteristic X-ray emissions are sufficiently strong to be measured with an instrument of modest size. Recent developments in X-ray detector technologies and electronic miniaturization have made possible space-flight X-ray imaging and nonimaging spectrometers of high sensitivity and excellent energy resolution that are rugged enough to survive long-duration space missions. Depending on the application, such instruments are capable of resolving elemental abundances of elements from carbon through iron. At the same time, by measuring the bremsstrahlung intensity and energy spectrum, the characteristics of the source electron flux can be determined. We will discuss these concepts, including estimated source strengths, and will describe a small instrument capable of providing this unique channel of information for future planetary missions. We propose to build this instrument using innovative electronics packaging methods to minimize size and weight.

Chenette, D. L.↗

Visualizing energy transfer between redox-active colloids

Redox-active colloids (RACs) represent a novel class of energy carriers that exchange electrical energy upon contact. Understanding contact-mediated electron transfer dynamics in RACs offers insights into physical contact events in colloidal suspensions and enables quantification of electrical energy transport in nonconjugated polymers. Redox-based electron transport was directly observed in monolayers of micron-sized RACs containing ethyl-viologen side groups via fluorescence microscopy through an unexpected nonlinear electrofluorochromism that is quantitatively coupled to the redox state of the colloid. Via imaging studies, using this electrofluorochromism, the apparent charge transfer diffusion coefficient D CT of the RAC was easily determined. The visualization of energy transport within suspensions of redox-active colloids was also demonstrated. Our work elucidates fundamental mechanisms of energy transport in colloidal systems, informs the development of next-generation redox flow batteries, and may inspire new designs of smart active soft matter including conductive polymers for applications ranging from electrochemical sensors and organic electronics to colloidal robotics.

Qu, Alan Subing [Univ. of Illinois at Urbana-Champ↗

Behavior of Windblown Sand on Mars: Results from Single-Particle Experiments

Experiments are investigating the behavior of individual sand grains in the high-energy martian aeolian regime. Energy partitioning during impact of a saltating grain determines grain longevity, but it also influences the way in which the bed becomes mobilized by reptation. When single grains of sand are fired into loose beds, the bed can absorb up to 90% of the impact energy by momentum transfer to other grains; it has been discovered that the impacting grains cause circular craters even at low impact angles. Hundreds of grains can be splashed by a single high-velocity (100 m/s) impact causing more bed disturbance through reptation than previously thought. The research is supported by NASA's PG&G Program. Because the martian aeolian environment in both high energy and of long duration, the most mobile fractions of windblown sand should have eradicated themselves by attrition, unless sand supply has kept pace with destruction. It is therefore important to understand the rate of grain attrition in order to make sense of the existence of vast dune fields on Mars. Attrition, has been addressed in other studies, but precise data for a single saltating grain striking a loose bed of sand have not been acquired -- the quintessential case to be understood for dunes on Mars. To acquire these data, we are employing a compound crossbow which has the bolt-firing mechanism replaced with a pneumatically-automated sabot system. The sabot can launch individual grains of sand of any size between several millimeters and about 50 microns, at velocities up to 100m/s. This is around the maximum velocity expected for saltating grains on Mars. The sabot sled is equipped with photoelectric sensors for measuring shot velocity. Baffling of the grain's exit orifice has enabled projection of single grains without significant aerodynamic effects from the sabot. Grains are fired into loose beds of sand at about 15 degrees from the horizontal (typical saltation trajectory at impact) while being filmed on high-speed video. High-intensity pulse illumination for the grains is triggered by the solenoid-operated bow trigger. A 45 degree mirror over the impact site provides simultaneous horizontal and vertical images of the impact on each video frame. UV fluorescence is enabling grain and grain-fragment recovery. At 100 m/s, grains of all sizes shatter into many fragments when the sand is replaced with a solid target. Kinetic energy of the grains at this velocity exceeds the critical energy for catastrophic failure of minerals. Although probably exceptional as a grain speed, it suggests that conditions on Mars might elevate materials into an attrition regime not encountered on other planets; individual grains blown across rock pavements on Mars will have short lifespans. When experimental grains impact loose (dune) sand, much, if not most of the kinetic energy is converted into momentum of other grains. Using high-speed filming, the energy involved in splashing grains at the impact site can be derived from the size of the crater, the speed of the splashed grains, and the rebound speed of the impactor. The amount of energy partitioned into material failure (as opposed to momentum) is too small a fraction of the total to be calculated under these circumstances. This does not necessarily mean that little damage occurs to the grains (the full extent of the damage has yet to be determined) because only a small fraction of the impact energy is required for inducing brittle fracture. Damage is orders of magnitude less than during impact against solid surfaces.

Marshall, J. R.↗

System and method for measuring sun-induced chlorophyll fluorescence

A chlorophyll fluorescence measuring system having at least one spectrometer coupled to a data logger. The data logger provides direct control of the spectrometer and includes on-board memory for storage of target and reference spectrum data obtained by the spectrometer. The data logger may be coupled to an external computer that receives and analyzes target and reference spectrum data to determine SIF using a spectral fitting algorithm. The system may include a spectrometer aiming system coupled to and controlled by the data logger. The system may also include one or more environmental sensors configured to measure environment variables. The environmental sensors may be coupled to the data logger for control and data storage. The environmental data may be communicated to the external computer for use in the spectral fitting algorithm. The data logger may be connected to a network for remote monitoring and control.

Gu, Lianhong↗

Laser-based conversion electron Mössbauer spectroscopy of 229 ThO 2

The exceptionally low-energy 229 Th nuclear isomeric state is expected to provide several new and powerful applications, including the construction of a robust and portable solid-state nuclear clock, perhaps contributing to a redefinition of the second, exploration of nuclear superradiance and tests of fundamental physics. Further, analogous to the capabilities of traditional Mössbauer spectroscopy, the sensitivity of the nucleus to its environment can be used to realize laser Mössbauer spectroscopy and, with it, new types of strain and temperature sensors and a new probe of the solid-state environment, all with excellent sensitivity. However, current models for examining the nuclear transition in a solid require the use of a high-bandgap, vacuum ultraviolet (VUV) transmissive host, severely limiting the applicability of these techniques. Here we report the first, to the authors’ knowledge, demonstration of laser-induced conversion electron Mössbauer spectroscopy (CEMS) of the 229 Th isomer in a thin ThO 2 sample whose bandgap (approximately 6 eV) is considerably smaller than the nuclear isomeric state energy (8.4 eV). Unlike fluorescence spectroscopy of the 229 Th isomeric transition, this technique is compatible with materials whose bandgap is less than the nuclear transition energy, opening a wider class of systems to study and the potential of a conversion-electron-based nuclear clock.

36 MATERIALS SCIENCE↗

Cyclically conjugated porphyrin trimers linked through benzo[4,5]imidazo[2,1- a ]isoindole bridges

Cyclically conjugated porphyrin trimers were prepared via a concise synthetic method. Zn-Trimer-1 displayed strong exciton coupling, suggesting the presence of effective electronic interactions. UV-Vis absorption and fluorescence spectra obtained through titration studies on the donor–acceptor adduct (Zn-Trimer-1–C60Im) indicate the occurrence of excited state photo-events. Inspired by nature's photosynthetic process of plants and bacteria, electronically interacting multiporphyrinic systems have long been investigated as synthetic models of natural photosynthetic systems. Remarkable successes have been achieved with cyclic porphyrin arrays, where porphyrins are arranged in a wheel-like structure or nanoring as photosynthetic antennae. Considerable efforts have also been devoted to constructing other porphyrin arrays such as molecular wires for potential applications in sensors, solar energy conversion, non-linear optical (NLO) materials, and more. Due to synthetic difficulties, cyclically conjugated porphyrin arrays in which porphyrins are arranged in planar geometry are very rare. Very recently, Anderson and coworkers have reported unprecedented β,β-directly linked porphyrin trimers and tetramers with different geometries, propeller and saddle, respectively, which result in distinct photophysical and electrochemical properties (Fig. 1a). In this paper, we present novel cyclically conjugated porphyrin trimers, which feature planar geometry with porphyrins assembled around the aromatic triphenylene at the center (Fig. 1b).

Han, Ting↗