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At least 883 records · Page 49

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NASA NTRS1993

Gas and dust in the pre-main-sequence multiple system GG Tauri

We present 1.4 and 2.7 mm aperture synthesis maps of the gas and dust continuum emission around GG Tauri, a very young component of a premain-sequence multiple star system; both GG Tau and its apparent companion, GG Tau/c, at 1500 AU separation, are themselves binaries. At 1.4 mm, dust continuum emission of about 750 AU in extent is associated with GG Tau, and a secondary peak is near GG Tau/c. Spectral line images reveal gaseous structure around GG Tau, elongated along the GG Tau-GG Tau/c axis. There is some suggestion that the gas associated with GG Tau/c alone is extended in a different direction. Marked changes in the morphology and velocity structure of the molecular emission near GG Tau/c also indicate that this system is differently oriented. Clumps between the two systems may be vestiges of a connecting bar. GG Tau and GG Tau/c appear to have originated in a common cloud; their different systemic orientations suggest that they formed from an initially prolate cloud rather than from an extensive and highly flattened disk.

Koerner, D. W.↗

NASA NTRS1995

Instantaneous flow measurements in a supersonic wind tunnel using spectrally resolved Rayleigh scattering

Results of a feasibility study to apply laser Rayleigh scattering to non-intrusively measure flow properties in a small supersonic wind tunnel are presented. The technique uses an injection seeded, frequency doubled Nd:YAG laser tuned to an absorption band of iodine. The molecular Rayleigh scattered light is filtered with an iodine cell to block light at the laser frequency. The Doppler-shifted Rayleigh scattered light that passes through the iodine cell is analyzed with a planar mirror Fabry-Perot interferometer used in a static imaging mode. An intensified CCD camera is used to record the images. The images are analyzed at several subregions, where the flow velocity is determined. Each image is obtained with a single laser pulse, giving instantaneous measurements.

Seasholtz, Richard G.↗

NASA NTRS2002

Pickup Ions at Dione and Enceladus

Voyager images of the icy satellites of Saturn, Dione and Enceladus, suggest they have been geologically active and are not only composed of ice. Recent observations by HST have shown the presence of ozone at both Dione and Rhea which also implies the presence of molecular oxygen at these bodies. The Cassini Plasma Spectrometer (CAPS) will provide the capability to determine the global composition of these bodies by measuring the pickup ions produced by the ionization of their sputter produced atmospheres. We will present a model of these atmospheres and associated pickup ions and demonstrate CAPS ability to distinguish the freshly produced picked up ions from the ambient plasma. Such ions are expected to form a ring distribution that will have a uniquely different energy-angle dependence than the ambient plasma ions. In the case of Dione we expect the potential for a moderate strength interaction for which both Voyager 1 and Pioneer 11 spacecraft measured ion cyclotron waves centered on the Dione L shell and near the equatorial plane. Since Enceladus may be the source of the E-ring, some surprises may be encountered during its close encounter with the Cassini spacecraft. In the case of Dione we will show that a wake pass at 500 km altitude is more than an order of magnitude better than an upstream pass at 500 km altitude. Pickup ion detection for minor ion species such as NH3+ is possible for 500 km altitude wake pass but not for a 500 km altitude upstream pass at closest approach. For navigation reasons a 100 km pass is not allowed and therefore it is essential to have a wake pass to maximize the science return for a targeted flyby with Dione. The CAPS observations when combined with magnetometer, plasma wave and energetic particle observations will allow us to estimate the source of ions into Saturn's magnetosphere due to these two bodies and to characterize the nature of the interaction with Saturn's magnetosphere.

Sittler, E.↗

NASA NTRS2004

Microscale Distribution of Hydrogen Isotopes in Two Carbonaceous Chondrites

Hydrogen isotopes are highly variable among primitive meteorites and interplanetary dust particles (IDPs) [1, 2]. In particular, many primitive objects exhibit D (and N-15) enrichments, relative to terrestrial values, thought to represent partial preservation of presolar material fractionated in molecular clouds. However, the diversity of D/H ratios among IDPs and chondrites indicates a complex history of processing in the solar nebula and on meteorite parent bodies. Deconvolving this record requires identification and characterization of the carriers of D enrichments in different objects. Isotopic imaging has proven to be a powerful method to quantitatively explore the distribution of D/H ratios on a one to several m scale in IDPs [2-4] and the CR chondrite Renazzo [5, 6]. In this study, we have used ion imaging to explore the microscale D/H distribution of two carbonaceous chondrites, Tagish Lake (unique) and Al Rais (CR2). Previous D/H measurements (on a ~tens of microns scale) of Tagish Lake matrix fragments by Messenger [7] and Engrand et al. [8] have found different results, most likely related to the analytical techniques used. Previous work has also shown a large range of D/H ratios in CR chondrites, including very large variations on a scale of a few microns [5, 6, 9].

Young, A. F.↗

NASA NTRSDate not supplied

Low-Temperature Production of Cosmic Grains Analogs From Gas-Phase Molecular Precursors with the NASA Ames' COSmIC Facility

Complex carbon molecules and ions are ubiquitous in space and form the building blocks of carbonaceous cosmic dust grains, ultimately contributing to the formation of planets. Here, we report the first experimental investigations of the low temperature chemical pathways leading to the production of cosmic grain analogs from gas phase molecular precursors in order to better understand the evolution of cosmic carbon. The experiments were performed using the COsmic SImulation Chamber (COSmIC) to generate and characterize grains formed from gas phase precursors under controlled conditions representative of astrophysical environments [1]. Using COSmIC, it is possible to investigate the evolution of cosmic carbon from the formation of neutral and ionized gas phase hydrocarbons and PAHs [2], to carbon grains [3, 4]. This is achieved by using a discharge nozzle to (1) produce an adiabatic jet expansion and cool down Ar-hydrocarbon/PAH gas mixtures to astrophysically relevant temperature (150 K) before inducing chemistry by generating a plasma discharge in the stream of the expansion. This plasma-induced chemistry results in the formation of complex molecules and solid particles, analogs of cosmic grains. Solid grains are produced in-situ in the plasma expansion, carried by the accelerated gas in the expansion, and collected on substrates placed a few centimeters downstream of the electrodes. The results of a preliminary solid phase ex-situ analysis of cosmic grain analogs produced at low temperature in COSmIC from gas mixtures including molecular precursors of cosmic carbon grains, i.e., such as CH4 and C2H2 found in circumstellar ejecta of late C stars [3] as well as PAHs ubiquitously detected in IR emission bands seen in galactic and extragalactic environments [4] will be discussed. Scanning Electron Microscopy imaging was used to provide insight on the morphology and growth structure of the grains produced in COSmIC, and to investigate how the precursors used to produce the grains affect these parameters [3]. Laser desorption mass spectrometry was used to identify the molecules making up the main structures within the condensed grains [4]. NIR to FIR optical properties are characterized to provide critical information (functional groups, optical constants) to the scientific community, for use in radiative transfer models in particular, and to help decipher and enhance the return data from space observations.

F. Salama↗

NASA NTRS1992

Systematic variations in the spectral properties of bright regions on Mars

The color and albedo of the martian surface define two basic surface units, dark gray material interpreted as relatively unaltered 'rock' and bright reddish material interpreted as weathered 'soil'. Understanding the processes contributing to soil formation first requires assessment of the soil's composition and compositional diversity. We report first results of an investigation of the character and variability of Fe- and water-bearing phases in bright reddish materials using ISM data. We also explore implications of these results for chemical evolution of martian soil. Information on the composition and distribution of bright reddish material comes from three major sources: Viking images, measurements by the XRF and GCMS instruments on the Viking Landers, and spectroscopic data. The XRF experiment found nearly identical, Fe-rich major-element compositions comparable to weathered basalt. Soil water, amounting to approximately 1-3 wt. percent as measured by the GCMS, was liberated mostly by heating to greater than or equal to 350 C, suggesting that it is present in a chemically bound form. Spectroscopic studies have detected ferric oxide, probably hematite, as well as molecular water. However, the identities of major silicate phases have been controversial, with conflicting evidence regarding phyllosilicates. Two main interpretations of this evidence have been proposed: Weathering of basaltic glasses by H2O and CO2 formed a mixture of oxides, salts, and metastable phyllosilicates such as montmorillonite and 'palagonite' formed when basaltic melt contacted ground ice or water. Palagonite is a hydrated basaltic glass containing dispersed ferric oxide, recrystallized in varying degrees to phyllosilicates. Typically it is aphanitic, although some examples contain phenocrysts. In either case, eolian redistribution is thought to have resulted in the material's global homogenization. Imaging spectroscopic data returned by the ISM instrument on Phobos 2 provide a powerful new basis for evaluating the composition and origin of martian soil because they are indicative of the presence and distribution of water- and Fe-bearing phases whose mineralogy is sensitive to the history of chemical weathering.

Murchie, Scott↗

DOE OSTI2026

Data‐Driven Engineering of Thermostable Collagen‐Mimetic Peptoid Triple Helices

Collagen-mimetic peptides (CMPs) are engineered molecules designed to replicate the triple-helical structure of natural collagen. A repeating x–y-Gly sequence is the defining motif of CMPs and is critical to their triple-helical structure and stability. Substitutions to the residues occupying the x and y positions present a means to modulate the CMP structure and properties. Peptoid residues—N-substituted glycine derivatives—present an attractive potential substitution due to their thermal stability, proteolytic resistance, biocompatibility, and diverse palette of non-natural side chains, but also tend to introduce a high degree of backbone flexibility that can diminish the stability of the triple helix. In this work, we report a computational active learning cycle comprising molecular dynamics simulation, Gaussian process regression, and Bayesian optimization to computationally identify a number of promising peptoid substitutions predicted to stabilize the desired quaternary structure through side chain interactions and produce stable peptoid-based collagen-like triple helices. To experimentally test the computational predictions, a top candidate identified by the screen was synthesized and imaged using scanning electron microscopy to resolve fibril-like bundles consistent with collagen-like triple helices. This work predicts a number of CMP peptoid substitutions capable of forming stable triple-helical structures, presents a generalizable design strategy for engineering desired peptoid structures, and opens new avenues for the design of peptoid-based biomimetic materials.

active learning↗

DOE OSTI2023

Discovery of Nanoscale Electric Field‐Induced Phase Transitions in ZrO 2

Abstract The emergence of ferroelectric and antiferroelectric properties in the semiconductor industry's most prominent high‐k dielectrics, HfO 2 and ZrO 2 , is leading to technology developments unanticipated a decade ago. Yet the failure to clearly distinguish ferroelectric from antiferroelectric behavior is impeding progress. Band‐excitation piezoresponse force microscopy and molecular dynamics are used to elucidate the nanoscale electric field‐induced phase transitions present in ZrO 2 ‐based antiferroelectrics. Antiferroelectric ZrO 2 is clearly distinguished from a closely resembling pinched La‐doped HfO 2 ferroelectric. Crystalline grains in the range of 3 – 20 nm are imaged independently undergoing reversible electric field induced phase transitions. The electrically accessible nanoscale phase transitions discovered in this study open up an unprecedented paradigm for the development of new nanoelectronic devices.

36 MATERIALS SCIENCE↗

DOE OSTI2021

The Steel Equivalency Workbook: An X-Ray Transmission Calculator

The Steel Equivalency Spreadsheet was created as a robust, user-friendly method for the following: 1. Determining if it is possible to image an object. 2. Reducing time needed when selecting equipment and preparing for field imaging activities. 3. Reducing the amount of equipment taken for field imaging activities. 4. Determining a starting point for exposure settings prior to imaging an object. 5. Reducing the amount of dose deposited to complete an imaging operation (in the spirit of As Low As Reasonably Achievable, ALARA). The spreadsheet was developed using various physical models to account for different phenomena. Future implementations aim to expand beyond the Digital Radiography and Computed Tomography Single Munition Scanner (DRCT SMS) in standard configuration to include high energy XRGs (Betatrons) and sub-MeV pulsed XRGs (XRS4). Alternative computational methods that compensate for incoherent scattering when calculating relative transmission are also being pursued. Ultimately the spreadsheet exceeded the developmental goal of having less than 10% average error when comparing calculations to real-world images.

36 MATERIALS SCIENCE↗

DOE OSTI2024

Spatiotemporal metabolic responses to water deficit stress in distinct leaf cell-types of poplar

The impact of water-deficit (WD) stress on plant metabolism has been predominantly studied at the whole tissue level. However, plant tissues are made of several distinct cell types with unique and differentiated functions, which limits whole tissue ‘omics’-based studies to determine only an averaged molecular signature arising from multiple cell types. Advancements in spatial omics technologies provide an opportunity to understand the molecular mechanisms underlying plant responses to WD stress at distinct cell-type levels. Here, we studied the spatiotemporal metabolic responses of two poplar ( Populus tremula× P. alba ) leaf cell types -palisade and vascular cells- to WD stress using matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI). We identified unique WD stress-mediated metabolic shifts in each leaf cell type when exposed to early and prolonged WD stresses and recovery from stress. During water-limited conditions, flavonoids and phenolic metabolites were exclusively accumulated in leaf palisade cells. However, vascular cells mainly accumulated sugars and fatty acids during stress and recovery conditions, respectively, highlighting the functional divergence of leaf cell types in response to WD stress. By comparing our MALDI-MSI metabolic data with whole leaf tissue gas chromatography-mass spectrometry (GC-MS)-based metabolic profile, we identified only a few metabolites including monosaccharides, hexose phosphates, and palmitic acid that showed a similar accumulation trend at both cell-type and whole leaf tissue levels. Overall, this work highlights the potential of the MSI approach to complement the whole tissue-based metabolomics techniques and provides a novel spatiotemporal understanding of plant metabolic responses to WD stress. This will help engineer specific metabolic pathways at a cellular level in strategic perennial trees like poplars to help withstand future aberrations in environmental conditions and to increase bioenergy sustainability.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS1995

Laser diagnostics for microgravity droplet studies

An instrument has been designed, built, and tested for performing laser diagnostic measurements of droplet combustion in low-gravity-flight aircraft. Nonintrusive measurements are of particular importance for droplet combustion (the simplest example of non-premixed combustion) and transport in microgravity environments, where physical contact would introduce an unacceptable level of perturbations. The resolution of these diagnostics can also isolate transport to length scales much smaller than the droplet diameter. These techniques can be configured to instantaneously map an entire flow field in two and three dimensions, providing either qualitative or quantitative information on the distribution of a desired scalar or vector quantity. Detailing the gas-phase flow field and position of the flame front can be achieved using planar laser-induced fluorescence (PLIF) of OH or another flame front marker. An alternative approach is to obtain LIF from a diagnostic seed included in the liquid phase fuel; it would be consumed at the flame front. The main advantage to this approach is that it is easier to choose the wavelength of the molecular absorption which coincides with convenient laser wavelengths rather than finding lasers which can be configured to access OH. Our present method uses a nitrogen-pumped dye laser tuned to a sodium absorption and addition of small concentrations of NaCl to the fuel. Particle image velocimetry (PIV) is a laser-based technique which has recently had its practicality greatly enhanced by the development of high-resolution CCD cameras and the increase in speed and capacity of computer systems. With this technique, a seeded flow is illuminated with a double-pulsed laser sheet to generate a double exposure image on a film or CCD camera. Computer analysis of the image is used to determine the particle velocity vectors and, thus, the gas velocity within the plane of the laser sheet. Our current experiment uses PIV for measuring relative droplet gas-phase velocities in a parabolic flight environment. The results show successful application of PLIF relative to luminosity and radical emission near 308 nm. PIV results are less successful due to gas thermal expansion deflecting the seed particles away from the measurement volume. PIV is applicable for measurements in the absence of combustion.

Winter, Michael↗

NASA NTRS1996

High Resolution UV Spectroscopy of H2 and N2 Applied to Observations of the Planets by Spacecraft

The next generation of high resolution UV imaging spacecraft are being prepared for studying the airglow and aurora of the Earth, the other terrestrial planets and the Jovian planets. To keep pace with these technological improvements we have developed a laboratory program to provide electron impact collision cross sections of the major molecular planetary gases (H2, N2, CO2, O2, and CO). Spectra under optically thin conditions have been measured with a high resolution (lambda/delta(lambda) = 50000) UV spectrometer in tandem with electron impact collision chamber. High resolution spectra of the Lyman and Wemer band systems of H2 have been obtained and modeled. Synthetic spectral intensities based on the J-dependent transition probabilities that include ro-vibronic perturbations are in very good agreement with experimental intensities. The kinetic energy distribution of H(2p,3p) atoms resulting from electron impact dissociation of H2 has been measured. The distribution is based on the first measurement of the H Lyman-alpha (H L(alpha)) and H Lyman-beta (H L(beta)) emission line Doppler profiles. Electron impact dissociation of H2 is believed to be one of the major mechanisms leading to the observed wide profile of H L-alpha from Jupiter aurora by the Hubble Space Telescope (HST). Analysis of the deconvolved line profile of H L-alpha reveals the existence of a narrow line peak (40 mA FWHM) and a broad pedestal base (240 mA FWHM). The band strengths of the electron excited N2 (C(sup 3) Pi(sub(upsilon) - B(sup 3)Pi(sub g)) second positive system have been measured in the middle ultraviolet. We report a quantitative measurement of the predissociation fraction 0.15 +/- 01(sup .045, sub .01) at 300 K in the N2 c'(sub )4 (1)sigma(sup +, sub g) - x(1)sigma(sup +, sub g)(00) band, with an experimental determination of rotational line strengths to be used to understand N2 EUV emission from Titan, Triton and the Earth.

Ajello, J.↗

NASA NTRS2001

AVIRIS Inflight Calibration Experiments, Analysis, and Results in 2000

The NASA Airborne Visible/Infrared Imaging Spectrometer (AVIRIS) measures spectra from 370 to 2500 nm with nominally 10-nm sampling and resolution. The spectra are acquired as images with an 11 km width and up to 800 km length from the ER-2 platform or 2.1 km width and 160 km length from the Twin Otter platform. AVIRIS measurements are used for a range of Earth science research and application objectives. The molecular absorption and particle scattering properties expressed in the calibrated AVIRIS measurements are used. For both science research and application objectives, calibration of the AVIRIS spectra is required to produce useful results. Each year prior to the flight season AVIRIS is calibrated in the laboratory. However, the temperature, pressure, vibration, and observational geometry, as well as mechanical, electrical, and operational interfaces of the laboratory are different than the environment on board the airborne platform. To validate the calibration of AVIRIS in the flight environment, an inflight calibration experiment is orchestrated at the beginning of each flight season. In most years additional inflight calibration experiments occur towards the middle and end of the flight season as well. For an inflight calibration experiment, AVIRIS acquires airborne data over a designated calibration target. In concert with the airborne data acquisition, surface and atmospheric properties at the calibration target are measured in situ. These in situ measurements are used to constrain a radiative transfer code and predict the radiance incident at the AVIRIS instrument from the calibration target. This prediction is compared with the AVIRIS-measured radiance to validate the calibration of AVIRIS in the flight environment. Additional properties (such as the AVIRIS inflight measurement precision) are determined as well. This paper presents measurements, analyses, and results from the inflight calibration experiment held on the dry lake bed surface of Rogers Dry Lake, California, on June 5, 2000.

Green, Robert O.↗

NASA NTRS1995

Polar distribution of annexin-like proteins during phytochrome-mediated initiation and growth of rhizoids in the ferns Dryopteris and Anemia

Although the calcium requirement of phytochrome-mediated fern spore germination and early rhizoid growth is well established, the calcium-binding proteins that serve as transducers for these responses are not known. Here we report the presence of annexin-like proteins in germinating spores of Dryopteris filix-mas (L.) Schott and Anemia phyllitidis (L.) Sw. and evidence that they may be important participants in early photomorphogenic changes in gametophytes. Immunolocalization and immunoblot assays of these proteins were carried out using polyclonal antibodies raised either against a 35-kDa annexin-like protein from pea or against anchorin CII from chicken. Western-blot analysis showed that crude protein extracts obtained from both species after red-light treatment contained two cross-reactive protein bands with molecular weights around 70 kDa. These proteins were annexin-like in that they bound to a phosphatidylserine affinity column in a calcium-dependent fashion. Using this column, two protein bands around 70 kDa, i.e. 67 and 73 kDa, were partially purified together with proteins at 36 kDa and a doublet at 54 kDa. Proteins of these latter molecular weights are suggested to be members of the annexin family, but no cross-reactivity could be found between these and the two antibodies used in our investigations. Immunodetectable levels of these proteins were observed only after light-mediated induction of spore germination. Imaging of the immuno-localization patterns observed with both antibodies showed that the annexin-like proteins are concentrated at the extreme tips of the rhizoids in D. filix-mas and A. phyllitidis during rhizoid initiation and all stages of elongation. We suggest that these proteins may play a major role in the tip-oriented exocytosis events that are critical for the initiation and growth of fern rhizoids.

NASA Discipline Number 40-50↗

NASA NTRS1995

High Resolution UV Spectroscopy of H2 and N2 Applied to Observations of the Planets by Spacecraft

The next generation of high resolution UV imaging Spacecraft are being prepared for studying the airglow and aurora of the Earth, the other terrestrial planets and the Jovian planets. To keep pace with these technological improvements we have developed a laboratory program to provide electron impact collision cross sections of the major molecular planetary gases. This program is described in detail.

Spectroscopy↗

DOE OSTI2021

Realization of AlSb in the Double-Layer Honeycomb Structure: A Robust Class of Two-Dimensional Material

Exploring new two-dimensional (2D) van der Waals (vdW) systems is at the forefront of materials physics. Here, through molecular beam epitaxy on graphene-covered SiC(0001), we report successful growth of AlSb in the double-layer honeycomb (DLHC) structure, a 2D vdW material which has no direct analogue to its 3D bulk and is predicted kinetically stable when freestanding. The structural morphology and electronic structure of the experimental 2D AlSb are characterized with spectroscopic imaging scanning tunneling microscopy and cross-sectional imaging scanning transmission electron microscopy, which compare well to the proposed DLHC structure. The 2D AlSb exhibits a bandgap of 0.93 eV versus the predicted 1.06 eV, which is substantially smaller than the 1.6 eV of bulk. We also attempt the less-stable InSb DLHC structure; however, it grows into bulk islands instead. Here, the successful growth of a DLHC material here opens the door for the realization of a large family of novel 2D DLHC traditional semiconductors with unique excitonic, topological, and electronic properties.

36 MATERIALS SCIENCE↗

NASA NTRS1999

Cameleon calcium indicator reports cytoplasmic calcium dynamics in Arabidopsis guard cells

Cytoplasmic free calcium ([Ca2+]cyt) acts as a stimulus-induced second messenger in plant cells and multiple signal transduction pathways regulate [Ca2+]cyt in stomatal guard cells. Measuring [Ca2+]cyt in guard cells has previously required loading of calcium-sensitive dyes using invasive and technically difficult micro-injection techniques. To circumvent these problems, we have constitutively expressed the pH-independent, green fluorescent protein-based calcium indicator yellow cameleon 2.1 in Arabidopsis thaliana (Miyawaki et al. 1999; Proc. Natl. Acad. Sci. USA 96, 2135-2140). This yellow cameleon calcium indicator was expressed in guard cells and accumulated predominantly in the cytoplasm. Fluorescence ratio imaging of yellow cameleon 2.1 allowed time-dependent measurements of [Ca2+]cyt in Arabidopsis guard cells. Application of extracellular calcium or the hormone abscisic acid (ABA) induced repetitive [Ca2+]cyt transients in guard cells. [Ca2+]cyt changes could be semi-quantitatively determined following correction of the calibration procedure for chloroplast autofluorescence. Extracellular calcium induced repetitive [Ca2+]cyt transients with peak values of up to approximately 1.5 microM, whereas ABA-induced [Ca2+]cyt transients had peak values up to approximately 0.6 microM. These values are similar to stimulus-induced [Ca2+]cyt changes previously reported in plant cells using ratiometric dyes or aequorin. In some guard cells perfused with low extracellular KCl concentrations, spontaneous calcium transients were observed. As yellow cameleon 2.1 was expressed in all guard cells, [Ca2+]cyt was measured independently in the two guard cells of single stomates for the first time. ABA-induced, calcium-induced or spontaneous [Ca2+]cyt increases were not necessarily synchronized in the two guard cells. Overall, these data demonstrate that that GFP-based cameleon calcium indicators are suitable to measure [Ca2+]cyt changes in guard cells and enable the pattern of [Ca2+]cyt dynamics to be measured with a high level of reproducibility in Arabidopsis cells. This technical advance in combination with cell biological and molecular genetic approaches will become an invaluable tool in the dissection of plant cell signal transduction pathways.

Non-NASA Center↗

DOE OSTI2024

A red-emitting carborhodamine for monitoring and measuring membrane potential

Biological membrane potentials, or voltages, are a central facet of cellular life. Optical methods to visualize cellular membrane voltages with fluorescent indicators are an attractive complement to traditional electrode-based approaches, since imaging methods can be high throughput, less invasive, and provide more spatial resolution than electrodes. Recently developed fluorescent indicators for voltage largely report changes in membrane voltage by monitoring voltage-dependent fluctuations in fluorescence intensity. However, it would be useful to be able to not only monitor changes but also measure values of membrane potentials. This study discloses a fluorescent indicator which can address both. We describe the synthesis of a sulfonated tetramethyl carborhodamine fluorophore. When this carborhodamine is conjugated with an electron-rich, methoxy (-OMe) containing phenylenevinylene molecular wire, the resulting molecule, CRhOMe, is a voltage-sensitive fluorophore with red/far-red fluorescence. Using CRhOMe, changes in cellular membrane potential can be read out using fluorescence intensity or lifetime. In fluorescence intensity mode, CRhOMe tracks fast-spiking neuronal action potentials (APs) with greater signal-to-noise than state-of-the-art BeRST 1 (another voltage-sensitive fluorophore). CRhOMe can also measure values of membrane potential. The fluorescence lifetime of CRhOMe follows a single exponential decay, substantially improving the quantification of membrane potential values using fluorescence lifetime imaging microscopy (FLIM). The combination of red-shifted excitation and emission, mono-exponential decay, and high voltage sensitivity enable fast FLIM recording of APs in cardiomyocytes. The ability to both monitor and measure membrane potentials with red light using CRhOMe makes it an important approach for studying biological voltages.

59 BASIC BIOLOGICAL SCIENCES↗