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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 199 records · Page 11
All sourcesNASA NTRSDOE OSTI
DOE OSTI2025

Volumetric imaging of the 3D orientation of cellular structures with a polarized fluorescence light-sheet microscope

Polarized fluorescence microscopy is a valuable tool for measuring molecular orientations in biological samples, but techniques for recovering three-dimensional orientations and positions of fluorescent ensembles are limited. We report a polarized dual-view light-sheet system for determining the diffraction-limited three-dimensional distribution of the orientations and positions of ensembles of fluorescent dipoles that label biological structures. We share a set of visualization, histogram, and profiling tools for interpreting these positions and orientations. We model the distributions based on the polarization-dependent efficiency of excitation and detection of emitted fluorescence, using coarse-grained representations we call orientation distribution functions (ODFs). We apply ODFs to create physics-informed models of image formation with spatio-angular point-spread and transfer functions. We use theory and experiment to conclude that light-sheet tilting is a necessary part of our design for recovering all three-dimensional orientations. We use our system to extend known two-dimensional results to three dimensions in FM1-43-labeled giant unilamellar vesicles, fast-scarlet-labeled cellulose in xylem cells, and phalloidin-labeled actin in U2OS cells. Additionally, we observe phalloidin-labeled actin in mouse fibroblasts grown on grids of labeled nanowires and identify correlations between local actin alignment and global cell-scale orientation, indicating cellular coordination across length scales.

Science & Technology - Other Topics↗
DOE OSTI2025

State-dependent motion of a genetically encoded fluorescent biosensor

Genetically encoded biosensors can measure biochemical properties such as small-molecule concentrations with single-cell resolution, even in vivo. Despite their utility, these sensors are “black boxes”: Very little is known about the structures of their low- and high-fluorescence states or what features are required to transition between them. We used LiLac, a lactate biosensor with a quantitative fluorescence-lifetime readout, as a model system to address these questions. X-ray crystal structures and engineered high-affinity metal bridges demonstrate that LiLac exhibits a large interdomain twist motion that pulls the fluorescent protein away from a “sealed,” high-lifetime state in the absence of lactate to a “cracked,” low-lifetime state in its presence. Understanding the structures and dynamics of LiLac will help to think about and engineer other fluorescent biosensors.

Rosen, Paul C. (ORCID:000000017414454X)↗
DOE OSTI2022

X-ray fluorescence microscopy methods for biological tissues

Abstract Synchrotron-based X-ray fluorescence microscopy is a flexible tool for identifying the distribution of trace elements in biological specimens across a broad range of sample sizes. The technique is not particularly limited by sample type and can be performed on ancient fossils, fixed or fresh tissue specimens, and in some cases even live tissue and live cells can be studied. The technique can also be expanded to provide chemical specificity to elemental maps, either at individual points of interest in a map or across a large field of view. While virtually any sample type can be characterized with X-ray fluorescence microscopy, common biological sample preparation methods (often borrowed from other fields, such as histology) can lead to unforeseen pitfalls, resulting in altered element distributions and concentrations. A general overview of sample preparation and data-acquisition methods for X-ray fluorescence microscopy is presented, along with outlining the general approach for applying this technique to a new field of investigation for prospective new users. Considerations for improving data acquisition and quality are reviewed as well as the effects of sample preparation, with a particular focus on soft tissues. The effects of common sample pretreatment steps as well as the underlying factors that govern which, and to what extent, specific elements are likely to be altered are reviewed along with common artifacts observed in X-ray fluorescence microscopy data.

Pushie, M. Jake (ORCID:0000000174945427)↗
DOE OSTI2022

A new route to obtain fluorescence X-ray absorption spectra of compounds and to remove the self-absorption induced nonlinearity in the spectra

A new route to obtain fluorescence X-ray absorption spectra of compounds and to remove the self-absorption induced nonlinearity in the spectra is described. The fluorescent intensity If is linearly proportional to the absorption coefficient μ. For studies of surface structures around an element (κ) the fluorescence detection is often the mode of choice. However, the measurement may suffer from a self-absorption (SA) effect which nonlinearly distorts the spectra. The effect is severe when κ is concentrated or the measurements are carried out in certain geometries. Here, the correlations among emission events in compounds are examined following resonance X-ray core-electron excitation within κ. Under conditions leading to SA, If emitted from κ apparently has a conjugated relationship with the fluorescent intensities simultaneously emitted from other elements (ξ). Normalizing the former (κ) by the latter (ξ) will largely remove SA effects and reduce this nonlinear problem to a tractable linear problem. This does result in a moderate reduction of the spectral amplitude due to the so-called secondary emission from ξ excited by the emission from κ. Nonetheless, the resulting spectra will allow one to accurately determine bond distances and disorder and, in some respects, can be superior to spectra obtained via the absorption channel. For μξ < μκ and grazing incidence geometry, the amplitude reduction can be small and simple normalization is sufficient to restore the spectral integrity with remarkable accuracy. This has been instrumental in unravelling the surface and subsurface structures around cations in amorphous Ga–In–O and Zn–Sn–O films which are otherwise inaccessible due to severe SA effects. This method has also been applied to several samples with μξ ≃ μκ to examine its applicability. For these samples, the amplitude reduction is 12 ± 4% versus their standards for the data measured with the classical 45°/45° geometry. This experimental method is easy to implement. Since If from κ and ξ are measured by the same detector system, it is also superior to other methods in removing systematic errors such as detector system nonlinearity, electronic noise, and some beam instabilities, and in removing spectral imperfections due to, for example, SA effects, diffraction effects and sample inhomogeneity. The distortions resulting from the latter can be severe in the spectra measured in transmission mode.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2022

NIR fluorescence lifetime macroscopic imaging with a time-gated SPAD camera

The performance of SwissSPAD2 (SS2), a large scale, widefield time-gated CMOS SPAD imager developed for fluorescence lifetime imaging, has recently been described in the context of visible range and fluorescence lifetime imaging microscopy (FLIM) of dyes with lifetimes in the 2.5 – 4 ns range. Here, we explore its capabilities in the NIR regime relevant for small animal imaging, where its sensitivity is lower and typical NIR fluorescent dye lifetimes are much shorter (1 ns or less). We carry out this study in a simple macroscopic imaging setup based on a compact NIR picosecond pulsed laser, an engineered diffuser-based illumination optics, and NIR optimized imaging lens suitable for well-plate or small animal imaging. Because laser repetition rates can vary between models, but the synchronization signal frequency accepted by SS2 is fixed to 20 MHz, we first checked that a simple frequency-division scheme enables data recording for different laser repetition rates. Next, we acquired data using different time gate widths, including gates with duration longer than the laser period, and analyzed the resulting data using both standard nonlinear least-square fit (NLSF) and phasor analysis. We show that the fixed synchronization rate and large gate widths characterizing SS2 (10 ns and over) are not an obstacle to accurately extracting lifetime in the 1 ns range and to distinguishing between close lifetimes. As a result, SS2 and similar very large gated SPAD imagers appear as a versatile alternative to other widefield time-resolved detectors for NIR fluorescence lifetime imaging, including preclinical molecular applications.

FLIM↗
DOE OSTI2025

Chatbots can guide experimentalists to avoid many common mistakes in measuring fluorescence spectra

The concepts of fluorescence spectroscopy are well established, yet the experimental collection of a spectrum is susceptible to a range of errors. A recent study showed that chatbots – derived from substantial advances in artificial intelligence – can provide practical assistance to the experimentalist during collection of absorption spectra and thereby achieve improved quality. Fluorescence, while the complement of absorption, is prone to a far richer array of experimental errors encompassing environmental factors, instrumental settings, and experimentalist mistakes. Here, 10 chatbots (ChatGPT 5, Gemini 2.5 Pro, Microsoft Copilot, Meta AI Llama 4, Claude Sonnet 4.5, X.ai Grok 4, Perplexity Pro, DeepSeek V3.1, Z.ai GLM-4.6, and KIMI K2) have been evaluated for the suitability in providing quality-improvement advice to the experimentalist during acquisition of fluorescence spectra. Significant opportunities for practical assistance are noted, although intervention by a domain expert is often required. Here, if combined with image recognition and/or signal processing, chatbots embedded in instruments may enable real-time guidance during fluorescence data acquisition.

Artificial intelligence↗
DOE OSTI2022

Real-time image denoising of mixed Poisson–Gaussian noise in fluorescence microscopy images using ImageJ

Fluorescence microscopy imaging speed is fundamentally limited by the measurement signal-to-noise ratio (SNR). To improve image SNR for a given image acquisition rate, computational denoising techniques can be used to suppress noise. However, common techniques to estimate a denoised image from a single frame either are computationally expensive or rely on simple noise statistical models. These models assume Poisson or Gaussian noise statistics, which are not appropriate for many fluorescence microscopy applications that contain quantum shot noise and electronic Johnson–Nyquist noise, therefore a mixture of Poisson and Gaussian noise. In this paper, we show convolutional neural networks (CNNs) trained on mixed Poisson and Gaussian noise images to overcome the limitations of existing image denoising methods. The trained CNN is presented as an open-source ImageJ plugin that performs real-time image denoising (within tens of milliseconds) with superior performance (SNR improvement) compared to conventional fluorescence microscopy denoising methods. The method is validated on external datasets with out-of-distribution noise, contrast, structure, and imaging modalities from the training data and consistently achieves high-performance ( > <#comment/> 8 d B ) denoising in less time than other fluorescence microscopy denoising methods.

47 OTHER INSTRUMENTATION↗
DOE OSTI2023

Direct selection of functional fluorescent-protein antibody fusions by yeast display

Antibodies are important reagents for research, diagnostics, and therapeutics. Many examples of chimeric proteins combining the specific target recognition of antibodies with complementing functionalities such as fluorescence, toxicity or enzymatic activity have been described. However, antibodies selected solely on the basis of their binding specificities are not necessarily ideal candidates for the construction of chimeras. Here, we describe a high throughput method based on yeast display to directly select antibodies most suitable for conversion to fluorescent chimera. A library of scFv binders was converted to a fluorescent chimeric form, by cloning thermal green protein into the linker between VH and VL, and directly selecting for both binding and fluorescent functionality. This allowed us to directly identify antibodies functional in the single chain TGP format, that manifest higher protein expression, easier protein purification, and one-step binding assays.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2025

Measurement report: A comparative analysis of an intensive incursion of fluorescing African dust particles over Puerto Rico and another over Spain

Measurements during episodes of African dust, made with two wideband integrated bioaerosol spectrometers (WIBSs), one on the northeastern coast of Puerto Rico and the other in the city of León, Spain, show unmistakable, bioaerosol-like fluorescing aerosol particles (FAPs) that can be associated with these dust episodes. The Puerto Rico event occurred during a major incursion of African dust during June 2020. The León event occurred in the late winter and spring of 2022, when widespread, elevated layers of dust inundated the Iberian Peninsula. Satellite and back-trajectory analyses confirm that dust from northern Africa was the source of the particles during both events. The WIBSs measure the size of individual particles in the range from 0.5 to 30 µm, derive a shape factor, and classify seven types of fluorescence from the FAPs. In general, it is not possible to directly determine the specific biological identity from fluorescence signatures; however, measurements of these types of bioaerosols in laboratory studies allow us to compare ambient fluorescence patterns with whole microbial cells measured under controlled conditions. Here we introduce some new metrics that offer a more quantitative approach for comparing FAP characteristics derived from particles measured under different environmental conditions. The analysis highlights the similarities and differences at the two locations and reveals differences that can be attributed to the age and history of the dust plumes, e.g., the amount of time that the air masses were in the mixed layer and the frequency of precipitation along the air mass trajectory.

54 ENVIRONMENTAL SCIENCES↗
NASA NTRS1974

Multiplet effects on the L/sub 2,3/ fluorescence yield of multiply ionized Ar

The 2p fluorescence yield of Ar in the presence of zero to six 3p holes has been calculated by statistically averaging the fluorescence yields of initial states that consist of individual multiplet configurations, formed by coupling the 2p vacancy to the partially filled 3p shell. The L(sub 2,3) fluorescence yields for the (2p)-1 (3p)-n configurations of Ar are found to be 1.48, 17.97, 24.83, 37.84, 79.61, 112.16, and 121.48 x .0001 for n = 0, 1, 2, 3, 4, 5, and 6, respectively. Results agree reasonably well with experimental fluorescence yields deduced from ion-atom collision measurements.

Chen, M. H.↗
NASA NTRS1975

Photomultiplier window materials under electron irradiation - Fluorescence and phosphorescence

The fluorescence and phosphorescence of photomultiplier window materials under electron irradiation have been investigated using a Sr-90/Y-90 beta emitter as the electron source. Spectral emission curves of UV-grade, optical-grade, and electron-irradiated samples of MgF2 and LiF, and of CaF2, BaF2, sapphire, fused silica, and UV-transmitting glasses were obtained over the 200-650-nm spectral range. Fluorescence yields were determined on these materials utilizing photomultiplier tubes with cesium telluride, bialkali, and trialkali (S-20) photocathodes, respectively. Optical-grade MgF2 and LiF, as well as electron-irradiated UV-grade samples of these two materials, show enhanced fluorescence due to color-center formation and associated emission bands in the blue and red wavelength regions. Large variations in fluorescence intensities were found in UV-grade sapphire samples of different origins, particularly in the red end of the spectrum, presumably due to various amounts of chromium-ion content. Phosphorescence decay with time is best described by a sum of exponential terms, with time constants ranging from a few minutes to several days.

Viehmann, W.↗
NASA NTRS1976

Intensity and pressure dependence of resonance fluorescence of OH induced by a tunable UV laser

The intensity and pressure dependence of the fluorescence spectrum of OH in the presence of N2 and H2O molecules was studied. Saturation of the absorption transition was observed at low pressures, and the corresponding fluorescence signal was found to vary as the square root of the exciting intensity. This observed dependence agreed with the predicted dependence which took into account the presence of laser modes in the spectrum of the exciting radiation. With full laser power incident, a saturation parameter as high as 3 x 10 to the 5th was observed. The fluorescence spectrum was found to peak at 3145 and at 3090 A, with the relative peak intensities dependent upon gas pressures and upon the particular rotational electronic transition used for excitation. It is concluded that vibrational relaxation of the electronically excited OH due to water vapor in the system plays a dominant role in determining the observed fluorescence spectrum.

Killinger, D. K.↗
NASA NTRS1978

Saturation of fluorescence in flames with a Gaussian laser beam

The method of saturated fluorescence for measuring species concentrations in flames is usually performed with laser beams that do not provide a constant intensity distribution across the focal volume. Because of the intensity distribution across the beam, the fluorescence signal does not depend on laser power or intensity in the same manner as for uniform illumination. This leads to anomolous apparent saturation intensities. In the following, the effect is considered for atomic fluorescence. Relations for the fluorescence signal under two common excitation geometries are derived and uncertainty relations used to consider the benefits of high laser intensity.

Daily, J. W.↗
NASA NTRS1978

Detectability limit and uncertainty considerations for laser induced fluorescence spectroscopy in flames

Laser induced fluorescence spectroscopy of flames is discussed, and derived uncertainty relations are used to calculate detectability limits due to statistical errors. Interferences due to Rayleigh scattering from molecules as well as Mie scattering and incandescence from particles have been examined for their effect on detectability limits. Fluorescence trapping is studied, and some methods for reducing the effect are considered. Fluorescence trapping places an upper limit on the number density of the fluorescing species that can be measured without signal loss.

Daily, J. W.↗
NASA NTRS1978

Laser excited fluorescence in the cesium-xenon excimer and the cesium dimer

Argon ion laser lines are used to excite fluorescence in a mixture of cesium and xenon. Excimer band fluorescence is observed at higher pressures (about 1 atm) while at lower pressures (several torr) a diffuse fluorescence due to the cesium dimer is observed whose character changes with exciting wavelength. The excimer fluorescence is shown to be directly related to the location of the exciting wavelength within previously measured Cs/Xe line shapes. This fact suggests that the excimer systems may be efficiently pumped through these line shapes. Qualitative energy-level schemes are proposed to explain the observations in both the excimer and dimer systems.

Exton, R. J.↗
NASA NTRS1981

Quantitative absorption and fluorescence studies of NO between 1060 and 2000 A

Synchrotron radiation in the 1060 to 2000 A region was used to measure the average absorption and fluorescence cross sections of NO and to determine approximate photodissociation quantum yields. Several vibrational levels of the D(2) sigma(+), E(2) sigma(+), and B(2) delta states have high fluorescence quantum yields. The C(2) and B(2) states do not fluoresce when the excitation energies are above the first dissociation limit, in accord with previous experiments. In general, the fluorescence yields decrease with increasing photon energy. The quantitative measurements are compared with spectroscopic observations and are found to be reasonably consistent.

Guest, J. A.↗
NASA NTRS1981

Fluorescent radiation converter

A fluorescence radiation converter is described which includes a substantially undoped optically transparent substrate and a waveshifter coating deposited on at least one portion of the substrate for absorption of radiation and conversion of fluorescent radiation. The coating is formed to substantially 1000 g/liter of a solvent, 70 to 200 g/liter of an organic polymer, and 0.2 to 25 g/liter of at least one organic fluorescent dye. The incoming incident radiation impinges on the coating. Radiation is absorbed by the fluorescent dye and is re-emitted as a longer wavelength radiation. Radiation is trapped within the substrate and is totally internally reflected by the boundary surface. Emitted radiation leaves the substrate ends to be detected.

Viehmann, W.↗
NASA NTRS1982

Laser measurement of the spectral extinction coefficients of fluorescent, highly absorbing liquids

A conceptual method is developed to deduce rapidly the spectral extinction coefficient of fluorescent, highly absorbing liquids, such as crude or refined petroleum oils. The technique offers the advantage of only requiring one laser wavelength and a single experimental assembly and execution for any specific fluorescent liquid. The liquid is inserted into an extremely thin wedge-shaped cavity for stimulation by a laser from one side and flurescence measurement on the other side by a monochromator system. For each arbitrarily selected extinction wavelength, the wedge is driven slowly to increasing thicknesses until the fluorescence extinguishes. The fluorescence as a function of wedge thickness permits a determination of the extinction coefficient using an included theoretical model. When the monochromator is set to the laser emission wavelength, the extinction coefficient is determined using the usual on-wavelength signal extinction procedure.

Hoge, F. E.↗