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All sourcesNASA NTRSDOE OSTI
NASA NTRS2002

Rationally designed fluorescently labeled sulfate-binding protein mutants: evaluation in the development of a sensing system for sulfate

Periplasmic binding proteins from E. coli undergo large conformational changes upon binding their respective ligands. By attaching a fluorescent probe at rationally selected unique sites on the protein, these conformational changes in the protein can be monitored by measuring the changes in fluorescence intensity of the probe which allow the development of reagentless sensing systems for their corresponding ligands. In this work, we evaluated several sites on bacterial periplasmic sulfate-binding protein (SBP) for attachment of a fluorescent probe and rationally designed a reagentless sensing system for sulfate. Eight different mutants of SBP were prepared by employing the polymerase chain reaction (PCR) to introduce a unique cysteine residue at a specific location on the protein. The sites Gly55, Ser90, Ser129, Ala140, Leu145, Ser171, Val181, and Gly186 were chosen for mutagenesis by studying the three-dimensional X-ray crystal structure of SBP. An environment-sensitive fluorescent probe (MDCC) was then attached site-specifically to the protein through the sulfhydryl group of the unique cysteine residue introduced. Each fluorescent probe-conjugated SBP mutant was characterized in terms of its fluorescence properties and Ser171 was determined to be the best site for the attachment of the fluorescent probe that would allow for the development of a reagentless sensing system for sulfate. Three different environment-sensitive fluorescent probes (1,5-IAEDANS, MDCC, and acylodan) were studied with the SBP171 mutant protein. A calibration curve for sulfate was constructed using the labeled protein and relating the change in the fluorescence intensity with the amount of sulfate present in the sample. The detection limit for sulfate was found to be in the submicromolar range using this system. The selectivity of the sensing system was demonstrated by evaluating its response to other anions. A fast and selective sensing system with detection limits for sulfate in the submicromolar range was developed. Copyright 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 78: 517-526, 2002.

Evaluation Studies↗
NASA NTRS2013

Global Monitoring of Terrestrial Chlorophyll Fluorescence from Moderate-Spectral-Resolution Near-Infrared Satellite Measurements: Methodology, Simulations, and Application to GOME-2

Globally mapped terrestrial chlorophyll fluorescence retrievals are of high interest because they can provide information on the functional status of vegetation including light-use efficiency and global primary productivity that can be used for global carbon cycle modeling and agricultural applications. Previous satellite retrievals of fluorescence have relied solely upon the filling-in of solar Fraunhofer lines that are not significantly affected by atmospheric absorption. Although these measurements provide near-global coverage on a monthly basis, they suffer from relatively low precision and sparse spatial sampling. Here, we describe a new methodology to retrieve global far-red fluorescence information; we use hyperspectral data with a simplified radiative transfer model to disentangle the spectral signatures of three basic components: atmospheric absorption, surface reflectance, and fluorescence radiance. An empirically based principal component analysis approach is employed, primarily using cloudy data over ocean, to model and solve for the atmospheric absorption. Through detailed simulations, we demonstrate the feasibility of the approach and show that moderate-spectral-resolution measurements with a relatively high signal-to-noise ratio can be used to retrieve far-red fluorescence information with good precision and accuracy. The method is then applied to data from the Global Ozone Monitoring Instrument 2 (GOME-2). The GOME-2 fluorescence retrievals display similar spatial structure as compared with those from a simpler technique applied to the Greenhouse gases Observing SATellite (GOSAT). GOME-2 enables global mapping of far-red fluorescence with higher precision over smaller spatial and temporal scales than is possible with GOSAT. Near-global coverage is provided within a few days. We are able to show clearly for the first time physically plausible variations in fluorescence over the course of a single month at a spatial resolution of 0.5 0.5. We also show some significant differences between fluorescence and coincident normalized difference vegetation indices (NDVI) retrievals.

Fluorescence↗
NASA NTRS2013

Global Monitoring of Terrestrial Chlorophyll Fluorescence from Moderate-spectral-resolution Near-infrared Satellite Measurements: Methodology, Simulations, and Application to GOME-2

Globally mapped terrestrial chlorophyll fluorescence retrievals are of high interest because they can provide information on the functional status of vegetation including light-use efficiency and global primary productivity that can be used for global carbon cycle modeling and agricultural applications. Previous satellite retrievals of fluorescence have relied solely upon the filling-in of solar Fraunhofer lines that are not significantly affected by atmospheric absorption. Although these measurements provide near-global coverage on a monthly basis, they suffer from relatively low precision and sparse spatial sampling. Here, we describe a new methodology to retrieve global far-red fluorescence information; we use hyperspectral data with a simplified radiative transfer model to disentangle the spectral signatures of three basic components: atmospheric absorption, surface reflectance, and fluorescence radiance. An empirically based principal component analysis approach is employed, primarily using cloudy data over ocean, to model and solve for the atmospheric absorption. Through detailed simulations, we demonstrate the feasibility of the approach and show that moderate-spectral-resolution measurements with a relatively high signal-to-noise ratio can be used to retrieve far-red fluorescence information with good precision and accuracy. The method is then applied to data from the Global Ozone Monitoring Instrument 2 (GOME-2). The GOME-2 fluorescence retrievals display similar spatial structure as compared with those from a simpler technique applied to the Greenhouse gases Observing SATellite (GOSAT). GOME-2 enables global mapping of far-red fluorescence with higher precision over smaller spatial and temporal scales than is possible with GOSAT. Near-global coverage is provided within a few days. We are able to show clearly for the first time physically plausible variations in fluorescence over the course of a single month at a spatial resolution of 0.5 deg × 0.5 deg. We also show some significant differences between fluorescence and coincident normalized difference vegetation indices (NDVI) retrievals.

remote sensing↗
NASA NTRS1985

Laser-induced fluorescence of green plants. III - LIF spectral signatures of five major plant types

A technique amenable to remote sensing use which utilizes laser-induced fluorescence (LIF) properties of plants has been successfully used in the laboratory to identify five major plant types. These included herbaceous dicots, herbaceous monocots, conifers, hardwoods, and algae. Each of these plant types exhibited a characteristic LIF spectra when excited by a pulsed N2 laser emitting at 337 nm. Although monocots and dicots possess common fluorescence maxima at 440, 685, and 740 nm, they could be differentiated from one another by using the ratio of the square of the fluorescence intensity at 440 nm to the nonsquared intensity at 685 nm, i.e., (440)-squared/685. In all cases, monocots yielded a significantly higher ratio. Conifers have fluorescence maxima at 440, 525, and 740 nm but none at 685 nm. Hardwoods exhibited fluorescence at 440, 525, 685, and 740 nm. Algae had very low fluorescence at 440 nm, no fluorescence at 525 nm, and fluorescence maxima at 685 and 740 nm. For algae, the ratio of the fluorescence intensity at 685 nm to that at 740 nm was much greater than that for monocots, dicots, and hardwoods. The potential use of the LIF technique for individual species identification is suggested.

Chappelle, E. W.↗
NASA NTRS2000

Advances in Fluorescence Sensing Systems for the Remote Assessment of Nitrogen Supply in Field Corn

The studies described herein were conducted to better define changes in fluorescence properties of leaves from field grown corn (Zea mays L.) as they relate to varying levels of nitrogen (N) fertilization. This research was directed toward: 1) providing a remote non-destructive sensing technique to aid in the determination of optimal rates of N fertilization in corn crops and, 2) defining parameters for further development of fluorescence instrumentation to be operated remotely at field canopy levels. Fluorescence imaging bands centered in the blue (450 nm), green (525 nm), red (680 nm), and far-red (740 nm) and ratios of these bands were compared with the following plant parameters: rates of photosynthesis, N:C ratio, pigment concentrations, and grain yields. Both the fluorescence and physiological measures exhibited similar curvilinear responses to N fertilization level while significant linear correlations were obtained among fluorescence bands and band ratios to certain physiological measures of plant productivity. The red / blue, red / green, far-red / blue, far-red /green fluorescence ratios are well suited for remote observation and provided high correlations to grain yield, LAI, N:C, and chlorophyll contents. The results from this investigation indicate that fluorescence technology could aid in the determination of N fertilization requirements for corn. This discussion will also address design concepts and preliminary field trials of a mobile field-based Laser Induced Fluorescence Imaging System (LIFIS) capable of simultaneously acquiring images of four fluorescence emission bands from areas of plant canopies equaling 1 sq m and greater without interference of ambient solar radiation.

Corp, L. A.↗
NASA NTRS2007

Mosaic-Detector-Based Fluorescence Spectral Imager

A battery-powered, pen-sized, portable instrument for measuring molecular fluorescence spectra of chemical and biological samples in the field has been proposed. Molecular fluorescence spectroscopy is among the techniques used most frequently in laboratories to analyze compositions of chemical and biological samples. Heretofore, it has been possible to measure fluorescence spectra of molecular species at relative concentrations as low as parts per billion (ppb), with a few nm spectral resolution. The proposed instrument would include a planar array (mosaic) of detectors, onto which a fluorescence spectrum would be spatially mapped. Unlike in the larger laboratory-type molecular fluorescence spectrometers, mapping of wavelengths to spatial positions would be accomplished without use of relatively bulky optical parts. The proposed instrument is expected to be sensitive enough to enable measurement of spectra of chemical species at relative concentrations <1 ppb, with spectral resolution that could be tailored by design to be comparable to a laboratory molecular fluorescence spectrometer. The proposed instrument (see figure) would include a button-cell battery and a laser diode, which would generate the monochromatic ultraviolet light needed to excite fluorescence in a sample. The sample would be held in a cell bounded by far-ultraviolet-transparent quartz or optical glass. The detector array would be, more specifically, a complementary metal oxide/ semiconductor or charge-coupled- device imaging photodetector array, the photodetectors of which would be tailored to respond to light in the wavelength range of the fluorescence spectrum to be measured. The light-input face of the photodetector array would be covered with a matching checkerboard array of multilayer thin film interference filters, such that each pixel in the array would be sensitive only to light in a spectral band narrow enough so as not to overlap significantly with the band of an adjacent pixel. The wavelength interval between adjacent pixels (and, thus, the spectral resolution) would typically be chosen by design to be approximately equal to the width of the total fluorescence wavelength range of interest divided by the number of pixels. The unitary structure comprising the photodetector array overlaid with the matching filter array would be denoted a hyperspectral mosaic detector (HMD) array.

Son, Kyung-Ah↗
NASA NTRS2004

Phytoplankton-Fluorescence-Lifetime Vertical Profiler

A battery-operated optoelectronic instrument is designed to be lowered into the ocean to measure the intensity and lifetime of fluorescence of chlorophyll A in marine phytoplankton as a function of depth from 0 to 300 m. Fluorescence lifetimes are especially useful as robust measures of photosynthetic productivity of phytoplankton and of physical and chemical mechanisms that affect photosynthesis. The knowledge of photosynthesis in phytoplankton gained by use of this and related instruments is expected to contribute to understanding of global processes that control the time-varying fluxes of carbon and associated biogenic elements in the ocean. The concentration of chlorophyll in the ocean presents a major detection challenge because in order to obtain accurate values of photosynthetic parameters, the intensity of light used to excite fluorescence must be kept very low so as not to disturb the photosynthetic system. Several innovations in fluorometric instrumentation were made in order to make it possible to reach the required low detection limit. These innovations include a highly efficient optical assembly with an integrated flow-through sample interface, and a high-gain, low-noise electronic detection subsystem. The instrument also incorporates means for self-calibration during operation, and electronic hardware and software for control, acquisition and analysis of data, and communications. The electronic circuitry is highly miniaturized and designed to minimize power demand. The instrument is housed in a package that can withstand the water pressure at the maximum depth of 300 m. A light-emitting diode excites fluorescence in the sample flow cell, which is placed at one focal point of an ellipsoidal reflector. A photomultiplier tube is placed at the other focal point. This optical arrangement enables highly efficient collection of fluorescence emitted over all polar directions. Fluorescence lifetime is measured indirectly, by use of a technique based on the same principle as the one described in "Fluorometer for Analysis of Photosynthesis in Phytoplankton" (SSC-00110), NASA Tech Briefs, Vol. 24, No. 1 (November 2000), page 79. The excitation is modulated at a frequency of 70 MHz, and the phase shift between the excitation light and the emitted fluorescence is measured by a detection method in which the 70 MHz signal is down-converted to a 400 Hz signal. The fluorescence lifetime can be computed from the known relationship among the fluorescence lifetime, phase shift, and modulation frequency

Fernandez, Salvador M.↗
NASA NTRS1973

Detection of S02 and N02 in Stack Plume by Raman Scattering and Fluorescence

We have been studying laser-Raman radar which can be used as the remote detector of sulfur dioxide concentration in the stack plume of boiler exhaust gas, and some results have been published. In this paper, we report the interference of nitrogen dioxide fluorescence against sulfur dioxide Raman scattering and the measuring method of sulfur dioxide and nitrogen dioxide concentration. In a stack plume, high density dust and high concentration carbon dioxide are included, therefore very strong Mie back-scattering and carbon dioxide Raman scattering are observed. The separation of these scattering signals from sulfur dioxide Raman signal was the first problem for the laser-Raman radar. But, this problem was solved by using the filter which have high resolving power. It is well known that nitrogen dioxide can be excited to emit fluorescence of broad spectrum by blue-green light. The light source of the laser-Raman radar is SH of Nd:YAG laser, and boiler exhaust gas includes several tens ppm of nitrogen dioxide, then the interference of nitrogen dioxide fluorescence brings the error to sulfur dioxide measurement. 'rhe rejection of the interference can be achieved by the subtraction of the nitrogen dioxide fluorescence contribution from detected signal at sulfur dioxide Raman scattering wavelength. The nitrogen dioxide fluorescence contribution can be measured by two methods. The first is to convert the nitrogen dioxide fluorescence intensity which is measured at a different wavelength from sulfur dioxide Raman line into the one at sulfur dioxide Raman line. The second is to convert the detected signal intensity, which is obtained when the range gate is set just behind the plume, into the one obtained at the range of the plume using the difference of the time dependence between sulfur dioxide Raman scattering and nitrogen dioxide fluorescence. Nitrogen dioxide fluorescence has lifetime of about 300nS, while Raman scattering has none. By either of the two methods, the contribution of nitrogen dioxide fluorescence is determined. Then, we can measure the nitrogen dioxide and sulfur dioxide concentration.The minimum detectable concentration and the experimental results of the remote sensing of sulfur dioxide and nitrogen dioxide in stack plume will be discussed in detail.

Nakahara, S.↗
DOE OSTI2014

Three-dimensional x-ray fluorescence mapping of a gold nanoparticle-loaded phantom

Purpose : X-ray fluorescence (XRF) is a promising technique with sufficient specificity and sensitivity for identifying and quantifying features in small samples containing high atomic number (Z) materials such as iodine, gadolinium, and gold. In this study, the feasibility of applying XRF to early breast cancer diagnosis and treatment is studied using a novel approach for three-dimensional (3D) x-ray fluorescence mapping (XFM) of gold nanoparticle (GNP)-loaded objects in a physical phantom at the technical level. Methods : All the theoretical analysis and experiments are conducted under the condition of using x-ray pencil beam and a compactly integrated x-ray spectrometer. The penetrability of the fluorescence x-rays from GNPs is first investigated by adopting a combination of BR12 with 70 mm/50 mm in thickness on the excitation/emission path to mimic the possible position of tumor goldin vivo. Then, a physical phantom made of BR12 is designed to translate in 3D space with three precise linear stages and subsequently the step by step XFM scanning is performed. The experimental technique named as background subtraction is applied to isolate the gold fluorescence from each spectrum obtained by the spectrometer. Afterwards, the attenuations of both the incident primary x-ray beam with energies beyond the gold K-edge energy (80.725 keV) and the isolated gold K{sub α} fluorescence x-rays (65.99 –69.80 keV) acquired after background subtraction are well calibrated, and finally the unattenuated K{sub α} fluorescence counts are used to realize mapping reconstruction and to describe the linear relationship between gold fluorescence counts and corresponding concentration of gold solutions. Results : The penetration results show that the goldK{sub α} fluorescence x-rays have sufficient penetrability for this phantom study, and the reconstructed mapping results indicate that both the spatial distribution and relative concentration of GNPs within the designed BR12 phantom can be well identified and quantified. Conclusions : Although the XFM method in this investigation is still studied at the technical level and is not yet practical for routinein vivo mapping tasks with GNPs, the current penetrability measurements and phantom study strongly suggest the feasibility to establish and develop a 3D XFM system.

61 RADIATION PROTECTION AND DOSIMETRY↗
DOE OSTI2014

TU-F-18A-05: An X-Ray Fluorescence Technique for Energy Calibration of Photon-Counting Detectors

Purpose: To investigate the feasibility of energy response calibration of a Si strip photon-counting detector by using the x-ray fluorescence technique. Methods: X-ray fluorescence was generated by using a pencil beam from a tungsten anode x-ray tube with 2 mm Al filtration. Spectra were acquired at 90° from the primary beam direction with an energy-resolved photon-counting detector based on Si strips. The distances from the source to target and the target to detector were approximately 19 and 11 cm, respectively. Four different materials, containing Ag, I, Ba, and Gd, were placed in small plastic aliquots with a diameter of approximately 0.7 cm for x-ray fluorescence measurements. Linear regression analysis was performed to derive the gain and offset values for the correlation between the measured fluorescence peak center and the known energies for materials. The energy resolution was derived from the full width at half maximum (FWHM) of the fluorescence peaks. In addition, the angular dependence of the recorded fluorescence spectra was studied at 30°, 60°, and 120°. Results: Strong fluorescence signals of all four target materials were recorded with the investigated geometry for the Si strip detector. The recorded pulse height was calibrated with respect to photon energy and the gain and offset values were calculated to be 7.0 mV/keV and −69.3 mV, respectively. Negligible variation in energy calibration was observed among the four energy thresholds. The variation among different pixels was estimated to be approximately 1 keV. The energy resolution of the detector was estimated to be 7.9% within the investigated energy range. Conclusion: The performance of a spectral imaging system using energy-resolved photon-counting detectors is very dependent on the energy calibration of the detector. The proposed x-ray fluorescence technique provides an accurate and efficient way to calibrate the energy response of a photon-counting detector.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗
DOE OSTI2017

Fluorescence and confocal imaging of mammalian cells using conjugated oligoelectrolytes with phenylenevinylene core

Over the last few years, considerable efforts are taken, in order to find a molecular fluorescent probe fulfilling their applicability requirements. Due to a good optical properties and affinity to biological structures conjugated oligoelectrolytes (COEs) can be considered as a promising dyes for application in fluorescence-based bioimaging. In this work, we synthetized COEs with phenylenevinylene core (PV-COEs) and applied as fluorescent membranous-specific probes. Cytotoxicity effects of each COE were probed on cancerous and non-cancerous cell types and little to no toxicity effects were observed at the high range of concentrations. The intensity of cell fluorescence following the COE staining was determined by the photoluminescence analysis and fluorescence activated cell sorting method (FACS). Intercalation of tested COEs into mammalian cell membranes was revealed by fluorescent and confocal microscopy colocalization with commercial dyes specific for cellular structures including mitochondria, Golgi apparatus and endoplasmic reticulum. The phenylenevinylene conjugated oligoelectrolytes have been found to be suitable for fluorescent bioimaging of mammalian cells and membrane-rich organelles. Due to their water solubility coupled with spontaneous intercalation into cells, favorable photophysical features, ease of cell staining, low cytotoxicity and selectivity for membranous structures, PV-COEs can be applied as markers for fluorescence imaging of a variety of cell types.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2015

Latest methods of fluorescence-based protein crystal identification

Fluorescence, whether intrinsic or by using trace fluorescent labeling, can be a powerful aid in macromolecule crystallization. Its use in screening for crystals is discussed here. Successful protein crystallization screening experiments are dependent upon the experimenter being able to identify positive outcomes. The introduction of fluorescence techniques has brought a powerful and versatile tool to the aid of the crystal grower. Trace fluorescent labeling, in which a fluorescent probe is covalently bound to a subpopulation (<0.5%) of the protein, enables the use of visible fluorescence. Alternatively, one can avoid covalent modification and use UV fluorescence, exploiting the intrinsic fluorescent amino acids present in most proteins. By the use of these techniques, crystals that had previously been obscured in the crystallization drop can readily be identified and distinguished from amorphous precipitate or salt crystals. Additionally, lead conditions that may not have been obvious as such under white-light illumination can be identified. In all cases review of the screening plate is considerably accelerated, as the eye can quickly note objects of increased intensity.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗
DOE OSTI2015

Trace fluorescent labeling for protein crystallization

The presence of a covalently bound fluorescent probe at a concentration of <0.5% does not affect the outcome of macromolecule crystallization screening experiments. Additionally, the fluorescence can be used to determine new, not immediately apparent, lead crystallization conditions. Fluorescence can be a powerful tool to aid in the crystallization of proteins. In the trace-labeling approach, the protein is covalently derivatized with a high-quantum-yield visible-wavelength fluorescent probe. The final probe concentration typically labels ≤0.20% of the protein molecules, which has been shown to not affect the crystal nucleation or diffraction quality. The labeled protein is then used in a plate-screening experiment in the usual manner. As the most densely packed state of the protein is the crystalline form, then crystals show as the brightest objects in the well under fluorescent illumination. A study has been carried out on the effects of trace fluorescent labeling on the screening results obtained compared with nonlabeled protein, and it was found that considering the stochastic nature of the crystal nucleation process the presence of the probe did not affect the outcomes obtained. Other effects are realised when using fluorescence. Crystals are clearly seen even when buried in precipitate. This approach also finds ‘hidden’ leads, in the form of bright spots, with ∼30% of the leads found being optimized to crystals in a single-pass optimization trial. The use of visible fluorescence also enables the selection of colors that bypass interfering substances, and the screening materials do not have to be UV-transparent.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗
DOE OSTI2015

SU-C-303-05: Photosensitizer Determination for PDT Using Interstitial and Surface Measurements of Fluorescence

Purpose: Photosensitizer concentration during photodynamic therapy (PDT) is an important parameter for accurate dosimetry. Fluorescence signal can be used as a measure of photosensitizer concentration. Two methods of data acquisition were compared to an ex vivo study both for in vivo and phantom models. Methods: Fluorescence signal of commonly used photosensitizer benzoporphyrin derivative monoacid ring A (BPD) was obtained in phantoms and mouse tumors using an excitation light of 405 nm. Interstitial fluorescence signal was obtained using a side-cut fiber inserted into the tumor tissue of interest. Using a previously developed multi-fiber probe, tumor surface fluorescence measurements were also collected. Signals were calibrated according to optical phantoms with known sensitizer fluorescence. Optical properties for each sample were determined and the influence of different absorption and scattering properties on the fluorescence signals was investigated. Using single value decomposition of the spectra, the sensitizer concentration was determined using the two different measurement geometries. An ex vivo analysis was also performed for tumor samples to determine the sensitizer concentration. Results: The two fluorescence signals obtained from the surface multi-fiber probe and the interstitial measurements were compared and were corresponding for both phantoms and mouse models. The values obtained were comparable to the ex vivo measurements as well. Despite the difference in geometry, the surface probe measurements can still be used as a metric for determining the presence of sensitizer in small volume tumors. Conclusion: The multi-fiber contact probe can be used as a tool to measure fluorescence at the surface of the treatment area for PDT and predict sensitizer concentration throughout the tumor. This is advantageous in that the measurement does not damage any tissue. Future work will include investigating the dependence of these results on intratumor sensitizer distribution.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2019

Fluorescent Tissue Assessment of Colorectal Cancer Liver Metastases Ablation Zone: A Potential Real-Time Biomarker of Complete Tumor Ablation

Background: This study aimed to evaluate whether rapid fluorescent tissue examination immediately after colorectal cancer liver metastasis (CLM) ablation correlates with standard pathologic and immunohistochemical (IHC) assessments. Methods: This prospective, National Institutes of Health-supported study enrolled 34 consecutive patients with 53 CLMs ablated between January 2011 and December 2014. Immediately after ablation, core needle sampling of the ablation zone was performed. Tissue samples were evaluated with fluorescent viability (MitoTracker Red) and nuclear (Hoechst) stains. Confocal microscope imaging was performed within 30 min after ablation. The same samples were subsequently fixed and stained with hematoxylin and eosin (H&E). Identified tumor cells underwent IHC staining for proliferation (Ki67) and viability (OxPhos). The study pathologist, blinded to the H&E and IHC assessment, evaluated the fluorescent images separately to detect viable tumor cells. Sensitivity, specificity, and overall concordance of the fluorescent versus H&E and IHC assessments were calculated. Results: A total of 63 tissue samples were collected and processed. The overall concordance rate between the immediate fluorescent and the subsequent H&E and IHC assessments was 94% (59/63). The fluorescent assessment sensitivity and specificity for the identification of tumor cells were respectively 100% (18/18) and 91% (41/45). Conclusions: The study showed a high concordance rate between the immediate fluorescent assessment and the standard H&E and IHC assessment of the ablation zone. Given the documented prognostic value of ablation zone tissue characteristics for outcomes after ablation of CLM, the fluorescent assessment offers a potential intra-procedural biomarker of complete tumor ablation.

62 RADIOLOGY AND NUCLEAR MEDICINE↗
DOE OSTI2019

Polymer concentration detection method based on fluorescent polymer to evaluate its retention and percolation

ABSTRACT A new fluorescent polymer was prepared by copolymerization of acrylamide and a modified rhodamine B (RhB). Fourier transformation infrared spectrometer and nuclear magnetic resonance confirmed the successful modification of RhB and the synthesis of fluorescent polymer. The rheological and fluorescence properties were studied by fluorospectrophotometer and rotational rheometer. The adsorption retention and percolation properties were evaluated by sand pack displacement experiments. The results indicated that fluorescence intensity revealed a distinct linear relationship with polymer concentration within a certain concentration range. High temperature and strong acidic conditions could diminish fluorescence intensity whereas formation water ions exerted inconsiderable influence on it. Rheological results manifested that fluorescent polymer can be used as polymer flooding agent. The sand pack displacement results proved the feasibility of using fluorescent polymer to detect polymer concentration. By this means, polymer breakthrough time and concentration variations could be confirmed. Based on the curves of injection pressure and polymer concentration, the percolation state and retention of polymer were able to be analyzed. Significantly, this work can provide support and guidance for the practical applications of fluorescent polymer. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019 , 136 , 47468.

Kang, Wanli↗
DOE OSTI2026

A Chimeric LBT-GFP Biosensor Exhibits Antithetical Fluorescence Responses to Ca 2+ and Dy 3+ Binding

Rare earth elements (REEs) are critical components in emerging technologies, but their mining and refining processes are often laborious, costly, and environmentally damaging. Developing green and efficient separation methods for REEs is crucial. Biomolecular approaches using lanthanide-binding proteins and peptides show promise for selective REE extraction and separation. In this study, we present the design and characterization of a genetically encoded fluorescence indicator (GEFI) construct that combines a superfolder green fluorescent protein (sfGFP) with a dual lanthanide-binding tag (2×dLBT). The 2×dLBT insert induces conformational changes in sfGFP upon lanthanide binding, modulating the fluorescence intensity. The sfGFP-2×dLBT biosensor exhibited distinct fluorescence responses to different lanthanide ions, with the highest dynamic range observed for heavy REEs like dysprosium (Dy 3+ ). Interestingly, the sensor displayed an antithetical response, where low concentrations of lanthanides initially quenched the fluorescence, but higher concentrations led to a significant fluorescence increase (1.5-fold). The Ca 2+ ion on the other hand showed only a dose-dependent quenching of the fluorescence response. Based on these observations, the biphasic response of the biosensor to lanthanides was eliminated by pretreating the sensor with calcium, which further expanded the dynamic range up to 3-fold for Dy 3+ . The lanthanide-selective and concentration-dependent fluorescence changes of the sfGFP-2×dLBT biosensor demonstrate its potential as a platform for developing specific sensors for various REEs. These sensors could enable rapid and cost-effective determination of REE composition in complex mixtures, facilitating the separation and recovery of critical REEs from electronic waste and other REE-containing sources.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Fluorescent CQD-Doped Styrene Acrylic Emulsion Coating Film with Enhanced Optical Properties

Styrene acrylic emulsions (SAEs) have emerged as a promising material for water-based coatings. However, they are still limited by their own defects in practical applications, poor weatherability, and degradation of performance at lower or higher temperatures. Here, we introduce a facile approach to producing fluorescent carbon quantum dots (CQDs) from wood processing residues and fabricating fluorescent CQD/SAE coating films via emulsion-casting. The addition of the fluorescent CQDs enhanced the optical performance of the CQD/SAE coating films. The fluorescent CQDs were prepared via a hydrothermal approach and were obtained after heating at 180 °C for 6 h at a reaction concentration of 50 mg/mL. The synthesized CQDs resulted in a high fluorescence, and the CQDs had an average size of 1.63 nm. Various concentrations of the fluorescent CQDs were doped into the SAE coating film, which improved its optical properties. We also characterized and discussed the products and then explored their optical properties. This study presents the potential of fluorescent CQD/SAE coating films for applications in anti-counterfeiting coatings, fluorescent adhesives, and papermaking.

36 MATERIALS SCIENCE↗