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At least 37 records · Page 2

Multi Spectral Fluorescence Imager (MSFI)

Genetic transformation with in vivo reporter genes for fluorescent proteins can be performed on a variety of organisms to address fundamental biological questions. Model organisms that may utilize an ISS imager include unicellular organisms (Saccharomyces cerevisiae), plants (Arabidopsis thaliana), and invertebrates (Caenorhabditis elegans). The multispectral fluorescence imager (MSFI) will have the capability to accommodate 10 cm x 10 cm Petri plates, various sized multi-well culture plates, and other custom culture containers. Features will include programmable temperature and light cycles, ethylene scrubbing (less than 25 ppb), CO2 control (between 400 ppm and ISS-ambient levels in units of 100 ppm) and sufficient airflow to prevent condensation that would interfere with imaging.

Fluorescence↗

Retrieving images from tarnished daguerreotypes using X-ray fluorescence imaging with an X-ray micro beam with tunable energy

We report recent observations using a synchrotron X-ray micro-beam to retrieve images from tarnished 19th century daguerreotypes. We confirm that high quality image can always be retrieved from tarnished plates using Hg L α XRF as long as the bulk of the image particles and their distribution remains intact. We also report results from using tunable tender X-rays (2 - 7 keV) to conduct imaging in high vacuum at energy above the Ag L-edge and the Hg M-edge, extracting images using Ag L α and Hg M α , respectively among others (e.g., S to track corrosion). Images obtained with the surface sensitive total electron yield (TEY) and the bulk sensitive fluorescence yield (FLY) as well as corresponding micro-XANES are reported. Flux tolerance to high intensity X-beam is also explored. These results and their implications for cultural heritage research are discussed.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

PBT2 acts through a different mechanism of action than other 8-hydroxyquinolines: an X-ray fluorescence imaging study

8-Hydroxyquinolines (8HQs) comprise a family of metal-binding compounds that have been used or tested for use in numerous medicinal applications, including as treatments for bacterial infection, Alzheimer's disease, and cancer. Two key 8HQs, CQ (5-chloro-7-iodo-8-hydroxyquinoline) and PBT2 (2-(dimethylamino)methyl-5,7-dichloro-8-hydroxyquinoline), have drawn considerable interest and have been the focus of many studies investigating their in vivo properties. These drugs have been described as copper and zinc ionophores because they do not cause metal depletion, as would be expected for a chelation mechanism, but rather cellular accumulation of these ions. In studies of their anti-cancer properties, CQ has been proposed to elicit toxic intracellular copper accumulation and to trigger apoptotic cancer cell death through several possible pathways. In this study we used synchrotron X-ray fluorescence imaging, in combination with biochemical assays and light microscopy, to investigate 8HQ-induced alterations to metal ion homeostasis, as well as cytotoxicity and cell death. We used the bromine fluorescence from a bromine labelled CQ congener (5,7-dibromo-8-hydroxyquinoline; B2Q) to trace the intracellular localization of B2Q following treatment and found that B2Q crosses the cell membrane. We also found that 8HQ co-treatment with Cu(ii) results in significantly increased intracellular copper and significant cytotoxicity compared with 8HQ treatments alone. PBT2 was found to be more cytotoxic, but a weaker Cu(ii) ionophore than other 8HQs. Moreover, treatment of cells with copper in the presence of CQ or B2Q resulted in copper accumulation in the nuclei, while PBT2-guided copper was distributed near to the cell membrane. Furthermore, these results suggest that PBT2 may be acting through a different mechanism than that of other 8HQs to cause the observed cytotoxicity.

60 APPLIED LIFE SCIENCES↗

Excitation spectral microscopy for highly multiplexed fluorescence imaging and quantitative biosensing

The multiplexing capability of fluorescence microscopy is severely limited by the broad fluorescence spectral width. Spectral imaging offers potential solutions, yet typical approaches to disperse the local emission spectra notably impede the attainable throughput. Here we show that using a single, fixed fluorescence emission detection band, through frame-synchronized fast scanning of the excitation wavelength from a white lamp via an acousto-optic tunable filter, up to six subcellular targets, labeled by common fluorophores of substantial spectral overlap, can be simultaneously imaged in live cells with low (~1%) crosstalks and high temporal resolutions (down to ~10 ms). The demonstrated capability to quantify the abundances of different fluorophores in the same sample through unmixing the excitation spectra next enables us to devise novel, quantitative imaging schemes for both bi-state and Förster resonance energy transfer fluorescent biosensors in live cells. We thus achieve high sensitivities and spatiotemporal resolutions in quantifying the mitochondrial matrix pH and intracellular macromolecular crowding, and further demonstrate, for the first time, the multiplexing of absolute pH imaging with three additional target organelles/proteins to elucidate the complex, Parkin-mediated mitophagy pathway. Together, excitation spectral microscopy provides exceptional opportunities for highly multiplexed fluorescence imaging. The prospect of acquiring fast spectral images without the need for fluorescence dispersion or care for the spectral response of the detector offers tremendous potential.

59 BASIC BIOLOGICAL SCIENCES↗

Utilizing machine learning to improve the precision of fluorescence imaging of cavity-generated spin squeezed states

We present a supervised learning model to calibrate the photon collection rate during the fluorescence imaging of cold atoms. The linear regression model finds the collection rate at each location on the sensor such that the atomic population difference equals that of a highly precise optical cavity measurement. This 192 variable regression results in a measurement variance 27% smaller than our previous single variable regression calibration. The measurement variance is now in agreement with the theoretical limit due to other known noise sources. This model efficiently trains in less than a minute on a standard personal computer's CPU and requires less than 10 min of data collection. Furthermore, the model is applicable across a large change in population difference and across data collected on different days.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Ultrafast two-photon fluorescence imaging of cerebral blood circulation in the mouse brain in vivo

Characterizing blood flow dynamics in vivo is critical to understanding the function of the vascular network under physiological and pathological conditions. Existing methods for hemodynamic imaging have insufficient spatial and temporal resolution to monitor blood flow at the cellular level in large blood vessels. By using an ultrafast line-scanning module based on free-space angular chirped enhanced delay, we achieved two-photon fluorescence imaging of cortical blood flow at 1,000 two-dimensional (2D) frames and 1,000,000 one-dimensional line scans per second in the awake mouse. This orders-of-magnitude increase in temporal resolution allowed us to measure cerebral blood flow at up to 49 mm/s and observe pulsatile blood flow at harmonics of heart rate. Directly visualizing red blood cell (RBC) flow through vessels down to >800 µm in depth, we characterized cortical layer–dependent flow velocity distributions of capillaries, obtained radial velocity profiles and kilohertz 2D velocity mapping of multifile blood flow, and performed RBC flux measurements from penetrating blood vessels.

59 BASIC BIOLOGICAL SCIENCES↗

Computational microscopy for fast widefield deep-tissue fluorescence imaging using a commercial dual-cannula probe

A solid-glass cannula serves as a micro-endoscope that can deliver excitation light deep inside tissue while also collecting emitted fluorescence. Then, we utilize deep neural networks to reconstruct images from the collected intensity distributions. By using a commercially available dual-cannula probe, and training a separate deep neural network for each cannula, we effectively double the field of view compared to prior work. We demonstrated ex vivo imaging of fluorescent beads and brain slices and in vivo imaging from whole brains. We clearly resolved 4 µm beads, with FOV from each cannula of 0.2 mm (diameter), and produced images from a depth of ∼1.2 mm in the whole brain, currently limited primarily by the labeling. Since no scanning is required, fast widefield fluorescence imaging limited primarily by the brightness of the fluorophores, collection efficiency of our system, and the frame rate of the camera becomes possible.

42 ENGINEERING↗

Prostate Cancer Targeted X-Ray Fluorescence Imaging via Gold Nanoparticles Functionalized With Prostate-Specific Membrane Antigen (PSMA)

The gold nanoparticle (GNP) as a promising theranostic probe has been increasingly studied. The tumor-targeting efficiency of GNPs is crucial to increase the therapeutic ratio. In this study, we developed PSMA-targeted GNPs to enhance GNP uptake in prostate cancer and developed an x-ray fluorescence imaging system to noninvasively monitor and assess GNP delivery.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

A Planar-Fluorescence Imaging Technique for Studying Droplet-Turbulence Interactions in Vaporizing Sprays

Droplet turbulence interactions directly affect the vaporization and dispersion of droplets in liquid sprays and therefore play a major role in fuel oxidizer mixing in liquid fueled combustion systems. Proper characterization of droplet turbulence interactions in vaporizing sprays require measurement of droplet size velocity and size temperature correlations. A planar, fluorescence imaging technique is described which is being developed for simultaneously measuring the size, velocity, and temperature of individual droplets in vaporizing sprays. Preliminary droplet size velocity correlation measurements made with this technique are presented. These measurements are also compared to and show very good agreement with measurements made in the same spray using a phase Doppler particle analyzer.

Santavicca, Dom A.↗

Integration of EDWARD readout architecture in full-field fluorescence imaging detector

Data bandwidth, timing resolution and resource utilization in readouts of radiation detectors are a constant challenge. Event driven solutions are pushing against well-trenched framed solutions. The idea for an asynchronous readout architecture called EDWARD ( E vent- D riven W ith A ccess and R eset D ecoder) was presented at the TWEPP 2021 conference. Here we show the progress of our work which resulted in two chip prototypes. The first one, named 3FI65P1, is a full device with the analog pixel circuitry suited for full-field fluorescence imaging. It is already manufactured, and preliminary results are presented. Finally, the second chip, named EDWARD65P1, contains digital pulse generators with Poisson-exponential distribution in each pixel for extraction of the performance matrix of the EDWARD architecture alone.

47 OTHER INSTRUMENTATION↗

Early Career Award: Single Molecule Fluorescence Imaging for a Background-Free Neutrinoless Double Beta Decay Search

This project aimed to develop a suite of novel technologies targeted at detecting individual barium ions produced in neutrinoless double beta decays of 136 Xe, as part of the NEXT program of gaseous xenon detectors. We realized novel molecules which could sense Ba 2+ ions in dry conditions; new microscopy techniques operable in dry noble gases, including the required operating conditions of xenon at 10 bar pressure; and new radiofrequency ion transport methods (developed both theoretically and experimentally) that can transport ions in dense xenon gas. Together, these developments represent a major advance toward a background-free neutrinoless double beta decay experiment based on single molecule fluorescence imaging of Ba 2+ daughter ions.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Hohlraum x-ray preheat asymmetry measurement at the ICF capsule via Mo ball fluorescence imaging

In inertial confinement fusion, penetrating asymmetric hohlraum preheat radiation (>1.8 keV, which includes high temperature coronal M-band emission from laser spots) can lead to asymmetric ablation front and ablator–fuel interface hydrodynamic instability growth in the imploding capsule. First experiments to infer the preheat asymmetries at the capsule were performed on the National Ignition Facility for high density carbon (HDC) capsules in low density fill (0.3 mg/cc 4He) Au hohlraums by time resolved imaging of 2.3 keV fluorescence emission of a smaller Mo sphere placed inside the capsule. Measured Mo emission is pole hot (P2 > 0) since M-band is generated mainly by the outer laser beams as their irradiance at the hohlraum wall is 5× higher than for the inner beams. P2 has a large swing vs time, giving insight into the laser heated hohlraum dynamics. P4 asymmetry is small at the sphere due to efficient geometric smoothing of hohlraum P4 asymmetries at large hohlraum-to-capsule radii ratios. The asymmetry at the HDC capsule is inferred from the Mo emission asymmetry accounting for the Mo/HDC radius difference and HDC capsule opacity.

Dewald, E. L. (ORCID:0000000340546967)↗

Fluorescence Imaging Reveals Surface Contamination

In technique to detect surface contamination, object inspected illuminated by ultraviolet light to make contaminants fluoresce; low-light-level video camera views fluorescence. Image-processing techniques quantify distribution of contaminants. If fluorescence of material expected to contaminate surface is not intense, tagged with low concentration of dye.

Schirato, Richard↗

UV-Fluorescence Imaging of Silicon PV Modules After Outdoor Aging and Accelerated Stress Testing

Fast, non-destructive, outdoor-compatible methods for photovoltaic module characterization are essential for monitoring module quality without disrupting energy production. UV-fluorescence (UVF) imaging of the encapsulant layer can be used for crack detection in the field. We show that UVF patterns vary widely between modules and types of applied stress. We propose that combining accelerated stress testing (e.g. thermal cycling and damp heat) with field investigations can help build understanding of different types of UVF, which are formed and subsequently quenched at varying rates. Ultimately, such understanding could enable estimating when cracks were formed.

41 EE - Solar Energy Technologies Office (EE-4S)↗

Deep UV Native Fluorescence Imaging of Antarctic Cryptoendolithic Communities

An interdisciplinary team at the Jet Propulsion Laboratory Center for Life Detection has embarked on a project to provide in situ chemical and morphological characterization of Antarctic cryptoendolithic microbial communities. We present here in situ deep ultraviolet (UV) native fluorescence and environmental scanning electron microscopy images transiting 8.5 mm into a sandstone sample from the Antarctic Dry Valleys. The deep ultraviolet imaging system employs 224.3, 248.6, and 325 nm lasers to elicit differential fluorescence and resonance Raman responses from biomolecules and minerals. The 224.3 and 248.6 nm lasers elicit a fluorescence response from the aromatic amino and nucleic acids. Excitation at 325 nm may elicit activity from a variety of biomolecules, but is more likely to elicit mineral fluorescence. The resultant fluorescence images provide in situ chemical and morphological maps of microorganisms and the associated organic matrix. Visible broadband reflectance images provide orientation against the mineral background. Environmental scanning electron micrographs provided detailed morphological information. The technique has made possible the construction of detailed fluorescent maps extending from the surface of an Antarctic sandstone sample to a depth of 8.5 mm. The images detect no evidence of microbial life in the superficial 0.2 mm crustal layer. The black lichen component between 0.3 and 0.5 mm deep absorbs all wavelengths of both laser and broadband illumination. Filamentous deep ultraviolet native fluorescent activity dominates in the white layer between 0.6 mm and 5.0 mm from the surface. These filamentous forms are fungi that continue into the red (iron-rich) region of the sample extending from 5.0 to 8.5 mm. Using differential image subtraction techniques it is possible to identify fungal nuclei. The ultraviolet response is markedly attenuated in this region, apparently from the absorption of ultraviolet light by iron-rich particles coating the filaments. Below 8.5 mm the filamentous morphology of the upper layers gives way to punctate 1-2 micron particles evidencing fluorescent activity following excitation at both deep ultraviolet wavelengths.

Storrie-Lombardi, M. C.↗

Ultraviolet Fluorescence Imaging for Photovoltaic Module Metrology: Best Practices and Survey of Features Observed in Fielded Modules

As the photovoltaics (PV) industry grows in sophistication, so must the extent to which systems are characterized. UV Fluorescence (UVF) imaging is a valuable, easy-to-perform, high-throughput, nonintrusive technique for characterizing modules in the field and in the lab. However, UVF is still a relatively new technique, and many in the PV industry are still unaware of its potential. We provide a guideline for obtaining, processing, and interpreting UVF images. We have provided a list of considerations for imaging hardware and settings, a suggested pipeline for image processing, and details on a survey of features shown in UVF images. As a result, a new database with UVF images of 7190 modules and another database curated by BrightSpot Automation are publicly available.

14 SOLAR ENERGY↗

Laser-induced-fluorescence imaging of a spin-polarized ultracold neutral plasma in a magnetic field

In this work, we report Doppler-sensitive laser-induced-fluorescence (LIF) imaging of an ultracold neutral plasma in a magnetic field. Local values of ion density, hydrodynamic fluid velocity, temperature, and spin polarization are obtained using a fluorescence model based on velocity-resolved rate equations (REs) including the transfer of ions between states due to laser coupling and spontaneous emission. The RE approach captures optical pumping of ions into states that are not driven by the LIF excitation laser, and this is validated with experimental data. Combined molecular-dynamics and quantum-trajectories simulations verify that velocity-changing collisions have a negligible impact on the state population evolution for typical experimental conditions. Relative intensities of Zeeman components of the LIF spectra provide clear evidence that the ions are electron-spin-polarized when created by photoionization of magnetically trapped 88 Sr atoms. This probe opens many possibilities for studying thermal transport and the equilibration of neutral plasmas in overlapping regimes of strong coupling and magnetization.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗