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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 37 records · Page 2

A modular chemigenetic calcium indicator for multiplexed in vivo functional imaging

Abstract Genetically encoded fluorescent calcium indicators allow cellular-resolution recording of physiology. However, bright, genetically targetable indicators that can be multiplexed with existing tools in vivo are needed for simultaneous imaging of multiple signals. Here we describe WHaloCaMP, a modular chemigenetic calcium indicator built from bright dye-ligands and protein sensor domains. Fluorescence change in WHaloCaMP results from reversible quenching of the bound dye via a strategically placed tryptophan. WHaloCaMP is compatible with rhodamine dye-ligands that fluoresce from green to near-infrared, including several that efficiently label the brain in animals. When bound to a near-infrared dye-ligand, WHaloCaMP shows a 7× increase in fluorescence intensity and a 2.1-ns increase in fluorescence lifetime upon calcium binding. We use WHaloCaMP1a to image Ca 2+ responses in vivo in flies and mice, to perform three-color multiplexed functional imaging of hundreds of neurons and astrocytes in zebrafish larvae and to quantify Ca 2+ concentration using fluorescence lifetime imaging microscopy (FLIM).

Biochemistry & Molecular Biology↗

Seeing in with X-rays: 4D Strain and Thermometry Measurements for Thermal-Mechanical Testing

Understanding temperature-dependent material decomposition and structural deformation induced by combined thermal-mechanical environments is critical for safety qualification of hardware under accident scenarios. Seeing in with X-rays elucidated the physics necessary to develop X-ray strain and thermometry diagnostics for use in optically opaque environments. Two parallel thermometry schemes were explored: X-ray fluorescence and X-ray diffraction of inorganic doped ceramics– colloquially known as thermographic phosphors. Two parallel surface strain techniques–Path-Integrated Digital Image Correlation and Frequency Multiplexed Digital Image Correlation–were demonstrated. Finally, preliminary demonstration of time-resolved digital volume correlation was performed by taking advantage of limited view reconstruction techniques. Additionally, research into blended ceramic-metal coatings was critical to generating intrinsic thermographic patterns for the future combination of X-ray strain and thermometry measurements.

36 MATERIALS SCIENCE↗

Method of Stamping Surface-Enhance Raman Spectroscopy for Label-Free, Multiplexed, Molecular Sensing and Imaging

The present disclosure relates the use of a stamping surface enhanced Raman scattering (S-SERS) technique with nanoporous gold disk (NPGD) plasmonic substrates to produce a label-free, multiplexed molecular sensing and imaging technique. A NPGD SERS substrate is stamped onto a surface containing one or more target molecules, followed by SERS measurement of the target molecules located between the surface and SERS substrate. The target molecules may be deposited on the surface, which may be a carrier substrate such as polydimethylsiloxane (PDMS).

Shih, Wei-Chuan↗

Large CMOS imager using hadamard transform based multiplexing

We have developed a concept design for a large (~10k x 10k) CMOS imaging array whose elements are grouped in small subarrays with N pixels in each. The subarrays are code-division multiplexed using the Hadamard Transform (HT) based encoding. The Hadamard code improves the signal-to-noise (SNR) ratio to the reference of the read-out amplifier by a factor of N^1/2. This way of grouping pixels reduces the number of hybridization bumps by N. A single chip layout has been designed and the architecture of the imager has been developed to accommodate the HT base multiplexing into the existing CMOS technology. The imager architecture allows for a trade-off between the speed and the sensitivity. The envisioned imager would operate at a speed >100 fps with the pixel noise < 20 e-. The power dissipation would be ~100 pW/pixe1. The combination of the large format, high speed, high sensitivity and low power dissipation can be very attractive for space reconnaissance applications.

CMOS imager↗

Infrared Linear Imaging Array

Photodiodes, multiplexer, and preamp reside in hybrid ceramic package. Hybrid microcircuit produces 128-element line images of infrared scenes. Each picture element is 0.008-in. (0.2-mm) square. Even without image data processing, microcircuit resolves temperature differences as small as 0.05 degrees C in infrared images.

Bailey, G. C.↗

Single-cell spatial metabolomics with cell-type specific protein profiling for tissue systems biology

Metabolic reprogramming in cancer and immune cells occurs to support their increasing energy needs in biological tissues. Here we propose Single Cell Spatially resolved Metabolic (scSpaMet) framework for joint protein-metabolite profiling of single immune and cancer cells in male human tissues by incorporating untargeted spatial metabolomics and targeted multiplexed protein imaging in a single pipeline. We utilized the scSpaMet to profile cell types and spatial metabolomic maps of 19507, 31156, and 8215 single cells in human lung cancer, tonsil, and endometrium tissues, respectively. The scSpaMet analysis revealed cell type-dependent metabolite profiles and local metabolite competition of neighboring single cells in human tissues. Deep learning-based joint embedding revealed unique metabolite states within cell types. Trajectory inference showed metabolic patterns along cell differentiation paths. Here we show scSpaMet’s ability to quantify and visualize the cell-type specific and spatially resolved metabolic-protein mapping as an emerging tool for systems-level understanding of tissue biology.

59 BASIC BIOLOGICAL SCIENCES↗

Colorectal Cancer Metastases in the Liver Establish Immunosuppressive Spatial Networking between Tumor-Associated SPP1 + Macrophages and Fibroblasts

Abstract Purpose: The liver is the most frequent metastatic site for colorectal cancer. Its microenvironment is modified to provide a niche that is conducive for colorectal cancer cell growth. This study focused on characterizing the cellular changes in the metastatic colorectal cancer (mCRC) liver tumor microenvironment (TME). Experimental Design: We analyzed a series of microsatellite stable (MSS) mCRCs to the liver, paired normal liver tissue, and peripheral blood mononuclear cells using single-cell RNA sequencing (scRNA-seq). We validated our findings using multiplexed spatial imaging and bulk gene expression with cell deconvolution. Results: We identified TME-specific SPP1-expressing macrophages with altered metabolism features, foam cell characteristics, and increased activity in extracellular matrix (ECM) organization. SPP1+ macrophages and fibroblasts expressed complementary ligand–receptor pairs with the potential to mutually influence their gene-expression programs. TME lacked dysfunctional CD8 T cells and contained regulatory T cells, indicative of immunosuppression. Spatial imaging validated these cell states in the TME. Moreover, TME macrophages and fibroblasts had close spatial proximity, which is a requirement for intercellular communication and networking. In an independent cohort of mCRCs in the liver, we confirmed the presence of SPP1+ macrophages and fibroblasts using gene-expression data. An increased proportion of TME fibroblasts was associated with the worst prognosis in these patients. Conclusions: We demonstrated that mCRC in the liver is characterized by transcriptional alterations of macrophages in the TME. Intercellular networking between macrophages and fibroblasts supports colorectal cancer growth in the immunosuppressed metastatic niche in the liver. These features can be used to target immune-checkpoint–resistant MSS tumors.

60 APPLIED LIFE SCIENCES↗

Science Operations with the James Webb Space Telescope

The James Webb Space Telescope (JWST) will be a powerful space observatory whose four science instruments will deliver rich imaging and multiplexed spectroscopic datasets to the astronomical and planetary science communities. The ground segment for JWST, now being designed and built, will carry out JWST's science operations. The ground segment includes: (1) software that the scientific community will use to propose and specify new observations; (2) software that will schedule both science and calibration observations in a way that optimizes observing efficiency while managing the accumulation of momentum; (3) the infrastructure to regularly measure and maintain the telescope's wavefront; (4) orbit determination, ranging, and tracking; (5) communication via the Deep Space Network to command the observatory and retrieve scientific data; (6) onboard scripts that execute each observing program in an event-driven fashion, with occasional interruptions for targets of opportunity or time-critical observations; and (7) a system that processes and calibrates the data into science ready products, automatically recalibrates when calibrations improve, and archives the data for timely access by the principal investigator and later worldwide access by the scientific community. This ground system builds on experience from operating the Hubble Space Telescope, while solving challenges that are unique to the James Webb Space Telescope. In this paper, we describe the elements of the JWST ground system, how it will work operationally from the perspective of the observatory itself, and how a typical user will interact with the system to turn his/her idea into scientific discovery.

Rigby, Jane↗

Development and evaluation of a Hadamard transform imaging spectrometer and a Hadamard transform thermal imager

A spectrometric imager and a thermal imager, which achieve multiplexing by the use of binary optical encoding masks, were developed. The masks are based on orthogonal, pseudorandom digital codes derived from Hadamard matrices. Spatial and/or spectral data is obtained in the form of a Hadamard transform of the spatial and/or spectral scene; computer algorithms are then used to decode the data and reconstruct images of the original scene. The hardware, algorithms and processing/display facility are described. A number of spatial and spatial/spectral images are presented. The achievement of a signal-to-noise improvement due to the signal multiplexing was also demonstrated. An analysis of the results indicates both the situations for which the multiplex advantage may be gained, and the limitations of the technique. A number of potential applications of the spectrometric imager are discussed.

Harwit, M.↗

Cryosectioning-enhanced super-resolution microscopy for single-protein imaging across cells and tissues

DNA-points accumulation for imaging in nanoscale topography (DNA-PAINT) enables nanoscale imaging with virtually unlimited multiplexing and molecular counting. Here, we address challenges, such as variable imaging performance and target accessibility, that can limit its broader applicability. Specifically, we enhance its capacity for robust single-protein imaging and molecular counting by optimizing the integration of total internal reflection fluorescence microscopy with physical sectioning, in particular, Tokuyasu cryosectioning. Our method, tomographic and kinetically enhanced DNA-PAINT (tkPAINT), achieves 3 nm localization precision across diverse samples, enhanced imager binding, and improved cellular integrity. tkPAINT can facilitate molecular counting with DNA-PAINT inside the nucleus, as demonstrated through its quantification of the in situ abundance of RNA Polymerase II in both HeLa cells as well as mouse tissues. Anticipating that tkPAINT could become a versatile tool for the exploration of biomolecular organization and interactions across cells and tissues, we also demonstrate its capacity to support multiplexing, multimodal targeting of proteins and nucleic acids, and three-dimensional (3D) imaging.

Science & Technology - Other Topics↗

MOSAIC - A space-multiplexing technique for optical processing of large images

A technique for Fourier processing of images larger than the space-bandwidth products of conventional or smart spatial light modulators and two-dimensional detector arrays is described. The technique involves a spatial combination of subimages displayed on individual spatial light modulators to form a phase-coherent image, which is subsequently processed with Fourier optical techniques. Because of the technique's similarity with the mosaic technique used in art, the processor used is termed an optical MOSAIC processor. The phase accuracy requirements of this system were studied by computer simulation. It was found that phase errors of less than lambda/8 did not degrade the performance of the system and that the system was relatively insensitive to amplitude nonuniformities. Several schemes for implementing the subimage combination are described. Initial experimental results demonstrating the validity of the mosaic concept are also presented.

Athale, Ravindra A.↗

Hadamard transform imager and imaging spectrometer

An imager and a spectrometric imager, which achieve multiplexing by the use of binary optical encoding masks, have been built and tested. The masks are based on orthogonal, pseudorandom digital codes derived from Hadamard matrices. The spatial (and/or spectral) data are therefore obtained in the form of a Hadamard transform of the spatial (and/or spectral) scene. Computer algorithms are used to decode the data and reconstruct images of the original scene. The hardware, algorithms processing and display facility are described. A number of spatial and spatial/spectral images, obtained in the laboratory, are presented.

Swift, R. D.↗

Covalent Surface Modification Effects on Single‐Walled Carbon Nanotubes for Targeted Sensing and Optical Imaging

Abstract Optical nanoscale technologies often implement covalent or noncovalent strategies for the modification of nanoparticles, whereby both functionalizations are leveraged for multimodal applications but can affect the intrinsic fluorescence of nanoparticles. Specifically, single‐walled carbon nanotubes (SWCNTs) can enable real‐time imaging and cellular delivery; however, the introduction of covalent SWCNT sidewall functionalizations often attenuates SWCNT fluorescence. Recent advances in SWCNT covalent functionalization chemistries preserve the SWCNT's pristine graphitic lattice and intrinsic fluorescence, and here, such covalently functionalized SWCNTs maintain intrinsic fluorescence‐based molecular recognition of neurotransmitter and protein analytes. The covalently modified SWCNT nanosensor preserves its fluorescence response towards its analyte for certain nanosensors, presumably dependent on the intermolecular interactions between SWCNTs or the steric hindrance introduced by the covalent functionalization that hinders noncovalent interactions with the SWCNT surface. These SWCNT nanosensors are further functionalized via their covalent handles with a targeting ligand, biotin, to self‐assemble on passivated microscopy slides, and these dual‐functionalized SWCNT materials are explored for future use in multiplexed sensing and imaging applications.

Chio, Linda↗

Extreme ultraviolet imaging spectrometer for thermospheric emissions

One of the instruments that has been developed to fly on the Space Shuttle is the Imaging Spectrometric Observatory, an array of five imaging spectrometers to cover the 300-12,000-A wavelength range. In this paper the spectrometer designed to operate in the extreme ultraviolet is described. The instrument is intended for studies of the thermosphere and magnetosphere and support of various plasma experiments to be performed from the Shuttle. The design is modular so that various components such as gratings and detectors can be changed and optimized for a range of specific studies following the first survey mission. The detector is an intensified 2-D CCD which permits simultaneous spectral and spatial imaging. The spectral multiplexing gives the spectrometer a considerable speed advantage. The instrument has a small field of view and is thus capable of obtaining much needed scale height information on the atmospheric EUV emissions. Operating from the Shuttle, global and temporal coverage will be obtained, and the EUV data will be enhanced by the simultaneously acquired UV, visible, and near-IR observations.

Torr, M. R.↗

Advanced Spectroscopic and Thermal Imaging Instrumentation for Shock Tube and Ballistic Range Facilities

The Electric Arc Shock Tube (EAST) facility and Hypervelocity Free Flight Aerodynamic Facility (HFFAF, an aeroballistic range) at NASA Ames support basic research in aerothermodynamic phenomena of atmospheric entry, specifically shock layer radiation spectroscopy, convective and radiative heat transfer, and transition to turbulence. Innovative optical instrumentation has been developed and implemented to meet the challenges posed from obtaining such data in these impulse facilities. Spatially and spectrally resolved measurements of absolute radiance of a travelling shock wave in EAST are acquired using multiplexed, time-gated imaging spectrographs. Nearly complete spectral coverage from the vacuum ultraviolet to the near infrared is possible in a single experiment. Time-gated thermal imaging of ballistic range models in flight enables quantitative, global measurements of surface temperature. These images can be interpreted to determine convective heat transfer rates and reveal transition to turbulence due to isolated and distributed surface roughness at hypersonic velocities. The focus of this paper is a detailed description of the optical instrumentation currently in use in the EAST and HFFAF.

Grinstead, Jay H.↗