What are the advantages of ghost imaging? Multiplexing for x-ray and electron imaging
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47 OTHER INSTRUMENTATION↗
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NaroNet: Discovery of tumor microenvironment elements from highly multiplexed images
Understanding the spatial interactions between the elements of the tumor microenvironment -i.e. tumor cells. fibroblasts, immune cells- and how these interactions relate to the diagnosis or prognosis of a tumor is one of the goals of computational pathology. We present NaroNet, a deep learning framework that models the multi-scale tumor microenvironment from multiplex-stained cancer tissue images and provides patient-level interpretable predictions using a seamless end-to-end learning pipeline. Trained only with multiplex-stained tissue images and their corresponding patient-level clinical labels, NaroNet unsupervisedly learns which cell phenotypes, cell neighborhoods, and neighborhood interactions have the highest influence to predict the correct label. To this end, NaroNet incorporates several novel and state-of-the-art deep learning techniques, such as patch-level contrastive learning, multi-level graph embeddings, a novel max-sum pooling operation, or a metric that quantifies the relevance that each microenvironment element has in the individual predictions. We validate NaroNet using synthetic data simulating multiplex-immunostained images where a patient label is artificially associated to the -adjustable- probabilistic incidence of different microenvironment elements. We then apply our model to two sets of images of human cancer tissues: 336 seven-color multiplex-immunostained images from 12 high-grade endometrial cancer patients; and 382 35-plex mass cytometry images from 215 breast cancer patients. In both synthetic and real datasets, NaroNet provides outstanding predictions of relevant clinical information while associating those predictions to the presence of specific microenvironment elements.
60 APPLIED LIFE SCIENCES↗
Multiplex imaging with multiple-pinhole cameras
When making photographs in X rays or gamma rays with a multiple-pinhole camera, the individual images of an extended object such as the sun may be allowed to overlap. Then the situation is in many ways analogous to that in a multiplexing device such as a Fourier spectroscope. Some advantages and problems arising with such use of the camera are discussed, and expressions are derived to describe the relative efficacy of three exposure/postprocessing schemes using multiple-pinhole cameras.
Brown, C.↗
Multiplexed structured imaging of laser induced fluorescence at 10 GHz rate
The frame rate of conventional high-speed imaging devices is limited by exposure time and signal read-out time. Structured imaging is able to push the frame rate beyond the limit of the imaging device. This work demonstrates the feasibility of potential TeraHz rate structured imaging with a multiplexed structured imaging setup. In this work, a multiplexed structured image capture system was employed to image UV laser induced krypton-II emission lines at 10 GHz. The measured emission intensity of the 764 nm line over time suggests a temporal resolution of 97 ps. The temporal resolution is adjustable, and the frame rate can be further increased to the TeraHz level.
47 OTHER INSTRUMENTATION↗
Two-phase accurate multiplexed structured image capture (2pAc-MUSIC)
Multiplexed Structural Image Capture (MUSIC) technique uses modulation of light from a scene prior to imaging to encode the temporal evolution of target into spatial frequency shifts, with each shift corresponding to a unique time and resulting in individual distinct snapshots. Computational-processing of the multiplexed single-shot image can recover the nanosecond-resolution scene evolution. One of the limitations is the loss of spatial resolution of the recovered image due to limitations of the low-pass spatial frequency filtering depth. Here we demonstrated that two images with complementary spatial phases can be used to enhance the spatial resolutions. For this application, 2 Phase Accurate (2pAc)-MUSIC is implemented that a scene is encoded with two complementary phases. The increased spatial information captured reduces the lower frequency noise in the Fourier domain found in single-phase MUSIC. A recovery algorithm combines the 180-degree spatial frequency shifted images into a single scene and applies a spatial filter in the Fourier domain that has a larger depth than previously possible to achieve a scene with increased spatial resolution. The 2pAc-MUSIC can effectively enhance the spatial resolution by removing the overlapping zero-order spatial frequencies.
71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗
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↗
Dielectric image-resonator multiplexer
A two, three or four channel multiplexer can be operated in the TE 01δ or HE 11δ modes, each channel of the multiplexer has one bandpass filter, with each filter usually having a plurality of cavities. One cavity of each filter is a common cavity with all of the other filters of the multiplexer. The common cavity contains one independent dielectric quarter-cut or half-cut image resonator for each filter representing a channel- of the multiplexer. Each resonator is mounted on a suitable support the common cavity provides a common junction and contains a loop coupling to couple electromagnetic energy between the various· channels. The multiplexer has at least one input and at least one output. Previous microwave multiplexers have a plurality of bandpass filters arranged on a coaxial manifold. With the multiplexer of the present invention, the manifold can be eliminated. Further, at the S-band or L-band, substantial weight and volume savings can be achieved using the multiplexer of the present invention without sacrificing performance. This can be very important for space applications.
Wai-Cheung Tang↗
Optical Signal Processing: Poisson Image Restoration and Shearing Interferometry
Optical signal processing can be performed in either digital or analog systems. Digital computers and coherent optical systems are discussed as they are used in optical signal processing. Topics include: image restoration; phase-object visualization; image contrast reversal; optical computation; image multiplexing; and fabrication of spatial filters. Digital optical data processing deals with restoration of images degraded by signal-dependent noise. When the input data of an image restoration system are the numbers of photoelectrons received from various areas of a photosensitive surface, the data are Poisson distributed with mean values proportional to the illuminance of the incoherently radiating object and background light. Optical signal processing using coherent optical systems is also discussed. Following a brief review of the pertinent details of Ronchi's diffraction grating interferometer, moire effect, carrier-frequency photography, and achromatic holography, two new shearing interferometers based on them are presented. Both interferometers can produce variable shear.
Hong, Yie-Ming↗
In situ , in vivo , and in operando imaging and spectroscopy of liquids using microfluidics in vacuum
This review offers a succinct overview of the development of a vacuum-compatible microfluidic reactor system for analysis at the liquid vacuum interface (SALVI), and its diverse applications in in situ, in vivo, and in operando imaging of liquid surfaces as well as the air-liquid (a-l), liquid-liquid (l-l), and solid-liquid (s-l) interfaces in the past decade. SALVI is one of the first microfluidics-based reactors that has enabled direct analysis of volatile liquids in vacuum surface tools such as scanning electron microscopy (SEM) and time-of-flight secondary ion mass spectrometry (ToF-SIMS). Its integration into ambient and vacuum spectroscopy and microscopy is illustrated. Several applications are highlighted including (1) imaging nanoparticles in liquid using in situ SEM; (2) mapping the evolving l-l interface using in situ x-ray absorption spectroscopy and ToF-SIMS; (3) following complex a-l interfacial oxidation reaction products using in situ ToF-SIMS; (4) capturing biological interfaces of cells and microbes via in vivo multimodal and correlative imaging; and (5) monitoring the dynamic solid electrode and liquid electrolyte interface using in operando molecular imaging. Finally, outlook and recommendations are presented. Besides showing the holistic information volume obtained by real-time multiplexed imaging, this review intends to convey the importance of tool development in revolutionizing surface and interface analysis using vacuum platforms previously limited to solid surfaces. Microfluidics is manifested to be not limited to ambient conditions in many examples in this review. Moreover, fundamental interfacial phenomena underpinning mass and charge transfer can now be pursued in real time via innovated chemical imaging and spectroscopy.
42 ENGINEERING↗
Stereo Imaging Miniature Endoscope
Stereo imaging requires two different perspectives of the same object and, traditionally, a pair of side-by-side cameras would be used but are not feasible for something as tiny as a less than 4-mm-diameter endoscope that could be used for minimally invasive surgeries or geoexploration through tiny fissures or bores. The proposed solution here is to employ a single lens, and a pair of conjugated, multiple-bandpass filters (CMBFs) to separate stereo images. When a CMBF is placed in front of each of the stereo channels, only one wavelength of the visible spectrum that falls within the passbands of the CMBF is transmitted through at a time when illuminated. Because the passbands are conjugated, only one of the two channels will see a particular wavelength. These time-multiplexed images are then mixed and reconstructed to display as stereo images. The basic principle of stereo imaging involves an object that is illuminated at specific wavelengths, and a range of illumination wavelengths is time multiplexed. The light reflected from the object selectively passes through one of the two CMBFs integrated with two pupils separated by a baseline distance, and is focused onto the imaging plane through an objective lens. The passband range of CMBFs and the illumination wavelengths are synchronized such that each of the CMBFs allows transmission of only the alternate illumination wavelength bands. And the transmission bandwidths of CMBFs are complementary to each other, so that when one transmits, the other one blocks. This can be clearly understood if the wavelength bands are divided broadly into red, green, and blue, then the illumination wavelengths contain two bands in red (R1, R2), two bands in green (G1, G2), and two bands in blue (B1, B2). Therefore, when the objective is illuminated by R1, the reflected light enters through only the left-CMBF as the R1 band corresponds to the transmission window of the left CMBF at the left pupil. This is blocked by the right CMBF. The transmitted band is focused on the focal plane array (FPA).
Bae, Youngsam↗
NucD-25 Nuclear Diagnostics Development: NIS capability, Maximizing gamma ray dynamic range, (n,γ) measurements for carbon ρR [Slides]
The LANL Neutron Imaging team is preparing for NIS upgrades and novel capabilities with the advent of consistently higher yields at NIF. Novel scintillators, solid-state detectors, multiplexed imaging detectors, upgraded data acquisition systems, and camera testing are all areas in need of experimental study to further develop imaging capabilities and dynamic range at NIF.
46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗
Extending the Near-Infrared Emission Range of Indium Phosphide Quantum Dots for Multiplexed In Vivo Imaging
This report of the reddest emitting indium phosphide quantum dots (InP QDs) to date demonstrates tunable, near-infrared (NIR) photoluminescence (PL) as well as PL multiplexing in the first optical tissue window while avoiding toxic constituents. This synthesis overcomes the InP “growth bottleneck” and extends the emission peak of InP QDs deeper into the first optical tissue window using an inverted QD heterostructure, specifically ZnSe/InP/ZnS core/shell/shell nanoparticles. The QDs exhibit InP shell thickness-dependent tunable emission with peaks ranging from 515–845 nm. The high absorptivity of InP yields effective photoexcitation of the QDs with UV, visible, and NIR wavelengths. These nanoparticles extend the range of tunable direct-bandgap emission from InP-based nanostructures, effectively overcoming a synthetic barrier that has prevented InPbased QDs from reaching their full potential as NIR imaging agents. Further, multiplexed lymph node imaging in a mouse model demonstrates the potential of the NIR-emitting InP particles for in vivo imaging.
77 NANOSCIENCE AND NANOTECHNOLOGY↗
Introduction
The 2014 SPIE Sensing Technologies for Global Health, Military Medicine and Environmental Monitoring conference embraced a wealth of state-of-the-art information in basic and applied science. This event covered the latest developments in the following areas: Non-invasive Disease Diagnostics for Global Health- This opening series of two consecutive sessions focused on oral biospecimen based rapid assays and point-of-care devices for the detection of pathogens causing infectious diseases, biomarkers for cancer, and analytes for noncommunicable diseases such as diabetes. They also covered presentations on the human proteasome and microbiome with linkage to human diseases and diagnostic approaches. The sessions were built on the past experience and expertise of the National Institutes of Health, National Institutes of Dental and Craniofacial Research. Military Medicine I: Traumatic Brain Injury and PTSD-This assembly covered oral-biomarker based diagnostics for brain damage and TBI as well as prevention and rehabilitation technologies. Neurorehabilitation and noninvasive neuromodulation were also discussed as critical approaches for effective functioning. Military Medicine II: Physiology and Medicine of Extreme Environments and Spaceflight-This scientific segment showcased physiological, pharmacological and diagnostic sensing methodologies during spaceflight per the National Aeronautics and Space Administration as well as military-relevant toxicans and future sensing trends per the Department of Defense. It also included latest technologies to determine hydration status in warfighters, eye surgery using the latest laser technologies, and sensing tools for blood analysis. Sensing Technologies for Disease Diagnostics and Environmental Monitoring-This closing series of two consecutive sessions provided the venues to learn and discuss more results on the next generation of diagnostic tools and field technologies for diseases, including biomarker detection by digital imaging, multiplex technologies, capillary electrophoresis and molecular platforms serving as labs-on-chips. This conference allowed cross-fertilization of ideas, projects and collaborative work by a multidisciplinary audience of national and international colleagues from the academia, industry and federal government: The National Institutes of Health, National Aeronautics and Space Administration, and the Department of Defense.
Wotring, Virginia E.↗
A radiation-hard, low-background multiplexer design for spacecraft imager applications
A possible multiplexer design for the focal plane for the Cassini Visible and Infrared Mapping Spectrometer (VIMS) is reviewed. The instrument's requirements for the multiplexed array are summarized. The VIMS instrument has a modest radiation-hardness requirement due to the trajectory and planetary environments in which the instrument will be required to operate. The total ionizing dose hardness requirement is a few tens of kilorads. A thin-gate oxide of a few hundred angstroms thickness is to be used. Field hardness is to be achieved by guard bands or hardened dielectric isolation. The design is argued to meet the low-noise and radiation-hardness required for imaging at Saturn. The design is versatile enough to provide double-correlated and double-uncorrelated sampling, which is accomplished in the signal processing electronics outside the focal plane.
Staller, Craig↗
Real-time multiple-look synthetic aperture radar processor for spacecraft applications
A spaceborne synthetic aperture radar (SAR) having pipeline multiple-look data processing is described which makes use of excessive azimuth bandwidth in radar echo signals to produce multiple-looking images. Time multiplexed single-look image lines from an azimuth correlator go through an energy analyzer which analyzes the mean energy in each separate look to determine the radar antenna electric boresight for use in generating the correct reference functions for the production of high quality SAR images. The multiplexed single look image lines also go through a registration delay to produce multi-look images.
Wu, C.↗
Toward Large FOV High-Resolution X-Ray Imaging Spectrometer: Microwave Multiplexed Readout of 32 TES Microcalorimeters
We performed a small-scale demonstration at GSFC of high-resolution x-ray TES microcalorimeters read out using a microwave SQUID multiplexer. This work is part of our effort to develop detector and readout technologies for future space based x-ray instruments such as the microcalorimeter spectrometer envisaged for Lynx, a large mission concept under development for the Astro 2020 Decadal Survey. In this paper we describe our experiment, including details of a recently designed, microwave-optimized low-temperature setup that is thermally anchored to the 50 mK stage of our laboratory ADR. Using a ROACH2 FPGA at room temperature, we simultaneously read out 32 pixels of a GSFC-built detector array via a NIST-built multiplexer chip with Nb coplanar waveguide resonators coupled to RF SQUIDs. The resonators are spaced 6 MHz apart (at approx. 5.9 GHz) and have quality factors of approximately 15,000. Using flux-ramp modulation frequencies of 160 kHz we have achieved spectral resolutions of 3 eV FWHM on each pixel at 6 keV. We will present the measured system-level noise and maximum slew rates, and briefly describe the implications for future detector and readout design.
Yoon, Wonsik↗
Toward Large Field-Of-View High-Resolution X-Ray Imaging Spectrometers: Microwave Multiplexed Readout of 28 TES Microcalorimeters
We performed small-scale demonstrations at GSFC of high-resolution Xray TES microcalorimeters read out using a microwave SQUID multiplexer. This work is part of our effort to develop detector and readout technologies for future space-based X-ray instruments such as the microcalorimeter spectrometer envisaged for Lynx, a large mission concept under development for the Astro 2020 Decadal Survey. In this paper we describe our experiment, including details of a recently designed, microwave-optimized low-temperature setup that is thermally anchored to the 55mKstage of our laboratory ADR. Using aROACH2 FPGA at room temperature, we read out pixels of a GSFC-built detector array via a NIST-built multiplexer chip with Nb coplanar waveguide resonators coupled to rf-SQUIDs. The resonators are spaced 6 MHz apart (at ∼ 5.9 GHz) and have quality factors of ∼ 15,000. In our initial demonstration, we used flux-ramp modulation frequencies of 125 kHz to read out 5 pixels simultaneously and achieved spectral resolutions of 2.8-3.1 eV FWHM at 5.9 keV. Our subsequent work is ongoing: to-date we have achieved a median spectral resolution of 3.4 eV FWHM at 5.9 keV while reading out 28 pixels simultaneously with flux-ramp frequencies of 160 kHz. We present the measured system-level noise and maximum slew rates and briefly describe our future development work.
Yoon, W.↗