High Resolution Imaging with MUSTANG-2 on the GBT

NASA Astrophysics Data System (ADS)

Stanchfield, Sara; Ade, Peter; Aguirre, James; Brevik, Justus A.; Cho, Hsiao-Mei; Datta, Rahul; Devlin, Mark; Dicker, Simon R.; Dober, Bradley; Duff, Shannon M.; Egan, Dennis; Ford, Pam; Hilton, Gene; Hubmayr, Johannes; Irwin, Kent; Knowles, Kenda; Marganian, Paul; Mason, Brian Scott; Mates, John A. B.; McMahon, Jeff; Mello, Melinda; Mroczkowski, Tony; Romero, Charles; Sievers, Jonathon; Tucker, Carole; Vale, Leila R.; Vissers, Michael; White, Steven; Whitehead, Mark; Ullom, Joel; Young, Alexander

2018-01-01

We present early science results from MUSTANG-2, a 90 GHz feedhorn-coupled, microwave SQUID-multiplexed TES bolometer array operating on the Robert C. Byrd Green Bank Telescope (GBT). The feedhorn and waveguide-probe-coupled detector technology is a mature technology, which has been used on instruments such as the South Pole Telescope, the Atacama Cosmology Telescope, and the Atacama B-mode Search telescope. The microwave SQUID multiplexer-based readout system developed for MUSTANG-2 currently reads out 66 detectors with a single coaxial cable and will eventually allow thousands of detectors to be multiplexed. This microwave SQUID multiplexer combines the proven abilities of millimeter wave TES detectors with the multiplexing capabilities of KIDs with no degradation in noise performance of the detectors. Each multiplexing device is read out using warm electronics consisting of a commercially available ROACH board, a DAC/ADC card, and an Intermediate Frequency mixer circuit. The hardware was originally developed by the Collaboration for Astronomy Signal Processing and Electronic Research (CASPER) group, whose primary goal is to develop scalable FPGA-based hardware with the flexibility to be used in a wide range of radio signal processing applications. MUSTANG-2 is the first on-sky instrument to use microwave SQUID multiplexing and is available as a shared-risk/PI instrument on the GBT. In MUSTANG-2’s first season 7 separate proposals were awarded a total of 230 hours of telescope time.

Fluorescence-Raman Dual Modal Endoscopic System for Multiplexed Molecular Diagnostics

NASA Astrophysics Data System (ADS)

Jeong, Sinyoung; Kim, Yong-Il; Kang, Homan; Kim, Gunsung; Cha, Myeong Geun; Chang, Hyejin; Jung, Kyung Oh; Kim, Young-Hwa; Jun, Bong-Hyun; Hwang, Do Won; Lee, Yun-Sang; Youn, Hyewon; Lee, Yoon-Sik; Kang, Keon Wook; Lee, Dong Soo; Jeong, Dae Hong

2015-03-01

Optical endoscopic imaging, which was recently equipped with bioluminescence, fluorescence, and Raman scattering, allows minimally invasive real-time detection of pathologies on the surface of hollow organs. To characterize pathologic lesions in a multiplexed way, we developed a dual modal fluorescence-Raman endomicroscopic system (FRES), which used fluorescence and surface-enhanced Raman scattering nanoprobes (F-SERS dots). Real-time, in vivo, and multiple target detection of a specific cancer was successful, based on the fast imaging capability of fluorescence signals and the multiplex capability of simultaneously detected SERS signals using an optical fiber bundle for intraoperative endoscopic system. Human epidermal growth factor receptor 2 (HER2) and epidermal growth factor receptor (EGFR) on the breast cancer xenografts in a mouse orthotopic model were successfully detected in a multiplexed way, illustrating the potential of FRES as a molecular diagnostic instrument that enables real-time tumor characterization of receptors during routine endoscopic procedures.

Development of a Time Domain Fluorimeter for Fluorescent Lifetime Multiplexing Analysis

PubMed Central

Weissleder, Ralph; Mahmood, Umar

2009-01-01

We show that a portable, inexpensive USB-powered time domain fluorimeter (TDF) and analysis scheme were developed for use in evaluating a new class of fluorescent lifetime multiplexed dyes. Fluorescent proteins, organic dyes, and quantum dots allow the labeling of more and more individual features within biological systems, but the wide absorption and emission spectra of these fluorophores limit the number of distinct processes which may be simultaneously imaged using spectral separation alone. By additionally separating reporters in a second dimension, fluorescent lifetime multiplexing provides a means to multiply the number of available imaging channels. PMID:19830273

FIR Detector Sensitivity, Dynamic Range, and Multiplexing Requirements for the Origins Space Telescope (OST)

NASA Astrophysics Data System (ADS)

Staguhn, Johannes G.

2018-05-01

Spectroscopic, cold, space-based mid-to-far-infrared (FIR) missions, such as the Origins Space Telescope, will require large (tens of kilopixels), ultra-sensitive FIR detector arrays with sufficient dynamic range and high-density multiplexing schemes for the readout, in order to optimize the scientific return while staying within a realistic cost range. Issues like power consumption of multiplexers and their readout are significantly more important for space missions than they are for ground-based or suborbital applications. In terms of the detectors and their configuration into large arrays, significant development efforts are needed even for both of the most mature candidate superconducting detector technologies, namely transition edge sensors and (microwave) kinetic inductance detectors. Here we explore both practical and fundamental limits for those technologies in order to lay out a realistic path forward for both technologies. We conclude that beyond the need to enhance the detector sensitivities and pixel numbers by about an order of magnitude over currently existing devices, improved concepts for larger dynamic range and multiplexing density will be needed in order to optimize the scientific return of future cold FIR space missions. Background-limited, very high spectral resolution instruments will require photon-counting detectors.

Near-infrared laser-induced fluorescence detection in capillary electrophoresis.

PubMed

McWhorter, S; Soper, S A

2000-04-01

As capillary electrophoresis continues to focus on miniaturization, either through reducing column dimensions or situating entire electrophoresis systems on planar chips, advances in detection become necessary to meet the challenges posed by these electrophoresis platforms. The challenges result from the fact that miniaturization requires smaller load volumes, demanding highly sensitive detection. In addition, many times multiple targets must be analyzed simultaneously (multiplexed applications), further complicating detection. Near-infrared (NIR) fluorescence offers an attractive alternative to visible fluorescence for critical applications in capillary electrophoresis due to the impressive limits of detection that can be generated, in part resulting from the low background levels that are observed in the NIR. Advances in instrumentation and fluorogenic labels appropriate for NIR monitoring have led to a growing number of examples of the use of NIR fluorescence in capillary electrophoresis. In this review, we will cover instrumental components used to construct ultrasensitive NIR fluorescence detectors, including light sources and photon transducers. In addition, we will discuss various types of labeling dyes appropriate for NIR fluorescence and finally, we will present several applications that have used NIR fluorescence in capillary electrophoresis, especially for DNA sequencing and fragment analysis.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mukundan, Harshini; Xei, Hongshi; Anderson, Aaron S

We have developed a waveguide-based optical biosensor for the sensitive and specific detection of biomarkers associated with disease. Our technology combines the superior optical properties of single-mode planar waveguides, the robust nature of functionalized self-assembled monolayer sensing films and the specificity of fluorescence sandwich immunoassays to detect biomarkers in complex biological samples such as serum, urine and sputum. We have previously reported the adaptation of our technology to the detection of biomarkers associated with breast cancer and anthrax. However, these approaches primarily used phospholipid bilayers as the functional film and organic dyes (ex: AlexaFluors) as the fluorescence reporter. Organic dyesmore » are easily photodegraded and are not amenable to multiplexing because of their narrow Stokes' shift. Here we have developed strategies for conjugation of the detector antibodies with quantum dots for use in a multiplex detection platform. We have previously evaluated dihydroxylipoic acid quantum dots for the detection of a breast cancer biomarker. In this manuscript, we investigate the detection of the Bacillus anthracis protective antigen using antibodies conjugated with polymer-coated quantum dots. Kinetics of binding on the waveguide-based biosensor is reported. We compare the sensitivity of quantum dot labeled antibodies to those labeled with AlexaFluor and demonstrate the photostability of the former in our assay platform. In addition, we compare sulfydryl labeling of the antibody in the hinge region to that of nonspecific amine labeling. This is but the first step in developing a multiplex assay for such biomarkers on our waveguide platform.« less

Continuous-Flow Detector for Rapid Pathogen Identification

DOE Office of Scientific and Technical Information (OSTI.GOV)

Barrett, Louise M.; Skulan, Andrew J.; Singh, Anup K.

2006-09-01

This report describes the continued development of a low-power, portable detector for the rapid identification of pathogens such as B. anthracis and smallpox. Based on our successful demonstration of the continuous filter/concentrator inlet, we believe strongly that the inlet section will enable differentiation between viable and non-viable populations, between types of cells, and between pathogens and background contamination. Selective, continuous focusing of particles in a microstream enables highly selective and sensitive identification using fluorescently labeled antibodies and other receptors such as peptides, aptamers, or small ligands to minimize false positives. Processes such as mixing and lysing will also benefit frommore » the highly localized particle streams. The concentrator is based on faceted prisms to contract microfluidic flows while maintaining uniform flowfields. The resulting interfaces, capable of high throughput, serve as high-, low-, and band-pass filters to direct selected bioparticles to a rapid, affinity-based detection system. The proposed device is superior to existing array-based detectors as antibody-pathogen binding can be accomplished in seconds rather than tens of minutes or even hours. The system is being designed to interface with aerosol collectors under development by the National Laboratories or commercial systems. The focused stream is designed to be interrogated using diode lasers to differentiate pathogens by light scattering. Identification of particles is done using fluorescently labeled antibodies to tag the particles, followed by multiplexed laser-induced fluorescence (LIF) detection (achieved by labeling each antibody with a different dye).« less

Design and Performance of the Multiplexed SQUID/TES Array at Ninety Gigahertz

NASA Astrophysics Data System (ADS)

Stanchfield, Sara; Ade, Peter; Aguirre, James; Brevik, Justus A.; Cho, Hsiao-Mei; Datta, Rahul; Devlin, Mark; Dicker, Simon R.; Dober, Bradley; Duff, Shannon M.; Egan, Dennis; Ford, Pam; Hilton, Gene; Hubmayr, Johannes; Irwin, Kent; Knowles, Kenda; Marganian, Paul; Mason, Brian Scott; Mates, John A. B.; McMahon, Jeff; Mello, Melinda; Mroczkowski, Tony; Romero, Charles; Sievers, Jonathon; Tucker, Carole; Vale, Leila R.; Vissers, Michael; White, Steven; Whitehead, Mark; Ullom, Joel; Young, Alexander

2018-01-01

We present the array performance and astronomical images from early science results from MUSTANG-2, a 90 GHz feedhorn-coupled, microwave SQUID-multiplexed TES bolometer array operating on the Robert C. Byrd Green Bank Telescope (GBT). MUSTANG-2 was installed on the GBT on December 2, 2016 and immediately began commissioning efforts, followed by science observations, which are expected to conclude June 2017. The feedhorn and waveguide-probe-coupled detector technology is a mature technology, which has been used on instrument including the South Pole Telescope, the Atacama Cosmology Telescope, and the Atacama B-mode Search telescope. The microwave SQUID readout system developed for MUSTANG-2 currently reads out 66 detectors with a single coaxial cable and will eventually allow thousands of detectors to be multiplexed. This microwave SQUID multiplexer combines the proven abilities of millimeterwave TES detectors with the multiplexing capabilities of KIDs with no degradation in noise performance of the detectors. Each multiplexing device is read out using warm electronics consisting of a commercially available ROACH board, a DAC/ADC card, and an Intermediate Frequency mixer circuit. The hardware was originally developed by the UC Berkeley Collaboration for Astronomy Signal Processing and Electronic Research (CASPER) group, whose primary goal is to develop scalable FPGA-based hardware with the flexibility to be used in a wide range of radio signal processing applications. MUSTANG-2 is the first on-sky instrument to use microwave SQUID multiplexing and is available as a shared-risk/PI instrument on the GBT. In MUSTANG-2's first season 7 separate proposals were awarded a total of 230 hours of telescope time.

Analysis of Multiplexed Nanosensor Arrays Based on Near-Infrared Fluorescent Single-Walled Carbon Nanotubes.

PubMed

Dong, Juyao; Salem, Daniel P; Sun, Jessica H; Strano, Michael S

2018-04-24

The high-throughput, label-free detection of biomolecules remains an important challenge in analytical chemistry with the potential of nanosensors to significantly increase the ability to multiplex such assays. In this work, we develop an optical sensor array, printable from a single-walled carbon nanotube/chitosan ink and functionalized to enable a divalent ion-based proximity quenching mechanism for transducing binding between a capture protein or an antibody with the target analyte. Arrays of 5 × 6, 200 μm near-infrared (nIR) spots at a density of ≈300 spots/cm 2 are conjugated with immunoglobulin-binding proteins (proteins A, G, and L) for the detection of human IgG, mouse IgM, rat IgG2a, and human IgD. Binding kinetics are measured in a parallel, multiplexed fashion from each sensor spot using a custom laser scanning imaging configuration with an nIR photomultiplier tube detector. These arrays are used to examine cross-reactivity, competitive and nonspecific binding of analyte mixtures. We find that protein G and protein L functionalized sensors report selective responses to mouse IgM on the latter, as anticipated. Optically addressable platforms such as the one examined in this work have potential to significantly advance the real-time, multiplexed biomolecular detection of complex mixtures.

Thermal Design and Characterization of Transition-Edge Sensor (TES) Bolometers for Frequency-Domain Multiplexing

NASA Astrophysics Data System (ADS)

Lueker, Martin; Benson, Bradford A.; Chang, Clarence L.; Cho, Hsiao-Mei; Dobbs, Matt; Holzapfel, William L.; Lanting, Trevor; Lee, Adrian T.; Mehl, Jared; Plagge, Thomas; Shirokoff, Erik; Spieler, Helmuth G.; Vieira, Joaquin D.

2009-06-01

In contemporary cosmic microwave background experiments, bolometric detectors are often background limited, and in this case the sensitivity of instruments can only be improved by increasing the number of background-limited detectors, and so contemporary TES receivers contain as many pixels as possible. Frequency-domain multiplexing (fMUX) is one strategy for reading out many detectors with one SQUID. For any readout system, it is important to carefully evaluate the thermal design of detector, in conjunction with the readout bandwidth, in order to ensure stable electro-thermal feedback (ETF). We demonstrate a novel technique for characterizing the thermal circuit of our detectors, using am AC-bias and the fMUX electronics. This technique is used to study the internal thermal coupling of a TES bolometer. We illustrate how the insights gathered by this technique have been instrumental in improving the stability of our multiplexed detectors for the south pole telescope (SPT).

Optical delay encoding for fast timing and detector signal multiplexing in PET

DOE Office of Scientific and Technical Information (OSTI.GOV)

Grant, Alexander M.; Levin, Craig S., E-mail: cslevin@stanford.edu; Molecular Imaging Program at Stanford

2015-08-15

Purpose: The large number of detector channels in modern positron emission tomography (PET) scanners poses a challenge in terms of readout electronics complexity. Multiplexing schemes are typically implemented to reduce the number of physical readout channels, but often result in performance degradation. Novel methods of multiplexing in PET must be developed to avoid this data degradation. The preservation of fast timing information is especially important for time-of-flight PET. Methods: A new multiplexing scheme based on encoding detector interaction events with a series of extremely fast overlapping optical pulses with precise delays is demonstrated in this work. Encoding events in thismore » way potentially allows many detector channels to be simultaneously encoded onto a single optical fiber that is then read out by a single digitizer. A two channel silicon photomultiplier-based prototype utilizing this optical delay encoding technique along with dual threshold time-over-threshold is demonstrated. Results: The optical encoding and multiplexing prototype achieves a coincidence time resolution of 160 ps full width at half maximum (FWHM) and an energy resolution of 13.1% FWHM at 511 keV with 3 × 3 × 5 mm{sup 3} LYSO crystals. All interaction information for both detectors, including timing, energy, and channel identification, is encoded onto a single optical fiber with little degradation. Conclusions: Optical delay encoding and multiplexing technology could lead to time-of-flight PET scanners with fewer readout channels and simplified data acquisition systems.« less

Fluorescent genetic barcoding in mammalian cells for enhanced multiplexing capabilities in flow cytometry.

PubMed

Smurthwaite, Cameron A; Hilton, Brett J; O'Hanlon, Ryan; Stolp, Zachary D; Hancock, Bryan M; Abbadessa, Darin; Stotland, Aleksandr; Sklar, Larry A; Wolkowicz, Roland

2014-01-01

The discovery of the green fluorescent protein from Aequorea victoria has revolutionized the field of cell and molecular biology. Since its discovery a growing panel of fluorescent proteins, fluorophores and fluorescent-coupled staining methodologies, have expanded the analytical capabilities of flow cytometry. Here, we exploit the power of genetic engineering to barcode individual cells with genes encoding fluorescent proteins. For genetic engineering, we utilize retroviral technology, which allows for the expression of ectopic genetic information in a stable manner in mammalian cells. We have genetically barcoded both adherent and nonadherent cells with different fluorescent proteins. Multiplexing power was increased by combining both the number of distinct fluorescent proteins, and the fluorescence intensity in each channel. Moreover, retroviral expression has proven to be stable for at least a 6-month period, which is critical for applications such as biological screens. We have shown the applicability of fluorescent barcoded multiplexing to cell-based assays that rely themselves on genetic barcoding, or on classical staining protocols. Fluorescent genetic barcoding gives the cell an inherited characteristic that distinguishes it from its counterpart. Once cell lines are developed, no further manipulation or staining is required, decreasing time, nonspecific background associated with staining protocols, and cost. The increasing number of discovered and/or engineered fluorescent proteins with unique absorbance/emission spectra, combined with the growing number of detection devices and lasers, increases multiplexing versatility, making fluorescent genetic barcoding a powerful tool for flow cytometry-based analysis. © 2013 International Society for Advancement of Cytometry.

Development of 640 X 480 LWIR focal plane arrays

NASA Astrophysics Data System (ADS)

Shallcross, Frank V.; Meyerhofer, Dietrich; Dolny, Gary M.; Gilmartin, Harvey R.; Tower, John R.; Palfrey, Stephen L.

1992-08-01

The 640 X 480 MOS multiplexer developed for PtSi MWIR focal plane arrays has been adapted to LWIR operation. The multiplexer is very flexible and can be used in various operating modes. The MOS approach, with its high saturation capacity and low-temperature operating capability, is ideally suited for long-wavelength operation. In this paper applications of the multiplexer to IrSi Schottky detectors and SiGe heterojunction detectors are discussed.

Simultaneous off-axis multiplexed holography and regular fluorescence microscopy of biological cells.

PubMed

Nygate, Yoav N; Singh, Gyanendra; Barnea, Itay; Shaked, Natan T

2018-06-01

We present a new technique for obtaining simultaneous multimodal quantitative phase and fluorescence microscopy of biological cells, providing both quantitative phase imaging and molecular specificity using a single camera. Our system is based on an interferometric multiplexing module, externally positioned at the exit of an optical microscope. In contrast to previous approaches, the presented technique allows conventional fluorescence imaging, rather than interferometric off-axis fluorescence imaging. We demonstrate the presented technique for imaging fluorescent beads and live biological cells.

Side illuminated optical fiber as a multiplexing element for spectroscopic systems

NASA Astrophysics Data System (ADS)

Egalon, Claudio O.; Matta, Michael P.; Lavezzari, Delbert C.; Insley, Robert Y.; Jaring, Carolyn C.; Quiday, Marie F.

2013-09-01

A new type of colorimeter with multiple channels was demonstrated using a side illuminated optical fiber. When different spots of a properly modified fiber are side illuminated, multiple signals are generated and guided by the waveguide: the essence of multiplexing. This configuration is simple, low cost, does not require a sensitive coating and can analyze several samples along the fiber with a single detector: the most expensive component. Since regular colorimeters use one detector per sample, our new configuration considerably lowers the cost of analyzing multiple samples. This system consists of a fiber mounted over a support, three LEDs, an LED driver, a photo diode and a read-out: to increase the signal, the fiber was tapered. For calibration purposes, six solutions of different concentrations of food dye were prepared, placed inside cuvettes along the fiber length and illuminated by the LEDs. This light passes through the solution, strikes the fiber and is guided to the detector: the darker the solution the lower the signal intensity. Several calibration curves were obtained using different light intensities: it was found that the greater the intensity, the higher the colorimeter sensitivity. This simple capability can be used to easily control the device's sensitivity and its resolution. Although built for three samples only, this device can be modified to accommodate more. With cuvettes measuring 1 cm, it is possible to accommodate one sample per cm of fiber. Also, with minor modifications, this colorimeter can be used for fluorescence, scattering and index of refraction measurements.

Quantum detector tomography of a time-multiplexed superconducting nanowire single-photon detector at telecom wavelengths.

PubMed

Natarajan, Chandra M; Zhang, Lijian; Coldenstrodt-Ronge, Hendrik; Donati, Gaia; Dorenbos, Sander N; Zwiller, Val; Walmsley, Ian A; Hadfield, Robert H

2013-01-14

Superconducting nanowire single-photon detectors (SNSPDs) are widely used in telecom wavelength optical quantum information science applications. Quantum detector tomography allows the positive-operator-valued measure (POVM) of a single-photon detector to be determined. We use an all-fiber telecom wavelength detector tomography test bed to measure detector characteristics with respect to photon flux and polarization, and hence determine the POVM. We study the SNSPD both as a binary detector and in an 8-bin, fiber based, Time-Multiplexed (TM) configuration at repetition rates up to 4 MHz. The corresponding POVMs provide an accurate picture of the photon number resolving capability of the TM-SNSPD.

Polarization-multiplexing ghost imaging

NASA Astrophysics Data System (ADS)

Dongfeng, Shi; Jiamin, Zhang; Jian, Huang; Yingjian, Wang; Kee, Yuan; Kaifa, Cao; Chenbo, Xie; Dong, Liu; Wenyue, Zhu

2018-03-01

A novel technique for polarization-multiplexing ghost imaging is proposed to simultaneously obtain multiple polarimetric information by a single detector. Here, polarization-division multiplexing speckles are employed for object illumination. The light reflected from the objects is detected by a single-pixel detector. An iterative reconstruction method is used to restore the fused image containing the different polarimetric information by using the weighted sum of the multiplexed speckles based on the correlation coefficients obtained from the detected intensities. Next, clear images of the different polarimetric information are recovered by demultiplexing the fused image. The results clearly demonstrate that the proposed method is effective.

Fluorescence multiplexing with time-resolved and spectral discrimination using a near-IR detector.

PubMed

Zhu, Li; Stryjewski, Wieslaw; Lassiter, Suzanne; Soper, Steven A

2003-05-15

We report on the design and performance of a two-color, time-resolved detector for the acquisition of both steady-state and time-resolved fluorescence data acquired in real time during the capillary gel electrophoresis separation of DNA sequencing fragments. The detector consisted of a pair of pulsed laser diodes operating at 680 and 780 nm. The diode heads were coupled directly to single-mode fibers, which were terminated into a single fiber mounted via a FC/PC connector to the detector body. The detector contained a dichroic filter, which directed the dual-laser beams to an objective. The objective focused the laser light into a capillary gel column and also collected the resulting fluorescence emission. The dual-color emission was transmitted through the dichroic and focused onto a multimode fiber (core diameter 50 microm), which carried the luminescence to a pair of single-photon avalanche diodes (SPADs). The emission was sorted spectrally using a second dichroic onto one of two SPADs and isolated using appropriate interference filters (710- or 810-nm channel). The dual-color detector demonstrated a time response of 450 and 510 ps (fwhm) for the 710- and 810-nm channels, respectively. The mass detection limits for two near-IR dye-labeled sequencing primers electrophoresed in a capillary gel column were found to be 7.1 x 10(-21) and 3.2 x 10(-20) mol (SNR = 3) for the 710- and 810-nm detector channels, respectively. In addition, no leakage of luminescence excited at 680 nm was observed in the 810-nm channel or 780-nm excited luminescence into the 710-nm channel. An M13mp18 template was sequenced in a single capillary gel column using a two-color, two-lifetime format. The read length was found to be 650 base pairs for the test template at a calling accuracy of 95.1% using a linear poly(dimethylacrylamide) (POP6) gel column, with the read length determined primarily by the electrophoretic resolution produced by the sieving gel.

Demonstration of Time Domain Multiplexed Readout for Magnetically Coupled Calorimeters

NASA Technical Reports Server (NTRS)

Porst, J.-P.; Adams, J. S.; Balvin, M.; Bandler, S.; Beyer, J.; Busch, S. E.; Drung, D.; Seidel, G. M.; Smith, S. J.; Stevenson, T. R.

2012-01-01

Magnetically coupled calorimeters (MCC) have extremely high potential for x-ray applications due to the inherent high energy resolution capability and being non-dissipative. Although very high energy-resolution has been demonstrated, until now there has been no demonstration of multiplexed read-out. We report on the first realization of a time domain multiplexed (TDM) read-out. While this has many similarities with TDM of transition-edge-sensors (TES), for MGGs the energy resolution is limited by the SQUID read-out noise and requires the well established scheme to be altered in order to minimize degradation due to noise aliasing effects. In cur approach, each pixel is read out by a single first stage SQUID (SQ1) that is operated in open loop. The outputs of the SQ1 s are low-pass filtered with an array of low cross-talk inductors, then fed into a single-stage SQUID TD multiplexer. The multiplexer is addressed from room temperature and read out through a single amplifier channel. We present results achieved with a new detector platform. Noise performance is presented and compared to expectations. We have demonstrated multiplexed X-ray spectroscopy at 5.9keV with delta_FWHM=10eV. In an optimized setup, we show it is possible to multiplex 32 detectors without significantly degrading the Intrinsic detector resolution.

An integrated passive micromixer-magnetic separation-capillary electrophoresis microdevice for rapid and multiplex pathogen detection at the single-cell level.

PubMed

Jung, Jae Hwan; Kim, Gha-Young; Seo, Tae Seok

2011-10-21

Here we report an integrated microdevice consisting of an efficient passive mixer, a magnetic separation chamber, and a capillary electrophoretic microchannel in which DNA barcode assay, target pathogen separation, and barcode DNA capillary electrophoretic analysis were performed sequentially within 30 min for multiplex pathogen detection at the single-cell level. The intestine-shaped serpentine 3D micromixer provides a high mixing rate to generate magnetic particle-pathogenic bacteria-DNA barcode labelled AuNP complexes quantitatively. After magnetic separation and purification of those complexes, the barcode DNA strands were released and analyzed by the microfluidic capillary electrophoresis within 5 min. The size of the barcode DNA strand was controlled depending on the target bacteria (Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella typhimurium), and the different elution time of the barcode DNA peak in the electropherogram allows us to recognize the target pathogen with ease in the monoplex as well as in the multiplex analysis. In addition, the quantity of the DNA barcode strand (∼10(4)) per AuNP is enough to be observed in the laser-induced confocal fluorescence detector, thereby making single-cell analysis possible. This novel integrated microdevice enables us to perform rapid, sensitive, and multiplex pathogen detection with sample-in-answer-out capability to be applied for biosafety testing, environmental screening, and clinical trials.

Microgels for multiplex and direct fluorescence detection

NASA Astrophysics Data System (ADS)

Causa, Filippo; Aliberti, Anna; Cusano, Angela M.; Battista, Edmondo; Netti, Paolo A.

2015-05-01

Blood borne oligonucleotides fragments contain useful clinical information whose detection and monitoring represent the new frontier in liquid biopsy as they can transform the current diagnosis procedure. For instance, recent studies have identified a new class of circulating biomarkers such as s miRNAs, and demonstrated that changes in their concentration are closely associated with the development of cancer and other pathologies. However, direct detection of miRNAs in body fluids is particularly challenging and demands high sensitivity -concentration range between atto to femtomolarspecificity, and multiplexing Here we report on engineered multifunctional microgels and innovative probe design for a direct and multiplex detection of relevant clinical miRNAs in fluorescence by single particle assay. Polyethyleneglycol-based microgels have a coreshell architecture with two spectrally encoded fluorescent dyes for multiplex analyses and are endowed with fluorescent probes for miRNA detection. Encoding and detection fluorescence signals are distinguishable by not overlapping emission spectra. Tuneable fluorescence probe conjugation and corresponding emission confinement on single microgel allows for enhanced target detection. Such suspension array has indeed high selectivity and sensitivity with a detection limit of 10-15 M and a dynamic range from 10-9 to 10-15 M. We believe that sensitivity in the fM concentration range, signal background minimization, multiplexed capability and direct measurement of such microgels will translate into diagnostic benefits opening up new roots toward liquid biopsy in the context of point-of-care testing through an easy and fast detection of sensitive diagnostic biomarkers directly in serum.

Microwave SQUID Multiplexer for the Readout of Metallic Magnetic Calorimeters

NASA Astrophysics Data System (ADS)

Kempf, S.; Gastaldo, L.; Fleischmann, A.; Enss, C.

2014-06-01

We have realized a frequency-domain multiplexing technique for the readout of large metallic magnetic calorimeter detector arrays. It is based on non-hysteretic single-junction SQUIDs and allows for a simultaneous readout of hundreds or thousands of detectors by using a single cryogenic high electron mobility transistor amplifier and two coaxial cables that are routed from room-temperature to the detector array. We discuss the working principle of the multiplexer and present details about our prototype multiplexer design. We show that fabricated devices are fully operational and that characteristic SQUID parameters such as the input sensitivity of the SQUID or the resonance frequency of the readout circuit can be predicted with confidence. Our best device so far has shown a magnetic flux white noise level of 1.4 m which can in future be reduced by an optimization of the fabrication processes as well as an improved microwave readout system.

Photonic engineering for biological study

NASA Astrophysics Data System (ADS)

Wu, Fei

My dissertation focuses on designing and developing prototypes of optical tools in the laboratory that can facilitate practical medical therapies. More specifically, this dissertation examines two novel biophotonic techniques: (1) a frequency multiplexed confocal microscope with the potential to provide rational therapy of congestive heart failure (CHF), and (2) the "optical comb" with the potential to improve results of retina reattachment surgery and accelerate post surgical recovery. Next, I will discuss the background, design and initial experimental results of each study individually. Part I: The Frequency Multiplexed Confocal Microscope. To overcome the limitations of existing confocal microscope technology, this dissertation proposes a non-scanning, real-time, high resolution technique (a multi-point frequency multiplexed confocal microscope) to measure 3-D intracellular calcium ion concentration in a living cardiac myocyte. This method can be also applied to measure the intracellular sodium ion concentration, or other ions in which high quantum-yield fluorescent probes are available. The novelty of the proposed research lies in the introduction of carrier frequency multiplexing techniques which can differentiate fluorescence emitted at different spatial locations in cardiac myocyte by their modulated frequency. It therefore opens the possibility to visualize the transient dynamics of intracellular dynamics at multiple locations in cells simultaneously, which will shine a new light on our understanding of CHF. The procedure for frequency multiplexing proposed is described below. Multiple incident laser beams are focused onto different locations in an isolated rat cardiac myocyte with each beam modulated at a different carrier frequency. The fluorescence emission at each location therefore bears the same modulated frequency as the stimulation laser beam. Each fluorescence signal is sent to the photo multiplier tube (PMT) after being spatially filtered by a single mode fiber (functioning as a pinhole). Since each signal has a different carrier frequency, only one signal detector is required to collect multiple signal streams which eliminates the errors introduced by difference of multiple detectors. After taking the Fourier Transform of the collected data, multiple peaks can be found in the frequency domain. Each peak refers to a corresponding location in the sample. The temporal information of the fluorescence signal variation at each location can be obtained by demodulating the low frequency information from the carrier frequency, followed by an inverse Fourier transform. Part II: The "Optical Comb". Retinal detachment refers to separation of the inner layers of the retina from the underlying retinal pigment epithelium. It can cause degeneration of the retina and may lead to permanent vision loss if not promptly treated and hence is considered an ocular emergency. Currently, the only treatment available for retinal detachment is surgical reattachment. The idea of an "optical comb" is developed from the general working principle of the well known "optical tweezers" in the optical literature, which can pull micro-objects through the trapping force produced by a focused laser beam. If we can manage to incident the focused laser beam onto the misaligned photoreceptors and further scan it back and forth, trapping forces that produced may be able to "comb" the photoreceptors to be aligned, and thereby help with post surgery recovery. A series of experiments have been carried out to demonstrate the plausibility of this idea. First, several micro glass rods with size similar to human's photoreceptors (6 microns in diameter and 30 microns in length) were used. We observed that when the laser beam is focused close to one end of the micro rod originally laid on a glass coverslip, the rod is pulled to stand upright successfully, and we can manipulate the direction it faces by controlling its relative position to the laser beam. We are now experimenting with this combing technique with detached bovine retina samples to further verify its feasibility over live animal cells. (Abstract shortened by UMI.)

Enhanced capillary electrophoretic screening of Alzheimer based on direct apolipoprotein E genotyping and one-step multiplex PCR.

PubMed

Woo, Nain; Kim, Su-Kang; Sun, Yucheng; Kang, Seong Ho

2018-01-01

Human apolipoprotein E (ApoE) is associated with high cholesterol levels, coronary artery disease, and especially Alzheimer's disease. In this study, we developed an ApoE genotyping and one-step multiplex polymerase chain reaction (PCR) based-capillary electrophoresis (CE) method for the enhanced diagnosis of Alzheimer's. The primer mixture of ApoE genes enabled the performance of direct one-step multiplex PCR from whole blood without DNA purification. The combination of direct ApoE genotyping and one-step multiplex PCR minimized the risk of DNA loss or contamination due to the process of DNA purification. All amplified PCR products with different DNA lengths (112-, 253-, 308-, 444-, and 514-bp DNA) of the ApoE genes were analyzed within 2min by an extended voltage programming (VP)-based CE under the optimal conditions. The extended VP-based CE method was at least 120-180 times faster than conventional slab gel electrophoresis methods In particular, all amplified DNA fragments were detected in less than 10 PCR cycles using a laser-induced fluorescence detector. The detection limits of the ApoE genes were 6.4-62.0pM, which were approximately 100-100,000 times more sensitive than previous Alzheimer's diagnosis methods In addition, the combined one-step multiplex PCR and extended VP-based CE method was also successfully applied to the analysis of ApoE genotypes in Alzheimer's patients and normal samples and confirmed the distribution probability of allele frequencies. This combination of direct one-step multiplex PCR and an extended VP-based CE method should increase the diagnostic reliability of Alzheimer's with high sensitivity and short analysis time even with direct use of whole blood. Copyright © 2017 Elsevier B.V. All rights reserved.

Fluorescent microarray for multiplexed quantification of environmental contaminants in seawater samples

USDA-ARS?s Scientific Manuscript database

The development of a fluorescent multiplexed microarray platform able to detect and quantify a wide variety of pollutants in seawater is reported. The microarray platform has been manufactured by spotting 6 different bioconjugate competitors and it uses a cocktail of 6 monoclonal and polyclonal anti...

Performance of the SBRC 190, a cryogenic multiplexer for photoconductor arrays

NASA Technical Reports Server (NTRS)

Dotson, Jessie L.; Koerber, C. T.; Mason, C. G.; Simpson, J. P.; Moore, E. M.; Witteborn, F. C.; Farhoomand, J.; Erickson, E. F.; DeVincenzi, D. (Technical Monitor)

2002-01-01

The SBRC 190 cryogenic readouts were developed for use with far-infrared arrays of Ge:Sb and Ge:Ga photoconductor detectors. The SBRC 190 provides an AC-coupled CTIA (capacitance transimpedance amplifier) unit cell for each detector and multiplexes up to 32 detectors. This paper presents our test results characterizing and optimizing the performance of these novel devices. We will discuss their basic behavior in addition to describing the trade-offs inherent in different sampling strategies.

Lithographed Superconducting Resonator Development for Next-Generation Frequency Multiplexing Readout of Transition-Edge Sensors

NASA Astrophysics Data System (ADS)

Faramarzi, F.; De Haan, T.; Kusaka, A.; Lee, A.; Neuhauser, B.; Plambeck, R.; Raum, C.; Suzuki, A.; Westbrook, B.

2018-03-01

Ground-based cosmic microwave background (CMB) experiments are undergoing a period of exponential growth. Current experiments are observing with 1000-10,000 detectors, and the next-generation experiment (CMB stage 4) is proposing to deploy approximately 500,000 detectors. This order of magnitude increase in detector count will require a new approach for readout electronics. We have developed superconducting resonators for next-generation frequency-domain multiplexing (fMUX) readout architecture. Our goal is to reduce the physical size of resonators, such that resonators and detectors can eventually be integrated on a single wafer. To reduce the size of these resonators, we have designed spiral inductors and interdigitated capacitors that resonate around 10-100 MHz, an order of magnitude higher frequency compared to current fMUX readout systems. The higher frequency leads to a wider bandwidth and would enable higher multiplexing factor than the current ˜ 50 detectors per readout channel. We will report on the simulation, fabrication method, characterization technique, and measurement of quality factor of these resonators.

Highly multiplexed signal readout for a time-of-flight positron emission tomography detector based on silicon photomultipliers.

PubMed

Cates, Joshua W; Bieniosek, Matthew F; Levin, Craig S

2017-01-01

Maintaining excellent timing resolution in the generation of silicon photomultiplier (SiPM)-based time-of-flight positron emission tomography (TOF-PET) systems requires a large number of high-speed, high-bandwidth electronic channels and components. To minimize the cost and complexity of a system's back-end architecture and data acquisition, many analog signals are often multiplexed to fewer channels using techniques that encode timing, energy, and position information. With progress in the development SiPMs having lower dark noise, after pulsing, and cross talk along with higher photodetection efficiency, a coincidence timing resolution (CTR) well below 200 ps FWHM is now easily achievable in single pixel, bench-top setups using 20-mm length, lutetium-based inorganic scintillators. However, multiplexing the output of many SiPMs to a single channel will significantly degrade CTR without appropriate signal processing. We test the performance of a PET detector readout concept that multiplexes 16 SiPMs to two channels. One channel provides timing information with fast comparators, and the second channel encodes both position and energy information in a time-over-threshold-based pulse sequence. This multiplexing readout concept was constructed with discrete components to process signals from a [Formula: see text] array of SensL MicroFC-30035 SiPMs coupled to [Formula: see text] Lu 1.8 Gd 0.2 SiO 5 (LGSO):Ce (0.025 mol. %) scintillators. This readout method yielded a calibrated, global energy resolution of 15.3% FWHM at 511 keV with a CTR of [Formula: see text] FWHM between the 16-pixel multiplexed detector array and a [Formula: see text] LGSO-SiPM reference detector. In summary, results indicate this multiplexing scheme is a scalable readout technique that provides excellent coincidence timing performance.

Integrating IR detector imaging systems

NASA Technical Reports Server (NTRS)

Bailey, G. C. (Inventor)

1984-01-01

An integrating IR detector array for imaging is provided in a hybrid circuit with InSb mesa diodes in a linear array, a single J-FET preamplifier for readout, and a silicon integrated circuit multiplexer. Thin film conductors in a fan out pattern deposited on an Al2O3 substrate connect the diodes to the multiplexer, and thick film conductors also connect the reset switch and preamplifier to the multiplexer. Two phase clock pulses are applied with a logic return signal to the multiplexer through triax comprised of three thin film conductors deposited between layers. A lens focuses a scanned image onto the diode array for horizontal read out while a scanning mirror provides vertical scan.

Si:Bi switched photoconducttor infrared detector array

NASA Technical Reports Server (NTRS)

Eakin, C. E.

1983-01-01

A multiplexed infrared detector array is described. The small demonstration prototype consisted of two cryogenically cooled, bismuth doped silicon, extrinsic photoconductor pixels multiplexed onto a single output channel using an on focal plane switch integration sampling technique. Noise levels of the order of 400 to 600 rms electrons per sample were demonstrated for this chip and wire hybrid version.

Multispectral computational ghost imaging with multiplexed illumination

NASA Astrophysics Data System (ADS)

Huang, Jian; Shi, Dongfeng

2017-07-01

Computational ghost imaging has attracted wide attention from researchers in many fields over the last two decades. Multispectral imaging as one application of computational ghost imaging possesses spatial and spectral resolving abilities, and is very useful for surveying scenes and extracting detailed information. Existing multispectral imagers mostly utilize narrow band filters or dispersive optical devices to separate light of different wavelengths, and then use multiple bucket detectors or an array detector to record them separately. Here, we propose a novel multispectral ghost imaging method that uses one single bucket detector with multiplexed illumination to produce a colored image. The multiplexed illumination patterns are produced by three binary encoded matrices (corresponding to the red, green and blue colored information, respectively) and random patterns. The results of the simulation and experiment have verified that our method can be effective in recovering the colored object. Multispectral images are produced simultaneously by one single-pixel detector, which significantly reduces the amount of data acquisition.

Design, development and evaluation of a resistor-based multiplexing circuit for a 20×20 SiPM array

NASA Astrophysics Data System (ADS)

Wang, Zhonghai; Sun, Xishan; Lou, Kai; Meier, Joseph; Zhou, Rong; Yang, Chaowen; Zhu, Xiaorong; Shao, Yiping

2016-04-01

One technical challenge in developing a large-size scintillator detector with multiple Silicon Photomultiplier (SiPM) arrays is to read out a large number of detector output channels. To achieve this, different signal multiplexing circuits have been studied and applied with different performances and cost-effective tradeoffs. Resistor-based multiplexing circuits exhibit simplicity and signal integrity, but also present the disadvantage of timing shift among different channels. In this study, a resistor-based multiplexing circuit for a large-sized SiPM array readout was developed and evaluated by simulation and experimental studies. Similarly to a multiplexing circuit used for multi-anode PMT, grounding and branching resistors were connected to each SiPM output channel. The grounding resistor was used to simultaneously reduce the signal crosstalk among different channels and to improve timing performance. Both grounding and branching resistor values were optimized to maintain a balanced performance of the event energy, timing, and positioning. A multiplexing circuit was implemented on a compact PCB and applied for a flat-panel detector which consisted of a 32×32 LYSO scintillator crystals optically coupled to 5×5 SiPM arrays for a total 20×20 output channels. Test results showed excellent crystal identification for all 1024 LYSO crystals (each with 2×2×30 mm3 size) with 22Na flood-source irradiation. The measured peak-to-valley ratio from typical crystal map profile is around 3:1 to 6.6:1, an average single crystal energy resolution of about 17.3%, and an average single crystal timing resolution of about 2 ns. Timing shift among different crystals, as reported in some other resistor-based multiplexing circuit designs, was not observed. In summary, we have designed and implemented a practical resistor-based multiplexing circuit that can be readily applied for reading out a large SiPM array with good detector performance.

Preliminary Assessment of Microwave Readout Multiplexing Factor

DOE Office of Scientific and Technical Information (OSTI.GOV)

Croce, Mark Philip; Koehler, Katrina Elizabeth; Rabin, Michael W.

2017-01-23

Ultra-high resolution microcalorimeter gamma spectroscopy is a new non-destructive assay technology for measurement of plutonium isotopic composition, with the potential to reduce total measurement uncertainty to a level competitive with destructive analysis methods [1-4]. Achieving this level of performance in practical applications requires not only the energy resolution now routinely achieved with transition-edge sensor microcalorimeter arrays (an order of magnitude better than for germanium detectors) but also high throughput. Microcalorimeter gamma spectrometers have not yet achieved detection efficiency and count rate capability that is comparable to germanium detectors, largely because of limits from existing readout technology. Microcalorimeter detectors must bemore » operated at low temperature to achieve their exceptional energy resolution. Although the typical 100 mK operating temperatures can be achieved with reliable, cryogen-free systems, the cryogenic complexity and heat load from individual readout channels for large sensor arrays is prohibitive. Multiplexing is required for practical systems. The most mature multiplexing technology at present is time-division multiplexing (TDM) [3, 5-6]. In TDM, the sensor outputs are switched by applying bias current to one SQUID amplifier at a time. Transition-edge sensor (TES) microcalorimeter arrays as large as 256 pixels have been developed for X-ray and gamma-ray spectroscopy using TDM technology. Due to bandwidth limits and noise scaling, TDM is limited to a maximum multiplexing factor of approximately 32-40 sensors on one readout line [8]. Increasing the size of microcalorimeter arrays above the kilopixel scale, required to match the throughput of germanium detectors, requires the development of a new readout technology with a much higher multiplexing factor.« less

[A review of mixed gas detection system based on infrared spectroscopic technique].

PubMed

Dang, Jing-Min; Fu, Li; Yan, Zi-Hui; Zheng, Chuan-Tao; Chang, Yu-Chun; Chen, Chen; Wang, Yi-Din

2014-10-01

In order to provide the experiences and references to the researchers who are working on infrared (IR) mixed gas detection field. The proposed manuscript reviews two sections of the aforementioned field, including optical multiplexing structure and detection method. At present, the coherent light sources whose representative are quantum cascade laser (QCL) and inter-band cascade laser(ICL) become the mainstream light source in IR mixed gas detection, which replace the traditional non-coherent light source, such as IR radiation source and IR light emitting diode. In addition, the photon detector which has a super high detectivity and very short response time is gradually beyond thermal infrared detector, dominant in the field of infrared detector. The optical multiplexing structure is the key factor of IR mixed gas detection system, which consists of single light source multi-plexing detection structure and multi light source multiplexing detection structure. Particularly, single light source multiplexing detection structure is advantages of small volume and high integration, which make it a plausible candidate for the portable mixed gas detection system; Meanwhile, multi light source multiplexing detection structure is embodiment of time division multiplex, frequency division multiplexing and wavelength division multiplexing, and become the leading structure of the mixed gas detection system because of its wider spectral range, higher spectral resolution, etc. The detection method applied to IR mixed gas detection includes non-dispersive infrared (NDIR) spectroscopy, wavelength and frequency-modulation spectroscopy, cavity-enhanced spectroscopy and photoacoustic spectroscopy, etc. The IR mixed gas detection system designed by researchers after recognizing the whole sections of the proposed system, which play a significant role in industrial and agricultural production, environmental monitoring, and life science, etc.

Frequency multiplexed superconducting quantum interference device readout of large bolometer arrays for cosmic microwave background measurements.

PubMed

Dobbs, M A; Lueker, M; Aird, K A; Bender, A N; Benson, B A; Bleem, L E; Carlstrom, J E; Chang, C L; Cho, H-M; Clarke, J; Crawford, T M; Crites, A T; Flanigan, D I; de Haan, T; George, E M; Halverson, N W; Holzapfel, W L; Hrubes, J D; Johnson, B R; Joseph, J; Keisler, R; Kennedy, J; Kermish, Z; Lanting, T M; Lee, A T; Leitch, E M; Luong-Van, D; McMahon, J J; Mehl, J; Meyer, S S; Montroy, T E; Padin, S; Plagge, T; Pryke, C; Richards, P L; Ruhl, J E; Schaffer, K K; Schwan, D; Shirokoff, E; Spieler, H G; Staniszewski, Z; Stark, A A; Vanderlinde, K; Vieira, J D; Vu, C; Westbrook, B; Williamson, R

2012-07-01

A technological milestone for experiments employing transition edge sensor bolometers operating at sub-Kelvin temperature is the deployment of detector arrays with 100s-1000s of bolometers. One key technology for such arrays is readout multiplexing: the ability to read out many sensors simultaneously on the same set of wires. This paper describes a frequency-domain multiplexed readout system which has been developed for and deployed on the APEX-SZ and South Pole Telescope millimeter wavelength receivers. In this system, the detector array is divided into modules of seven detectors, and each bolometer within the module is biased with a unique ∼MHz sinusoidal carrier such that the individual bolometer signals are well separated in frequency space. The currents from all bolometers in a module are summed together and pre-amplified with superconducting quantum interference devices operating at 4 K. Room temperature electronics demodulate the carriers to recover the bolometer signals, which are digitized separately and stored to disk. This readout system contributes little noise relative to the detectors themselves, is remarkably insensitive to unwanted microphonic excitations, and provides a technology pathway to multiplexing larger numbers of sensors.

Laboratory Tests of Multiplex Detection of PCR Amplicons Using the Luminex 100 Flow Analyzer

DOE Office of Scientific and Technical Information (OSTI.GOV)

Venkateswaran, K.S.; Nasarabadi, S.; Langlois, R.G.

2000-05-05

Lawrence Livermore National Laboratory (LLNL) demonstrated the power of flow cytometry in detecting the biological agents simulants at JFT III. LLNL pioneered in the development of advanced nucleic acid analyzer (ANM) for portable real time identification. Recent advances in flow cytometry provide a means for multiplexed nucleic acid detection and immunoassay of pathogenic microorganisms. We are presently developing multiplexed immunoassays for the simultaneous detection of different simulants. Our goal is to build an integrated instrument for both nucleic acid analysis and immuno detection. In this study we evaluated the Luminex LX 100 for concurrent identification of more than one PCRmore » amplified product. ANAA has real-time Taqman fluorescent detection capability for rapid identification of field samples. However, its multiplexing ability is limited by the combination of available fluorescent labels. Hence integration of ANAA with flow cytometry can give the rapidity of ANAA amplification and the multiplex capability of flow cytometry. Multiplexed flow cytometric analysis is made possible using a set of fluorescent latex microsphere that are individually identified by their red and infrared fluorescence. A green fluorochrome is used as the assay signal. Methods were developed for the identification of specific nucleic acid sequences from Bacillus globigii (Bg), Bacillus thuringensis (Bt) and Erwinia herbicola (Eh). Detection sensitivity using different reporter fluorochromes was tested with the LX 100, and also different assay formats were evaluated for their suitability for rapid testing. A blind laboratory trial was carried out December 22-27, 1999 to evaluate bead assays for multiplex identification of Bg and Bt PCR products. This report summarizes the assay development, fluorochrome comparisons, and the results of the blind trial conducted at LLNL for the laboratory evaluation of the LX 100 flow analyzer.« less

Indium Hybridization of Large Format TES Bolometer Arrays to Readout Multiplexers for Far-Infrared Astronomy

NASA Technical Reports Server (NTRS)

Miller, Timothy M.; Costen, Nick; Allen, Christine

2007-01-01

This conference poster reviews the Indium hybridization of the large format TES bolometer arrays. We are developing a key technology to enable the next generation of detectors. That is the Hybridization of Large Format Arrays using Indium bonded detector arrays containing 32x40 elements which conforms to the NIST multiplexer readout architecture of 1135 micron pitch. We have fabricated and hybridized mechanical models with the detector chips bonded after being fully back-etched. The mechanical support consists of 30 micron walls between elements Demonstrated electrical continuity for each element. The goal is to hybridize fully functional array of TES detectors to NIST readout.

Far infrared through millimeter backshort-under-grid arrays

NASA Astrophysics Data System (ADS)

Allen, Christine A.; Abrahams, John; Benford, Dominic J.; Chervenak, James A.; Chuss, David T.; Staguhn, Johannes G.; Miller, Timothy M.; Moseley, S. Harvey; Wollack, Edward J.

2006-06-01

We are developing a large-format, versatile, bolometer array for a wide range of infrared through millimeter astronomical applications. The array design consists of three key components - superconducting transition edge sensor bolometer arrays, quarter-wave reflective backshort grids, and Superconducting Quantum Interference Device (SQUID) multiplexer readouts. The detector array is a filled, square grid of bolometers with superconducting sensors. The backshort arrays are fabricated separately and are positioned in the etch cavities behind the detector grid. The grids have unique three-dimensional interlocking features micromachined into the walls for positioning and mechanical stability. The ultimate goal of the program is to produce large-format arrays with background-limited sensitivity, suitable for a wide range of wavelengths and applications. Large-format (kilopixel) arrays will be directly indium bump bonded to a SQUID multiplexer circuit. We have produced and tested 8×8 arrays of 1 mm detectors to demonstrate proof of concept. 8×16 arrays of 2 mm detectors are being produced for a new Goddard Space Flight Center instrument. We have also produced models of a kilopixel detector grid and dummy multiplexer chip for bump bonding development. We present detector design overview, several unique fabrication highlights, and assembly technologies.

Performance of Multiplexed XY Resistive Micromegas detectors in a high intensity beam

NASA Astrophysics Data System (ADS)

Banerjee, D.; Burtsev, V.; Chumakov, A.; Cooke, D.; Depero, E.; Dermenev, A. V.; Donskov, S. V.; Dubinin, F.; Dusaev, R. R.; Emmenegger, S.; Fabich, A.; Frolov, V. N.; Gardikiotis, A.; Gninenko, S. N.; Hösgen, M.; Karneyeu, A. E.; Ketzer, B.; Kirsanov, M. M.; Konorov, I. V.; Kramarenko, V. A.; Kuleshov, S. V.; Levchenko, E.; Lyubovitskij, V. E.; Lysan, V.; Mamon, S.; Matveev, V. A.; Mikhailov, Yu. V.; Myalkovskiy, V. V.; Peshekhonov, V. D.; Peshekhonov, D. V.; Polyakov, V. A.; Radics, B.; Rubbia, A.; Samoylenko, V. D.; Tikhomirov, V. O.; Tlisov, D. A.; Toropin, A. N.; Vasilishin, B.; Arenas, G. Vasquez; Ulloa, P.; Crivelli, P.

2018-02-01

We present the performance of multiplexed XY resistive Micromegas detectors tested in the CERN SPS 100 GeV/c electron beam at intensities up to 3 . 3 × 105e- /(s ṡcm2) . So far, all studies with multiplexed Micromegas have only been reported for tests with radioactive sources and cosmic rays. The use of multiplexed modules in high intensity environments was not explored due to the effect of ambiguities in the reconstruction of the hit point caused by the multiplexing feature. For the specific mapping and beam intensities analyzed in this work with a multiplexing factor of five, more than 50% level of ambiguity is introduced due to particle pile-up as well as fake clusters due to the mapping feature. Our results prove that by using the additional information of cluster size and integrated charge from the signal clusters induced on the XY strips, the ambiguities can be reduced to a level below 2%. The tested detectors are used in the CERN NA64 experiment for tracking the incoming particles bending in a magnetic field in order to reconstruct their momentum. The average hit detection efficiency of each module was found to be ∼96% at the highest beam intensities. By using four modules a tracking resolution of 1.1% was obtained with ∼85% combined tracking efficiency.

Upconversion Nanoparticles-Encoded Hydrogel Microbeads-Based Multiplexed Protein Detection

NASA Astrophysics Data System (ADS)

Shikha, Swati; Zheng, Xiang; Zhang, Yong

2018-06-01

Fluorescently encoded microbeads are in demand for multiplexed applications in different fields. Compared to organic dye-based commercially available Luminex's xMAP technology, upconversion nanoparticles (UCNPs) are better alternatives due to their large anti-Stokes shift, photostability, nil background, and single wavelength excitation. Here, we developed a new multiplexed detection system using UCNPs for encoding poly(ethylene glycol) diacrylate (PEGDA) microbeads as well as for labeling reporter antibody. However, to prepare UCNPs-encoded microbeads, currently used swelling-based encapsulation leads to non-uniformity, which is undesirable for fluorescence-based multiplexing. Hence, we utilized droplet microfluidics to obtain encoded microbeads of uniform size, shape, and UCNPs distribution inside. Additionally, PEGDA microbeads lack functionality for probe antibodies conjugation on their surface. Methods to functionalize the surface of PEGDA microbeads (acrylic acid incorporation, polydopamine coating) reported thus far quench the fluorescence of UCNPs. Here, PEGDA microbeads surface was coated with silica followed by carboxyl modification without compromising the fluorescence intensity of UCNPs. In this study, droplet microfluidics-assisted UCNPs-encoded microbeads of uniform shape, size, and fluorescence were prepared. Multiple color codes were generated by mixing UCNPs emitting red and green colors at different ratios prior to encapsulation. UCNPs emitting blue color were used to label the reporter antibody. Probe antibodies were covalently immobilized on red UCNPs-encoded microbeads for specific capture of human serum albumin (HSA) as a model protein. The system was also demonstrated for multiplexed detection of both human C-reactive protein (hCRP) and HSA protein by immobilizing anti-hCRP antibodies on green UCNPs.

PbS-PbSe IR detector arrays

NASA Technical Reports Server (NTRS)

Barrett, John R. (Inventor)

1986-01-01

A silicon wafer is provided which does not employ individually bonded leads between the IR sensitive elements and the input stages of multiplexers. The wafer is first coated with lead selenide in a first detector array area and is thereafter coated with lead sulfide within a second detector array area. The described steps result in the direct chemical deposition of lead selenide and lead sulfide upon the silicon wafer to eliminate individual wire bonding, bumping, flip chipping, planar interconnecting methods of connecting detector array elements to silicon chip circuitry, e.g., multiplexers, to enable easy fabrication of very long arrays. The electrode structure employed, produces an increase in the electrical field gradient between the electrodes for a given volume of detector material, relative to conventional electrode configurations.

Development of a unit cell for a Ge:Ga detector array

NASA Technical Reports Server (NTRS)

1988-01-01

Two modules of gallium-doped germanium (Ge:Ga) infrared detectors with integrated multiplexing readouts and supporting drive electronics were designed and tested. This development investigated the feasibility of producing two-dimensional Ge:Ga arrays by stacking linear modules in a housing capable of providing uniaxial stress for enhanced long-wavelength response. Each module includes 8 detectors (1x1x2 mm) mounted to a sapphire board. The element spacing is 12 microns. The back faces of the detector elements are beveled with an 18 deg angle, which was proved to significantly enhance optical absorption. Each module includes a different silicon metal-oxide semiconductor field effect transistor (MOSFET) readout. The first circuit was built from discrete MOSFET components; the second incorporated devices taken from low-temperature integrated circuit multiplexers. The latter circuit exhibited much lower stray capacitance and improved stability. Using these switched-FET circuits, it was demonstrated that burst readout, with multiplexer active only during the readout period, could successfully be implemented at approximately 3.5 K.

TiO2 Nanolayer-Enhanced Fluorescence for Simultaneous Multiplex Mycotoxin Detection by Aptamer Microarrays on a Porous Silicon Surface.

PubMed

Liu, Rui; Li, Wei; Cai, Tingting; Deng, Yang; Ding, Zhi; Liu, Yan; Zhu, Xuerui; Wang, Xin; Liu, Jie; Liang, Baowen; Zheng, Tiesong; Li, Jianlin

2018-05-02

A new aptamer microarray method on the TiO 2 -porous silicon (PSi) surface was developed to simultaneously screen multiplex mycotoxins. The TiO 2 nanolayer on the surface of PSi can enhance the fluorescence intensity 14 times than that of the thermally oxidized PSi. The aptamer fluorescence signal recovery principle was performed on the TiO 2 -PSi surface by hybridization duplex strand DNA from the mycotoxin aptamer and antiaptamer, respectively, labeled with fluorescence dye and quencher. The aptamer microarray can simultaneously screen for multiplex mycotoxins with a dynamic linear detection range of 0.1-10 ng/mL for ochratoxin A (OTA), 0.01-10 ng/mL for aflatoxins B 1 (AFB 1 ), and 0.001-10 ng/mL for fumonisin B 1 (FB 1 ) and limits of detection of 15.4, 1.48, and 0.21 pg/mL for OTA, AFB 1 , and FB 1 , respectively. The newly developed method shows good specificity and recovery rates. This method can provide a simple, sensitive, and cost-efficient platform for simultaneous screening of multiplex mycotoxins and can be easily expanded to the other aptamer-based protocol.

Hyperspectral fluorescence imaging with multi wavelength LED excitation

NASA Astrophysics Data System (ADS)

Luthman, A. Siri; Dumitru, Sebastian; Quirós-Gonzalez, Isabel; Bohndiek, Sarah E.

2016-04-01

Hyperspectral imaging (HSI) can combine morphological and molecular information, yielding potential for real-time and high throughput multiplexed fluorescent contrast agent imaging. Multiplexed readout from targets, such as cell surface receptors overexpressed in cancer cells, could improve both sensitivity and specificity of tumor identification. There remains, however, a need for compact and cost effective implementations of the technology. We have implemented a low-cost wide-field multiplexed fluorescence imaging system, which combines LED excitation at 590, 655 and 740 nm with a compact commercial solid state HSI system operating in the range 600 - 1000 nm. A key challenge for using reflectance-based HSI is the separation of contrast agent fluorescence from the reflectance of the excitation light. Here, we illustrate how it is possible to address this challenge in software, using two offline reflectance removal methods, prior to least-squares spectral unmixing. We made a quantitative comparison of the methods using data acquired from dilutions of contrast agents prepared in well-plates. We then established the capability of our HSI system for non-invasive in vivo fluorescence imaging in small animals using the optimal reflectance removal method. The HSI presented here enables quantitative unmixing of at least four fluorescent contrast agents (Alexa Fluor 610, 647, 700 and 750) simultaneously in living mice. A successful unmixing of the four fluorescent contrast agents was possible both using the pure contrast agents and with mixtures. The system could in principle also be applied to imaging of ex vivo tissue or intraoperative imaging in a clinical setting. These data suggest a promising approach for developing clinical applications of HSI based on multiplexed fluorescence contrast agent imaging.

Highly multiplexed signal readout for a time-of-flight positron emission tomography detector based on silicon photomultipliers

PubMed Central

Cates, Joshua W.; Bieniosek, Matthew F.; Levin, Craig S.

2017-01-01

Abstract. Maintaining excellent timing resolution in the generation of silicon photomultiplier (SiPM)-based time-of-flight positron emission tomography (TOF-PET) systems requires a large number of high-speed, high-bandwidth electronic channels and components. To minimize the cost and complexity of a system’s back-end architecture and data acquisition, many analog signals are often multiplexed to fewer channels using techniques that encode timing, energy, and position information. With progress in the development SiPMs having lower dark noise, after pulsing, and cross talk along with higher photodetection efficiency, a coincidence timing resolution (CTR) well below 200 ps FWHM is now easily achievable in single pixel, bench-top setups using 20-mm length, lutetium-based inorganic scintillators. However, multiplexing the output of many SiPMs to a single channel will significantly degrade CTR without appropriate signal processing. We test the performance of a PET detector readout concept that multiplexes 16 SiPMs to two channels. One channel provides timing information with fast comparators, and the second channel encodes both position and energy information in a time-over-threshold-based pulse sequence. This multiplexing readout concept was constructed with discrete components to process signals from a 4×4 array of SensL MicroFC-30035 SiPMs coupled to 2.9×2.9×20  mm3 Lu1.8Gd0.2SiO5 (LGSO):Ce (0.025 mol. %) scintillators. This readout method yielded a calibrated, global energy resolution of 15.3% FWHM at 511 keV with a CTR of 198±2  ps FWHM between the 16-pixel multiplexed detector array and a 2.9×2.9×20  mm3 LGSO-SiPM reference detector. In summary, results indicate this multiplexing scheme is a scalable readout technique that provides excellent coincidence timing performance. PMID:28382312

Optofluidic wavelength division multiplexing for single-virus detection

PubMed Central

Ozcelik, Damla; Parks, Joshua W.; Wall, Thomas A.; Stott, Matthew A.; Cai, Hong; Parks, Joseph W.; Hawkins, Aaron R.; Schmidt, Holger

2015-01-01

Optical waveguides simultaneously transport light at different colors, forming the basis of fiber-optic telecommunication networks that shuttle data in dozens of spectrally separated channels. Here, we reimagine this wavelength division multiplexing (WDM) paradigm in a novel context––the differentiated detection and identification of single influenza viruses on a chip. We use a single multimode interference (MMI) waveguide to create wavelength-dependent spot patterns across the entire visible spectrum and enable multiplexed single biomolecule detection on an optofluidic chip. Each target is identified by its time-dependent fluorescence signal without the need for spectral demultiplexing upon detection. We demonstrate detection of individual fluorescently labeled virus particles of three influenza A subtypes in two implementations: labeling of each virus using three different colors and two-color combinatorial labeling. By extending combinatorial multiplexing to three or more colors, MMI-based WDM provides the multiplexing power required for differentiated clinical tests and the growing field of personalized medicine. PMID:26438840

Updates on the Transition-Edge Sensors and Multiplexed Readout for HOLMES

NASA Astrophysics Data System (ADS)

Puiu, A.; Becker, D.; Bennett, D.; Biasotti, M.; Borghesi, M.; Ceriale, V.; De Gerone, M.; Faverzani, M.; Ferri, E.; Fowler, J.; Gallucci, G.; Gard, J.; Hays-Wehle, J.; Hilton, G.; Giachero, A.; Mates, J.; Nucciotti, A.; Orlando, A.; Pessina, G.; Schmidt, D.; Swetz, D.; Ullom, J.; Vale, L.

2018-05-01

Measuring the neutrino mass is one of the most compelling issues in particle physics. HOLMES is an experiment for a direct measurement of the neutrino mass. HOLMES will perform a precise measurement of the end point of the electron capture decay spectrum of ^{163}Ho in order to extract information on the neutrino mass with a sensitivity as low as 1 eV. HOLMES, in its final configuration, will deploy a 1000-pixel array of low-temperature microcalorimeters: each calorimeter is made of an absorber, where the Ho atoms will be implanted, coupled to a transition-edge sensor (TES) thermometer. The detectors will be operated at the working temperature of 100 mK provided by a dilution refrigerator. In order to read out the 1000-detector array of HOLMES, a multiplexing system is necessary: the choice is to couple the transition-edge sensors to a multiplexed rf-SQUID. In this contribution we outline the progress made towards the final configuration of HOLMES regarding both the performances of the TES detectors and the characteristics of the multiplexing system.

Automated hybridization/imaging device for fluorescent multiplex DNA sequencing

DOEpatents

Weiss, R.B.; Kimball, A.W.; Gesteland, R.F.; Ferguson, F.M.; Dunn, D.M.; Di Sera, L.J.; Cherry, J.L.

1995-11-28

A method is disclosed for automated multiplex sequencing of DNA with an integrated automated imaging hybridization chamber system. This system comprises an hybridization chamber device for mounting a membrane containing size-fractionated multiplex sequencing reaction products, apparatus for fluid delivery to the chamber device, imaging apparatus for light delivery to the membrane and image recording of fluorescence emanating from the membrane while in the chamber device, and programmable controller apparatus for controlling operation of the system. The multiplex reaction products are hybridized with a probe, the enzyme (such as alkaline phosphatase) is bound to a binding moiety on the probe, and a fluorogenic substrate (such as a benzothiazole derivative) is introduced into the chamber device by the fluid delivery apparatus. The enzyme converts the fluorogenic substrate into a fluorescent product which, when illuminated in the chamber device with a beam of light from the imaging apparatus, excites fluorescence of the fluorescent product to produce a pattern of hybridization. The pattern of hybridization is imaged by a CCD camera component of the imaging apparatus to obtain a series of digital signals. These signals are converted by the controller apparatus into a string of nucleotides corresponding to the nucleotide sequence an automated sequence reader. The method and apparatus are also applicable to other membrane-based applications such as colony and plaque hybridization and Southern, Northern, and Western blots. 9 figs.

Automated hybridization/imaging device for fluorescent multiplex DNA sequencing

DOEpatents

Weiss, Robert B.; Kimball, Alvin W.; Gesteland, Raymond F.; Ferguson, F. Mark; Dunn, Diane M.; Di Sera, Leonard J.; Cherry, Joshua L.

1995-01-01

A method is disclosed for automated multiplex sequencing of DNA with an integrated automated imaging hybridization chamber system. This system comprises an hybridization chamber device for mounting a membrane containing size-fractionated multiplex sequencing reaction products, apparatus for fluid delivery to the chamber device, imaging apparatus for light delivery to the membrane and image recording of fluorescence emanating from the membrane while in the chamber device, and programmable controller apparatus for controlling operation of the system. The multiplex reaction products are hybridized with a probe, then an enzyme (such as alkaline phosphatase) is bound to a binding moiety on the probe, and a fluorogenic substrate (such as a benzothiazole derivative) is introduced into the chamber device by the fluid delivery apparatus. The enzyme converts the fluorogenic substrate into a fluorescent product which, when illuminated in the chamber device with a beam of light from the imaging apparatus, excites fluorescence of the fluorescent product to produce a pattern of hybridization. The pattern of hybridization is imaged by a CCD camera component of the imaging apparatus to obtain a series of digital signals. These signals are converted by the controller apparatus into a string of nucleotides corresponding to the nucleotide sequence an automated sequence reader. The method and apparatus are also applicable to other membrane-based applications such as colony and plaque hybridization and Southern, Northern, and Western blots.

Multi-channel detector readout method and integrated circuit

DOEpatents

Moses, William W.; Beuville, Eric; Pedrali-Noy, Marzio

2006-12-12

An integrated circuit which provides multi-channel detector readout from a detector array. The circuit receives multiple signals from the elements of a detector array and compares the sampled amplitudes of these signals against a noise-floor threshold and against one another. A digital signal is generated which corresponds to the location of the highest of these signal amplitudes which exceeds the noise floor threshold. The digital signal is received by a multiplexing circuit which outputs an analog signal corresponding the highest of the input signal amplitudes. In addition a digital control section provides for programmatic control of the multiplexer circuit, amplifier gain, amplifier reset, masking selection, and test circuit functionality on each input thereof.

Multi-channel detector readout method and integrated circuit

DOEpatents

Moses, William W.; Beuville, Eric; Pedrali-Noy, Marzio

2004-05-18

An integrated circuit which provides multi-channel detector readout from a detector array. The circuit receives multiple signals from the elements of a detector array and compares the sampled amplitudes of these signals against a noise-floor threshold and against one another. A digital signal is generated which corresponds to the location of the highest of these signal amplitudes which exceeds the noise floor threshold. The digital signal is received by a multiplexing circuit which outputs an analog signal corresponding the highest of the input signal amplitudes. In addition a digital control section provides for programmatic control of the multiplexer circuit, amplifier gain, amplifier reset, masking selection, and test circuit functionality on each input thereof.

Multi-Channel Hyperspectral Fluorescence Detection Excited by Coupled Plasmon-Waveguide Resonance

PubMed Central

Du, Chan; Liu, Le; Zhang, Lin; Guo, Jun; Guo, Jihua; Ma, Hui; He, Yonghong

2013-01-01

We propose in this paper a biosensor scheme based on coupled plasmon-waveguide resonance (CPWR) excited fluorescence spectroscopy. A symmetrical structure that offers higher surface electric field strengths, longer surface propagation lengths and depths is developed to support guided waveguide modes for the efficient excitation of fluorescence. The optimal parameters for the sensor films are theoretically and experimentally investigated, leading to a detection limit of 0.1 nM (for a Cy5 solution). Multiplex analysis possible with the fluorescence detection is further advanced by employing the hyperspectral fluorescence technique to record the full spectra for every pixel on the sample plane. We demonstrate experimentally that highly overlapping fluorescence (Cy5 and Dylight680) can be distinguished and ratios of different emission sources can be determined accurately. This biosensor shows great potential for multiplex detections of fluorescence analytes. PMID:24129023

Chopped molecular beam multiplexing system

NASA Technical Reports Server (NTRS)

Adams, Billy R. (Inventor)

1986-01-01

The integration of a chopped molecular beam mass spectrometer with a time multiplexing system is described. The chopping of the molecular beam is synchronized with the time intervals by a phase detector and a synchronous motor. Arithmetic means are generated for phase shifting the chopper with respect to the multiplexer. A four channel amplifier provides the capacity to independently vary the baseline and amplitude in each channel of the multiplexing system.

Development and potential applications of microarrays based on fluorescent nanocrystal-encoded beads for multiplexed cancer diagnostics

NASA Astrophysics Data System (ADS)

Brazhnik, Kristina; Grinevich, Regina; Efimov, Anton E.; Nabiev, Igor; Sukhanova, Alyona

2014-05-01

Advanced multiplexed assays have recently become an indispensable tool for clinical diagnostics. These techniques provide simultaneous quantitative determination of multiple biomolecules in a single sample quickly and accurately. The development of multiplex suspension arrays is currently of particular interest for clinical applications. Optical encoding of microparticles is the most available and easy-to-use technique. This technology uses fluorophores incorporated into microbeads to obtain individual optical codes. Fluorophore-encoded beads can be rapidly analyzed using classical flow cytometry or microfluidic techniques. We have developed a new generation of highly sensitive and specific diagnostic systems for detection of cancer antigens in human serum samples based on microbeads encoded with fluorescent quantum dots (QDs). The designed suspension microarray system was validated for quantitative detection of (1) free and total prostate specific antigen (PSA) in the serum of patients with prostate cancer and (2) carcinoembryonic antigen (CEA) and cancer antigen 15-3 (CA 15-3) in the serum of patients with breast cancer. The serum samples from healthy donors were used as a control. The antigen detection is based on the formation of an immune complex of a specific capture antibody (Ab), a target antigen (Ag), and a detector Ab on the surface of the encoded particles. The capture Ab is bound to the polymer shell of microbeads via an adapter molecule, for example, protein A. Protein A binds a monoclonal Ab in a highly oriented manner due to specific interaction with the Fc-region of the Ab molecule. Each antigen can be recognized and detected due to a specific microbead population carrying the unique fluorescent code. 100 and 231 serum samples from patients with different stages of prostate cancer and breast cancer, respectively, and those from healthy donors were examined using the designed suspension system. The data were validated by comparing with the results of the "gold standard" enzyme-linked immunosorbent assay (ELISA). They have shown that our approach is a good alternative to the diagnostics of cancer markers using conventional assays, especially in early diagnostic applications.

Method of fabricating a PbS-PbSe IR detector array

NASA Technical Reports Server (NTRS)

Barrett, John R. (Inventor)

1987-01-01

A silicon wafer is provided which does not employ individually bonded leads between the IR sensitive elements and the input stages of multiplexers. The wafer is first coated with lead selenide in a first detector array area and is thereafter coated with lead sulfide within a second detector array area. The described steps result in the direct chemical deposition of lead selenide and lead sulfide upon the silicon wafer to eliminate individual wire bonding, bumping, flip chiping, planar interconnecting methods of connecting detector array elements to silicon chip circuitry, e.g., multiplexers, to enable easy fabrication of very long arrays. The electrode structure employed, produces an increase in the electrical field gradient between the electrodes for a given volume of detector material, relative to conventional electrode configurations.

A portable time-domain LED fluorimeter for nanosecond fluorescence lifetime measurements

NASA Astrophysics Data System (ADS)

Wang, Hongtao; Qi, Ying; Mountziaris, T. J.; Salthouse, Christopher D.

2014-05-01

Fluorescence lifetime measurements are becoming increasingly important in chemical and biological research. Time-domain lifetime measurements offer fluorescence multiplexing and improved handling of interferers compared with the frequency-domain technique. In this paper, an all solid-state, filterless, and highly portable light-emitting-diode based time-domain fluorimeter (LED TDF) is reported for the measurement of nanosecond fluorescence lifetimes. LED based excitation provides more wavelengths options compared to laser diode based excitation, but the excitation is less effective due to the uncollimated beam, less optical power, and longer latency in state transition. Pulse triggering and pre-bias techniques were implemented in our LED TDF to improve the peak optical power to over 100 mW. The proposed pulsing circuit achieved an excitation light fall time of less than 2 ns. Electrical resetting technique realized a time-gated photo-detector to remove the interference of the excitation light with fluorescence. These techniques allow the LED fluorimeter to accurately measure the fluorescence lifetime of fluorescein down to concentration of 0.5 μM. In addition, all filters required in traditional instruments are eliminated for the non-attenuated excitation/emission light power. These achievements make the reported device attractive to biochemical laboratories seeking for highly portable lifetime detection devices for developing sensors based on fluorescence lifetime changes. The device was initially validated by measuring the lifetimes of three commercial fluorophores and comparing them with reported lifetime data. It was subsequently used to characterize a ZnSe quantum dot based DNA sensor.

Frequency-multiplexed bias and readout of a 16-pixel superconducting nanowire single-photon detector array

NASA Astrophysics Data System (ADS)

Doerner, S.; Kuzmin, A.; Wuensch, S.; Charaev, I.; Boes, F.; Zwick, T.; Siegel, M.

2017-07-01

We demonstrate a 16-pixel array of microwave-current driven superconducting nanowire single-photon detectors with an integrated and scalable frequency-division multiplexing architecture, which reduces the required number of bias and readout lines to a single microwave feed line. The electrical behavior of the photon-sensitive nanowires, embedded in a resonant circuit, as well as the optical performance and timing jitter of the single detectors is discussed. Besides the single pixel measurements, we also demonstrate the operation of a 16-pixel array with a temporal, spatial, and photon-number resolution.

Multiplex fluorescent PCR for noninvasive prenatal detection of fetal-derived paternally inherited diseases using circulatory fetal DNA in maternal plasma.

PubMed

Tang, Dong-ling; Li, Yan; Zhou, Xin; Li, Xia; Zheng, Fang

2009-05-01

To develop a fluorescent polymerase chain reaction (PCR) assay for the detection of circulating fetal DNA in maternal plasma and use the established multiplex in noninvasive prenatal genetic diagnosis and its further applications in forensic casework. The DNA template was extracted from 47 pregnant women and the whole blood samples from the stated biological fathers were used to detect genotype. Using multiplex fluorescent PCR at 16 different polymorphic short tandem repeat (STR) loci, maternal DNA extracted from plasma samples at early pregnancy, medium pregnancy and late pregnancy were used to detect genotype. Their husbands' DNA was also used for fetal genotype ascertainment. Multiplex fluorescent PCR with 16 polymorphic short tandem repeats revealed the presence of fetal DNA in all cases. Every pregnant women/husband pair was informative in at least 3 of 16 loci. The chances of detecting paternally inherited fetal alleles ranged from 66.67 to 94.12%. They are 66.67% in early pregnancy, 85.71% in medium pregnancy and 94.12% in late pregnancy. The accuracy of Multiplex PCR assay to detect fetal DNA was 100%. Circulating fetal DNA analysis can be used as a possible alternative tool in routine laboratory prenatal diagnosis in the near future; this highly polymorphic STR multiplex has greatly improved the chances of detecting paternally inherited fetal alleles compared with other fetal DNA detection systems that use fetus-derived Y sequences to detect only male fetal DNA in maternal plasma. Our proposed technique can be applied to both female and male fetuses, which provides a sensitive, accurate and efficient method for noninvasive prenatal genetic diagnosis and forensic casework.

Capillary Array Waveguide Amplified Fluorescence Detector for mHealth

PubMed Central

Balsam, Joshua; Bruck, Hugh Alan; Rasooly, Avraham

2013-01-01

Mobile Health (mHealth) analytical technologies are potentially useful for carrying out modern medical diagnostics in resource-poor settings. Effective mHealth devices for underserved populations need to be simple, low cost, and portable. Although cell phone cameras have been used for biodetection, their sensitivity is a limiting factor because currently it is too low to be effective for many mHealth applications, which depend on detection of weak fluorescent signals. To improve the sensitivity of portable phones, a capillary tube array was developed to amplify fluorescence signals using their waveguide properties. An array configured with 36 capillary tubes was demonstrated to have a ~100X increase in sensitivity, lowering the limit of detection (LOD) of mobile phones from 1000 nM to 10 nM for fluorescein. To confirm that the amplification was due to waveguide behavior, we coated the external surfaces of the capillaries with silver. The silver coating interfered with the waveguide behavior and diminished the fluorescence signal, thereby proving that the waveguide behavior was the main mechanism for enhancing optical sensitivity. The optical configuration described here is novel in several ways. First, the use of capillaries waveguide properties to improve detection of weak florescence signal is new. Second we describe here a three dimensional illumination system, while conventional angular laser waveguide illumination is spot (or line), which is functionally one-dimensional illumination, can illuminate only a single capillary or a single column (when a line generator is used) of capillaries and thus inherently limits the multiplexing capability of detection. The planar illumination demonstrated in this work enables illumination of a two dimensional capillary array (e.g. x columns and y rows of capillaries). In addition, the waveguide light propagation via the capillary wall provides a third dimension for illumination along the axis of the capillaries. Such an array can potentially be used for sensitive analysis of multiple fluorescent detection assays simultaneously. The simple phone based capillary array approach presented in this paper is capable of amplifying weak fluorescent signals thereby improving the sensitivity of optical detectors based on mobile phones. This may allow sensitive biological assays to be measured with low sensitivity detectors and may make mHealth practical for many diagnostics applications, especially in resource-poor and global health settings. PMID:24039345

Linkage analysis with multiplexed short tandem repeat polymorphisms using infrared fluorescence and M13 tailed primers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Oetting, W.S.; Lee, H.K.; Flanders, D.J.

The use of short tandem repeat polymorphisms (STRPs) as marker loci for linkage analysis is becoming increasingly important due to their large numbers in the human genome and their high degree of polymorphism. Fluorescence-based detection of the STRP pattern with an automated DNA sequencer has improved the efficiency of this technique by eliminating the need for radioactivity and producing a digitized autoradiogram-like image that can be used for computer analysis. In an effort to simplify the procedure and to reduce the cost of fluorescence STRP analysis, we have developed a technique known as multiplexing STRPs with tailed primers (MSTP) usingmore » primers that have a 19-bp extension, identical to the sequence of an M13 sequencing primer, on the 5{prime} end of the forward primer in conjunction with multiplexing several primer pairs in a single polymerase chain reaction (PCR) amplification. The banding pattern is detected with the addition of the M13 primer-dye conjugate as the sole primer conjugated to the fluorescent dye, eliminating the need for direct conjugation of the infrared fluorescent dye to the STRP primers. The use of MSTP for linkage analysis greatly reduces the number of PCR reactions. Up to five primer pairs can be multiplexed together in the same reaction. At present, a set of 148 STRP markers spaced at an average genetic distance of 28 cM throughout the autosomal genome can be analyzed in 37 sets of multiplexed amplification reactions. We have automated the analysis of these patterns for linkage using software that both detects the STRP banding pattern and determines their sizes. This information can then be exported in a user-defined format from a database manager for linkage analysis. 15 refs., 2 figs., 4 tabs.« less

Homo-FRET Based Biosensors and Their Application to Multiplexed Imaging of Signalling Events in Live Cells

PubMed Central

Warren, Sean C.; Margineanu, Anca; Katan, Matilda; Dunsby, Chris; French, Paul M. W.

2015-01-01

Multiplexed imaging of Förster Resonance Energy Transfer (FRET)-based biosensors potentially presents a powerful approach to monitoring the spatio-temporal correlation of signalling pathways within a single live cell. Here, we discuss the potential of homo-FRET based biosensors to facilitate multiplexed imaging. We demonstrate that the homo-FRET between pleckstrin homology domains of Akt (Akt-PH) labelled with mCherry may be used to monitor 3′-phosphoinositide accumulation in live cells and show how global analysis of time resolved fluorescence anisotropy measurements can be used to quantify this accumulation. We further present multiplexed imaging readouts of calcium concentration, using fluorescence lifetime measurements of TN-L15-a CFP/YFP based hetero-FRET calcium biosensor-with 3′-phosphoinositide accumulation. PMID:26133241

Task-based design of a synthetic-collimator SPECT system used for small animal imaging.

PubMed

Lin, Alexander; Kupinski, Matthew A; Peterson, Todd E; Shokouhi, Sepideh; Johnson, Lindsay C

2018-05-07

In traditional multipinhole SPECT systems, image multiplexing - the overlapping of pinhole projection images - may occur on the detector, which can inhibit quality image reconstructions due to photon-origin uncertainty. One proposed system to mitigate the effects of multiplexing is the synthetic-collimator SPECT system. In this system, two detectors, a silicon detector and a germanium detector, are placed at different distances behind the multipinhole aperture, allowing for image detection to occur at different magnifications and photon energies, resulting in higher overall sensitivity while maintaining high resolution. The unwanted effects of multiplexing are reduced by utilizing the additional data collected from the front silicon detector. However, determining optimal system configurations for a given imaging task requires efficient parsing of the complex parameter space, to understand how pinhole spacings and the two detector distances influence system performance. In our simulation studies, we use the ensemble mean-squared error of the Wiener estimator (EMSE W ) as the figure of merit to determine optimum system parameters for the task of estimating the uptake of an 123 I-labeled radiotracer in three different regions of a computer-generated mouse brain phantom. The segmented phantom map is constructed by using data from the MRM NeAt database and allows for the reduction in dimensionality of the system matrix which improves the computational efficiency of scanning the system's parameter space. To contextualize our results, the Wiener estimator is also compared against a region of interest estimator using maximum-likelihood reconstructed data. Our results show that the synthetic-collimator SPECT system outperforms traditional multipinhole SPECT systems in this estimation task. We also find that image multiplexing plays an important role in the system design of the synthetic-collimator SPECT system, with optimal germanium detector distances occurring at maxima in the derivative of the percent multiplexing function. Furthermore, we report that improved task performance can be achieved by using an adaptive system design in which the germanium detector distance may vary with projection angle. Finally, in our comparative study, we find that the Wiener estimator outperforms the conventional region of interest estimator. Our work demonstrates how this optimization method has the potential to quickly and efficiently explore vast parameter spaces, providing insight into the behavior of competing factors, which are otherwise very difficult to calculate and study using other existing means. © 2018 American Association of Physicists in Medicine.

Large resistive 2D Micromegas with genetic multiplexing and some imaging applications

NASA Astrophysics Data System (ADS)

Bouteille, S.; Attié, D.; Baron, P.; Calvet, D.; Magnier, P.; Mandjavidze, I.; Procureur, S.; Riallot, M.

2016-10-01

The performance of the first large resistive Micromegas detectors with 2D readout and genetic multiplexing is presented. These detectors have a 50 × 50cm2 active area and are equipped with 1024 strips both in X- and Y-directions. The same genetic multiplexing pattern is applied on both coordinates, resulting in the compression of signals on 2 × 61 readout channels. Four such detectors have been built at CERN, and extensively tested with cosmics. The resistive strip film allows for very high gain operation, compensating for the charge spread on the 2 dimensions as well as the S / N loss due to the huge, 1 nF input capacitance. This film also creates a significantly different signal shape in the X- and Y-coordinates due to the charge evacuation along the resistive strips. All in all a detection efficiency above 95% is achieved with a 1 cm drift gap. Though not yet optimal, the measured 300 μm spatial resolution allows for very precise imaging in the field of muon tomography, and some applications of these detectors are presented.

Digital frequency domain multiplexing readout electronics for the next generation of millimeter telescopes

NASA Astrophysics Data System (ADS)

Bender, Amy N.; Cliche, Jean-François; de Haan, Tijmen; Dobbs, Matt A.; Gilbert, Adam J.; Montgomery, Joshua; Rowlands, Neil; Smecher, Graeme M.; Smith, Ken; Wilson, Andrew

2014-07-01

Frequency domain multiplexing (fMux) is an established technique for the readout of transition-edge sensor (TES) bolometers in millimeter-wavelength astrophysical instrumentation. In fMux, the signals from multiple detectors are read out on a single pair of wires reducing the total cryogenic thermal loading as well as the cold component complexity and cost of a system. The current digital fMux system, in use by POLARBEAR, EBEX, and the South Pole Telescope, is limited to a multiplexing factor of 16 by the dynamic range of the Superconducting Quantum Interference Device pre-amplifier and the total system bandwidth. Increased multiplexing is key for the next generation of large format TES cameras, such as SPT-3G and POLARBEAR2, which plan to have on the of order 15,000 detectors. Here, we present the next generation fMux readout, focusing on the warm electronics. In this system, the multiplexing factor increases to 64 channels per module (2 wires) while maintaining low noise levels and detector stability. This is achieved by increasing the system bandwidth, reducing the dynamic range requirements though active feedback, and digital synthesis of voltage biases with a novel polyphase filter algorithm. In addition, a version of the new fMux readout includes features such as low power consumption and radiation-hard components making it viable for future space-based millimeter telescopes such as the LiteBIRD satellite.

Design, Fabrication, and Testing of Lumped Element Kinetic inductance Detectors for 3 mm CMB Observations

NASA Technical Reports Server (NTRS)

Lowitz, Amy E.; Brown, Ari David; Stevenson, Thomas R.; Timbie, Peter T.; Wollack, Edward J.

2014-01-01

Kinetic inductance detectors (KIDs) are a promising technology for low-noise, highly-multiplexible mm- and submm-wave detection. KIDs have a number of advantages over other detector technologies, which make them an appealing option in the cosmic microwave background B-mode anisotropy search, including passive frequency domain multiplexing and relatively simple fabrication, but have suffered from challenges associated with noise control. Here we describe design and fabrication of a 20-pixel prototype array of lumped element molybdenum KIDs. We show Q, frequency and temperature measurements from the array under dark conditions. We also present evidence for a double superconducting gap in molybdenum.

The Microwave SQUID Multiplexer

NASA Astrophysics Data System (ADS)

Mates, John Arthur Benson

2011-12-01

This thesis describes a multiplexer of Superconducting Quantum Interference Devices (SQUIDs) with low-noise, ultra-low power dissipation, and great scalability. The multiplexer circuit measures the magnetic flux in a large number of unshunted rf SQUIDs by coupling each SQUID to a superconducting microwave resonator tuned to a unique resonance frequency and driving the resonators from a common feedline. A superposition of microwave tones measures each SQUID simultaneously using only two coaxial cables between the cryogenic device and room temperature. This multiplexer will enable the instrumentation of arrays with hundreds of thousands of low-temperature detectors for new applications in cosmology, materials analysis, and nuclear non-proliferation. The driving application of the Microwave SQUID Multiplexer is the readout of large arrays of superconducting transition-edge sensors, by some figures of merit the most sensitive detectors of electromagnetic signals over a span of more than nine orders of magnitude in energy, from 40 GHz microwaves to 200 keV gamma rays. Modern transition-edge sensors have noise-equivalent power as low as 10-20 W / Hz1/2 and energy resolution as good as 2 eV at 6 keV. These per-pixel sensitivities approach theoretical limits set by the underlying signals, motivating a rapid increase in pixel count to access new science. Compelling applications, like the non-destructive assay of nuclear material for treaty verification or the search for primordial gravity waves from inflation use arrays of these detectors to increase collection area or tile a focal plane. We developed three generations of SQUID multiplexers, optimizing the first for flux noise 0.17 muPhi0 / Hz1/2, the second for input current noise 19 pA / Hz1/2, and the last for practical multiplexing of large arrays of cosmic microwave background polarimeters based on transition-edge sensors. Using the last design we demonstrated multiplexed readout of prototype polarimeters with the performance required for the future development of a large-scale astronomical instrument.

Development of a multiplexed readout with high position resolution for positron emission tomography

NASA Astrophysics Data System (ADS)

Lee, Sangwon; Choi, Yong; Kang, Jihoon; Jung, Jin Ho

2017-04-01

Detector signals for positron emission tomography (PET) are commonly multiplexed to reduce the number of digital processing channels so that the system can remain cost effective while also maintaining imaging performance. In this work, a multiplexed readout combining Anger position estimation algorithm and position decoder circuit (PDC) was developed to reduce the number of readout channels by a factor of 24, 96-to-4. The data acquisition module consisted of a TDC (50 ps resolution), 4-channel ADCs (12 bit, 105 MHz sampling rate), 2 GB SDRAM and USB3.0. The performance of the multiplexed readout was assessed with a high-resolution PET detector block composed of 2×3 detector modules, each consisting of an 8×8 array of 1.52×1.52×6 mm3 LYSO, a 4×4 array of 3×3 mm2 silicon photomultiplier (SiPM) and 13.4×13.4 mm2 light guide with 0.7 mm thickness. The acquired flood histogram showed that all 384 crystals could be resolved. The average energy resolution at 511 keV was 13.7±1.6% full-width-at-half-maximum (FWHM) and the peak-to-valley ratios of the flood histogram on the horizontal and vertical lines were 18.8±0.8 and 22.8±1.3, respectively. The coincidence resolving time of a pair of detector blocks was 6.2 ns FWHM. The reconstructed phantom image showed that rods down to a diameter of 1.6 mm could be resolved. The results of this study indicate that the multiplexed readout would be useful in developing a PET with a spatial resolution less than the pixel size of the photosensor, such as a SiPM array.

Using the Wiener estimator to determine optimal imaging parameters in a synthetic-collimator SPECT system used for small animal imaging

NASA Astrophysics Data System (ADS)

Lin, Alexander; Johnson, Lindsay C.; Shokouhi, Sepideh; Peterson, Todd E.; Kupinski, Matthew A.

2015-03-01

In synthetic-collimator SPECT imaging, two detectors are placed at different distances behind a multi-pinhole aperture. This configuration allows for image detection at different magnifications and photon energies, resulting in higher overall sensitivity while maintaining high resolution. Image multiplexing the undesired overlapping between images due to photon origin uncertainty may occur in both detector planes and is often present in the second detector plane due to greater magnification. However, artifact-free image reconstruction is possible by combining data from both the front detector (little to no multiplexing) and the back detector (noticeable multiplexing). When the two detectors are used in tandem, spatial resolution is increased, allowing for a higher sensitivity-to-detector-area ratio. Due to variability in detector distances and pinhole spacings found in synthetic-collimator SPECT systems, a large parameter space must be examined to determine optimal imaging configurations. We chose to assess image quality based on the task of estimating activity in various regions of a mouse brain. Phantom objects were simulated using mouse brain data from the Magnetic Resonance Microimaging Neurological Atlas (MRM NeAt) and projected at different angles through models of a synthetic-collimator SPECT system, which was developed by collaborators at Vanderbilt University. Uptake in the different brain regions was modeled as being normally distributed about predetermined means and variances. We computed the performance of the Wiener estimator for the task of estimating activity in different regions of the mouse brain. Our results demonstrate the utility of the method for optimizing synthetic-collimator system design.

Superconducting Bolometer Array Architectures

NASA Technical Reports Server (NTRS)

Benford, Dominic; Chervenak, Jay; Irwin, Kent; Moseley, S. Harvey; Shafer, Rick; Staguhn, Johannes; Wollack, Ed; Oegerle, William (Technical Monitor)

2002-01-01

The next generation of far-infrared and submillimeter instruments require large arrays of detectors containing thousands of elements. These arrays will necessarily be multiplexed, and superconducting bolometer arrays are the most promising present prospect for these detectors. We discuss our current research into superconducting bolometer array technologies, which has recently resulted in the first multiplexed detections of submillimeter light and the first multiplexed astronomical observations. Prototype arrays containing 512 pixels are in production using the Pop-Up Detector (PUD) architecture, which can be extended easily to 1000 pixel arrays. Planar arrays of close-packed bolometers are being developed for the GBT (Green Bank Telescope) and for future space missions. For certain applications, such as a slewed far-infrared sky survey, feedhorncoupling of a large sparsely-filled array of bolometers is desirable, and is being developed using photolithographic feedhorn arrays. Individual detectors have achieved a Noise Equivalent Power (NEP) of -10(exp 17) W/square root of Hz at 300mK, but several orders of magnitude improvement are required and can be reached with existing technology. The testing of such ultralow-background detectors will prove difficult, as this requires optical loading of below IfW. Antenna-coupled bolometer designs have advantages for large format array designs at low powers due to their mode selectivity.

Graphene-based aptamer logic gates and their application to multiplex detection.

PubMed

Wang, Li; Zhu, Jinbo; Han, Lei; Jin, Lihua; Zhu, Chengzhou; Wang, Erkang; Dong, Shaojun

2012-08-28

In this work, a GO/aptamer system was constructed to create multiplex logic operations and enable sensing of multiplex targets. 6-Carboxyfluorescein (FAM)-labeled adenosine triphosphate binding aptamer (ABA) and FAM-labeled thrombin binding aptamer (TBA) were first adsorbed onto graphene oxide (GO) to form a GO/aptamer complex, leading to the quenching of the fluorescence of FAM. We demonstrated that the unique GO/aptamer interaction and the specific aptamer-target recognition in the target/GO/aptamer system were programmable and could be utilized to regulate the fluorescence of FAM via OR and INHIBIT logic gates. The fluorescence changed according to different input combinations, and the integration of OR and INHIBIT logic gates provided an interesting approach for logic sensing applications where multiple target molecules were present. High-throughput fluorescence imagings that enabled the simultaneous processing of many samples by using the combinatorial logic gates were realized. The developed logic gates may find applications in further development of DNA circuits and advanced sensors for the identification of multiple targets in complex chemical environments.

Multivariate optical element platform for compressed detection of fluorescence markers

NASA Astrophysics Data System (ADS)

Priore, Ryan J.; Swanstrom, Joseph A.

2014-05-01

The success of a commercial fluorescent diagnostic assay is dependent on the selection of a fluorescent biomarker; due to the broad nature of fluorescence biomarker emission profiles, only a small number of fluorescence biomarkers may be discriminated from each other as a function of excitation source. Multivariate Optical Elements (MOEs) are thin-film devices that encode a broad band, spectroscopic pattern allowing a simple broadband detector to generate a highly sensitive and specific detection for a target analyte. MOEs have historically been matched 1:1 to a discrete analyte or class prediction; however, MOE filter sets are capable of sensing projections of the original sparse spectroscopic space enabling a small set of MOEs to discriminate a multitude of target analytes. This optical regression can offer real-time measurements with relatively high signal-to-noise ratios that realize the advantages of multiplexed detection and pattern recognition in a simple optical instrument. The specificity advantage of MOE-based sensors allows fluorescent biomarkers that were once incapable of discrimination from one another via optical band pass filters to be employed in a common assay panel. A simplified MOE-based sensor may ultimately reduce the requirement for highly trained operators as well as move certain life science applications like disease prognostication from the laboratory to the point of care. This presentation will summarize the design and fabrication of compressed detection MOE filter sets for detecting multiple fluorescent biomarkers simultaneously with strong spectroscopic interference as well as comparing the detection performance of the MOE sensor with traditional optical band pass filter methodologies.

A portable time-domain LED fluorimeter for nanosecond fluorescence lifetime measurements

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang, Hongtao; Salthouse, Christopher D., E-mail: salthouse@ecs.umass.edu; Center for Personalized Health Monitoring, University of Massachusetts, Amherst, Massachusetts 01003

2014-05-15

Fluorescence lifetime measurements are becoming increasingly important in chemical and biological research. Time-domain lifetime measurements offer fluorescence multiplexing and improved handling of interferers compared with the frequency-domain technique. In this paper, an all solid-state, filterless, and highly portable light-emitting-diode based time-domain fluorimeter (LED TDF) is reported for the measurement of nanosecond fluorescence lifetimes. LED based excitation provides more wavelengths options compared to laser diode based excitation, but the excitation is less effective due to the uncollimated beam, less optical power, and longer latency in state transition. Pulse triggering and pre-bias techniques were implemented in our LED TDF to improve themore » peak optical power to over 100 mW. The proposed pulsing circuit achieved an excitation light fall time of less than 2 ns. Electrical resetting technique realized a time-gated photo-detector to remove the interference of the excitation light with fluorescence. These techniques allow the LED fluorimeter to accurately measure the fluorescence lifetime of fluorescein down to concentration of 0.5 μM. In addition, all filters required in traditional instruments are eliminated for the non-attenuated excitation/emission light power. These achievements make the reported device attractive to biochemical laboratories seeking for highly portable lifetime detection devices for developing sensors based on fluorescence lifetime changes. The device was initially validated by measuring the lifetimes of three commercial fluorophores and comparing them with reported lifetime data. It was subsequently used to characterize a ZnSe quantum dot based DNA sensor.« less

A 90GHz Bolometer Camera Detector System for the Green Bank Telescope

NASA Technical Reports Server (NTRS)

Benford, Dominic J.; Allen, Christine A.; Buchanan, Ernest D.; Chen, Tina C.; Chervenak, James A.; Devlin, Mark J.; Dicker, Simon R.; Forgione, Joshua B.

2004-01-01

We describe a close-packed, two-dimensional imaging detector system for operation at 90GHz (3.3mm) for the 100 m Green Bank Telescope (GBT) This system will provide high sensitivity (<1mjy in 1s rapid imaging (15'x15' to 250 microJy in 1 hr) at the world's largest steerable aperture. The heart of this camera is an 8x8 close packed, Nyquist-sampled array of superconducting transition edge sensor bolometers. We have designed and are producing a functional superconducting bolometer array system using a monolithic planar architecture and high-speed multiplexed readout electronics. With an NEP of approx. 2.10(exp 17) W/square root Hz, the TES bolometers will provide fast linear sensitive response for high performance imaging. The detectors are read out by and 8x8 time domain SQUID multiplexer. A digital/analog electronics system has been designed to enable read out by SQUID multiplexers. First light for this instrument on the GBT is expected within a year.

A 90GHz Bolometer Camera Detector System for the Green

NASA Technical Reports Server (NTRS)

Benford, Dominic J.; Allen, Christine A.; Buchanan, Ernest; Chen, Tina C.; Chervenak, James A.; Devlin, Mark J.; Dicker, Simon R.; Forgione, Joshua B.

2004-01-01

We describe a close-packed, two-dimensional imaging detector system for operation at 90GHz (3.3 mm) for the 100m Green Bank Telescope (GBT). This system will provide high sensitivity (less than 1mJy in 1s) rapid imaging (15'x15' to 150 micron Jy in 1 hr) at the world's largest steerable aperture. The heart of this camera is an 8x8 close-packed, Nyquist-sampled array of superconducting transition edge sensor (TES) bolometers. We have designed and are producing a functional superconducting bolometer array system using a monolithic planar architecture and high-speed multiplexed readout electronics. With an NEP of approximately 2 x 10(exp -17) W/square root of Hz, the TES bolometers will provide fast, linear, sensitive response for high performance imaging. The detectors are read out by an 8x8 time domain SQUID multiplexer. A digital/analog electronics system has been designed to enable read out by SQUID multiplexers. First light for this instrument on the GBT is expected within a year.

High-speed bioimaging with frequency-division-multiplexed fluorescence confocal microscopy

NASA Astrophysics Data System (ADS)

Mikami, Hideharu; Harmon, Jeffrey; Ozeki, Yasuyuki; Goda, Keisuke

2017-04-01

We present methods of fluorescence confocal microscopy that enable unprecedentedly high frame rate of > 10,000 fps. The methods are based on a frequency-division multiplexing technique, which was originally developed in the field of communication engineering. Specifically, we achieved a broad bandwidth ( 400 MHz) of detection signals using a dual- AOD method and overcame limitations in frame rate, due to a scanning device, by using a multi-line focusing method, resulting in a significant increase in frame rate. The methods have potential biomedical applications such as observation of sub-millisecond dynamics in biological tissues, in-vivo three-dimensional imaging, and fluorescence imaging flow cytometry.

Electrical delay line multiplexing for pulsed mode radiation detectors

NASA Astrophysics Data System (ADS)

Vinke, Ruud; Yeom, Jung Yeol; Levin, Craig S.

2015-04-01

Medical imaging systems are composed of a large number of position sensitive radiation detectors to provide high resolution imaging. For example, whole-body Positron Emission Tomography (PET) systems are typically composed of thousands of scintillation crystal elements, which are coupled to photosensors. Thus, PET systems greatly benefit from methods to reduce the number of data acquisition channels, in order to reduce the system development cost and complexity. In this paper we present an electrical delay line multiplexing scheme that can significantly reduce the number of readout channels, while preserving the signal integrity required for good time resolution performance. We experimented with two 4 × 4 LYSO crystal arrays, with crystal elements having 3 mm × 3 mm × 5 mm and 3 mm × 3 mm × 20 mm dimensions, coupled to 16 Hamamatsu MPPC S10931-050P SiPM elements. Results show that each crystal could be accurately identified, even in the presence of scintillation light sharing and inter-crystal Compton scatter among neighboring crystal elements. The multiplexing configuration degraded the coincidence timing resolution from ∼243 ps FWHM to ∼272 ps FWHM when 16 SiPM signals were combined into a single channel for the 4 × 4 LYSO crystal array with 3 mm × 3 mm × 20 mm crystal element dimensions, in coincidence with a 3 mm × 3 mm × 5 mm LYSO crystal pixel. The method is flexible to allow multiplexing configurations across different block detectors, and is scalable to an entire ring of detectors.

A centrifugal direct recombinase polymerase amplification (direct-RPA) microdevice for multiplex and real-time identification of food poisoning bacteria.

PubMed

Choi, Goro; Jung, Jae Hwan; Park, Byung Hyun; Oh, Seung Jun; Seo, Ji Hyun; Choi, Jong Seob; Kim, Do Hyun; Seo, Tae Seok

2016-06-21

In this study, we developed a centrifugal direct recombinase polymerase amplification (direct-RPA) microdevice for multiplex and real-time identification of food poisoning bacteria contaminated milk samples. The microdevice was designed to contain identical triplicate functional units and each unit has four reaction chambers, thereby making it possible to perform twelve direct-RPA reactions simultaneously. The integrated microdevice consisted of two layers: RPA reagents were injected in the top layer, while spiked milk samples with food poisoning bacteria were loaded into sample reservoirs in the bottom layer. For multiplex bacterial detection, the target gene-specific primers and probes were dried in each reaction chamber. The introduced samples and reagents could be equally aliquoted and dispensed into each reaction chamber by centrifugal force, and then the multiplex direct-RPA reaction was executed. The target genes of bacteria spiked in milk could be amplified at 39 °C without a DNA extraction step by using the direct-RPA cocktails, which were a combination of a direct PCR buffer and RPA enzymes. As the target gene amplification proceeded, the increased fluorescence signals coming from the reaction chambers were recorded in real-time at an interval of 2 min. The entire process, including the sample distribution, the direct-RPA reaction, and the real-time analysis, was accomplished with a custom-made portable genetic analyzer and a miniaturized optical detector. Monoplex, duplex, and triplex food poisoning bacteria (Salmonella enterica, Escherichia coli O157:H7, and Vibrio parahaemolyticus) detection was successfully performed with a detection sensitivity of 4 cells per 3.2 μL of milk samples within 30 min. By implementing the direct-PRA on the miniaturized centrifugal microsystem, the on-site food poisoning bacteria analysis would be feasible with high speed, sensitivity, and multiplicity.

Genetic barcoding with fluorescent proteins for multiplexed applications.

PubMed

Smurthwaite, Cameron A; Williams, Wesley; Fetsko, Alexandra; Abbadessa, Darin; Stolp, Zachary D; Reed, Connor W; Dharmawan, Andre; Wolkowicz, Roland

2015-04-14

Fluorescent proteins, fluorescent dyes and fluorophores in general have revolutionized the field of molecular cell biology. In particular, the discovery of fluorescent proteins and their genes have enabled the engineering of protein fusions for localization, the analysis of transcriptional activation and translation of proteins of interest, or the general tracking of individual cells and cell populations. The use of fluorescent protein genes in combination with retroviral technology has further allowed the expression of these proteins in mammalian cells in a stable and reliable manner. Shown here is how one can utilize these genes to give cells within a population of cells their own biosignature. As the biosignature is achieved with retroviral technology, cells are barcoded 'indefinitely'. As such, they can be individually tracked within a mixture of barcoded cells and utilized in more complex biological applications. The tracking of distinct populations in a mixture of cells is ideal for multiplexed applications such as discovery of drugs against a multitude of targets or the activation profile of different promoters. The protocol describes how to elegantly develop and amplify barcoded mammalian cells with distinct genetic fluorescent markers, and how to use several markers at once or one marker at different intensities. Finally, the protocol describes how the cells can be further utilized in combination with cell-based assays to increase the power of analysis through multiplexing.

SERS-fluorescence joint spectral encoded magnetic nanoprobes for multiplex cancer cell separation.

PubMed

Wang, Zhuyuan; Zong, Shenfei; Chen, Hui; Wang, Chunlei; Xu, Shuhong; Cui, Yiping

2014-11-01

A new kind of cancer cell separation method is demonstrated, using surface-enhanced Raman scattering (SERS) and fluorescence dual-encoded magnetic nanoprobes. The designed nanoprobes can realize SERS-fluorescence joint spectral encoding (SFJSE) and greatly improve the multiplexing ability. The nanoprobes have four main components, that is, the magnetic core, SERS generator, fluorescent agent, and targeting antibody. These components are assembled with a multi-layered structure to form the nanoprobes. Specifically, silica-coated magnetic nanobeads (MBs) are used as the inner core. Au core-Ag shell nanorods (Au@Ag NRs) are employed as the SERS generators and attached on the silica-coated MBs. After burying these Au@Ag NRs with another silica layer, CdTe quantum dots (QDs), that is, the fluorescent agent, are anchored onto the silica layer. Finally, antibodies are covalently linked to CdTe QDs. SFJSE is fulfilled by using different Raman molecules and QDs with different emission wavelengths. By utilizing four human cancer cell lines and one normal cell line as the model cells, the nanoprobes can specifically and simultaneously separate target cancer cells from the normal ones. This SFJSE-based method greatly facilitates the multiplex, rapid, and accurate cancer cell separation, and has a prosperous potential in high-throughput analysis and cancer diagnosis. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Design and performance of dual-polarization lumped-element kinetic inductance detectors for millimeter-wave polarimetry

NASA Astrophysics Data System (ADS)

McCarrick, H.; Jones, G.; Johnson, B. R.; Abitbol, M. H.; Ade, P. A. R.; Bryan, S.; Day, P.; Essinger-Hileman, T.; Flanigan, D.; Leduc, H. G.; Limon, M.; Mauskopf, P.; Miller, A.; Tucker, C.

2018-02-01

Aims: Lumped-element kinetic inductance detectors (LEKIDs) are an attractive technology for millimeter-wave observations that require large arrays of extremely low-noise detectors. We designed, fabricated and characterized 64-element (128 LEKID) arrays of horn-coupled, dual-polarization LEKIDs optimized for ground-based CMB polarimetry. Our devices are sensitive to two orthogonal polarizations in a single spectral band centered on 150 GHz with Δν/ν = 0.2. The 65 × 65 mm square arrays are designed to be tiled into the focal plane of an optical system. We demonstrate the viability of these dual-polarization LEKIDs with laboratory measurements. Methods: The LEKID modules are tested with an FPGA-based readout system in a sub-kelvin cryostat that uses a two-stage adiabatic demagnetization refrigerator. The devices are characterized using a blackbody and a millimeter-wave source. The polarization properties are measured with a cryogenic stepped half-wave plate. We measure the resonator parameters and the detector sensitivity, noise spectrum, dynamic range, and polarization response. Results: The resonators have internal quality factors approaching 1 × 106. The detectors have uniform response between orthogonal polarizations and a large dynamic range. The detectors are photon-noise limited above 1 pW of absorbed power. The noise-equivalent temperatures under a 3.4 K blackbody load are <100 μK √s. The polarization fractions of detectors sensitive to orthogonal polarizations are >80%. The entire array is multiplexed on a single readout line, demonstrating a multiplexing factor of 128. The array and readout meet the requirements for 4 arrays to be read out simultaneously for a multiplexing factor of 512. Conclusions: This laboratory study demonstrates the first dual-polarization LEKID array optimized specifically for CMB polarimetry and shows the readiness of the detectors for on-sky observations.

Microwave SQUID Multiplexing of Metallic Magnetic Calorimeters: Status of Multiplexer Performance and Room-Temperature Readout Electronics Development

NASA Astrophysics Data System (ADS)

Wegner, M.; Karcher, N.; Krömer, O.; Richter, D.; Ahrens, F.; Sander, O.; Kempf, S.; Weber, M.; Enss, C.

2018-02-01

To our present best knowledge, microwave SQUID multiplexing (μ MUXing) is the most suitable technique for reading out large-scale low-temperature microcalorimeter arrays that consist of hundreds or thousands of individual pixels which require a large readout bandwidth per pixel. For this reason, the present readout strategy for metallic magnetic calorimeter (MMC) arrays combining an intrinsic fast signal rise time, an excellent energy resolution, a large energy dynamic range, a quantum efficiency close to 100% as well as a highly linear detector response is based on μ MUXing. Within this paper, we summarize the state of the art in MMC μ MUXing and discuss the most recent results. This particularly includes the discussion of the performance of a 64-pixel detector array with integrated, on-chip microwave SQUID multiplexer, the progress in flux ramp modulation of MMCs as well as the status of the development of a software-defined radio-based room-temperature electronics which is specifically optimized for MMC readout.

Contact-lens type of micromachined hydrogenated amorphous Si fluorescence detector coupled with microfluidic electrophoresis devices

NASA Astrophysics Data System (ADS)

Kamei, Toshihiro; Wada, Takehito

2006-09-01

A 5.8-μm-thick SiO2/Ta2O5 multilayer optical interference filter was monolithically integrated and micromachined on a hydrogenated amorphous Si (a-Si :H) pin photodiode to form a fluorescence detector. A microfluidic electrophoresis device was mounted on a detection platform comprising a fluorescence-collecting half-ball lens and the micromachined fluorescence detector. The central aperture of the fluorescence detector allows semiconductor laser light to pass up through the detector and to irradiate an electrophoretic separation channel. The limit of detection is as low as 7nM of the fluorescein solution, and high-speed DNA fragment sizing can be achieved with high separation efficiency. The micromachined a-Si :H fluorescence detector exhibits high sensitivity for practical fluorescent labeling dyes as well as integration flexibility on various substances, making it ideal for application to portable microfluidic bioanalysis devices.

Surface-Enhanced Raman Scattering Based Nonfluorescent Probe for Multiplex DNA Detection

PubMed Central

Sun, Lan; Yu, Chenxu; Irudayaraj, Joseph

2008-01-01

To provide rapid and accurate detection of DNA markers in a straightforward, inexpensive and multiplex format, an alternative surface enhanced Raman scattering (SERS) based probe was designed and fabricated to covalently attach both DNA probing sequence and non-fluorescent Raman tags to the surface of gold nanoparticles (DNA-AuP-RTag). The intensity of Raman signal of the probes could be controlled through the surface coverage of the non-fluorescent Raman tags (RTags). Detection sensitivity of these probes could be optimized by fine-tuning the amount of DNA molecules and RTags on the probes. Long-term stability of the DNA-AuP-RTag probes was found to be good (over 3 months). Excellent multiplexing capability of the DNA-AuP-RTag scheme was demonstrated by simultaneous identification of up to eight probes in a mixture. Detection of hybridization of single-stranded DNA (ssDNA) to its complementary targets was successfully accomplished with a long-term goal to use non-fluorescent RTags in a Raman-based DNA microarray platform. PMID:17465531

Energy response calibration of photon-counting detectors using x-ray fluorescence: a feasibility study.

PubMed

Cho, H-M; Ding, H; Ziemer, B P; Molloi, S

2014-12-07

Accurate energy calibration is critical for the application of energy-resolved photon-counting detectors in spectral imaging. The aim of this study is to investigate the feasibility of energy response calibration and characterization of a photon-counting detector using x-ray fluorescence. A comprehensive Monte Carlo simulation study was performed using Geant4 Application for Tomographic Emission (GATE) to investigate the optimal technique for x-ray fluorescence calibration. Simulations were conducted using a 100 kVp tungsten-anode spectra with 2.7 mm Al filter for a single pixel cadmium telluride (CdTe) detector with 3 × 3 mm(2) in detection area. The angular dependence of x-ray fluorescence and scatter background was investigated by varying the detection angle from 20° to 170° with respect to the beam direction. The effects of the detector material, shape, and size on the recorded x-ray fluorescence were investigated. The fluorescent material size effect was considered with and without the container for the fluorescent material. In order to provide validation for the simulation result, the angular dependence of x-ray fluorescence from five fluorescent materials was experimentally measured using a spectrometer. Finally, eleven of the fluorescent materials were used for energy calibration of a CZT-based photon-counting detector. The optimal detection angle was determined to be approximately at 120° with respect to the beam direction, which showed the highest fluorescence to scatter ratio (FSR) with a weak dependence on the fluorescent material size. The feasibility of x-ray fluorescence for energy calibration of photon-counting detectors in the diagnostic x-ray energy range was verified by successfully calibrating the energy response of a CZT-based photon-counting detector. The results of this study can be used as a guideline to implement the x-ray fluorescence calibration method for photon-counting detectors in a typical imaging laboratory.

Energy response calibration of photon-counting detectors using x-ray fluorescence: a feasibility study

NASA Astrophysics Data System (ADS)

Cho, H.-M.; Ding, H.; Ziemer, BP; Molloi, S.

2014-12-01

Accurate energy calibration is critical for the application of energy-resolved photon-counting detectors in spectral imaging. The aim of this study is to investigate the feasibility of energy response calibration and characterization of a photon-counting detector using x-ray fluorescence. A comprehensive Monte Carlo simulation study was performed using Geant4 Application for Tomographic Emission (GATE) to investigate the optimal technique for x-ray fluorescence calibration. Simulations were conducted using a 100 kVp tungsten-anode spectra with 2.7 mm Al filter for a single pixel cadmium telluride (CdTe) detector with 3  ×  3 mm2 in detection area. The angular dependence of x-ray fluorescence and scatter background was investigated by varying the detection angle from 20° to 170° with respect to the beam direction. The effects of the detector material, shape, and size on the recorded x-ray fluorescence were investigated. The fluorescent material size effect was considered with and without the container for the fluorescent material. In order to provide validation for the simulation result, the angular dependence of x-ray fluorescence from five fluorescent materials was experimentally measured using a spectrometer. Finally, eleven of the fluorescent materials were used for energy calibration of a CZT-based photon-counting detector. The optimal detection angle was determined to be approximately at 120° with respect to the beam direction, which showed the highest fluorescence to scatter ratio (FSR) with a weak dependence on the fluorescent material size. The feasibility of x-ray fluorescence for energy calibration of photon-counting detectors in the diagnostic x-ray energy range was verified by successfully calibrating the energy response of a CZT-based photon-counting detector. The results of this study can be used as a guideline to implement the x-ray fluorescence calibration method for photon-counting detectors in a typical imaging laboratory.

Energy response calibration of photon-counting detectors using X-ray fluorescence: a feasibility study

PubMed Central

Cho, H-M; Ding, H; Ziemer, BP; Molloi, S

2014-01-01

Accurate energy calibration is critical for the application of energy-resolved photon-counting detectors in spectral imaging. The aim of this study is to investigate the feasibility of energy response calibration and characterization of a photon-counting detector using X-ray fluorescence. A comprehensive Monte Carlo simulation study was performed using Geant4 Application for Tomographic Emission (GATE) to investigate the optimal technique for X-ray fluorescence calibration. Simulations were conducted using a 100 kVp tungsten-anode spectra with 2.7 mm Al filter for a single pixel cadmium telluride (CdTe) detector with 3 × 3 mm2 in detection area. The angular dependence of X-ray fluorescence and scatter background was investigated by varying the detection angle from 20° to 170° with respect to the beam direction. The effects of the detector material, shape, and size on the recorded X-ray fluorescence were investigated. The fluorescent material size effect was considered with and without the container for the fluorescent material. In order to provide validation for the simulation result, the angular dependence of X-ray fluorescence from five fluorescent materials was experimentally measured using a spectrometer. Finally, eleven of the fluorescent materials were used for energy calibration of a CZT-based photon-counting detector. The optimal detection angle was determined to be approximately at 120° with respect to the beam direction, which showed the highest fluorescence to scatter ratio (FSR) with a weak dependence on the fluorescent material size. The feasibility of X-ray fluorescence for energy calibration of photon-counting detectors in the diagnostic X-ray energy range was verified by successfully calibrating the energy response of a CZT-based photon-counting detector. The results of this study can be used as a guideline to implement the X-ray fluorescence calibration method for photon-counting detectors in a typical imaging laboratory. PMID:25369288

[Detection of large deletions in X linked Alport syndrome using competitive multiplex fluorescence polymerase chain reaction].

PubMed

Wang, F; Zhang, Y Q; Ding, J; Yu, L X

2017-10-18

To evaluate the ability of multiplex competitive fluorescence polymerase chain reaction in detection of large deletion and duplication genotypes of X-linked Alport syndrome. Clinical diagnosis of X-linked Alport syndrome was based on either abnormal staining of type IV collagen α5 chain in the epidermal basement membrane alone or with abnormal staining of type IV collagen α5 chain in the glomerular basement membrane and Bowman's capsule/ultrastructural changes in the glomerular basement membrane typical of Alport syndrome. A total of 20 unrelated Chinese patients (13 males and 7 females) clinically diagnosed as X-linked Alport syndrome were included in the study. Their genotypes were unknown. Control subjects included a male patient with other renal disease and two patients who had large deletions in COL4A5 gene detected by multiplex ligation-dependent probe amplification. Genomic DNA was isolated from peripheral blood leukocytes in all the participants. Multiplex competitive fluorescence polymerase chain reaction was used to coamplify 53 exons of COL4A5 gene and four reference genes in a single reaction. When a deletion removed exon 1 of COL4A5 gene was identified, the same method was used to coamplify the first 4 exons of COL4A5 and COL4A6 genes, a promoter shared by COL4A5 and COL4A6 genes, and three reference genes in a single reaction. Any copy number loss suggested by this method was verified by electrophoresis of corresponding polymerase chain reaction amplified products or DNA sequencing to exclude possible DNA variations in the primer regions. Genotypes of two positive controls identified by multiplex competitive fluorescence polymerase chain reaction were consistent with those detected by multiplex ligation-dependent probe amplification. Deletions were identified in 6 of the 20 patients, including two large deletions removing the 5' part of both COL4A5 and COL4A6 genes with the breakpoint located in the second intron of COL4A6, two large deletions removing more than 30 exons of COL4A5 gene, one large deletion removing at least 1 exon of COL4A5 gene, and one small deletion involving 13 bps. No duplication was found. Our results show that multiplex competitive fluorescence polymerase chain reaction is a good alternative to classical techniques for large deletion genotyping in X-linked Alport syndrome.

Frequency-encoded laser-induced fluorescence for multiplexed detection in infrared-mediated quantitative PCR

PubMed Central

Schrell, Adrian M.; Roper, Michael G.

2014-01-01

A frequency-modulated fluorescence encoding method was used as a means to increase the number of fluorophores monitored during infrared-mediated polymerase chain reaction. Laser lines at 488-nm and 561-nm were modulated at 73- and 137-Hz, respectively, exciting fluorescence from the dsDNA intercalating dye, EvaGreen, and the temperature insensitive dye, ROX. Emission was collected in a color-blind manner using a single photomultiplier tube for detection and demodulated by frequency analysis. The resulting frequency domain signal resolved the contribution from the two fluorophores as well as the background from the IR lamp. The detection method was successfully used to measure amplification of DNA samples containing 104 – 107 starting copies of template producing an amplification efficiency of 96%. The utility of this methodology was further demonstrated by simultaneous amplification of two genes from human genomic DNA using different color TaqMan probes. This method of multiplexing fluorescence detection with IR-qPCR is ideally suited as it allowed isolation of the signals of interest from the background in the frequency domain and is expected to further reduce the complexity of multiplexed microfluidic IR-qPCR instrumentation. PMID:24448431

The readout chain for the bar PANDA MVD strip detector

NASA Astrophysics Data System (ADS)

Schnell, R.; Brinkmann, K.-Th.; Di Pietro, V.; Kleines, H.; Goerres, A.; Riccardi, A.; Rivetti, A.; Rolo, M. D.; Sohlbach, H.; Zaunick, H.-G.

2015-02-01

The bar PANDA (antiProton ANnihilation at DArmstadt) experiment will study the strong interaction in annihilation reactions between an antiproton beam and a stationary gas jet target. The detector will comprise different sub-detectors for tracking, particle identification and calorimetry. The Micro-Vertex Detector (MVD) as the innermost part of the tracking system will allow precise tracking and detection of secondary vertices. For the readout of the double-sided silicon strip sensors a custom-made ASIC is being developed, employing the Time-over-Threshold (ToT) technique for digitization and utilize time-to-digital converters (TDC) to provide a high-precision time stamp of the hit. A custom-made Module Data Concentrator ASIC (MDC) will multiplex the data of all front-ends of one sensor towards the CERN-developed GBT chip set (GigaBit Transceiver). The MicroTCA-based MVD Multiplexer Board (MMB) at the off-detector site will receive and concentrate the data from the GBT links and transfer it to FPGA-based compute nodes for global event building.

Progress Towards High-Sensitivity Arrays of Detectors of Sub-mm Radiation using Superconducting Tunnel Junctions with Radio-Frequency Single-Electron Transistors

NASA Technical Reports Server (NTRS)

Stevenson, T. R.; Hsieh, W.-T.; Li, M. J.; Stahle, C. M.; Wollack, E. J.; Schoelkopf, R. J.; Krebs, Carolyn (Technical Monitor)

2002-01-01

The science drivers for the SPIRIT/SPECS missions demand sensitive, fast, compact, low-power, large-format detector arrays for high resolution imaging and spectroscopy in the far infrared and submillimeter. Detector arrays with 10,000 pixels and sensitivity less than 10(exp 20)-20 W/Hz(exp 20)0.5 are needed. Antenna-coupled superconducting tunnel junction detectors with integrated rf single-electron transistor readout amplifiers have the potential for achieving this high level of sensitivity, and can take advantage of an rf multiplexing technique when forming arrays. The device consists of an antenna structure to couple radiation into a small superconducting volume and cause quasiparticle excitations, and a single-electron transistor to measure currents through tunnel junction contacts to the absorber volume. We will describe optimization of device parameters, and recent results on fabrication techniques for producing devices with high yield for detector arrays. We will also present modeling of expected saturation power levels, antenna coupling, and rf multiplexing schemes.

Protein Multiplexed Immunoassay Analysis with R.

PubMed

Breen, Edmond J

2017-01-01

Plasma samples from 177 control and type 2 diabetes patients collected at three Australian hospitals are screened for 14 analytes using six custom-made multiplex kits across 60 96-well plates. In total 354 samples were collected from the patients, representing one baseline and one end point sample from each patient. R methods and source code for analyzing the analyte fluorescence response obtained from these samples by Luminex Bio-Plex ® xMap multiplexed immunoassay technology are disclosed. Techniques and R procedures for reading Bio-Plex ® result files for statistical analysis and data visualization are also presented. The need for technical replicates and the number of technical replicates are addressed as well as plate layout design strategies. Multinomial regression is used to determine plate to sample covariate balance. Methods for matching clinical covariate information to Bio-Plex ® results and vice versa are given. As well as methods for measuring and inspecting the quality of the fluorescence responses are presented. Both fixed and mixed-effect approaches for immunoassay statistical differential analysis are presented and discussed. A random effect approach to outlier analysis and detection is also shown. The bioinformatics R methodology present here provides a foundation for rigorous and reproducible analysis of the fluorescence response obtained from multiplexed immunoassays.

Multiplexed electronically programmable multimode ionization detector for chromatography

DOEpatents

Wise, M.B.; Buchanan, M.V.

1988-05-19

Method and apparatus for detecting and differentiating organic compounds based on their electron affinity. An electron capture detector cell (ECD) is operated in a plurality of multiplexed electronically programmable operating modes to alter the detector response during a single sampling cycle to acquire multiple simultaneous chromatograms corresponding to each of the different operating modes. The cell is held at a constant subatmospheric pressure while the electron collection bias voltage applied to the cell is modulated electronically to allow acquisition of multiple chromatograms for a single sample elution from a chromatograph representing three distinctly different response modes. A system is provided which automatically controls the programmed application of bias pulses at different intervals and/or amplitudes to switch the detector from an ionization mode to the electron capture mode and various degrees therebetween to provide an improved means of tuning an ECD for multimode detection and improved specificity. 6 figs.

Multiplexed electronically programmable multimode ionization detector for chromatography

DOEpatents

Wise, Marcus B.; Buchanan, Michelle V.

1989-01-01

Method and apparatus for detecting and differentiating organic compounds based on their electron affinity. An electron capture detector cell (ECD) is operated in a plurality of multiplexed electroncially programmable operating modes to alter the detector response during a single sampling cycle to acquire multiple simultaneous chromatograms corresponding to each of the different operating modes. The cell is held at a constant subatmospheric pressure while the electron collection bias voltage applied to the cell is modulated electronically to allow acquisition of multiple chromatograms for a single sample elution from a chromatograph representing three distinctly different response modes. A system is provided which automatically controls the programmed application of bias pulses at different intervals and/or amplitudes to switch the detector from an ionization mode to the electron capture mode and various degrees therebetween to provide an improved means of tuning an ECD for multimode detection and improved specificity.

Functionalization of optical nanotip arrays with an electrochemical microcantilever for multiplexed DNA detection.

PubMed

Descamps, Emeline; Duroure, Nathalie; Deiss, Frédérique; Leichlé, Thierry; Adam, Catherine; Mailley, Pascal; Aït-Ikhlef, Ali; Livache, Thierry; Nicu, Liviu; Sojic, Neso

2013-08-07

Optical nanotip arrays fabricated on etched fiber bundles were functionalized with DNA spots. Such unconventional substrates (3D and non-planar) are difficult to pattern with standard microfabrication techniques but, using an electrochemical cantilever, up to 400 spots were electrodeposited on the nanostructured optical surface in 5 min. This approach allows each spot to be addressed individually and multiplexed fluorescence detection is demonstrated. Finally, remote fluorescence detection was performed by imaging through the optical fiber bundle itself after hybridisation with the complementary sequence.

Detector arrays for low-background space infrared astronomy

NASA Technical Reports Server (NTRS)

Mccreight, C. R.; Mckelvey, M. E.; Goebel, J. H.; Anderson, G. M.; Lee, J. H.

1986-01-01

The status of development and characterization tests of integrated infrared detector array technology for astronomy applications is described. The devices under development include intrinsic, extrinsic silicon, and extrinsic germanium detectors, with hybrid silicon multiplexers. Laboratory test results and successful astronomy imagery have established the usefulness of integrated arrays in low-background astronomy applications.

Detector arrays for low-background space infrared astronomy

NASA Technical Reports Server (NTRS)

Mccreight, C. R.; Mckelvey, M. E.; Goebel, J. H.; Anderson, G. M.; Lee, J. H.

1986-01-01

The status of development and characterization tests of integrated infrared detector array technology for astronomy applications is described. The devices under development include intrinsic, extrinsic silicon, and extrinsic germanium detectors, with hybrid silicon multiplexers. Laboratary test results and successful astronomy imagery have established the usefulness of integrated arrays in low-background astronomy applications.

Integrated detector array technology for infrared astronomy

NASA Technical Reports Server (NTRS)

Mccreight, c. R.; Goebel, J. H.; Mckelvey, M. E.; Stafford, P. S.; Lee, J. H.

1984-01-01

The status of laboratory and telescope tests of integrated infrared detector array technology for astronomical applications is described. The devices tested represent a number of extrinsic and intrinsic detector materials and various multiplexer designs. Infrared arrays have now been used in successful astronomical applications. These have shown that device sensitivities can be comparable to those of discrete detector systems and excellent astronomical imagery can be produced.

Multiplexing readout channels in proportional counters

NASA Technical Reports Server (NTRS)

Caristi, James

1991-01-01

Proportional counters are important instruments used in sensing hard x-rays. The possibility is described of doubling the number of readout channels in the detector without increasing the electronics needed to amplify channel signals. This suggests that it should be possible, conversely, to reduce the number of amplifiers, thereby reducing the weight and energy budget of the instrument. Various numerical multiplexing schemes are analyzed, and a computer program is presented that can reconstruct multiplexed channel outputs with very good accuracy.

Fiber optic wavelength division multiplexing: Principles and applications in telecommunications and spectroscopy

NASA Technical Reports Server (NTRS)

Erdmann, R. K.; Walton, B. D.

1988-01-01

Design and fabrication tradeoffs of wavelength division multiplexers are discussed and performance parameters are given. The same multiplexer construction based on prism gratings has been used in spectroscopic applications, in the wavelength region from 450 to 1600 nm. For shorter wavelengths down to 200 nm, a similar instrument based on longer fibers (500 to 1000 micrometer) has been constructed and tested with both a fiber array and a photodiode detector array at the output.

Characterization of energy response for photon-counting detectors using x-ray fluorescence

PubMed Central

Ding, Huanjun; Cho, Hyo-Min; Barber, William C.; Iwanczyk, Jan S.; Molloi, Sabee

2014-01-01

Purpose: To investigate the feasibility of characterizing a Si strip photon-counting detector using x-ray fluorescence. 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 an edge illuminated Si strip detector. The distances from the source to target and the target to detector were approximately 19 and 11 cm, respectively. Four different materials, containing silver (Ag), iodine (I), barium (Ba), and gadolinium (Gd), were placed in small plastic containers 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 fluorescence energies. The energy resolutions and charge-sharing fractions were also obtained from analytical fittings of the recorded fluorescence spectra. An analytical model, which employed four parameters that can be determined from the fluorescence calibration, was used to estimate the detector response function. Results: Strong fluorescence signals of all four target materials were recorded with the investigated geometry for the Si strip detector. The average gain and offset of all pixels for detector energy calibration were determined to be 6.95 mV/keV and −66.33 mV, respectively. The detector’s energy resolution remained at approximately 2.7 keV for low energies, and increased slightly at 45 keV. The average charge-sharing fraction was estimated to be 36% within the investigated energy range of 20–45 keV. The simulated detector output based on the proposed response function agreed well with the experimental measurement. Conclusions: 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 offers an accurate and efficient way to calibrate the energy response of a photon-counting detector. PMID:25471962

Rapid detection of Wuchereria bancrofti and Brugia malayi in mosquito vectors (Diptera: Culicidae) using a real-time fluorescence resonance energy transfer multiplex PCR and melting curve analysis.

PubMed

Intapan, Pewpan M; Thanchomnang, Tongjit; Lulitanond, Viraphong; Maleewong, Wanchai

2009-01-01

We developed a single-step real-time fluorescence resonance energy transfer (FRET) multiplex polymerase chain reaction (PCR) merged with melting curve analysis for the detection of Wuchereria bancrofti and Brugia malayi DNA in blood-fed mosquitoes. Real-time FRET multiplex PCR is based on fluorescence melting curve analysis of a hybrid of amplicons generated from two families of repeated DNA elements: the 188 bp SspI repeated sequence, specific to W. bancrofti, and the 153-bp HhaI repeated sequence, specific to the genus Brugia and two pairs of specific fluorophore-labeled probes. Both W. bancrofti and B. malayi can be differentially detected in infected vectors by this process through their different fluorescence channel and melting temperatures. The assay could distinguish both human filarial DNAs in infected vectors from the DNAs of Dirofilaria immitis- and Plasmodium falciparum-infected human red blood cells and noninfected mosquitoes and human leukocytes. The technique showed 100% sensitivity and specificity and offers a rapid and reliable procedure for differentially identifying lymphatic filariasis. The introduced real-time FRET multiplex PCR can reduce labor time and reagent costs and is not prone to carry over contamination. The test can be used to screen mosquito vectors in endemic areas and therefore should be a useful diagnostic tool for the evaluation of infection rate of the mosquito populations and for xenomonitoring in the community after eradication programs such as the Global Program to Eliminate Lymphatic Filariasis.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ding, Huanjun; Cho, Hyo-Min; Molloi, Sabee, E-mail: symolloi@uci.edu

Purpose: To investigate the feasibility of characterizing a Si strip photon-counting detector using x-ray fluorescence. 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 an edge illuminated Si strip detector. The distances from the source to target and the target to detector were approximately 19 and 11 cm, respectively. Four different materials, containing silver (Ag), iodine (I), barium (Ba), and gadolinium (Gd), were placed in small plastic containers with a diametermore » 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 fluorescence energies. The energy resolutions and charge-sharing fractions were also obtained from analytical fittings of the recorded fluorescence spectra. An analytical model, which employed four parameters that can be determined from the fluorescence calibration, was used to estimate the detector response function. Results: Strong fluorescence signals of all four target materials were recorded with the investigated geometry for the Si strip detector. The average gain and offset of all pixels for detector energy calibration were determined to be 6.95 mV/keV and −66.33 mV, respectively. The detector’s energy resolution remained at approximately 2.7 keV for low energies, and increased slightly at 45 keV. The average charge-sharing fraction was estimated to be 36% within the investigated energy range of 20–45 keV. The simulated detector output based on the proposed response function agreed well with the experimental measurement. Conclusions: 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 offers an accurate and efficient way to calibrate the energy response of a photon-counting detector.« less

Highly Tunable Aptasensing Microarrays with Graphene Oxide Multilayers

NASA Astrophysics Data System (ADS)

Jung, Yun Kyung; Lee, Taemin; Shin, Eeseul; Kim, Byeong-Su

2013-11-01

A highly tunable layer-by-layer (LbL)-assembled graphene oxide (GO) array has been devised for high-throughput multiplex protein sensing. In this array, the fluorescence of different target-bound aptamers labeled with dye is efficiently quenched by GO through fluorescence resonance energy transfer (FRET), and simultaneous multiplex target detection is performed by recovering the quenched fluorescence caused by specific binding between an aptamer and a protein. Thin GO films consisting of 10 bilayers displayed a high quenching ability, yielding over 85% fluorescence quenching with the addition of a 2 μM dye-labeled aptamer. The limit for human thrombin detection in the 6- and 10-bilayered GO array is estimated to be 0.1 and 0.001 nM, respectively, indicating highly tunable nature of LbL assembled GO multilayers in controlling the sensitivity of graphene-based FRET aptasensor. Furthermore, the GO chip could be reused up to four times simply by cleaning it with distilled water.

Multimode fiber optic wavelength division multiplexing

NASA Technical Reports Server (NTRS)

Spencer, J. L.

1982-01-01

Optical wavelength division multiplexing (WDM) systems, with signals transmitted on different wavelengths through a single optical fiber, can have increased bandwidth and fault isolation properties over single wavelength optical systems. Two WDM system designs that might be used with multimode fibers are considered and a general description of the components which could be used to implement the system are given. The components described are sources, multiplexers, demultiplexers, and detectors. Emphasis is given to the demultiplexer technique which is the major developmental component in the WDM system.

Reduced γ-γ time walk to below 50 ps using the multiplexed-start and multiplexed-stop fast-timing technique with LaBr3(Ce) detectors

NASA Astrophysics Data System (ADS)

Régis, J.-M.; Saed-Samii, N.; Rudigier, M.; Ansari, S.; Dannhoff, M.; Esmaylzadeh, A.; Fransen, C.; Gerst, R.-B.; Jolie, J.; Karayonchev, V.; Müller-Gatermann, C.; Stegemann, S.

2016-07-01

The electronic γ-γ fast-timing technique using arrays consisting of many LaBr3(Ce) detectors is a powerful method to determine lifetimes of nuclear excited states with a lower limit of about 5 ps. This method requires the determination of the energy-dependent time walk of the zero time which is represented by the centroid of a prompt γ-γ time distribution. The full-energy peak versus full-energy peak prompt response difference which represents the linearly combined mean γ-γ time walk of a fast-timing array consisting of 8 LaBr3(Ce) detectors was measured using a standard 152Eu γ-ray source for the energy region of 40-1408 keV. The data were acquired using a "multiplexed-start and multiplexed-stop" analogue electronics circuitry and analysed by employing the generalized centroid difference method. Concerning the cylindrical 1.5 in.×1.5 in. LaBr3(Ce) crystals which are coupled to the Hamamatsu R9779 photomultiplier tubes, the best fast-timing array time resolution of 202(3) ps is obtained for the two prompt γ lines of 60Co by using the leading-edge timing principle. When using the zero-crossover timing principle the time resolution is degraded by up to 30%, dependent on the energy and the shaping delay time of the constant fraction discriminator model Ortec 935. The smallest γ-γ time walk to below 50 ps is obtained by using a shaping delay time of about 17 ns and an optimum "time-walk adjustment" needed for detector output pulses with amplitudes smaller than 400 mV.

Two-wavelength quadrature multipoint detection of partial discharge in power transformers using fiber Fabry-Perot acoustic sensors

NASA Astrophysics Data System (ADS)

Dong, Bo; Han, Ming; Wang, Anbo

2012-06-01

A reliable and low-cost two-wavelength quadrature interrogating method has been developed to demodulate optical signals from diaphragm-based Fabry-Perot interferometric fiber optic sensors for multipoint partial discharge detection in power transformers. Commercial available fused-silica parts (a wafer, a fiber ferrule, and a mating sleeve) and a cleaved optical single mode fiber were bonded together to form an extrinsic Fabry-Perot acoustic sensor. Two lasers with center wavelengths separated by a quarter of the period of sensor interference fringes were used to probe acousticwave- induced diaphragm vibration. A coarse wavelength-division multiplexing (CWDM) add/drop multiplexer was used to separate the reflected two wavelengths before two photo detectors. Optical couplers were used to distribute mixed laser light to each sensor-detector module for multiplexing purpose. Sensor structure, detection system design and experiment results are presented.

[Determination of eleven fluorescent whitening agents in paper food packaging materials by UPLC-FLD/PDA with series double-detector].

PubMed

Wang, Tianjiao; Wu, Pinggu; Hu, Zhengyan; Wang, Liyuan; Tang, Jun; Jiang, Wei; Wang, Zhiyuan

2016-07-01

To establish a new qualitative and quantitative ultraperformance liquid chromatography-fluorescence detector / photodiode array detector with series double-detector method for the determination of eleven fluorescent whitening agents in paper food packaging materials. The sample was extracted with 40%acetonitrile water solution, separated by Waters ACQUITY UPLC BEH C_(18)column( 1. 7μm, 2. 1 mm × 100 mm) and eluted gradient. The excitation wavelength and emission wavelength of fluorescence detector( FLD) were 350 nm and 430 nm, and the wavelength of photodiode array detector( PDA) was 350 nm. The detectors were used in series to achieve qualitative and quantitative detection. In the substrates of paper cups, paper bowls, paper trays and paper boxes, those eleven fluorescent whitening agents were separated properly. For both detectors, in the linear range of 25- 1000 ng / m L, the correlation coefficient was greater than 0. 99, and the recoveries of spiked recoveries were between 82. 2%- 104. 1% with the RSD less than 10%( n = 6). The detection limits ofthose eleven fluorescent whitening agents were 0. 20- 0. 28 mg / kg for FLD and 1. 4- 2. 5mg / kg for PDA. The eleven fluorescent whitening agents could be separated properly with complete separation, good shapes and high recovery rate. This method is easy to operate also. Thus it's an effective method to detect the fluorescent whitening agents in paper food packaging materials.

Non-Destructive Optical Monitoring of Grape Maturation by Proximal Sensing

PubMed Central

Ben Ghozlen, Naïma; Cerovic, Zoran G.; Germain, Claire; Toutain, Sandrine; Latouche, Gwendal

2010-01-01

A new, commercial, fluorescence-based optical sensor for plant constituent assessment was recently introduced. This sensor, called the Multiplex® (FORCE-A, Orsay, France), was used to monitor grape maturation by specifically monitoring anthocyanin accumulation. We derived the empirical anthocyanin content calibration curves for Champagne red grape cultivars, and we also propose a general model for the influence of the proportion of red berries, skin anthocyanin content and berry size on Multiplex® indices. The Multiplex® was used on both berry samples in the laboratory and on intact clusters in the vineyard. We found that the inverted and log-transformed far-red fluorescence signal called the FERARI index, although sensitive to sample size and distance, is potentially the most widely applicable. The more robust indices, based on chlorophyll fluorescence excitation ratios, showed three ranges of dependence on anthocyanin content. We found that up to 0.16 mg cm−2, equivalent to approximately 0.6 mg g−1, all indices increase with accumulation of skin anthocyanin content. Excitation ratio-based indices decrease with anthocyanin accumulation beyond 0.27 mg cm−2. We showed that the Multiplex® can be advantageously used in vineyards on intact clusters for the non-destructive assessment of anthocyanin content of vine blocks and can now be tested on other fruits and vegetables based on the same model. PMID:22163456

TCSPC based approaches for multiparameter detection in living cells

NASA Astrophysics Data System (ADS)

Jahn, Karolina; Buschmann, Volker; Koberling, Felix; Hille, Carsten

2014-03-01

In living cells a manifold of processes take place simultaneously. This implies a precise regulation of intracellular ion homeostasis. In order to understand their spatio-temporal pattern comprehensively, the development of multiplexing concepts is essential. Due to the multidimensional characteristics of fluorescence dyes (absorption and emission spectra, decay time, anisotropy), the highly sensitive and non-invasive fluorescence microscopy is a versatile tool for realising multiplexing concepts. A prerequisite are analyte-specific fluorescence dyes with low cross-sensitivity to other dyes and analytes, respectively. Here, two approaches for multiparameter detection in living cells are presented. Insect salivary glands are well characterised secretory active tissues which were used as model systems to evaluate multiplexing concepts. Salivary glands secrete a KCl-rich or NaCl-rich fluid upon stimulation which is mainly regulated by intracellular Ca2+ as second messenger. Thus, pairwise detection of intracellular Na+, Cl- and Ca2+ with the fluorescent dyes ANG2, MQAE and ACR were tested. Therefore, the dyes were excited simultaneously (2-photon excitation) and their corresponding fluorescence decay times were recorded within two spectral ranges using time-correlated singlephoton counting (TCSPC). A second approach presented here is based on a new TCSPC-platform covering decay time detection from picoseconds to milliseconds. Thereby, nanosecond decaying cellular fluorescence and microsecond decaying phosphorescence of Ruthenium-complexes, which is quenched by oxygen, were recorded simultaneously. In both cases changes in luminescence decay times can be linked to changes in analyte concentrations. In consequence of simultaneous excitation as well as detection, it is possible to get a deeper insight into spatio-temporal pattern in living tissues.

On-Chip Synthesis of Protein Microarrays from DNA Microarrays Via Coupled In Vitro Transcription and Translation for Surface Plasmon Resonance Imaging Biosensor Applications

PubMed Central

Seefeld, Ting H.; Halpern, Aaron R.; Corn, Robert M.

2012-01-01

Protein microarrays are fabricated from double-stranded DNA (dsDNA) microarrays by a one-step, multiplexed enzymatic synthesis in an on-chip microfluidic format and then employed for antibody biosensing measurements with surface plasmon resonance imaging (SPRI). A microarray of dsDNA elements (denoted as generator elements) that encode either a His-tagged green fluorescent protein (GFP) or a His-tagged luciferase protein is utilized to create multiple copies of messenger RNA (mRNA) in a surface RNA polymerase reaction; the mRNA transcripts are then translated into proteins by cell-free protein synthesis in a microfluidic format. The His-tagged proteins diffuse to adjacent Cu(II)-NTA microarray elements (denoted as detector elements) and are specifically adsorbed. The net result is the on-chip, cell-free synthesis of a protein microarray that can be used immediately for SPRI protein biosensing. The dual element format greatly reduces any interference from the nonspecific adsorption of enzyme or proteins. SPRI measurements for the detection of the antibodies anti-GFP and anti-luciferase were used to verify the formation of the protein microarray. This convenient on-chip protein microarray fabrication method can be implemented for multiplexed SPRI biosensing measurements in both clinical and research applications. PMID:22793370

The Micro-X Imaging X-Ray Microcalorimeter Sounding Rocket Payload: Final Design and Performance Tests

NASA Astrophysics Data System (ADS)

Rutherford, John; Micro-X Collaboration

2011-09-01

The first operating set of transition edge sensors (TES) microcalorimeters in space will launch on a sounding rocket carrying the Micro-X imaging X-ray telescope in 2012. We present the final instrument flight design, as well as the results from initial performance tests. A spectral resolution of 2 eV is targeted across the science band of 0.3-2.5 keV. The 12x12 spectrometer array contains 128 active pixels on a 600 micron pitch, consisting of Au/Bi absorbers and Mo/Au bilayer TESs with a transition temperature of 100 mK. A SQUID time-division multiplexer will read out the array at 30 kHz, which is limited by the rocket telemetry. The TESs have been engineered with a 2 ms time constant to match the multiplexer. The detector array and two SQUID stages of the TDM readout system are accommodated in a lightweight Mg enclosure, which is mounted to the 50 mK stage of an adiabatic demagnetization refrigerator. A third SQUID amplification stage is located on the 1.6 K liquid He stage of the cryostat. An on-board 55-Fe source will fluoresce a Ca target, providing 3.7 and 4.0 keV calibration lines that will not interfere with the scientifically interesting energy band.

The Statistical Value of Raw Fluorescence Signal in Luminex xMAP Based Multiplex Immunoassays

PubMed Central

Breen, Edmond J.; Tan, Woei; Khan, Alamgir

2016-01-01

Tissue samples (plasma, saliva, serum or urine) from 169 patients classified as either normal or having one of seven possible diseases are analysed across three 96-well plates for the presences of 37 analytes using cytokine inflammation multiplexed immunoassay panels. Censoring for concentration data caused problems for analysis of the low abundant analytes. Using fluorescence analysis over concentration based analysis allowed analysis of these low abundant analytes. Mixed-effects analysis on the resulting fluorescence and concentration responses reveals a combination of censoring and mapping the fluorescence responses to concentration values, through a 5PL curve, changed observed analyte concentrations. Simulation verifies this, by showing a dependence on the mean florescence response and its distribution on the observed analyte concentration levels. Differences from normality, in the fluorescence responses, can lead to differences in concentration estimates and unreliable probabilities for treatment effects. It is seen that when fluorescence responses are normally distributed, probabilities of treatment effects for fluorescence based t-tests has greater statistical power than the same probabilities from concentration based t-tests. We add evidence that the fluorescence response, unlike concentration values, doesn’t require censoring and we show with respect to differential analysis on the fluorescence responses that background correction is not required. PMID:27243383

Backshort-Under-Grid arrays for infrared astronomy

NASA Astrophysics Data System (ADS)

Allen, C. A.; Benford, D. J.; Chervenak, J. A.; Chuss, D. T.; Miller, T. M.; Moseley, S. H.; Staguhn, J. G.; Wollack, E. J.

2006-04-01

We are developing a kilopixel, filled bolometer array for space infrared astronomy. The array consists of three individual components, to be merged into a single, working unit; (1) a transition edge sensor bolometer array, operating in the milliKelvin regime, (2) a quarter-wave backshort grid, and (3) superconducting quantum interference device multiplexer readout. The detector array is designed as a filled, square grid of suspended, silicon bolometers with superconducting sensors. The backshort arrays are fabricated separately and will be positioned in the cavities created behind each detector during fabrication. The grids have a unique interlocking feature machined into the walls for positioning and mechanical stability. The spacing of the backshort beneath the detector grid can be set from ˜30 300 μm, by independently adjusting two process parameters during fabrication. The ultimate goal is to develop a large-format array architecture with background-limited sensitivity, suitable for a wide range of wavelengths and applications, to be directly bump bonded to a multiplexer circuit. We have produced prototype two-dimensional arrays having 8×8 detector elements. We present detector design, fabrication overview, and assembly technologies.

Iterative color-multiplexed, electro-optical processor.

PubMed

Psaltis, D; Casasent, D; Carlotto, M

1979-11-01

A noncoherent optical vector-matrix multiplier using a linear LED source array and a linear P-I-N photodiode detector array has been combined with a 1-D adder in a feedback loop. The resultant iterative optical processor and its use in solving simultaneous linear equations are described. Operation on complex data is provided by a novel color-multiplexing system.

Frequency Domain Multiplexing for Use With NaI[Tl] Detectors

NASA Astrophysics Data System (ADS)

Belling, Samuel; Coherent Collaboration

2017-09-01

A process used in many forms of signal communication known as multiplexing is adapted for the purpose of combining signals from NaI[Tl] detectors so that fewer digitizer channels can be used to process the signal information from large experiments within the COHERENT collaboration. Each signal is passed through a ringing circuit to modulate it with a characteristic frequency. Information about the signal can be extracted from its amplitude, frequency, and phase. Simulations in LTSpice show that an operational amplifier circuit with a parallel LRC feedback loop can serve as the modulating circuit. Several such circuits can be constructed and housed compactly in a unit, and fed to an inverting, summing amplifier with tunable gain, such that the signals are carried by one cable. The signals are analyzed based on a Fourier transform after being digitized. The results show that the energy, channel, and time of the original interaction can be recovered by this process. In some cases it is possible through filtering and deconvolution to recover the shape of the original signal. The effort is ongoing, but with the design presented it is possible to multiplex 10 detectors into a single digitizer channel. NSF REU Program at Duke University.

Toward Large FOV High-Resolution X-Ray Imaging Spectrometer: Microwave Multiplexed Readout of 32 TES Microcalorimeters

NASA Technical Reports Server (NTRS)

Yoon, Wonsik; Adams, Joseph S.; Bandler, Simon R.; Chervenak, James A.; Datesman, Aaron M.; Eckart, Megan E.; Finkbeiner, Fred M.; Kelley, Richard L.; Kilbourne, Caroline A.; Miniussi, Antoine R.;

Horn-coupled, commercially-fabricated aluminum lumped-element kinetic inductance detectors for millimeter wavelengths

DOE Office of Scientific and Technical Information (OSTI.GOV)

McCarrick, H., E-mail: hlm2124@columbia.edu; Flanigan, D.; Jones, G.

We discuss the design, fabrication, and testing of prototype horn-coupled, lumped-element kinetic inductance detectors (LEKIDs) designed for cosmic microwave background studies. The LEKIDs are made from a thin aluminum film deposited on a silicon wafer and patterned using standard photolithographic techniques at STAR Cryoelectronics, a commercial device foundry. We fabricated 20-element arrays, optimized for a spectral band centered on 150 GHz, to test the sensitivity and yield of the devices as well as the multiplexing scheme. We characterized the detectors in two configurations. First, the detectors were tested in a dark environment with the horn apertures covered, and second, themore » horn apertures were pointed towards a beam-filling cryogenic blackbody load. These tests show that the multiplexing scheme is robust and scalable, the yield across multiple LEKID arrays is 91%, and the measured noise-equivalent temperatures for a 4 K optical load are in the range 26±6 μK√(s)« less

Messenger RNA Detection in Leukemia Cell lines by Novel Metal-Tagged in situ Hybridization using Inductively Coupled Plasma Mass Spectrometry.

PubMed

Ornatsky, Olga I; Baranov, Vladimir I; Bandura, Dmitry R; Tanner, Scott D; Dick, John

2006-01-01

Conventional gene expression profiling relies on using fluorescent detection of hybridized probes. Physical characteristics of fluorophores impose limitations on achieving a highly multiplex gene analysis of single cells. Our work demonstrates the feasibility of using metal-tagged in situ hybridization for mRNA detection by inductively coupled plasma mass spectrometry (ICP-MS). ICP-MS as an analytical detector has a number of unique and relevant properties: 1) metals and their stable isotopes generate non-overlapping distinct signals that can be detected simultaneously; 2) these signals can be measured over a wide dynamic range; 3) ICP-MS is quantitative and very sensitive. We used commercial antibodies conjugated to europium (Eu) and gold together with biotinylated oligonucleotide probes reacted with terbium-labeled streptavidin to demonstrate simultaneous mRNA and protein detection by ICP-MS in leukemia cells.

Messenger RNA Detection in Leukemia Cell lines by Novel Metal-Tagged in situ Hybridization using Inductively Coupled Plasma Mass Spectrometry

PubMed Central

Ornatsky, Olga I.; Baranov, Vladimir I.; Bandura, Dmitry R.; Tanner, Scott D.; Dick, John

2006-01-01

Conventional gene expression profiling relies on using fluorescent detection of hybridized probes. Physical characteristics of fluorophores impose limitations on achieving a highly multiplex gene analysis of single cells. Our work demonstrates the feasibility of using metal-tagged in situ hybridization for mRNA detection by inductively coupled plasma mass spectrometry (ICP-MS). ICP-MS as an analytical detector has a number of unique and relevant properties: 1) metals and their stable isotopes generate non-overlapping distinct signals that can be detected simultaneously; 2) these signals can be measured over a wide dynamic range; 3) ICP-MS is quantitative and very sensitive. We used commercial antibodies conjugated to europium (Eu) and gold together with biotinylated oligonucleotide probes reacted with terbium-labeled streptavidin to demonstrate simultaneous mRNA and protein detection by ICP-MS in leukemia cells. PMID:23662035

Automated high-throughput flow-through real-time diagnostic system

DOEpatents

Regan, John Frederick

2012-10-30

An automated real-time flow-through system capable of processing multiple samples in an asynchronous, simultaneous, and parallel fashion for nucleic acid extraction and purification, followed by assay assembly, genetic amplification, multiplex detection, analysis, and decontamination. The system is able to hold and access an unlimited number of fluorescent reagents that may be used to screen samples for the presence of specific sequences. The apparatus works by associating extracted and purified sample with a series of reagent plugs that have been formed in a flow channel and delivered to a flow-through real-time amplification detector that has a multiplicity of optical windows, to which the sample-reagent plugs are placed in an operative position. The diagnostic apparatus includes sample multi-position valves, a master sample multi-position valve, a master reagent multi-position valve, reagent multi-position valves, and an optical amplification/detection system.

State-of-the-art survey of multimode fiber optic wavelength division multiplexing

NASA Astrophysics Data System (ADS)

Spencer, J. L.

1983-05-01

Optical wavelength division multiplexing (WDM) systems, with signals transmitted on different wavelengths through a single fiber, can have increased information capacity and fault isolation properties over single wavelength optical systems. This paper describes a typical WDM system. Also, a state-of-the-art survey of optical multimode components which could be used to implement the system is made. The components to be surveyed are sources, multiplexers, and detectors. Emphasis is given to the demultiplexer techniques which are the major development components in the WDM system.

Integrated multiplexed capillary electrophoresis system

DOEpatents

Yeung, Edward S.; Tan, Hongdong

2002-05-14

The present invention provides an integrated multiplexed capillary electrophoresis system for the analysis of sample analytes. The system integrates and automates multiple components, such as chromatographic columns and separation capillaries, and further provides a detector for the detection of analytes eluting from the separation capillaries. The system employs multiplexed freeze/thaw valves to manage fluid flow and sample movement. The system is computer controlled and is capable of processing samples through reaction, purification, denaturation, pre-concentration, injection, separation and detection in parallel fashion. Methods employing the system of the invention are also provided.

Indium Hybridization of Large Format TES Bolometer Arrays to Readout Multiplexers for Far-Infrared Astronomy

NASA Technical Reports Server (NTRS)

Miller, Timothy M.; Costen, Nick; Allen, Christine

2007-01-01

The advance of new detector technologies combined with enhanced fabrication methods has resulted in an increase in development of large format arrays. The next generation of scientific instruments will utilize detectors containing hundreds to thousands of elements providing a more efficient means to conduct large area sky surveys. Some notable detectors include a 32x32 x-ray microcalorimeter for Constellation-X, an infrared bolometer called SAFIRE to fly on the airborne observatory SOFIA, and the sub-millimeter bolometer SCUBA-2 to be deployed at the JCMT which will use more than 10,000 elements for two colors, each color using four 32x40 arrays. Of these detectors, SCUBA-2 is farthest along in development and uses indium hybridization to multiplexers for readout of the large number of elements, a technology that will be required to enable the next generation of large format arrays. Our current efforts in working toward large format arrays have produced GISMO, the Goddard IRAM Superconducting 2-Millimeter observer. GISMO is a far infrared instrument to be field tested later this year at the IRAM 30 meter telescope in Spain. GISMO utilizes transition edge sensor (TES) technology in an 8x16 filled array format that allows for typical fan-out wiring and wire-bonding to four 1x32 NIST multiplexers. GISMO'S electrical wiring is routed along the tops of 30 micron walls which also serve as the mechanical framework for the array. This architecture works well for the 128 element array, but is approaching the limit for routing the necessary wires along the surface while maintaining a high fill factor. Larger format arrays will benefit greatly from making electrical connections through the wafer to the backside, where they can be hybridized to a read-out substrate tailored to handling the wiring scheme. The next generation array we are developing is a 32x40 element array on a pitch of 1135 microns that conforms to the NIST multiplexer, already developed for the SCUBA-2 instrument This architecture will utilize electrical connections that route from the TES to the support frame and through the wafer. The detector chip will then be hybridized to the NIST multiplexer via indium bump bonding. In our development scheme we are using substrates that allow for diagnostic testing of electrical continuity across the entire array and we are testing our process to minimize or eliminate any contact resistance at metal interfaces. Our goal is hybridizing a fully functional 32x40 array of TES bolometers to a NIST multiplexer. The following work presents our current progress toward enabling this technology.

Optimising the multiplex factor of the frequency domain multiplexed readout of the TES-based microcalorimeter imaging array for the X-IFU instrument on the Athena x-ray observatory

NASA Astrophysics Data System (ADS)

van der Kuur, J.; Gottardi, L. G.; Akamatsu, H.; van Leeuwen, B. J.; den Hartog, R.; Haas, D.; Kiviranta, M.; Jackson, B. J.

2016-07-01

Athena is a space-based X-ray observatory intended for exploration of the hot and energetic universe. One of the science instruments on Athena will be the X-ray Integrated Field Unit (X-IFU), which is a cryogenic X-ray spectrometer, based on a large cryogenic imaging array of Transition Edge Sensors (TES) based microcalorimeters operating at a temperature of 100mK. The imaging array consists of 3800 pixels providing 2.5 eV spectral resolution, and covers a field of view with a diameter of of 5 arc minutes. Multiplexed readout of the cryogenic microcalorimeter array is essential to comply with the cooling power and complexity constraints on a space craft. Frequency domain multiplexing has been under development for the readout of TES-based detectors for this purpose, not only for the X-IFU detector arrays but also for TES-based bolometer arrays for the Safari instrument of the Japanese SPICA observatory. This paper discusses the design considerations which are applicable to optimise the multiplex factor within the boundary conditions as set by the space craft. More specifically, the interplay between the science requirements such as pixel dynamic range, pixel speed, and cross talk, and the space craft requirements such as the power dissipation budget, available bandwidth, and electromagnetic compatibility will be discussed.

Parallel separations using capillary electrophoresis on a multilane microchip with multiplexed laser induced fluorescence detection

PubMed Central

Nikcevic, Irena; Piruska, Aigars; Wehmeyer, Kenneth R.; Seliskar, Carl J.; Limbach, Patrick A.; Heineman, William R.

2010-01-01

Parallel separations using capillary electrophoresis on a multilane microchip with multiplexed laser induced fluorescence detection is demonstrated. The detection system was developed to simultaneously record data on all channels using an expanded laser beam for excitation, a camera lens to capture emission, and a CCD camera for detection. The detection system enables monitoring of each channel continuously and distinguishing individual lanes without significant crosstalk between adjacent lanes. Multiple analytes can be analyzed on parallel lanes within a single microchip in a single run, leading to increased sample throughput. The pKa determination of small molecule analytes is demonstrated with the multilane microchip. PMID:20737446

The Detector and Readout Systems of the Micro-X High Resolution Microcalorimeter X-Ray Imaging Rocket

NASA Astrophysics Data System (ADS)

Wikus, P.; Doriese, W. B.; Eckart, M. E.; Adams, J. S.; Bandler, S. R.; Brekosky, R. P.; Chervenak, J. A.; Ewin, A. J.; Figueroa-Feliciano, E.; Finkbeiner, F. M.; Galeazzi, M.; Hilton, G.; Irwin, K. D.; Kelley, R. L.; Kilbourne, C. A.; Leman, S. W.; McCammon, D.; Porter, F. S.; Reintsema, C. D.; Rutherford, J. M.; Trowbridge, S. N.

2009-12-01

The Micro-X sounding rocket experiment will deploy an imaging transition-edge-sensor (TES) microcalorimeter spectrometer to observe astrophysical sources in the 0.2-3.0 keV band. The instrument has been designed at a systems level, and the first items of flight hardware are presently being built. In the first flight, planned for January 2011, the spectrometer will observe a recently discovered Silicon knot in the Puppis-A supernova remnant. Here we describe the design of the Micro-X science instrument, focusing on the instrument's detector and detector assembly. The current design of the 2-dimensional spectrometer array contains 128 close-packed pixels with a pitch of 600 μm. The conically approximated Wolter-1 mirror will map each of these pixels to a 0.95 arcmin region on the sky; the field of view will be 11.4 arcmin. Targeted energy resolution of the TESs is about 2 eV over the full observing band. A SQUID time-division multiplexer (TDM) will read out the array. The detector time constants will be engineered to approximately 2 ms to match the TDM, which samples each pixel at 32.6 kHz, limited only by the telemetry system of the rocket. The detector array and two SQUID stages of the TDM readout system are accommodated in a lightweight Mg enclosure, which is mounted to the 50 mK stage of an adiabatic demagnetization refrigerator. A third SQUID amplification stage is located on the 1.6 K liquid He stage of the cryostat. An on-board 55-Fe source will fluoresce a Ca target, providing 3.69 and 4.01 keV calibration lines that will not interfere with the scientifically interesting energy band.

The energy spectrum of cosmic rays above 1017.2 eV measured by the fluorescence detectors of the Telescope Array experiment in seven years

NASA Astrophysics Data System (ADS)

Abbasi, R. U.; Abe, M.; Abu-Zayyad, T.; Allen, M.; Azuma, R.; Barcikowski, E.; Belz, J. W.; Bergman, D. R.; Blake, S. A.; Cady, R.; Cheon, B. G.; Chiba, J.; Chikawa, M.; Cho, W. R.; Fujii, T.; Fukushima, M.; Goto, T.; Hanlon, W.; Hayashi, Y.; Hayashida, N.; Hibino, K.; Honda, K.; Ikeda, D.; Inoue, N.; Ishii, T.; Ishimori, R.; Ito, H.; Ivanov, D.; Jui, C. C. H.; Kadota, K.; Kakimoto, F.; Kalashev, O.; Kasahara, K.; Kawai, H.; Kawakami, S.; Kawana, S.; Kawata, K.; Kido, E.; Kim, H. B.; Kim, J. H.; Kim, J. H.; Kitamura, S.; Kitamura, Y.; Kuzmin, V.; Kwon, Y. J.; Lan, J.; Lundquist, J. P.; Machida, K.; Martens, K.; Matsuda, T.; Matsuyama, T.; Matthews, J. N.; Minamino, M.; Mukai, Y.; Myers, I.; Nagasawa, K.; Nagataki, S.; Nakamura, T.; Nonaka, T.; Nozato, A.; Ogio, S.; Ogura, J.; Ohnishi, M.; Ohoka, H.; Oki, K.; Okuda, T.; Ono, M.; Oshima, A.; Ozawa, S.; Park, I. H.; Pshirkov, M. S.; Rodriguez, D. C.; Rubtsov, G.; Ryu, D.; Sagawa, H.; Sakurai, N.; Scott, L. M.; Shah, P. D.; Shibata, F.; Shibata, T.; Shimodaira, H.; Shin, B. K.; Shin, H. S.; Smith, J. D.; Sokolsky, P.; Springer, R. W.; Stokes, B. T.; Stratton, S. R.; Stroman, T. A.; Suzawa, T.; Takamura, M.; Takeda, M.; Takeishi, R.; Taketa, A.; Takita, M.; Tameda, Y.; Tanaka, H.; Tanaka, K.; Tanaka, M.; Thomas, S. B.; Thomson, G. B.; Tinyakov, P.; Tkachev, I.; Tokuno, H.; Tomida, T.; Troitsky, S.; Tsunesada, Y.; Tsutsumi, K.; Uchihori, Y.; Udo, S.; Urban, F.; Vasiloff, G.; Wong, T.; Yamane, R.; Yamaoka, H.; Yamazaki, K.; Yang, J.; Yashiro, K.; Yoneda, Y.; Yoshida, S.; Yoshii, H.; Zollinger, R.; Zundel, Z.

2016-07-01

The Telescope Array (TA) experiment is the largest detector to observe ultra-high-energy cosmic rays in the northern hemisphere. The fluorescence detectors at two stations of TA are newly constructed and have now completed seven years of steady operation. One advantage of monocular analysis of the fluorescence detectors is a lower energy threshold for cosmic rays than that of other techniques like stereoscopic observations or coincidences with the surface detector array, allowing the measurement of an energy spectrum covering three orders of magnitude in energy. Analyzing data collected during those seven years, we report the energy spectrum of cosmic rays covering a broad range of energies above 1017.2eV measured by the fluorescence detectors and a comparison with previously published results.

Visible and infrared linear detector arrays for the Airborne Visible/Infrared Imaging Spectrometer (AVIRIS)

NASA Technical Reports Server (NTRS)

Bailey, Gary C.

1987-01-01

The Airborne Visible/Infrared Imaging Spectrometer (AVIRIS) instrument uses four separate focal plane assemblies consisting of line array detectors that are multiplexed to a common J-FET preamp using a FET switch multiplexing (MUX) technique. A 32-element silicon line array covers the spectral range from 0.41 to 0.70 microns. Three additional 64-element indium antimonide (InSb) line arrays cover the spectral range from 0.68 to 2.45 microns. The spectral sampling interval per detector element is nominally 9.8 nm, giving a total of 224 spectral channels. All focal planes operate at liquid nitrogen temperature and are housed in separate dewars. Electrical performance characteristics include a read noise of less than 1000 e(-) in all channels, response and dark nonuniformity of 5 percent peak to peak, and quantum efficiency of greater than 60 percent.

Progress Towards High-Sensitivity Arrays of Detectors of Sub-mm Radiation Using Superconducting Tunnel Junctions with Integrated Radio Frequency Single-Electron Transistors

NASA Technical Reports Server (NTRS)

Stevenson, T. R.; Hsieh, W.-T.; Li, M. J.; Prober, D. E.; Rhee, K. W.; Schoelkopf, R. J.; Stahle, C. M.; Teufel, J.; Wollack, E. J.

2004-01-01

For high resolution imaging and spectroscopy in the FIR and submillimeter, space observatories will demand sensitive, fast, compact, low-power detector arrays with 104 pixels and sensitivity less than 10(exp -20) W/Hz(sup 0.5). Antenna-coupled superconducting tunnel junctions with integrated rf single-electron transistor readout amplifiers have the potential for achieving this high level of sensitivity, and can take advantage of an rf multiplexing technique. The device consists of an antenna to couple radiation into a small superconducting volume and cause quasiparticle excitations, and a single-electron transistor to measure current through junctions contacting the absorber. We describe optimization of device parameters, and results on fabrication techniques for producing devices with high yield for detector arrays. We also present modeling of expected saturation power levels, antenna coupling, and rf multiplexing schemes.

Multiplex and high-throughput DNA detection using surface plasmon mediated fluorescence

NASA Astrophysics Data System (ADS)

Mei, Zhong

The overall objective of this research project was to develop a user-friendly and sensitive biosensor for nucleic acid aptamers with multiplexing and high-throughput capability. The sensing was based on the fluorescence signals emitted by the fluorophores coupling with plamonic nanoparticle (gold nanorod) deposited on a patterned substrate. Gold nanorods (GNRs) were synthesized using a binary mixture of hexadecyltrimethylammonium bromide (CTAB) and sodium oleate (NaOL) in seed mediated growth method. Polytetrafluoroethylene (PTFE) printed glass slides were selectively coated with a gold thin-film to define hydrophilic areas for GNR deposition. Due to the wettablity contrast, GNR solution dropped on the slide was induced to assemble exclusively in the hydrophilic spots. By controlling temperature and humidity of the evaporation process, vertically-standing GNR arrays were achieved on the pattered slide. Fluorescence was conjugated to GNR surface via DNA double strand with tunable length. Theoretical simulation predicted a flat layer ( 30 nm thick) of uniform "hot spots" presented on the GNR tips, which could modify the nearby fluorescence. Experimentally, the vertical GNR arrays yielded metallic enhanced fluorescence (MEF) effect, which was dependent on the spectrum overlap and GNR-fluorophore distance. Specifically, the maximum enhancement of Quasar 670 and Alexa 750 was observed when it was coupled with GNR664 (plasmonic wavelength 664 nm) and GNR778 respectively at a distance of 16 nm, while the carboxyfluorescein (FAM) was at maximal intensity when attached to gold nanosphere520. This offers an opportunity for multiplexed DNA sensing. Based on this, we developed a novel GNR mediated fluorescence biosensor for DNA detection. Fluorescence labeled haipin-DNA probes were introduced to designated spots of GNR array with the matching LSPR wavelengths on the substrate. The fluorescence was quenched originally because of Forster resonance energy transfer (FRET) effect. Upon hybridization with their complimentary target DNAs, hairpin structures were opened and the fluorescence enhancement from each GNR sensing spot was measured by fluorescence scanning. We demonstrated multiple DNA sequences were simultaneously detected at a picomolar level with high-throughput capability using the ordered GNR array biochip.

High-sensitivity detection of biological amines using fast Hadamard transform CE coupled with photolytic optical gating.

PubMed

Braun, Kevin L; Hapuarachchi, Suminda; Fernandez, Facundo M; Aspinwall, Craig A

2007-08-01

Here, we report the first utilization of Hadamard transform CE (HTCE), a high-sensitivity, multiplexed CE technique, with photolytic optical gating sample injection of caged fluorescent labels for the detection of biologically important amines. Previous implementations of HTCE have relied upon photobleaching optical gating sample injection of fluorescent dyes. Photolysis of caged fluorescent labels reduces the fluorescence background, providing marked enhancements in sensitivity compared to photobleaching. Application of fast Hadamard transform CE (fHTCE) for fluorescein-based dyes yields a ten-fold higher sensitivity for photolytic injections compared to photobleaching injections, due primarily to the reduced fluorescent background provided by caged fluorescent dyes. Detection limits as low as 5 pM (ca. 18 molecules per injection event) were obtained with on-column LIF detection using fHTCE in less than 25 s, with the capacity for continuous, online separations. Detection limits for glutamate and aspartate below 150 pM (1-2 amol/injection event) were obtained using photolytic sample injection, with separation efficiencies exceeding 1 x 10(6) plates/m and total multiplexed separation times as low as 8 s. These results strongly support the feasibility of this approach for high-sensitivity dynamic chemical monitoring applications.

Digitally synthesized beat frequency-multiplexed fluorescence lifetime spectroscopy

PubMed Central

Chan, Jacky C. K.; Diebold, Eric D.; Buckley, Brandon W.; Mao, Sien; Akbari, Najva; Jalali, Bahram

2014-01-01

Frequency domain fluorescence lifetime imaging is a powerful technique that enables the observation of subtle changes in the molecular environment of a fluorescent probe. This technique works by measuring the phase delay between the optical emission and excitation of fluorophores as a function of modulation frequency. However, high-resolution measurements are time consuming, as the excitation modulation frequency must be swept, and faster low-resolution measurements at a single frequency are prone to large errors. Here, we present a low cost optical system for applications in real-time confocal lifetime imaging, which measures the phase vs. frequency spectrum without sweeping. Deemed Lifetime Imaging using Frequency-multiplexed Excitation (LIFE), this technique uses a digitally-synthesized radio frequency comb to drive an acousto-optic deflector, operated in a cat’s-eye configuration, to produce a single laser excitation beam modulated at multiple beat frequencies. We demonstrate simultaneous fluorescence lifetime measurements at 10 frequencies over a bandwidth of 48 MHz, enabling high speed frequency domain lifetime analysis of single- and multi-component sample mixtures. PMID:25574449

RNA Imaging with Multiplexed Error Robust Fluorescence in situ Hybridization

PubMed Central

Moffitt, Jeffrey R.; Zhuang, Xiaowei

2016-01-01

Quantitative measurements of both the copy number and spatial distribution of large fractions of the transcriptome in single-cells could revolutionize our understanding of a variety of cellular and tissue behaviors in both healthy and diseased states. Single-molecule Fluorescence In Situ Hybridization (smFISH)—an approach where individual RNAs are labeled with fluorescent probes and imaged in their native cellular and tissue context—provides both the copy number and spatial context of RNAs but has been limited in the number of RNA species that can be measured simultaneously. Here we describe Multiplexed Error Robust Fluorescence In Situ Hybridization (MERFISH), a massively parallelized form of smFISH that can image and identify hundreds to thousands of different RNA species simultaneously with high accuracy in individual cells in their native spatial context. We provide detailed protocols on all aspects of MERFISH, including probe design, data collection, and data analysis to allow interested laboratories to perform MERFISH measurements themselves. PMID:27241748

Broadband Direct Detection Submillimeter Spectrometer with Multiplexed Superconducting Transition Edge Thermometer Bolometers

NASA Technical Reports Server (NTRS)

Benford, D. J.; Ames, T. A.; Chervenak, J. A.; Moseley, S. H.; Shafer, R. A.; Staguhn, J. G.; Voellmer, G. M.; Pajot, F.; Rioux, C.; Phillips, T. G.;

Enhancements to a Superconducting Quantum Interference Device (SQUID) Multiplexer Readout and Control System

NASA Technical Reports Server (NTRS)

Forgione, J.; Benford, D. J.; Buchanan, E. D.; Moseley, S. H.; Rebar, J.; Shafer, R. A.

2004-01-01

Far-infrared detector arrays such as the 16x32 superconducting bolometer array for the SAFIRE instrument (flying on the SOFIA airborne observatory) require systems of readout and control electronics to provide translation between a user-driven, digital PC and the cold, analog world of the cryogenic detector. In 2001, the National Institute of Standards and Technology (NIST) developed their Mark III electronics for purposes of control and readout of their 1x32 SQUID Multiplexer chips. We at NASA s Goddard Space Flight Center acquired a Mark 111 system and subsequently designed upgrades to suit our and our collaborators purposes. We developed an arbitrary, programmable multiplexing system that allows the user to cycle through rows in a SQUID array in an infinite number of combinations. We provided hooks in the Mark III system to allow readout of signals from outside the Mark 111 system, such as telescope status information. Finally, we augmented the heart of the system with a new feedback algorithm implementation, flexible diagnostic tools, and informative telemetry.

Enhancements to a superconducting quantum interference device (SQUID) multiplexer readout and control system

NASA Astrophysics Data System (ADS)

Forgione, Joshua B.; Benford, Dominic J.; Buchanan, Ernest D.; Moseley, S. H., Jr.; Rebar, Joyce; Shafer, Richard A.

2004-10-01

Far-infrared detector arrays such as the 16x32 superconducting bolometer array for the SAFIRE instrument (flying on the SOFIA airborne observatory) require systems of readout and control electronics to provide translation between a user-driven, digital PC and the cold, analog world of the cryogenic detector. In 2001, the National Institute of Standards and Technology (NIST) developed their Mark III electronics for purposes of control and readout of their 1x32 SQUID Multiplexer chips. We at NASA's Goddard Space Flight Center acquired a Mark III system and subsequently designed upgrades to suit our and our collaborators' purposes. We developed an arbitrary, programmable multiplexing system that allows the user to cycle through rows in a SQUID array in an infinite number of combinations. We provided 'hooks' in the Mark III system to allow readout of signals from outside the Mark III system, such as telescope status information. Finally, we augmented the heart of the system with a new feedback algorithm implementation, flexible diagnostic tools, and informative telemetry.

Development of capacitive multiplexing circuit for SiPM-based time-of-flight (TOF) PET detector

NASA Astrophysics Data System (ADS)

Choe, Hyeok-Jun; Choi, Yong; Hu, Wei; Yan, Jianhua; Jung, Jin Ho

2017-04-01

There has been great interest in developing a time-of-flight (TOF) PET to improve the signal-to-noise ratio of PET image relative to that of non-TOF PET. Silicon photomultiplier (SiPM) arrays have attracted attention for use as a fast TOF PET photosensor. Since numerous SiPM arrays are needed to construct a modern human PET, a multiplexing method providing both good timing performance and high channel reduction capability is required to develop a SiPM-based TOF PET. The purpose of this study was to develop a capacitive multiplexing circuit for the SiPM-based TOF PET. The proposed multiplexing circuit was evaluated by measuring the coincidence resolving time (CRT) and the energy resolution as a function of the overvoltage using three different capacitor values of 15, 30, and 51 pF. A flood histogram was also obtained and quantitatively assessed. Experiments were performed using a 4× 4 array of 3× 3 mm2 SiPMs. Regarding the capacitor values, the multiplexing circuit using a smaller capacitor value showed the best timing performance. On the other hand, the energy resolution and flood histogram quality of the multiplexing circuit deteriorated as the capacitor value became smaller. The proposed circuit was able to achieve a CRT of 260+/- 4 ps FWHM and an energy resolution of 17.1 % with a pair of 2× 2× 20 mm3 LYSO crystals using a capacitor value of 30 pF at an overvoltage of 3.0 V. It was also possible to clearly resolve a 6× 6 array of LYSO crystals in the flood histogram using the multiplexing circuit. The experiment results indicate that the proposed capacitive multiplexing circuit is useful to obtain an excellent timing performance and a crystal-resolving capability in the flood histogram with a minimal degradation of the energy resolution, as well as to reduce the number of the readout channels of the SiPM-based TOF PET detector.

Hole-Impeded-Doping-Superlattice LWIR Detectors

NASA Technical Reports Server (NTRS)

Maserjian, Joseph

1991-01-01

Hole-Impeded-Doping-Superlattice (HIDS) InAs devices proposed for use as photoconductive or photovoltaic detectors of radiation in long-wavelength infrared (LWIR) range of 8 to 17 micrometers. Array of HIDS devices fabricated on substrates GaAs or Si. Radiation incident on black surface, metal contacts for picture elements serve as reactors, effectively doubling optical path and thereby increasing absorption of photons. Photoconductive detector offers advantages of high gain and high impedance; photovoltaic detector offers lower noise and better interface to multiplexer readouts.

NASA AMES infrared detector assemblies

NASA Technical Reports Server (NTRS)

1979-01-01

Silicon: Gallium infrared detector assemblies were designed, fabricated, and tested using techniques representative of those employed for hybrid arrays to determine the suitability of this candidate technology for infrared astronomical detector array applications. Both the single channel assembly and the assembly using a 32 channel CMOS multiplexer are considered. The detector material was certified to have a boron background of less than 10 to the 13th power atoms/sq cm counter doped with phosphorus. The gallium concentration is 2 x 10 to the 16th power atoms/cu cm.

Demonstration of a scalable frequency-domain readout of metallic magnetic calorimeters by means of a microwave SQUID multiplexer

NASA Astrophysics Data System (ADS)

Kempf, Sebastian; Wegner, Mathias; Fleischmann, Andreas; Gastaldo, Loredana; Herrmann, Felix; Papst, Maximilian; Richter, Daniel; Enss, Christian

2017-01-01

We report on the first demonstration of a scalable GHz frequency-domain readout of metallic magnetic calorimeters (MMCs) using a 64 pixel detector array that is read out by an integrated, on-chip microwave SQUID multiplexer. The detector array is optimized for detecting soft X-ray photons and the multiplexer is designed to provide a signal rise time τrise<400 ns and an intrinsic energy sensitivity ɛ <30 h . This results in an expected energy resolution Δ EFWHM <10 eV . We measured a signal rise time τrise as low as 90 ns and an energy resolution Δ EFWHM as low as 50 eV for 5.9 keV photons. The rise time is about an order of magnitude faster compared to other multiplexed low-temperature microcalorimeters and close to the intrinsic value set by the coupling between electron and spins. The energy resolution is degraded with respect to our design value due to a rather low intrinsic quality factor of the microwave resonators that is caused by the quality of the Josephson junction of the associated rf-SQUID as well as an elevated chip temperature as compared to the heat bath. Though the achieved energy resolution is not yet compatible with state-of-the-art single-channel MMCs, this demonstration of a scalable readout approach for MMCs in combination with the full understanding of the device performance showing ways how to improve represents an important milestone for the development of future large-scale MMC detector arrays.

Externally-Modulated Electro-Optically Coupled Detector Architecture for Nuclear Physics Instrumentation

NASA Astrophysics Data System (ADS)

Xi, Wenze; McKisson, J. E.; Weisenberger, Andrew G.; Zhang, Shukui; Zorn, Carl

2014-06-01

A new laser-based externally-modulated electro-optically coupled detector (EOCD) architecture is being developed to enable high-density readout for radiation detectors with accurate analog radiation pulse shape and timing preservation. Unlike digital conversion before electro-optical modulation, the EOCD implements complete analog optical signal modulation and multiplexing in its detector front-end. The result is a compact, high performance detector readout that can be both radiation tolerant and immune to magnetic fields. In this work, the feasibility of EOCD was explored by constructing a two-wavelength laser-based externally-modulated EOCD, and testing analog pulse shape preservation and wavelength-division multiplexing (WDM) crosstalk. Comparisons were first made between the corresponding initial pulses and the electro-optically coupled analog pulses. This confirmed an excellent analog pulse preservation over 29% of the modulator's switching voltage range. Optical spectrum analysis revealed less than -14 dB crosstalk with 1.2 nm WDM wavelength bandgap, and provided insight on experimental conditions that could lead to increased inter-wavelength crosstalk. Further discussions and previous research on the radiation tolerance and magnetic field immunity of the candidate materials were also given, and quantitative device testing is proposed in the future.

Multiplex detection of IgG and IgM to Rift Valley fever virus nucleoprotein, nonstructural proteins, and glycoprotein in ovine and bovine

USDA-ARS?s Scientific Manuscript database

A multiplex fluorescence microsphere immunoassay (FMIA) was used to detect bovine and ovine IgM and IgG antibodies to several Rift Valley fever virus (RVFV) proteins, including the major surface glycoprotein, Gn; the nonstructural proteins, NSs and NSm; and the nucleoprotein, N. Target antigens were...

A Portable Immunoassay Platform for Multiplexed Detection of Biotoxins in Clinical and Environmental Samples

DOE Office of Scientific and Technical Information (OSTI.GOV)

Koh, Chung-Yan; Piccini, Matthew Ernest; Schaff, Ulrich Y.

Multiple cases of attempted bioterrorism events using biotoxins have highlighted the urgent need for tools capable of rapid screening of suspect samples in the field (e.g., mailroom and public events). We present a portable microfluidic device capable of analyzing environmental (e.g., white powder), food (e.g., milk) and clinical (e.g., blood) samples for multiplexed detection of biotoxins. The device is rapid (<15-30 min sample-to-answer), sensitive (< 0.08 pg/mL detection limit for botulinum toxin), multiplexed (up to 64 parallel assays) and capable of analyzing small volume samples (< 20 μL total sample input). The immunoassay approach (SpinDx) is based on binding ofmore » toxins in a sample to antibody-laden capture particles followed by sedimentation of particles through a density-media in a microfluidic disk and quantification using a laser-induced fluorescence detector. A direct, blinded comparison with a gold standard ELISA revealed a 5-fold more sensitive detection limit for botulinum toxin while requiring 250-fold less sample volume and a 30 minute assay time with a near unity correlation. A key advantage of the technique is its compatibility with a variety of sample matrices with no additional sample preparation required. Ultrasensitive quantification has been demonstrated from direct analysis of multiple clinical, environmental and food samples, including white powder, whole blood, saliva, salad dressing, whole milk, peanut butter, half and half, honey, and canned meat. We believe that this device can met an urgent need in screening both potentially exposed people as well as suspicious samples in mail-rooms, airports, public sporting venues and emergency rooms. The general-purpose immunodiagnostics device can also find applications in screening of infectious and systemic diseases or serve as a lab device for conducting rapid immunoassays.« less

SearchLight: a freely available web-based quantitative spectral analysis tool (Conference Presentation)

NASA Astrophysics Data System (ADS)

Prabhat, Prashant; Peet, Michael; Erdogan, Turan

2016-03-01

In order to design a fluorescence experiment, typically the spectra of a fluorophore and of a filter set are overlaid on a single graph and the spectral overlap is evaluated intuitively. However, in a typical fluorescence imaging system the fluorophores and optical filters are not the only wavelength dependent variables - even the excitation light sources have been changing. For example, LED Light Engines may have a significantly different spectral response compared to the traditional metal-halide lamps. Therefore, for a more accurate assessment of fluorophore-to-filter-set compatibility, all sources of spectral variation should be taken into account simultaneously. Additionally, intuitive or qualitative evaluation of many spectra does not necessarily provide a realistic assessment of the system performance. "SearchLight" is a freely available web-based spectral plotting and analysis tool that can be used to address the need for accurate, quantitative spectral evaluation of fluorescence measurement systems. This tool is available at: http://searchlight.semrock.com/. Based on a detailed mathematical framework [1], SearchLight calculates signal, noise, and signal-to-noise ratio for multiple combinations of fluorophores, filter sets, light sources and detectors. SearchLight allows for qualitative and quantitative evaluation of the compatibility of filter sets with fluorophores, analysis of bleed-through, identification of optimized spectral edge locations for a set of filters under specific experimental conditions, and guidance regarding labeling protocols in multiplexing imaging assays. Entire SearchLight sessions can be shared with colleagues and collaborators and saved for future reference. [1] Anderson, N., Prabhat, P. and Erdogan, T., Spectral Modeling in Fluorescence Microscopy, http://www.semrock.com (2010).

Fiber optic detector

DOE Office of Scientific and Technical Information (OSTI.GOV)

Partin, J.K.; Ward, T.E.; Grey, A.E.

1990-12-31

This invention is comprised of a portable fiber optic detector that senses the presence of specific target chemicals by exchanging the target chemical for a fluorescently-tagged antigen that is bound to an antibody which is in turn attached to an optical fiber. Replacing the fluorescently-tagged antigen reduces the fluorescence so that a photon sensing detector records the reduced light level and activates an appropriate alarm or indicator.

Fiber optic detector

NASA Astrophysics Data System (ADS)

Partin, Judy K.; Ward, Thomas E.; Grey, Alan E.

1990-04-01

This invention is comprised of a portable fiber optic detector that senses the presence of specific target chemicals by exchanging the target chemical for a fluorescently-tagged antigen that is bound to an antibody which is in turn attached to an optical fiber. Replacing the fluorescently-tagged antigen reduces the fluorescence so that a photon sensing detector records the reduced light level and activates an appropriate alarm or indicator.

Sensitive Detection of Protein and miRNA Cancer Biomarkers using Silicon-Based Photonic Crystals and A Resonance Coupling Laser Scanning Platform

PubMed Central

George, Sherine; Chaudhery, Vikram; Lu, Meng; Takagi, Miki; Amro, Nabil; Pokhriyal, Anusha; Tan, Yafang; Ferreira, Placid; Cunningham, Brian T.

2013-01-01

Enhancement of the fluorescent output of surface-based fluorescence assays by performing them upon nanostructured photonic crystal (PC) surfaces has been demonstrated to increase signal intensities by >8000×. Using the multiplicative effects of optical resonant coupling to the PC in increasing the electric field intensity experienced by fluorescent labels (“enhanced excitation”) and the spatially biased funneling of fluorophore emissions through coupling to PC resonances (“enhanced extraction”), PC enhanced fluorescence (PCEF) can be adapted to reduce the limits of detection of disease biomarker assays, and to reduce the size and cost of high sensitivity detection instrumentation. In this work, we demonstrate the first silicon-based PCEF detection platform for multiplexed biomarker assay. The sensor in this platform is a silicon-based PC structure, comprised of a SiO2 grating that is overcoated with a thin film of high refractive index TiO2 and is produced in a semiconductor foundry for low cost, uniform, and reproducible manufacturing. The compact detection instrument that completes this platform was designed to efficiently couples fluorescence excitation from a semiconductor laser to the resonant optical modes of the PC, resulting in elevated electric field strength that is highly concentrated within the region <100 nm from the PC surface. This instrument utilizes a cylindrically focused line to scan a microarray in <1 minute. To demonstrate the capabilities of this sensor-detector platform, microspot fluorescent sandwich immunoassays using secondary antibodies labeled with Cy5 for two cancer biomarkers (TNF-α and IL-3) were performed. Biomarkers were detected at concentrations as low as 0.1 pM. In a fluorescent microarray for detection of a breast cancer miRNA biomarker miR-21, the miRNA was detectable at a concentration of 0.6 pM. PMID:23963502

Fiber optics for the future - wavelength division multiplexing

NASA Technical Reports Server (NTRS)

Spencer, J. L.

1982-01-01

Optical wavelength division multiplexing (WDM) systems, with signals transmitted on different wavelengths through a single fiber, can have increased information capacity and fault isolation properties over single wavelength optical systems. This paper describes a typical WDM system. The applicability of future standards to such a system are discussed. Also, a state-of-the-art survey of optical multimode components which could be used to implement the system are made. The components to be surveyed are sources, multiplexers, and detectors. Emphasis is given to the demultiplexer techniques which are the major developmental components in the WDM system.

High speed, long distance, data transmission multiplexing circuit

DOEpatents

Mariotti, Razvan

1991-01-01

A high speed serial data transmission multiplexing circuit, which is operable to accurately transmit data over long distances (up to 3 Km), and to multiplex, select and continuously display real time analog signals in a bandwidth from DC to 100 Khz. The circuit is made fault tolerant by use of a programmable flywheel algorithm, which enables the circuit to tolerate one transmission error before losing synchronization of the transmitted frames of data. A method of encoding and framing captured and transmitted data is used which has a low overhead and prevents some particular transmitted data patterns from locking an included detector/decoder circuit.

Superconducting Digital Multiplexers for Sensor Arrays

NASA Technical Reports Server (NTRS)

Kadin, Alan M.; Brock, Darren K.; Gupta, Deepnarayan

2004-01-01

Arrays of cryogenic microbolometers and other cryogenic detectors are being developed for infrared imaging. If the signal from each sensor is amplified, multiplexed, and digitized using superconducting electronics, then this data can be efficiently read out to ambient temperature with a minimum of noise and thermal load. HYPRES is developing an integrated system based on SQUID amplifiers, a high-resolution analog-to-digital converter (ADC) based on RSFQ (rapid single flux quantum) logic, and a clocked RSFQ multiplexer. The ADC and SQUIDs have already been demonstrated for other projects, so this paper will focus on new results of a digital multiplexer. Several test circuits have been fabricated using Nb Josephson technology and are about to be tested at T = 4.2 K, with a more complete prototype in preparation.

Fluorescent Cell Barcoding for Multiplex Flow Cytometry

PubMed Central

Krutzik, Peter O.; Clutter, Matthew R.; Trejo, Angelica; Nolan, Garry P.

2011-01-01

Fluorescent Cell Barcoding (FCB) enables high throughput, i.e. high content flow cytometry by multiplexing samples prior to staining and acquisition on the cytometer. Individual cell samples are barcoded, or labeled, with unique signatures of fluorescent dyes so that they can be mixed together, stained, and analyzed as a single sample. By mixing samples prior to staining, antibody consumption is typically reduced 10 to 100-fold. In addition, data robustness is increased through the combination of control and treated samples, which minimizes pipetting error, staining variation, and the need for normalization. Finally, speed of acquisition is enhanced, enabling large profiling experiments to be run with standard cytometer hardware. In this unit, we outline the steps necessary to apply the FCB method to cell lines as well as primary peripheral blood samples. Important technical considerations such as choice of barcoding dyes, concentrations, labeling buffers, compensation, and software analysis are discussed. PMID:21207359

Digital barcodes of suspension array using laser induced breakdown spectroscopy

PubMed Central

He, Qinghua; Liu, Yixi; He, Yonghong; Zhu, Liang; Zhang, Yilong; Shen, Zhiyuan

2016-01-01

We show a coding method of suspension array based on the laser induced breakdown spectroscopy (LIBS), which promotes the barcodes from analog to digital. As the foundation of digital optical barcodes, nanocrystals encoded microspheres are prepared with self-assembly encapsulation method. We confirm that digital multiplexing of LIBS-based coding method becomes feasible since the microsphere can be coded with direct read-out data of wavelengths, and the method can avoid fluorescence signal crosstalk between barcodes and analyte tags, which lead to overall advantages in accuracy and stability to current fluorescent multicolor coding method. This demonstration increases the capability of multiplexed detection and accurate filtrating, expanding more extensive applications of suspension array in life science. PMID:27808270

Measuring partial fluorescence yield using filtered detectors.

PubMed

Boyko, T D; Green, R J; Moewes, A; Regier, T Z

2014-07-01

Typically, X-ray absorption near-edge structure measurements aim to probe the linear attenuation coefficient. These measurements are often carried out using partial fluorescence yield techniques that rely on detectors having photon energy discrimination improving the sensitivity and the signal-to-background ratio of the measured spectra. However, measuring the partial fluorescence yield in the soft X-ray regime with reasonable efficiency requires solid-state detectors, which have limitations due to the inherent dead-time while measuring. Alternatively, many of the available detectors that are not energy dispersive do not suffer from photon count rate limitations. A filter placed in front of one of these detectors will make the energy-dependent efficiency non-linear, thereby changing the responsivity of the detector. It is shown that using an array of filtered X-ray detectors is a viable method for measuring soft X-ray partial fluorescence yield spectra without dead-time. The feasibility of this technique is further demonstrated using α-Fe2O3 as an example and it is shown that this detector technology could vastly improve the photon collection efficiency at synchrotrons and that these detectors will allow experiments to be completed with a much lower photon flux reducing X-ray-induced damage.

Optimization and Standardization of Fluorescent Cell Barcoding for Multiplexed Flow Cytometric Phenotyping

PubMed Central

Giudice, Valentina; Feng, Xingmin; Kajigaya, Sachiko; Young, Neal S.; Biancotto, Angélique

2017-01-01

Fluorescent cell barcoding (FCB) is a cell-based multiplexing technique for high-throughput flow cytometry. Barcoded samples can be stained and acquired collectively, minimizing staining variability and antibody consumption, and decreasing required sample volumes. Combined with functional measurements, FCB can be used for drug screening, signaling profiling, and cytokine detection, but technical issues are present. We optimized the FCB technique for routine utilization using DyLight 350, DyLight 800, Pacific Orange, and CBD500 for barcoding six, nine, or 36 human peripheral blood specimens. Working concentrations of FCB dyes ranging from 0 to 500 μg/ml were tested, and viability dye staining was optimized to increase robustness of data. A five-color staining with surface markers for Vβ usage analysis in CD4+ and CD8+ T cells was achieved in combination with nine sample barcoding. We provide improvements of the FCB technique that should be useful for multiplex drug screening and for lymphocyte characterization and perturbations in the diagnosis and during the course of disease. PMID:28692789

Protein detection using biobarcodes.

PubMed

Müller, Uwe R

2006-10-01

Over the past 50 years the development of assays for the detection of protein analytes has been driven by continuing demands for higher levels of sensitivity and multiplexing. The result has been a progression of sandwich-type immunoassays, starting with simple radioisotopic, colorimetric, or fluorescent labeling systems to include various enzymatic or nanostructure-based signal amplification schemes, with a concomitant sensitivity increase of over 1 million fold. Multiplexing of samples and tests has been enabled by microplate and microarray platforms, respectively, or lately by various molecular barcoding systems. Two different platforms have emerged as the current front-runners by combining a nucleic acid amplification step with the standard two-sided immunoassay. In both, the captured protein analyte is replaced by a multiplicity of oligonucleotides that serve as surrogate targets. One of these platforms employs DNA or RNA polymerases for the amplification step, while detection is by fluorescence. The other is based on gold nanoparticles for both amplification as well as detection. The latter technology, now termed Biobarcode, is completely enzyme-free and offers potentially much higher multiplexing power.

Surface elastic wave detectors

NASA Technical Reports Server (NTRS)

Lawson, R. L.

1971-01-01

The potential applications of acoustic surface wave technology to multiplex communication systems such as data-bus, are examined. The goals are primarily to characterize certain aspects of surface wave trapped delay lines, surface wave modulation techniques, and surface wave applications that are relevant to the evaluation of surface wave devices in multiplex systems. The results indicate that there is a potential for the application of surface wave technology in data-bus type systems.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ding, H; Cho, H; Molloi, S

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 approximatelymore » 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.« less

Dual-excitation upconverting nanoparticle and quantum dot aptasensor for multiplexed food pathogen detection.

PubMed

Kurt, Hasan; Yüce, Meral; Hussain, Babar; Budak, Hikmet

2016-07-15

In this report, a dual-excitation sensing method was developed using aptamer-functionalized quantum dots and upconverting nanoparticles, exhibiting Stokes and anti-Stokes type excitation profiles, respectively. Conjugation of the aptamer-functionalized luminescent nanoparticles with the magnetic beads, comprising short DNA sequences that were partially complementary to the aptamer sequences, enabled facile separation of the analyte-free conjugates for fluorescent measurement. UV-Visible spectroscopy, Circular Dichroism spectroscopy, Dynamic Light Scattering and Polyacrylamide Gel Electrophoresis techniques were used to characterize the aptamer probes developed. The target-specific luminescent conjugates were applied for multiplex detection of model food pathogens, Salmonella typhimurium, and Staphylococcus aureus, in which the fluorescent emission spectra were obtained under UV excitation at 325nm for quantum dots and NIR excitation at 980nm for upconverting nanoparticles, respectively. The dual-excitation strategy was aimed to minimize cross-talk between the luminescent signals for multiplexed detection, and yielded limit of detection values of 16 and 28cfumL(-1) for Staphylococcus aureus, and Salmonella typhimurium, respectively. By employing a greater number of quantum dots and upconverting nanoparticles with non-overlapping fluorescent emissions, the proposed methodology might be exploited further to detect several analytes, simultaneously. Copyright © 2016 Elsevier B.V. All rights reserved.

A novel lab-on-chip platform with integrated solid phase PCR and Supercritical Angle Fluorescence (SAF) microlens array for highly sensitive and multiplexed pathogen detection.

PubMed

Hung, Tran Quang; Chin, Wai Hoe; Sun, Yi; Wolff, Anders; Bang, Dang Duong

2017-04-15

Solid-phase PCR (SP-PCR) has become increasingly popular for molecular diagnosis and there have been a few attempts to incorporate SP-PCR into lab-on-a-chip (LOC) devices. However, their applicability for on-line diagnosis is hindered by the lack of sensitive and portable on-chip optical detection technology. In this paper, we addressed this challenge by combining the SP-PCR with super critical angle fluorescence (SAF) microlens array embedded in a microchip. We fabricated miniaturized SAF microlens array as part of a microfluidic chamber in thermoplastic material and performed multiplexed SP-PCR directly on top of the SAF microlens array. Attribute to the high fluorescence collection efficiency of the SAF microlens array, the SP-PCR assay on the LOC platform demonstrated a high sensitivity of 1.6 copies/µL, comparable to off-chip detection using conventional laser scanner. The combination of SP-PCR and SAF microlens array allows for on-chip highly sensitive and multiplexed pathogen detection with low-cost and compact optical components. The LOC platform would be widely used as a high-throughput biosensor to analyze food, clinical and environmental samples. Copyright © 2016 Elsevier B.V. All rights reserved.

A nucleic acid strand displacement system for the multiplexed detection of tuberculosis-specific mRNA using quantum dots

NASA Astrophysics Data System (ADS)

Gliddon, H. D.; Howes, P. D.; Kaforou, M.; Levin, M.; Stevens, M. M.

2016-05-01

The development of rapid, robust and high performance point-of-care diagnostics relies on the advancement and combination of various areas of research. We have developed an assay for the detection of multiple mRNA molecules that combines DNA nanotechnology with fluorescent nanomaterials. The core switching mechanism is toehold-mediated strand displacement. We have used fluorescent quantum dots (QDs) as signal transducers in this assay, as they bring many benefits including bright fluorescence and multiplexing abilities. The resulting assay is capable of multiplexed detection of long RNA targets against a high concentration of background non-target RNA, with high sensitivity and specificity and limits of detection in the nanomolar range using only a standard laboratory plate reader. We demonstrate the utility of our QD-based system for the detection of two genes selected from a microarray-derived tuberculosis-specific gene expression signature. Levels of up- and downregulated gene transcripts comprising this signature can be combined to give a disease risk score, making the signature more amenable for use as a diagnostic marker. Our QD-based approach to detect these transcripts could pave the way for novel diagnostic assays for tuberculosis.The development of rapid, robust and high performance point-of-care diagnostics relies on the advancement and combination of various areas of research. We have developed an assay for the detection of multiple mRNA molecules that combines DNA nanotechnology with fluorescent nanomaterials. The core switching mechanism is toehold-mediated strand displacement. We have used fluorescent quantum dots (QDs) as signal transducers in this assay, as they bring many benefits including bright fluorescence and multiplexing abilities. The resulting assay is capable of multiplexed detection of long RNA targets against a high concentration of background non-target RNA, with high sensitivity and specificity and limits of detection in the nanomolar range using only a standard laboratory plate reader. We demonstrate the utility of our QD-based system for the detection of two genes selected from a microarray-derived tuberculosis-specific gene expression signature. Levels of up- and downregulated gene transcripts comprising this signature can be combined to give a disease risk score, making the signature more amenable for use as a diagnostic marker. Our QD-based approach to detect these transcripts could pave the way for novel diagnostic assays for tuberculosis. Electronic supplementary information (ESI) available: Base pair mismatch tuning of CProbes. Binding capacity of the QDs. Theoretical limit of detection (LOD) for the monoplex systems. Kinetics of strand displacement. Kinetics of QProbe-CProbe binding. LOD and saturation point calculations. See DOI: 10.1039/c6nr00484a

Laser-induced fluorescence imaging of subsurface tissue structures with a volume holographic spatial-spectral imaging system.

PubMed

Luo, Yuan; Gelsinger-Austin, Paul J; Watson, Jonathan M; Barbastathis, George; Barton, Jennifer K; Kostuk, Raymond K

2008-09-15

A three-dimensional imaging system incorporating multiplexed holographic gratings to visualize fluorescence tissue structures is presented. Holographic gratings formed in volume recording materials such as a phenanthrenquinone poly(methyl methacrylate) photopolymer have narrowband angular and spectral transmittance filtering properties that enable obtaining spatial-spectral information within an object. We demonstrate this imaging system's ability to obtain multiple depth-resolved fluorescence images simultaneously.

Bessel beam fluorescence lifetime tomography of live embryos (Conference Presentation)

NASA Astrophysics Data System (ADS)

Xu, Dongli; Peng, Leilei

2016-03-01

Optical tomography allows isotropic 3D imaging of embryos. Scanning-laser optical tomography (SLOT) has superior light collecting efficiency than wide-field optical tomography, making it ideal for fluorescence imaging of live embryos. We previously reported an imaging system that combines SLOT with a novel Fourier-multiplexed fluorescence lifetime imaging (FmFLIM) technique named FmFLIM-SLOT. FmFLIM-SLOT performs multiplexed FLIM-FRET readout of multiple FRET sensors in live embryos. Here we report a recent effort on improving the spatial resolution of the FmFLIM-SLOT system in order to image complex biochemical processes in live embryos at the cellular level. Optical tomography has to compromise between resolution and the depth of view. In SLOT, the commonly-used focused Gaussian beam diverges quickly from the focal plane, making it impossible to achieve high resolution imaging in a large volume specimen. We thus introduce Bessel beam laser-scanning tomography, which illuminates the sample with a spatial-light-modulator-generated Bessel beam that has an extended focal depth. The Bessel beam is scanned across the whole specimen. Fluorescence projection images are acquired at equal angular intervals as the sample rotates. Reconstruction artifacts due to annular-rings of the Bessel beam are removed by a modified 3D filtered back projection algorithm. Furthermore, in combination of Fourier-multiplexing fluorescence lifetime imaging (FmFLIM) method, the Bessel FmFLIM-SLOT system is capable of perform 3D lifetime imaging of live embryos at cellular resolution. The system is applied to in-vivo imaging of transgenic Zebrafish embryos. Results prove that Bessel FmFLIM-SLOT is a promising imaging method in development biology research.

Retrospective Species Identification of Microsporidian Spores in Diarrheic Fecal Samples from Human Immunodeficiency Virus/AIDS Patients by Multiplexed Fluorescence In Situ Hybridization▿

PubMed Central

Graczyk, Thaddeus K.; Johansson, Michael A.; Tamang, Leena; Visvesvara, Govinda S.; Moura, Laci S.; DaSilva, Alexandre J.; Girouard, Autumn S.; Matos, Olga

2007-01-01

In order to assess the applicability of multiplexed fluorescence in situ hybridization (FISH) assay for the clinical setting, we conducted retrospective analysis of 110 formalin-stored diarrheic stool samples from human immunodeficiency virus (HIV)/AIDS patients with intestinal microsporidiosis collected between 1992 and 2003. The multiplexed FISH assay identified microsporidian spores in 94 of 110 (85.5%) samples: 49 (52.1%) were positive for Enterocytozoon bieneusi, 43 (45.8%) were positive for Encephalitozoon intestinalis, 2 (2.1%) were positive for Encephalitozoon hellem, and 9 samples (9.6%) contained both E. bieneusi and E. intestinalis spores. Quantitative spore counts per ml of stool yielded concentration values from 3.5 × 103 to 4.4 × 105 for E. bieneusi (mean, 8.8 × 104/ml), 2.3 × 102 to 7.8 × 104 (mean, 1.5 × 104/ml) for E. intestinalis, and 1.8 × 102 to 3.6 × 102 for E. hellem (mean, 2.7 × 102/ml). Identification of microsporidian spores by multiplex FISH assay was more sensitive than both Chromotrope-2R and CalcoFluor White M2R stains; 85.5% versus 72.7 and 70.9%, respectively. The study demonstrated that microsporidian coinfection in HIV/AIDS patients with intestinal microsporidiosis is not uncommon and that formalin-stored fecal samples older than 10 years may not be suitable for retrospective analysis by techniques targeting rRNA. Multiplexed FISH assay is a reliable, quantitative fluorescence microscopy method for the simultaneous identification of E. bieneusi, E. intestinalis, and E. hellem, as well as Encephalitozoon cuniculi, spores in fecal samples and is a useful tool for assessing spore shedding intensity in intestinal microsporidiosis. The method can be used for epidemiological investigations and applied in clinical settings. PMID:17287331

A multiplexed fluorescent assay for independent second-messenger systems: decoding GPCR activation in living cells.

PubMed

Tewson, Paul H; Quinn, Anne Marie; Hughes, Thomas E

2013-08-01

There is a growing need in drug discovery and basic research to measure multiple second-messenger components of cell signaling pathways in real time and in relevant tissues and cell types. Many G-protein-coupled receptors activate the heterotrimeric protein, Gq, which in turn activates phospholipase C (PLC). PLC cleaves phosphatidylinositol 4,5-bisphosphate (PIP2) to produce two second messengers: diacylglycerol (DAG), which remains in the plasma membrane, and inositol triphosphate (IP3), which diffuses through the cytosol to release stores of intracellular calcium ions (Ca(2+)). Our goal was to create a series of multiplex sensors that would make it possible to simultaneously measure two different components of the Gq pathway in living cells. Here we describe new fluorescent sensors for DAG and PIP2 that produce robust changes in green or red fluorescence and can be combined with one another, or with existing Ca(2+) sensors, in a live-cell assay. These assays can detect multiple components of Gq signaling, simultaneously in real time, on standard fluorescent plate readers or live-cell imaging systems.

Photon-Counting H33D Detector for Biological Fluorescence Imaging

PubMed Central

Michalet, X.; Siegmund, O.H.W.; Vallerga, J.V.; Jelinsky, P.; Millaud, J.E.; Weiss, S.

2010-01-01

We have developed a photon-counting High-temporal and High-spatial resolution, High-throughput 3-Dimensional detector (H33D) for biological imaging of fluorescent samples. The design is based on a 25 mm diameter S20 photocathode followed by a 3-microchannel plate stack, and a cross delay line anode. We describe the bench performance of the H33D detector, as well as preliminary imaging results obtained with fluorescent beads, quantum dots and live cells and discuss applications of future generation detectors for single-molecule imaging and high-throughput study of biomolecular interactions. PMID:20151021

Specification and Design of the SBRC-190: A Cryogenic Multiplexer for Far Infrared Photoconductor Detectors

NASA Technical Reports Server (NTRS)

Erickson, E. F.; Young, E. T.; Wolf, J.; Asbrock, J. F.; Lum, N.; DeVincenzi, D. (Technical Monitor)

2002-01-01

Arrays of far-infrared photoconductor detectors operate at a few degrees Kelvin and require electronic amplifiers in close proximity. For the electronics, a cryogenic multiplexer is ideal to avoid the large number of wires associated with individual amplifiers for each pixel, and to avoid adverse effects of thermal and radiative heat loads from the circuitry. For low background applications, the 32 channel CRC 696 CMOS device was previously developed for SIRTF, the cryogenic Space Infrared Telescope Facility. For higher background applications, we have developed a similar circuit, featuring several modifications: (a) an AC coupled, capacitive feedback transimpedence unit cell, to minimize input offset effects, thereby enabling low detector biases, (b) selectable feedback capacitors to enable operation over a wide range of backgrounds, and (c) clamp and sample & hold output circuits to improve sampling efficiency, which is a concern at the high readout rates required. We describe the requirements for and design of the new device.

Method and apparatus for signal processing in a sensor system for use in spectroscopy

DOEpatents

O'Connor, Paul [Bellport, NY; DeGeronimo, Gianluigi [Nesconset, NY; Grosholz, Joseph [Natrona Heights, PA

2008-05-27

A method for processing pulses arriving randomly in time on at least one channel using multiple peak detectors includes asynchronously selecting a non-busy peak detector (PD) in response to a pulse-generated trigger signal, connecting the channel to the selected PD in response to the trigger signal, and detecting a pulse peak amplitude. Amplitude and time of arrival data are output in first-in first-out (FIFO) sequence. An apparatus includes trigger comparators to generate the trigger signal for the pulse-receiving channel, PDs, a switch for connecting the channel to the selected PD, and logic circuitry which maintains the write pointer. Also included, time-to-amplitude converters (TACs) convert time of arrival to analog voltage and an analog multiplexer provides FIFO output. A multi-element sensor system for spectroscopy includes detector elements, channels, trigger comparators, PDs, a switch, and a logic circuit with asynchronous write pointer. The system includes TACs, a multiplexer and analog-to-digital converter.

Multiplexed single-mode wavelength-to-time mapping of multimode light

PubMed Central

Chandrasekharan, Harikumar K; Izdebski, Frauke; Gris-Sánchez, Itandehui; Krstajić, Nikola; Walker, Richard; Bridle, Helen L.; Dalgarno, Paul A.; MacPherson, William N.; Henderson, Robert K.; Birks, Tim A.; Thomson, Robert R.

2017-01-01

When an optical pulse propagates along an optical fibre, different wavelengths travel at different group velocities. As a result, wavelength information is converted into arrival-time information, a process known as wavelength-to-time mapping. This phenomenon is most cleanly observed using a single-mode fibre transmission line, where spatial mode dispersion is not present, but the use of such fibres restricts possible applications. Here we demonstrate that photonic lanterns based on tapered single-mode multicore fibres provide an efficient way to couple multimode light to an array of single-photon avalanche detectors, each of which has its own time-to-digital converter for time-correlated single-photon counting. Exploiting this capability, we demonstrate the multiplexed single-mode wavelength-to-time mapping of multimode light using a multicore fibre photonic lantern with 121 single-mode cores, coupled to 121 detectors on a 32 × 32 detector array. This work paves the way to efficient multimode wavelength-to-time mapping systems with the spectral performance of single-mode systems. PMID:28120822

Frequency division multiplexed readout of TES detectors with baseband feedback

NASA Astrophysics Data System (ADS)

den Hartog, R.; Audley, M. D.; Beyer, J.; Bruijn, M. P.; de Korte, P.; Gottardi, L.; Hijmering, R.; Jackson, B.; Nieuwenhuizen, A.; van der Kuur, J.; van Leeuwen, B.-J.; Van Loon, D.

2012-09-01

SRON is developing an electronic system for the multiplexed read-out of an array of transition edge sensors (TES) by combining the techniques of frequency domain multiplexing (FDM) with base-band feedback (BBFB). The astronomical applications are the read-out of soft X-ray microcalorimeters and the far-infrared bolometers for the SAFARI instrument on the Japanese mission SPICA. In this paper we derive the requirements for the read-out system regarding noise and dynamic range in the context of the SAFARI instrument, and demonstrate that the current experimental prototype is capable of simultaneously locking 57 channels and complies with these requirements.

Low-picomolar limits of detection using high-power light-emitting diodes for fluorescence.

PubMed

de Jong, Ebbing P; Lucy, Charles A

2006-05-01

Fluorescence detectors are ever more frequently being used with light-emitting diodes (LEDs) as the light source. Technological advances in the solid-state lighting industry have produced LEDs which are also suitable tools in analytical measurements. LEDs are now available which deliver 700 mW of radiometric power. While this greater light power can increase the fluorescence signal, it is not trivial to make proper use of this light. This new generation of LEDs has a large emitting area and a highly divergent beam. This presents a classic problem in optics where one must choose between either a small focused light spot, or high light collection efficiency. We have selected for light collection efficiency, which yields a light spot somewhat larger than the emitting area of the LED. This light is focused onto a flow cell. Increasing the detector cell internal diameter (i.d.) produces gains in (sensitivity)3. However, since the detector cell i.d. is smaller than the LED spot size, scattering of excitation light towards the detector remains a significant source of background signal. This can be minimized through the use of spectral filters and spatial filters in the form of pinholes. The detector produced a limit of detection (LOD) of 3 pM, which is roughly three orders of magnitude lower than other reports of LED-based fluorescence detectors. Furthermore, this LOD comes within a factor of six of much more expensive laser-based fluorescence systems. This detector has been used to monitor a separation from a gel filtration column of fluorescently labeled BSA from residual labeling reagent. The LOD of fluorescently labeled BSA is 25 pM.

On-chip multiplexed solid-phase nucleic acid hybridization assay using spatial profiles of immobilized quantum dots and fluorescence resonance energy transfer.

PubMed

Noor, M Omair; Tavares, Anthony J; Krull, Ulrich J

2013-07-25

A microfluidic based solid-phase assay for the multiplexed detection of nucleic acid hybridization using quantum dot (QD) mediated fluorescence resonance energy transfer (FRET) is described herein. The glass surface of hybrid glass-polydimethylsiloxane (PDMS) microfluidic channels was chemically modified to assemble the biorecognition interface. Multiplexing was demonstrated using a detection system that was comprised of two colors of immobilized semi-conductor QDs and two different oligonucleotide probe sequences. Green-emitting and red-emitting QDs were paired with Cy3 and Alexa Fluor 647 (A647) labeled oligonucleotides, respectively. The QDs served as energy donors for the transduction of dye labeled oligonucleotide targets. The in-channel assembly of the biorecognition interface and the subsequent introduction of oligonucleotide targets was accomplished within minutes using a combination of electroosmotic flow and electrophoretic force. The concurrent quantification of femtomole quantities of two target sequences was possible by measuring the spatial coverage of FRET sensitized emission along the length of the channel. In previous reports, multiplexed QD-FRET hybridization assays that employed a ratiometric method for quantification had challenges associated with lower analytical sensitivity arising from both donor and acceptor dilution that resulted in reduced energy transfer pathways as compared to single-color hybridization assays. Herein, a spatial method for quantification that is based on in-channel QD-FRET profiles provided higher analytical sensitivity in the multiplexed assay format as compared to single-color hybridization assays. The selectivity of the multiplexed hybridization assays was demonstrated by discrimination between a fully-complementary sequence and a 3 base pair sequence at a contrast ratio of 8 to 1. Copyright © 2013 Elsevier B.V. All rights reserved.

Species-specific differentiation of variola, monkeypox, and varicella-zoster viruses by multiplex real-time PCR assay.

PubMed

Maksyutov, Rinat A; Gavrilova, Elena V; Shchelkunov, Sergei N

2016-10-01

A method of one-stage rapid detection and differentiation of epidemiologically important variola virus (VARV), monkeypox virus (MPXV), and varicella-zoster virus (VZV) utilizing multiplex real-time TaqMan PCR assay was developed. Four hybridization probes with various fluorescent dyes and the corresponding fluorescence quenchers were simultaneously used for the assay. The hybridization probes specific for the VARV sequence contained FAM/BHQ1 as a dye/quencher pair; MPXV-specific, JOE/BHQ1; VZV-specific, TAMRA/BHQ2; and internal control-specific, Cy5/BHQ3. The specificity and sensitivity of the developed method were assessed by analyzing DNA of 32 strains belonging to orthopoxvirus and herpesvirus species. Copyright © 2016 Elsevier B.V. All rights reserved.

Simultaneous laser-induced fluorescence and scattering detection of individual particles separated by capillary electrophoresis.

PubMed

Andreyev, Dmitry; Arriaga, Edgar A

2007-07-15

This technical note describes a detector capable of simultaneously monitoring scattering and fluorescence signals of individual particles separated by capillary electrophoresis. Due to its nonselective nature, scattering alone is not sufficient to identify analyte particles. However, when the analyte particles are fluorescent, the detector described here is able to identify simultaneously occurring scattering and fluorescent signals, even when contaminating particles (i.e., nonfluorescent) are present. Both fluorescent polystyrene particles and 10-nonyl acridine orange (NAO)-labeled mitochondria were used as models. Fluorescence versus scattering (FVS) plots made it possible to identify two types of particles and a contaminant in a mixture of polystyrene particles. We also analyzed NAO-labeled mitochondria before and after cryogenic storage; the mitochondria FVS plots changed with storage, which suggests that the detector reported here is suitable for monitoring subtle changes in mitochondrial morphology that would not be revealed by monitoring only fluorescence or scattering signals.

Dual-detector X-ray fluorescence imaging of ancient artifacts with surface relief

PubMed Central

Smilgies, Detlef-M.; Powers, Judson A.; Bilderback, Donald H.; Thorne, Robert E.

2012-01-01

Interpretation of X-ray fluorescence images of archeological artifacts is complicated by the presence of surface relief and roughness. Using two symmetrically arranged fluorescence detectors in a back-reflection geometry, the proper X-ray fluorescence yield can be distinguished from intensity variations caused by surface topography. This technique has been applied to the study of Roman inscriptions on marble. PMID:22713888

Superconducting Detector Arrays for Astrophysics

NASA Technical Reports Server (NTRS)

Chervenak, James

2008-01-01

The next generation of astrophysics instruments will feature an order of magnitude more photon sensors or sensors that have an order of magnitude greater sensitivity. Since detector noise scales with temperature, a number of candidate technologies have been developed that use the intrinsic advantages of detector systems that operate below 1 Kelvin. Many of these systems employ of the superconducting phenomena that occur in metals at these temperatures to build ultrasensitive detectors and low-noise, low-power readout architectures. I will present one such system in use today to meet the needs of the astrophysics community at millimeter and x-ray wavelengths. Our group at NASA in collaboration with Princeton, NIST, Boulder and a number of other groups is building large format arrays of superconducting transition edge sensors (TES) read out with multiplexed superconducting quantum interference devices (SQUID). I will present the high sensitivity we have achieved in multiplexed x-ray sensors with the TES technology and describe the construction of a 1000-sensor TES/SQUID array for microwave measurements. With our collaboration's deployment of a kilopixel TES array for 2 mm radiation at the Atacarna Cosmology Telescope in November 2007, we have first images of the lensed Cosmic Microwave Background at fine angular scales.

Fabricating a UV-Vis and Raman Spectroscopy Immunoassay Platform.

PubMed

Hanson, Cynthia; Israelsen, Nathan D; Sieverts, Michael; Vargis, Elizabeth

2016-11-10

Immunoassays are used to detect proteins based on the presence of associated antibodies. Because of their extensive use in research and clinical settings, a large infrastructure of immunoassay instruments and materials can be found. For example, 96- and 384-well polystyrene plates are available commercially and have a standard design to accommodate ultraviolet-visible (UV-Vis) spectroscopy machines from various manufacturers. In addition, a wide variety of immunoglobulins, detection tags, and blocking agents for customized immunoassay designs such as enzyme-linked immunosorbent assays (ELISA) are available. Despite the existing infrastructure, standard ELISA kits do not meet all research needs, requiring individualized immunoassay development, which can be expensive and time-consuming. For example, ELISA kits have low multiplexing (detection of more than one analyte at a time) capabilities as they usually depend on fluorescence or colorimetric methods for detection. Colorimetric and fluorescent-based analyses have limited multiplexing capabilities due to broad spectral peaks. In contrast, Raman spectroscopy-based methods have a much greater capability for multiplexing due to narrow emission peaks. Another advantage of Raman spectroscopy is that Raman reporters experience significantly less photobleaching than fluorescent tags 1 . Despite the advantages that Raman reporters have over fluorescent and colorimetric tags, protocols to fabricate Raman-based immunoassays are limited. The purpose of this paper is to provide a protocol to prepare functionalized probes to use in conjunction with polystyrene plates for direct detection of analytes by UV-Vis analysis and Raman spectroscopy. This protocol will allow researchers to take a do-it-yourself approach for future multi-analyte detection while capitalizing on pre-established infrastructure.

A fully sealed plastic chip for multiplex PCR and its application in bacteria identification.

PubMed

Xu, Youchun; Yan, He; Zhang, Yan; Jiang, Kewei; Lu, Ying; Ren, Yonghong; Wang, Hui; Wang, Shan; Xing, Wanli

2015-07-07

Multiplex PCR is an effective tool for simultaneous multiple target detection but is limited by the intrinsic interference and competition among primer pairs when it is performed in one reaction tube. Dividing a multiplex PCR into many single PCRs is a simple strategy to overcome this issue. Here, we constructed a plastic, easy-to-use, fully sealed multiplex PCR chip based on reversible centrifugation for the simultaneous detection of 63 target DNA sequences. The structure of the chip is quite simple, which contains sine-shaped infusing channels and a number of reaction chambers connecting to one side of these channels. Primer pairs for multiplex PCR were sequentially preloaded in the different reaction chambers, and the chip was enclosed with PCR-compatible adhesive tape. For usage, the PCR master mix containing a DNA template is pipetted into the infusing channels and centrifuged into the reaction chambers, leaving the infusing channels filled with air to avoid cross-contamination of the different chambers. Then, the chip is sealed and placed on a flat thermal cycler for PCR. Finally, amplification products can be detected in situ using a fluorescence scanner or recovered by reverse centrifugation for further analyses. Therefore, our chip possesses two functions: 1) it can be used for multi-target detection based on end-point in situ fluorescence detection; and 2) it can work as a sample preparation unit for analyses that need multiplex PCR such as hybridization and target sequencing. The performance of this chip was carefully examined and further illustrated in the identification of 8 pathogenic bacterial genomic DNA samples and 13 drug-resistance genes. Due to simplicity of its structure and operation, accuracy and generality, high-throughput capacity, and versatile functions (i.e., for in situ detection and sample preparation), our multiplex PCR chip has great potential in clinical diagnostics and nucleic acid-based point-of-care testing.

A single-electrode electrochemical system for multiplex electrochemiluminescence analysis based on a resistance induced potential difference.

PubMed

Gao, Wenyue; Muzyka, Kateryna; Ma, Xiangui; Lou, Baohua; Xu, Guobao

2018-04-28

Developing low-cost and simple electrochemical systems is becoming increasingly important but still challenged for multiplex experiments. Here we report a single-electrode electrochemical system (SEES) using only one electrode not only for a single experiment but also for multiplex experiments based on a resistance induced potential difference. SEESs for a single experiment and multiplex experiments are fabricated by attaching a self-adhesive label with a hole and multiple holes onto an ITO electrode, respectively. This enables multiplex electrochemiluminescence analysis with high sensitivity at a very low safe voltage using a smartphone as a detector. For the multiplex analysis, the SEES using a single electrode is much simpler, cheaper and more user-friendly than conventional electrochemical systems and bipolar electrochemical systems using electrode arrays. Moreover, SEESs are free from the electrochemiluminescent background problem from driving electrodes in bipolar electrochemical systems. Since numerous electrodes and cover materials can be used to fabricate SEESs readily and electrochemistry is being extensively used, SEESs are very promising for broad applications, such as drug screening and high throughput analysis.

Image formation of volume holographic microscopy using point spread functions

NASA Astrophysics Data System (ADS)

Luo, Yuan; Oh, Se Baek; Kou, Shan Shan; Lee, Justin; Sheppard, Colin J. R.; Barbastathis, George

2010-04-01

We present a theoretical formulation to quantify the imaging properties of volume holographic microscopy (VHM). Volume holograms are formed by exposure of a photosensitive recording material to the interference of two mutually coherent optical fields. Recently, it has been shown that a volume holographic pupil has spatial and spectral sectioning capability for fluorescent samples. Here, we analyze the point spread function (PSF) to assess the imaging behavior of the VHM with a point source and detector. The coherent PSF of the VHM is derived, and the results are compared with those from conventional microscopy, and confocal microscopy with point and slit apertures. According to our analysis, the PSF of the VHM can be controlled in the lateral direction by adjusting the parameters of the VH. Compared with confocal microscopes, the performance of the VHM is comparable or even potentially better, and the VHM is also able to achieve real-time and three-dimensional (3D) imaging due to its multiplexing ability.

Ultra-high density optical data storage in common transparent plastics.

PubMed

Kallepalli, Deepak L N; Alshehri, Ali M; Marquez, Daniela T; Andrzejewski, Lukasz; Scaiano, Juan C; Bhardwaj, Ravi

2016-05-25

The ever-increasing demand for high data storage capacity has spurred research on development of innovative technologies and new storage materials. Conventional GByte optical discs (DVDs and Bluray) can be transformed into ultrahigh capacity storage media by encoding multi-level and multiplexed information within the three dimensional volume of a recording medium. However, in most cases the recording medium had to be photosensitive requiring doping with photochromic molecules or nanoparticles in a multilayer stack or in the bulk material. Here, we show high-density data storage in commonly available plastics without any special material preparation. A pulsed laser was used to record data in micron-sized modified regions. Upon excitation by the read laser, each modified region emits fluorescence whose intensity represents 32 grey levels corresponding to 5 bits. We demonstrate up to 20 layers of embedded data. Adjusting the read laser power and detector sensitivity storage capacities up to 0.2 TBytes can be achieved in a standard 120 mm disc.

Fluorescent Quantification of DNA Based on Core-Shell Fe3O4@SiO2@Au Nanocomposites and Multiplex Ligation-Dependent Probe Amplification.

PubMed

Fan, Jing; Yang, Haowen; Liu, Ming; Wu, Dan; Jiang, Hongrong; Zeng, Xin; Elingarami, Sauli; Ll, Zhiyang; Li, Song; Liu, Hongna; He, Nongyue

2015-02-01

In this research, a novel method for relative fluorescent quantification of DNA based on Fe3O4@SiO2@Au gold-coated magnetic nanocomposites (GMNPs) and multiplex ligation- dependent probe amplification (MLPA) has been developed. With the help of self-assembly, seed-mediated growth and chemical reduction method, core-shell Fe3O4@SiO2@Au GMNPs were synthesized. Through modified streptavidin on the GMNPs surface, we obtained a bead chip which can capture the biotinylated probes. Then we designed MLPA probes which were tagged with biotin or Cy3 and target DNA on the basis of human APP gene sequence. The products from the thermostable DNA ligase induced ligation reactions and PCR amplifications were incubated with SA-GMNPs. After washing, magnetic separation, spotting, the fluorescent scanning results showed our method can be used for the relative quantitative analysis of the target DNA in the concentration range of 03004~0.5 µM.

Real-time, multiplexed electrochemical DNA detection using an active complementary metal-oxide-semiconductor biosensor array with integrated sensor electronics.

PubMed

Levine, Peter M; Gong, Ping; Levicky, Rastislav; Shepard, Kenneth L

2009-03-15

Optical biosensing based on fluorescence detection has arguably become the standard technique for quantifying extents of hybridization between surface-immobilized probes and fluorophore-labeled analyte targets in DNA microarrays. However, electrochemical detection techniques are emerging which could eliminate the need for physically bulky optical instrumentation, enabling the design of portable devices for point-of-care applications. Unlike fluorescence detection, which can function well using a passive substrate (one without integrated electronics), multiplexed electrochemical detection requires an electronically active substrate to analyze each array site and benefits from the addition of integrated electronic instrumentation to further reduce platform size and eliminate the electromagnetic interference that can result from bringing non-amplified signals off chip. We report on an active electrochemical biosensor array, constructed with a standard complementary metal-oxide-semiconductor (CMOS) technology, to perform quantitative DNA hybridization detection on chip using targets conjugated with ferrocene redox labels. A 4 x 4 array of gold working electrodes and integrated potentiostat electronics, consisting of control amplifiers and current-input analog-to-digital converters, on a custom-designed 5 mm x 3 mm CMOS chip drive redox reactions using cyclic voltammetry, sense DNA binding, and transmit digital data off chip for analysis. We demonstrate multiplexed and specific detection of DNA targets as well as real-time monitoring of hybridization, a task that is difficult, if not impossible, with traditional fluorescence-based microarrays.

Multiplexed phase-space imaging for 3D fluorescence microscopy.

PubMed

Liu, Hsiou-Yuan; Zhong, Jingshan; Waller, Laura

2017-06-26

Optical phase-space functions describe spatial and angular information simultaneously; examples of optical phase-space functions include light fields in ray optics and Wigner functions in wave optics. Measurement of phase-space enables digital refocusing, aberration removal and 3D reconstruction. High-resolution capture of 4D phase-space datasets is, however, challenging. Previous scanning approaches are slow, light inefficient and do not achieve diffraction-limited resolution. Here, we propose a multiplexed method that solves these problems. We use a spatial light modulator (SLM) in the pupil plane of a microscope in order to sequentially pattern multiplexed coded apertures while capturing images in real space. Then, we reconstruct the 3D fluorescence distribution of our sample by solving an inverse problem via regularized least squares with a proximal accelerated gradient descent solver. We experimentally reconstruct a 101 Megavoxel 3D volume (1010×510×500µm with NA 0.4), demonstrating improved acquisition time, light throughput and resolution compared to scanning aperture methods. Our flexible patterning scheme further allows sparsity in the sample to be exploited for reduced data capture.

A bright cyan-excitable orange fluorescent protein facilitates dual-emission microscopy and enhances bioluminescence imaging in vivo

PubMed Central

Chu, Jun; Oh, Young-Hee; Sens, Alex; Ataie, Niloufar; Dana, Hod; Macklin, John J.; Laviv, Tal; Welf, Erik S.; Dean, Kevin M.; Zhang, Feijie; Kim, Benjamin B.; Tang, Clement Tran; Hu, Michelle; Baird, Michelle A.; Davidson, Michael W.; Kay, Mark A.; Fiolka, Reto; Yasuda, Ryohei; Kim, Douglas S.; Ng, Ho-Leung; Lin, Michael Z.

2016-01-01

Orange-red fluorescent proteins (FPs) are widely used in biomedical research for multiplexed epifluorescence microscopy with GFP-based probes, but their different excitation requirements make multiplexing with new advanced microscopy methods difficult. Separately, orange-red FPs are useful for deep-tissue imaging in mammals due to the relative tissue transmissibility of orange-red light, but their dependence on illumination limits their sensitivity as reporters in deep tissues. Here we describe CyOFP1, a bright engineered orange-red FP that is excitable by cyan light. We show that CyOFP1 enables single-excitation multiplexed imaging with GFP-based probes in single-photon and two-photon microscopy, including time-lapse imaging in light-sheet systems. CyOFP1 also serves as an efficient acceptor for resonance energy transfer from the highly catalytic blue-emitting luciferase NanoLuc. An optimized fusion of CyOFP1 and NanoLuc, called Antares, functions as a highly sensitive bioluminescent reporter in vivo, producing substantially brighter signals from deep tissues than firefly luciferase and other bioluminescent proteins. PMID:27240196

Multicolor-based discrimination of 21 short tandem repeats and amelogenin using four fluorescent universal primers.

PubMed

Asari, Masaru; Okuda, Katsuhiro; Hoshina, Chisato; Omura, Tomohiro; Tasaki, Yoshikazu; Shiono, Hiroshi; Matsubara, Kazuo; Shimizu, Keiko

2016-02-01

The aim of this study was to develop a cost-effective genotyping method using high-quality DNA for human identification. A total of 21 short tandem repeats (STRs) and amelogenin were selected, and fluorescent fragments at 22 loci were simultaneously amplified in a single-tube reaction using locus-specific primers with 24-base universal tails and four fluorescent universal primers. Several nucleotide substitutions in universal tails and fluorescent universal primers enabled the detection of specific fluorescent fragments from the 22 loci. Multiplex polymerase chain reaction (PCR) produced intense FAM-, VIC-, NED-, and PET-labeled fragments ranging from 90 to 400 bp, and these fragments were discriminated using standard capillary electrophoretic analysis. The selected 22 loci were also analyzed using two commercial kits (the AmpFLSTR Identifiler Kit and the PowerPlex ESX 17 System), and results for two loci (D19S433 and D16S539) were discordant between these kits due to mutations at the primer binding sites. All genotypes from the 100 samples were determined using 2.5 ng of DNA by our method, and the expected alleles were completely recovered. Multiplex 22-locus genotyping using four fluorescent universal primers effectively reduces the costs to less than 20% of genotyping using commercial kits, and our method would be useful to detect silent alleles from commercial kit analysis. Copyright © 2015 Elsevier Inc. All rights reserved.

Hybrid charge division multiplexing method for silicon photomultiplier based PET detectors

NASA Astrophysics Data System (ADS)

Park, Haewook; Ko, Guen Bae; Lee, Jae Sung

2017-06-01

Silicon photomultiplier (SiPM) is widely utilized in various positron emission tomography (PET) detectors and systems. However, the individual recording of SiPM output signals is still challenging owing to the high granularity of the SiPM; thus, charge division multiplexing is commonly used in PET detectors. Resistive charge division method is well established for reducing the number of output channels in conventional multi-channel photosensors, but it degrades the timing performance of SiPM-based PET detectors by yielding a large resistor-capacitor (RC) constant. Capacitive charge division method, on the other hand, yields a small RC constant and provides a faster timing response than the resistive method, but it suffers from an output signal undershoot. Therefore, in this study, we propose a hybrid charge division method which can be implemented by cascading the parallel combination of a resistor and a capacitor throughout the multiplexing network. In order to compare the performance of the proposed method with the conventional methods, a 16-channel Hamamatsu SiPM (S11064-050P) was coupled with a 4  ×  4 LGSO crystal block (3  ×  3  ×  20 mm3) and a 9  ×  9 LYSO crystal block (1.2  ×  1.2  ×  10 mm3). In addition, we tested a time-over-threshold (TOT) readout using the digitized position signals to further demonstrate the feasibility of the time-based readout of multiplexed signals based on the proposed method. The results indicated that the proposed method exhibited good energy and timing performance, thus inheriting only the advantages of conventional resistive and capacitive methods. Moreover, the proposed method showed excellent pulse shape uniformity that does not depend on the position of the interacted crystal. Accordingly, we can conclude that the hybrid charge division method is useful for effectively reducing the number of output channels of the SiPM array.

21 CFR 872.1745 - Laser fluorescence caries detection device.

Code of Federal Regulations, 2013 CFR

2013-04-01

... fluorescence detector housed in a dental handpiece, and a control console that performs device calibration, as...) Properly sterilize the emitter-detector handpick before each use, and (iv) Properly maintain and handle the...

21 CFR 872.1745 - Laser fluorescence caries detection device.

Code of Federal Regulations, 2012 CFR

2012-04-01

... fluorescence detector housed in a dental handpiece, and a control console that performs device calibration, as...) Properly sterilize the emitter-detector handpick before each use, and (iv) Properly maintain and handle the...

Enhanced speed in fluorescence imaging using beat frequency multiplexing

NASA Astrophysics Data System (ADS)

Mikami, Hideharu; Kobayashi, Hirofumi; Wang, Yisen; Hamad, Syed; Ozeki, Yasuyuki; Goda, Keisuke

2016-03-01

Fluorescence imaging using radiofrequency-tagged emission (FIRE) is an emerging technique that enables higher imaging speed (namely, temporal resolution) in fluorescence microscopy compared to conventional fluorescence imaging techniques such as confocal microscopy and wide-field microscopy. It works based on the principle that it uses multiple intensity-modulated fields in an interferometric setup as excitation fields and applies frequency-division multiplexing to fluorescence signals. Unfortunately, despite its high potential, FIRE has limited imaging speed due to two practical limitations: signal bandwidth and signal detection efficiency. The signal bandwidth is limited by that of an acousto-optic deflector (AOD) employed in the setup, which is typically 100-200 MHz for the spectral range of fluorescence excitation (400-600 nm). The signal detection efficiency is limited by poor spatial mode-matching between two interfering fields to produce a modulated excitation field. Here we present a method to overcome these limitations and thus to achieve higher imaging speed than the prior version of FIRE. Our method achieves an increase in signal bandwidth by a factor of two and nearly optimal mode matching, which enables the imaging speed limited by the lifetime of the target fluorophore rather than the imaging system itself. The higher bandwidth and better signal detection efficiency work synergistically because higher bandwidth requires higher signal levels to avoid the contribution of shot noise and amplifier noise to the fluorescence signal. Due to its unprecedentedly high-speed performance, our method has a wide variety of applications in cancer detection, drug discovery, and regenerative medicine.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cho, H; Ding, H; Ziemer, B

Purpose: To investigate the feasibility of energy calibration and energy response characterization of a photon counting detector using x-ray fluorescence. Methods: A comprehensive Monte Carlo simulation study was done to investigate the influence of various geometric components on the x-ray fluorescence measurement. Different materials, sizes, and detection angles were simulated using Geant4 Application for Tomographic Emission (GATE) Monte Carlo package. Simulations were conducted using 100 kVp tungsten-anode spectra with 2 mm Al filter for a single pixel cadmium telluride (CdTe) detector with 3 × 3 mm2 in detection area. The fluorescence material was placed 300 mm away from both themore » x-ray source and the detector. For angular dependence measurement, the distance was decreased to 30 mm to reduce the simulation time. Compound materials, containing silver, barium, gadolinium, hafnium, and gold in cylindrical shape, were simulated. The object size varied from 5 to 100 mm in diameter. The angular dependence of fluorescence and scatter were simulated from 20° to 170° with an incremental step of 10° to optimize the fluorescence to scatter ratio. Furthermore, the angular dependence was also experimentally measured using a spectrometer (X-123CdTe, Amptek Inc., MA) to validate the simulation results. Results: The detection angle between 120° to 160° resulted in more optimal x-ray fluorescence to scatter ratio. At a detection angle of 120°, the object size did not have a significant effect on the fluorescence to scatter ratio. The experimental results of fluorescence angular dependence are in good agreement with the simulation results. The Kα and Kβ peaks of five materials could be identified. Conclusion: The simulation results show that the x-ray fluorescence procedure has the potential to be used for detector energy calibration and detector response characteristics by using the optimal system geometry.« less

Multiplexed DNA detection using spectrally encoded porous SiO2 photonic crystal particles.

PubMed

Meade, Shawn O; Chen, Michelle Y; Sailor, Michael J; Miskelly, Gordon M

2009-04-01

A particle-based multiplexed DNA assay based on encoded porous SiO(2) photonic crystal disks is demonstrated. A "spectral barcode" is generated by electrochemical etch of a single-crystal silicon wafer using a programmed current-time waveform. A lithographic procedure is used to isolate cylindrical microparticles 25 microm in diameter and 10 microm thick, which are then oxidized, modified with a silane linker, and conjugated to various amino-functionalized oligonucleotide probes via cyanuric chloride. It is shown that the particles can be decoded based on their reflectivity spectra and that a multiple analyte assay can be performed in a single sample with a modified fluorescence microscope. The homogeneity of the reflectivity and fluorescence spectra, both within and across the microparticles, is also reported.

Recent advances in superconducting nanowire single photon detectors for single-photon imaging

NASA Astrophysics Data System (ADS)

Verma, V. B.; Allman, M. S.; Stevens, M.; Gerrits, T.; Horansky, R. D.; Lita, A. E.; Marsili, F.; Beyer, A.; Shaw, M. D.; Stern, J. A.; Mirin, R. P.; Nam, S. W.

2016-05-01

We demonstrate a 64-pixel free-space-coupled array of superconducting nanowire single photon detectors optimized for high detection efficiency in the near-infrared range. An integrated, readily scalable, multiplexed readout scheme is employed to reduce the number of readout lines to 16. The cryogenic, optical, and electronic packaging to read out the array, as well as characterization measurements are discussed.

Fluorescence lifetime microscopy with a time- and space-resolved single-photon counting detector

PubMed Central

Michalet, X.; Siegmund, O.H.W.; Vallerga, J.V.; Jelinsky, P.; Pinaud, F. F.; Millaud, J.E.; Weiss, S.

2017-01-01

We have recently developed a wide-field photon-counting detector (the H33D detector) having high-temporal and high-spatial resolutions and capable of recording up to 500,000 photons per sec. Its temporal performance has been previously characterized using solutions of fluorescent materials with different lifetimes, and its spatial resolution using sub-diffraction objects (beads and quantum dots). Here we show its application to fluorescence lifetime imaging of live cells and compare its performance to a scanning confocal TCSPC approach. With the expected improvements in photocathode sensitivity and increase in detector throughput, this technology appears as a promising alternative to the current lifetime imaging solutions. PMID:29449756

Integrated infrared detector arrays for low-background astronomy

NASA Technical Reports Server (NTRS)

Mccreight, C. R.

1979-01-01

Existing integrated infrared detector array technology is being evaluated under low-background conditions to determine its applicability in orbiting astronomical applications where extended integration times and photometric accuracy are of interest. Preliminary performance results of a 1 x 20 elements InSb CCD array under simulated astronomical conditions are presented. Using the findings of these tests, improved linear- and area-array technology will be developed for use in NASA programs such as the Shuttle Infrared Telescope Facility. For wavelengths less than 30 microns, extrinsic silicon and intrinsic arrays with CCD readout will be evaluated and improved as required, while multiplexed arrays of Ge:Ga for wavelengths in the range 30 to 120 microns will be developed as fundamental understanding of this material improves. Future efforts will include development of improved drive and readout circuitry, and consideration of alternate multiplexing schemes.

Single line-of-sight dual energy backlighter for mix width experiments

DOE Office of Scientific and Technical Information (OSTI.GOV)

Baker, K. L., E-mail: baker7@llnl.gov; Glendinning, S. G.; Martinez, D.

2014-11-15

We present a diagnostic technique used to spatially multiplex two x-ray radiographs of an object onto a detector along a single line-of-sight. This technique uses a thin, <2 μm, cosputtered backlighter target to simultaneously produce both Ni and Zn He{sub α} emission. A Ni picket fence filter, 500 μm wide bars and troughs, is then placed in front of the detector to pass only the Ni He{sub α} emission in the bar region and both energies in the trough region thereby spatially multiplexing the two radiographs on a single image. Initial experimental results testing the backlighter spectrum are presented alongmore » with simulated images showing the calculated radiographic images though the nickel picket fence filter which are used to measure the mix width in an accelerated nickel foam.« less

Development of a Massive, Highly Multiplexible, Phonon-Mediated Particle Detector Using Kinetic Inductance Detectors

NASA Astrophysics Data System (ADS)

Chang, Y.-Y.; Cornell, B.; Aralis, T.; Bumble, B.; Golwala, S. R.

2018-04-01

We present a status update on the development of a phonon-mediated particle detector using kinetic inductance detector (KID). The design is intended for O(1) kg substrate, using O(102) KIDs on a single readout line, to image the athermal phonon distribution at < 1 mm position resolution and O(10) eV energy resolution. The design specification is set by the need to improve position reconstruction fidelity while maintaining low energy threshold for future rare-event searches such as for low-mass dark matter. We report on the design, which shows negligible crosstalk and > 95% inductor current uniformity, using the coplanar waveguide feedline, ground shield, and a new class of KIDs with symmetric coplanar stripline (sCPS) inductor. The multiplexing is designed upon the frequency-geometry relation we develop for the sCPS KIDs. We introduce the fabrications of the Nb RF assessment prototypes and the high phonon collection efficiency Al-Nb devices. We achieve ≲ 0.07% frequency displacement on a 80-KID RF assessment prototype, and the result indicates that we may place more than 180 resonances in our 0.4 GHz readout band with minimal frequency misordering. The coupling quality factors are ˜ 105 as designed. Finally, we update our work in progress in fabricating the O(102) KID, bi-material, O(1) kg detectors, and the expected position and energy resolutions.

Multiplexed aberration measurement for deep tissue imaging in vivo

PubMed Central

Wang, Chen; Liu, Rui; Milkie, Daniel E.; Sun, Wenzhi; Tan, Zhongchao; Kerlin, Aaron; Chen, Tsai-Wen; Kim, Douglas S.; Ji, Na

2014-01-01

We describe a multiplexed aberration measurement method that modulates the intensity or phase of light rays at multiple pupil segments in parallel to determine their phase gradients. Applicable to fluorescent-protein-labeled structures of arbitrary complexity, it allows us to obtain diffraction-limited resolution in various samples in vivo. For the strongly scattering mouse brain, a single aberration correction improves structural and functional imaging of fine neuronal processes over a large imaging volume. PMID:25128976

Multiplexed capillary electrophoresis system

DOEpatents

Yeung, Edward S.; Li, Qingbo; Lu, Xiandan

1998-04-21

The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification ("base calling") is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations.

Multiplexed capillary electrophoresis system

DOEpatents

Yeung, Edward S.; Chang, Huan-Tsang; Fung, Eliza N.; Li, Qingbo; Lu, Xiandan

1996-12-10

The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification ("base calling") is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations.

Multiplexed capillary electrophoresis system

DOEpatents

Yeung, E.S.; Li, Q.; Lu, X.

1998-04-21

The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification (``base calling``) is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations. 19 figs.

Multiplexed capillary electrophoresis system

DOEpatents

Yeung, E.S.; Chang, H.T.; Fung, E.N.; Li, Q.; Lu, X.

1996-12-10

The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification (``base calling``) is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations. 19 figs.

Phage-protease-peptide: a novel trifecta enabling multiplex detection of viable bacterial pathogens.

PubMed

Alcaine, S D; Tilton, L; Serrano, M A C; Wang, M; Vachet, R W; Nugen, S R

2015-10-01

Bacteriophages represent rapid, readily targeted, and easily produced molecular probes for the detection of bacterial pathogens. Molecular biology techniques have allowed researchers to make significant advances in the bioengineering of bacteriophage to further improve speed and sensitivity of detection. Despite their host specificity, bacteriophages have not been meaningfully leveraged in multiplex detection of bacterial pathogens. We propose a proof-of-principal phage-based scheme to enable multiplex detection. Our scheme involves bioengineering bacteriophage to carry a gene for a specific protease, which is expressed during infection of the target cell. Upon lysis, the protease is released to cleave a reporter peptide, and the signal detected. Here we demonstrate the successful (i) modification of T7 bacteriophage to carry tobacco etch virus (TEV) protease; (ii) expression of TEV protease by Escherichia coli following infection by our modified T7, an average of 2000 units of protease per phage are produced during infection; and (iii) proof-of-principle detection of E. coli in 3 h after a primary enrichment via TEV protease activity using a fluorescent peptide and using a designed target peptide for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis (MALDI-TOF MS) analysis. This proof-of-principle can be translated to other phage-protease-peptide combinations to enable multiplex bacterial detection and readily adopted on multiple platforms, like MALDI-TOF MS or fluorescent readers, commonly found in labs.

Performance studies of X3 silicon detectors for the future ELISSA array at ELI-NP

NASA Astrophysics Data System (ADS)

Chesnevskaya, S.; Balabanski, D. L.; Choudhury, D.; Constantin, P.; Filipescu, D. M.; Ghita, D. G.; Guardo, G. L.; Lattuada, D.; Matei, C.; Rotaru, A.; State, A.

2018-05-01

ELISSA is an array of silicon strip detectors under construction at the ELI-NP facility for measurements of photodissociation reactions using high-brilliance, quasi monoenergetic gamma beams. The detection system consists of 35 single-sided position-sensitive X3 detectors arranged in a cylindrical configuration and eight QQQ3 detectors as end-caps. A batch of forty X3 detectors have been tested at ELI-NP. The energy and position resolution, ballistic deficit, leakage currents, and depletion voltage were measured and analyzed. Measurements of the energy resolution were carried out using two read-out electronic chains, one based on multichannel preamplifiers and another based on multiplexers.

Fluorescence decay data analysis correcting for detector pulse pile-up at very high count rates

NASA Astrophysics Data System (ADS)

Patting, Matthias; Reisch, Paja; Sackrow, Marcus; Dowler, Rhys; Koenig, Marcelle; Wahl, Michael

2018-03-01

Using time-correlated single photon counting for the purpose of fluorescence lifetime measurements is usually limited in speed due to pile-up. With modern instrumentation, this limitation can be lifted significantly, but some artifacts due to frequent merging of closely spaced detector pulses (detector pulse pile-up) remain an issue to be addressed. We propose a data analysis method correcting for this type of artifact and the resulting systematic errors. It physically models the photon losses due to detector pulse pile-up and incorporates the loss in the decay fit model employed to obtain fluorescence lifetimes and relative amplitudes of the decay components. Comparison of results with and without this correction shows a significant reduction of systematic errors at count rates approaching the excitation rate. This allows quantitatively accurate fluorescence lifetime imaging at very high frame rates.

The Slow Control System of the Auger Fluorescence Detectors

NASA Astrophysics Data System (ADS)

Barenthien, N.; Bethge, C.; Daumiller, K.; Gemmeke, H.; Kampert, K.-H.; Wiebusch, C.

2003-07-01

The fluorescence detector (FD) of the Pierre Auger experiment [1] comprises 24 telescopes that will be situated in 4 remote buildings in the Pampa Amarilla. It is planned to run the fluorescence detectors in absence of operators on site. Therefore, the main task of the Slow Control System (SCS) is to ensure a secure remote operation of the FD system. The Slow Control System works autonomously and continuously monitors those parameters which may disturb a secure operation. Commands from the data-acquisition system or the remote operator are accepted only if they do not violate safety rules that depend on the actual experimental conditions (e.g. high-voltage, wind-sp eed, light, etc.). In case of malfunctions (power failure, communication breakdown, ...) the SCS performs an orderly shutdown and subsequent startup of the fluorescence detector system. The concept and the implementation of the Slow Control System are presented.

Simultaneous detection of antibodies to five Actinobacillus pleuropneumoniae serovars using bead-based multiplex analysis.

PubMed

Berger, Sanne Schou; Lauritsen, Klara Tølbøll; Boas, Ulrik; Lind, Peter; Andresen, Lars Ole

2017-11-01

We developed and made a preliminary validation of a bead-based multiplexed immunoassay for simultaneous detection of porcine serum antibodies to Actinobacillus pleuropneumoniae serovars 1, 2, 6, 7, and 12. Magnetic fluorescent beads were coupled with A. pleuropneumoniae antigens and tested with a panel of serum samples from experimentally infected pigs and with serum samples from uninfected and naturally infected pigs. The multiplex assay was compared to in-house ELISAs and complement fixation (CF) tests, which have been used for decades as tools for herd classification in the Danish Specific Pathogen Free system. Assay specificities and sensitivities as well as the corresponding cutoff values were determined using receiver operating characteristic (ROC) curve analysis, and the A. pleuropneumoniae multiplex assay showed good correlation with the in-house ELISAs and CF tests with areas under ROC curves ≥ 0.988. Benefits of multiplexed assays compared to ELISAs and CF tests include reduced serum sample volumes needed for analysis, less labor, and shorter assay time.

Gas electron multiplier (GEM) enhanced ionization chamber for fluorescence detector

NASA Astrophysics Data System (ADS)

Shaban, E. H.; Siddons, D. P.; Kuczewski, A.

2007-11-01

Detecting dilute elements in thin materials using extended X-ray absorption fluorescence spectroscopy (EXAFS) method requires a detector capable of high count rate and low noise. For detection of dilute elements, the fluorescence signal amplitude is often overcome by the presence of noise or background interference. In this paper we have used a gas ionization chamber enhanced by a gas electron multiplier (GEM) to amplify the primary ionized electrons due to the X-ray fluorescence of a dilute element. The GEM provides an essentially noise free electron amplification of the signal primary photoelectrons. It provides a larger output current prior to the electronic amplification, allowing a lower gain amplifier with lower electronic circuit noise contribution and hence improved S/ N ratio. In addition, since the signal is produced only by electrons, and not from ion motion, the detector is capable of recording rapidly changing signals. Iron in an arbitrary tree leaf was used as a test sample. This sample was measured using our detector SUBRSAB, and also with Lytle and passivated implanted planar silicon (PIPS) detectors. An improvement in the signal amplitude by a factor of 20 and a factor of 2 are recorded for the proposed detector with respect to the Lytle and PIPS detectors, respectively. Although the gain in signal over the PIPS detector is small for this detector, its lack of sensitivity to light and its low and temperature-independent dark current are further advantages.

Frequency-domain cascading microwave superconducting quantum interference device multiplexers; beyond limitations originating from room-temperature electronics

NASA Astrophysics Data System (ADS)

Kohjiro, Satoshi; Hirayama, Fuminori

2018-07-01

A novel approach, frequency-domain cascading microwave multiplexers (MW-Mux), has been proposed and its basic operation has been demonstrated to increase the number of pixels multiplexed in a readout line U of MW-Mux for superconducting detector arrays. This method is an alternative to the challenging development of wideband, large power, and spurious-free room-temperature (300 K) electronics. The readout system for U pixels consists of four main parts: (1) multiplexer chips connected in series those contain U superconducting resonators in total. (2) A cryogenic high-electron-mobility transistor amplifier (HEMT). (3) A 300 K microwave frequency comb generator based on N(≡U/M) parallel units of digital-to-analog converters (DAC). (4) N parallel units of 300 K analog-to-digital converters (ADC). Here, M is the number of tones each DAC produces and each ADC handles. The output signal of U detectors multiplexed at the cryogenic stage is transmitted through a cable to the room temperature and divided into N processors where each handles M pixels. Due to the reduction factor of 1/N, U is not anymore dominated by the 300 K electronics but can be increased up to the potential value determined by either the bandwidth or the spurious-free power of the HEMT. Based on experimental results on the prototype system with N = 2 and M = 3, neither excess inter-pixel crosstalk nor excess noise has been observed in comparison with conventional MW-Mux. This indicates that the frequency-domain cascading MW-Mux provides the full (100%) usage of the HEMT band by assigning N 300 K bands on the frequency axis without inter-band gaps.

The exposure of the hybrid detector of the Pierre Auger Observatory

NASA Astrophysics Data System (ADS)

Abreu, P.; Aglietta, M.; Ahn, E. J.; Allard, D.; Allekotte, I.; Allen, J.; Alvarez Castillo, J.; Alvarez-Muñiz, J.; Ambrosio, M.; Aminaei, A.; Anchordoqui, L.; Andringa, S.; Antičić, T.; Anzalone, A.; Aramo, C.; Arganda, E.; Arisaka, K.; Arqueros, F.; Asorey, H.; Assis, P.; Aublin, J.; Ave, M.; Avenier, M.; Avila, G.; Bäcker, T.; Badagnani, D.; Balzer, M.; Barber, K. B.; Barbosa, A. F.; Bardenet, R.; Barroso, S. L. C.; Baughman, B.; Beatty, J. J.; Becker, B. R.; Becker, K. H.; Bellétoile, A.; Bellido, J. A.; Benzvi, S.; Berat, C.; Bergmann, T.; Bertou, X.; Biermann, P. L.; Billoir, P.; Blanco, F.; Blanco, M.; Bleve, C.; Blümer, H.; Boháčová, M.; Boncioli, D.; Bonifazi, C.; Bonino, R.; Borodai, N.; Brack, J.; Brogueira, P.; Brown, W. C.; Bruijn, R.; Buchholz, P.; Bueno, A.; Burton, R. E.; Busca, N. G.; Caballero-Mora, K. S.; Caramete, L.; Caruso, R.; Castellina, A.; Catalano, O.; Cataldi, G.; Cazon, L.; Cester, R.; Chauvin, J.; Chiavassa, A.; Chinellato, J. A.; Chou, A.; Chudoba, J.; Clay, R. W.; Colombo, E.; Coluccia, M. R.; Conceição, R.; Contreras, F.; Cook, H.; Cooper, M. J.; Coppens, J.; Cordier, A.; Cotti, U.; Coutu, S.; Covault, C. E.; Creusot, A.; Criss, A.; Cronin, J.; Curutiu, A.; Dagoret-Campagne, S.; Dallier, R.; Dasso, S.; Daumiller, K.; Dawson, B. R.; de Almeida, R. M.; de Domenico, M.; de Donato, C.; de Jong, S. J.; de La Vega, G.; de Mello Junior, W. J. M.; de Mello Neto, J. R. T.; de Mitri, I.; de Souza, V.; de Vries, K. D.; Decerprit, G.; Del Peral, L.; Deligny, O.; Della Selva, A.; Dembinski, H.; Denkiewicz, A.; di Giulio, C.; Diaz, J. C.; Díaz Castro, M. L.; Diep, P. N.; Dobrigkeit, C.; D'Olivo, J. C.; Dong, P. N.; Dorofeev, A.; Dos Anjos, J. C.; Dova, M. T.; D'Urso, D.; Dutan, I.; Ebr, J.; Engel, R.; Erdmann, M.; Escobar, C. O.; Etchegoyen, A.; Facal San Luis, P.; Falcke, H.; Farrar, G.; Fauth, A. C.; Fazzini, N.; Ferguson, A. P.; Ferrero, A.; Fick, B.; Filevich, A.; Filipčič, A.; Fleck, I.; Fliescher, S.; Fracchiolla, C. E.; Fraenkel, E. D.; Fröhlich, U.; Fuchs, B.; Fulgione, W.; Gamarra, R. F.; Gambetta, S.; García, B.; García Gámez, D.; Garcia-Pinto, D.; Garrido, X.; Gascon, A.; Gelmini, G.; Gemmeke, H.; Gesterling, K.; Ghia, P. L.; Giaccari, U.; Giller, M.; Glass, H.; Gold, M. S.; Golup, G.; Gomez Albarracin, F.; Gómez Berisso, M.; Gonçalves, P.; Gonzalez, D.; Gonzalez, J. G.; Gookin, B.; Góra, D.; Gorgi, A.; Gouffon, P.; Gozzini, S. R.; Grashorn, E.; Grebe, S.; Grigat, M.; Grillo, A. F.; Guardincerri, Y.; Guarino, F.; Guedes, G. P.; Hague, J. D.; Hansen, P.; Harari, D.; Harmsma, S.; Harton, J. L.; Haungs, A.; Hebbeker, T.; Heck, D.; Herve, A. E.; Hojvat, C.; Holmes, V. C.; Homola, P.; Hörandel, J. R.; Horneffer, A.; Hrabovský, M.; Huege, T.; Insolia, A.; Ionita, F.; Italiano, A.; Jiraskova, S.; Kadija, K.; Kaducak, M.; Kampert, K. H.; Karhan, P.; Karova, T.; Kasper, P.; Kégl, B.; Keilhauer, B.; Keivani, A.; Kelley, J. L.; Kemp, E.; Kieckhafer, R. M.; Klages, H. O.; Kleifges, M.; Kleinfeller, J.; Knapp, J.; Koang, D.-H.; Kotera, K.; Krohm, N.; Krömer, O.; Kruppke-Hansen, D.; Kuehn, F.; Kuempel, D.; Kulbartz, J. K.; Kunka, N.; La Rosa, G.; Lachaud, C.; Lautridou, P.; Leão, M. S. A. B.; Lebrun, D.; Lebrun, P.; Leigui de Oliveira, M. A.; Lemiere, A.; Letessier-Selvon, A.; Lhenry-Yvon, I.; Link, K.; López, R.; Lopez Agüera, A.; Louedec, K.; Lozano Bahilo, J.; Lucero, A.; Ludwig, M.; Lyberis, H.; Maccarone, M. C.; Macolino, C.; Maldera, S.; Mandat, D.; Mantsch, P.; Mariazzi, A. G.; Marin, V.; Maris, I. C.; Marquez Falcon, H. R.; Marsella, G.; Martello, D.; Martin, L.; Martínez Bravo, O.; Mathes, H. J.; Matthews, J.; Matthews, J. A. J.; Matthiae, G.; Maurizio, D.; Mazur, P. O.; McEwen, M.; Medina-Tanco, G.; Melissas, M.; Melo, D.; Menichetti, E.; Menshikov, A.; Meurer, C.; Mičanović, S.; Micheletti, M. I.; Miller, W.; Miramonti, L.; Mollerach, S.; Monasor, M.; Monnier Ragaigne, D.; Montanet, F.; Morales, B.; Morello, C.; Moreno, E.; Moreno, J. C.; Morris, C.; Mostafá, M.; Mueller, S.; Muller, M. A.; Münchmeyer, M.; Mussa, R.; Navarra, G.; Navarro, J. L.; Navas, S.; Necesal, P.; Nellen, L.; Nhung, P. T.; Nierstenhoefer, N.; Nitz, D.; Nosek, D.; Nožka, L.; Nyklicek, M.; Oehlschläger, J.; Olinto, A.; Oliva, P.; Olmos-Gilbaja, V. M.; Ortiz, M.; Pacheco, N.; Pakk Selmi-Dei, D.; Palatka, M.; Pallotta, J.; Palmieri, N.; Parente, G.; Parizot, E.; Parra, A.; Parrisius, J.; Parsons, R. D.; Pastor, S.; Paul, T.; Pavlidou, V.; Payet, K.; Pech, M.; PeĶala, J.; Pelayo, R.; Pepe, I. M.; Perrone, L.; Pesce, R.; Petermann, E.; Petrera, S.; Petrinca, P.; Petrolini, A.; Petrov, Y.; Petrovic, J.; Pfendner, C.; Phan, N.; Piegaia, R.; Pierog, T.; Pimenta, M.; Pirronello, V.; Platino, M.; Ponce, V. H.; Pontz, M.; Privitera, P.; Prouza, M.; Quel, E. J.; Rautenberg, J.; Ravel, O.; Ravignani, D.; Revenu, B.; Ridky, J.; Riggi, S.; Risse, M.; Ristori, P.; Rivera, H.; Rivière, C.; Rizi, V.; Robledo, C.; Rodriguez, G.; Rodriguez Martino, J.; Rodriguez Rojo, J.; Rodriguez-Cabo, I.; Rodríguez-Frías, M. D.; Ros, G.; Rosado, J.; Rossler, T.; Roth, M.; Rouillé-D'Orfeuil, B.; Roulet, E.; Rovero, A. C.; Salamida, F.; Salazar, H.; Salina, G.; Sánchez, F.; Santander, M.; Santo, C. E.; Santos, E.; Santos, E. M.; Sarazin, F.; Sarkar, S.; Sato, R.; Scharf, N.; Scherini, V.; Schieler, H.; Schiffer, P.; Schmidt, A.; Schmidt, F.; Schmidt, T.; Scholten, O.; Schoorlemmer, H.; Schovancova, J.; Schovánek, P.; Schroeder, F.; Schulte, S.; Schüssler, F.; Schuster, D.; Sciutto, S. J.; Scuderi, M.; Segreto, A.; Semikoz, D.; Settimo, M.; Shadkam, A.; Shellard, R. C.; Sidelnik, I.; Sigl, G.; Śmiałkowski, A.; Šmída, R.; Snow, G. R.; Sommers, P.; Sorokin, J.; Spinka, H.; Squartini, R.; Stapleton, J.; Stasielak, J.; Stephan, M.; Strazzeri, E.; Stutz, A.; Suarez, F.; Suomijärvi, T.; Supanitsky, A. D.; Šuša, T.; Sutherland, M. S.; Swain, J.; Szadkowski, Z.; Tamashiro, A.; Tapia, A.; Tarutina, T.; Taşcău, O.; Tcaciuc, R.; Tcherniakhovski, D.; Tegolo, D.; Thao, N. T.; Thomas, D.; Tiffenberg, J.; Timmermans, C.; Tiwari, D. K.; Tkaczyk, W.; Todero Peixoto, C. J.; Tomé, B.; Tonachini, A.; Travnicek, P.; Tridapalli, D. B.; Tristram, G.; Trovato, E.; Tueros, M.; Ulrich, R.; Unger, M.; Urban, M.; Valdés Galicia, J. F.; Valiño, I.; Valore, L.; van den Berg, A. M.; Vargas Cárdenas, B.; Vázquez, J. R.; Vázquez, R. A.; Veberič, D.; Venters, T.; Verzi, V.; Videla, M.; Villaseñor, L.; Wahlberg, H.; Wahrlich, P.; Wainberg, O.; Warner, D.; Watson, A. A.; Weidenhaupt, K.; Weindl, A.; Westerhoff, S.; Whelan, B. J.; Wieczorek, G.; Wiencke, L.; Wilczyńska, B.; Wilczyński, H.; Will, M.; Williams, C.; Winchen, T.; Winders, L.; Winnick, M. G.; Wommer, M.; Wundheiler, B.; Yamamoto, T.; Younk, P.; Yuan, G.; Yushkov, A.; Zamorano, B.; Zas, E.; Zavrtanik, D.; Zavrtanik, M.; Zaw, I.; Zepeda, A.; Ziolkowski, M.; Pierre Auger Collaboration

2011-01-01

The Pierre Auger Observatory is a detector for ultra-high energy cosmic rays. It consists of a surface array to measure secondary particles at ground level and a fluorescence detector to measure the development of air showers in the atmosphere above the array. The "hybrid" detection mode combines the information from the two subsystems. We describe the determination of the hybrid exposure for events observed by the fluorescence telescopes in coincidence with at least one water-Cherenkov detector of the surface array. A detailed knowledge of the time dependence of the detection operations is crucial for an accurate evaluation of the exposure. We discuss the relevance of monitoring data collected during operations, such as the status of the fluorescence detector, background light and atmospheric conditions, that are used in both simulation and reconstruction.

The exposure of the hybrid detector of the Pierre Auger Observatory

DOE Office of Scientific and Technical Information (OSTI.GOV)

Not Available

2010-06-01

The Pierre Auger Observatory is a detector for ultra-high energy cosmic rays. It consists of a surface array to measure secondary particles at ground level and a fluorescence detector to measure the development of air showers in the atmosphere above the array. The 'hybrid' detection mode combines the information from the two subsystems. We describe the determination of the hybrid exposure for events observed by the fluorescence telescopes in coincidence with at least one water-Cherenkov detector of the surface array. A detailed knowledge of the time dependence of the detection operations is crucial for an accurate evaluation of the exposure.more » We discuss the relevance of monitoring data collected during operations, such as the status of the fluorescence detector, background light and atmospheric conditions, that are used in both simulation and reconstruction.« less

A current-assisted CMOS photonic sampler with two taps for fluorescence lifetime sensing

NASA Astrophysics Data System (ADS)

Ingelberts, H.; Kuijk, M.

2016-04-01

Imaging based on fluorescence lifetime is becoming increasingly important in medical and biological applications. State-of- the-art fluorescence lifetime microscopes either use bulky and expensive gated image intensifiers coupled to a CCD or single-photon detectors in a slow scanning setup. Numerous attempts are being made to create compact, cost-effective all- CMOS imagers for fluorescence lifetime sensing. Single-photon avalanche diode (SPAD) imagers can have very good timing resolution and noise characteristics but have low detection efficiency. Another approach is to use CMOS imagers based on demodulation detectors. These imagers can be either very fast or very efficient but it remains a challenge to combine both characteristics. Recently we developed the current-assisted photonic sampler (CAPS) to tackle these problems and in this work, we present a new CAPS with two detection taps that can sample a fluorescence decay in two time windows. In the case of mono-exponential decays, two windows provide enough information to resolve the lifetime. We built an electro-optical setup to characterize the detector and use it for fluorescence lifetime measurements. It consists of a supercontinuum pulsed laser source, an optical system to focus light into the detector and picosecond timing electronics. We describe the structure and operation of the two-tap CAPS and provide basic characterization of the speed performance at multiple wavelengths in the visible and near-infrared spectrum. We also record fluorescence decays of different visible and NIR fluorescent dyes and provide different methods to resolve the fluorescence lifetime.

Read-noise characterization of focal plane array detectors via mean-variance analysis.

PubMed

Sperline, R P; Knight, A K; Gresham, C A; Koppenaal, D W; Hieftje, G M; Denton, M B

2005-11-01

Mean-variance analysis is described as a method for characterization of the read-noise and gain of focal plane array (FPA) detectors, including charge-coupled devices (CCDs), charge-injection devices (CIDs), and complementary metal-oxide-semiconductor (CMOS) multiplexers (infrared arrays). Practical FPA detector characterization is outlined. The nondestructive readout capability available in some CIDs and FPA devices is discussed as a means for signal-to-noise ratio improvement. Derivations of the equations are fully presented to unify understanding of this method by the spectroscopic community.

Lead sulfide - Silicon MOSFET infrared focal plane development

NASA Technical Reports Server (NTRS)

Barrett, J. R.; Jhabvala, M. D.

1983-01-01

A process for directly integrating photoconductive lead sulfide (PbS) infrared detector material with silicon MOS integrated circuits has been developed primarily for application in long (greater than 10,000 detector elements) linear arrays for pushbroom scanning applications. The processing technology is based on the conventional PMOS and CMOS technologies with a variation in the metallization. Results and measurements on a fully integrated eight-element multiplexer are shown.

Automatic neutron dosimetry system based on fluorescent nuclear track detector technology.

PubMed

Akselrod, M S; Fomenko, V V; Bartz, J A; Haslett, T L

2014-10-01

For the first time, the authors are describing an automatic fluorescent nuclear track detector (FNTD) reader for neutron dosimetry. FNTD is a luminescent integrating type of detector made of aluminium oxide crystals that does not require electronics or batteries during irradiation. Non-destructive optical readout of the detector is performed using a confocal laser scanning fluorescence imaging with near-diffraction limited resolution. The fully automatic table-top reader allows one to load up to 216 detectors on a tray, read their engraved IDs using a CCD camera and optical character recognition, scan and process simultaneously two types of images in fluorescent and reflected laser light contrast to eliminate false-positive tracks related to surface and volume crystal imperfections. The FNTD dosimetry system allows one to measure neutron doses from 0.1 mSv to 20 Sv and covers neutron energies from thermal to 20 MeV. The reader is characterised by a robust, compact optical design, fast data processing electronics and user-friendly software. © The Author 2013. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Improved Optical-Fiber Temperature Sensors

NASA Technical Reports Server (NTRS)

Rogowski, Robert S.; Egalon, Claudio O.

1993-01-01

In optical-fiber temperature sensors of proposed type, phosphorescence and/or fluorescence in temperature-dependent coating layers coupled to photodetectors. Phosphorescent and/or fluorescent behavior(s) of coating material(s) depend on temperature; coating material or mixture of materials selected so one can deduce temperature from known temperature dependence of phosphorescence and/or fluorescence spectrum, and/or characteristic decay of fluorescence. Basic optical configuration same as that of optical-fiber chemical detectors described in "Making Optical-Fiber Chemical Detectors More Sensitive" (LAR-14525).

Capillaries for use in a multiplexed capillary electrophoresis system

DOEpatents

Yeung, Edward S.; Chang, Huan-Tsang; Fung, Eliza N.

1997-12-09

The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification ("base calling") is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations.

Capillaries for use in a multiplexed capillary electrophoresis system

DOEpatents

Yeung, E.S.; Chang, H.T.; Fung, E.N.

1997-12-09

The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification (``base calling``) is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations. 19 figs.

Fiber optic detector for immuno-testing

DOEpatents

Partin, Judy K.; Ward, Thomas E.; Grey, Alan E.

1992-01-01

A portable fiber optic detector that senses the presence of specific target chemicals in air or a gas by exchanging the target chemical for a fluoroescently-tagged antigen that is bound to an antibody which is in turn attached to an optical fiber. Replacing the fluorescently-tagged antigen reduces the fluorescence so that a photon sensing detector records the reduced light level and activates an appropriate alarm or indicator.

Parallel multiplex laser feedback interferometry

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhang, Song; Tan, Yidong; Zhang, Shulian, E-mail: zsl-dpi@mail.tsinghua.edu.cn

2013-12-15

We present a parallel multiplex laser feedback interferometer based on spatial multiplexing which avoids the signal crosstalk in the former feedback interferometer. The interferometer outputs two close parallel laser beams, whose frequencies are shifted by two acousto-optic modulators by 2Ω simultaneously. A static reference mirror is inserted into one of the optical paths as the reference optical path. The other beam impinges on the target as the measurement optical path. Phase variations of the two feedback laser beams are simultaneously measured through heterodyne demodulation with two different detectors. Their subtraction accurately reflects the target displacement. Under typical room conditions, experimentalmore » results show a resolution of 1.6 nm and accuracy of 7.8 nm within the range of 100 μm.« less

Incorporation of antigens from whole cell lysates and purified virions from MP12 into fluorescence microsphere immunoassays for the detection of antibodies against Rift Valley fever virus

USDA-ARS?s Scientific Manuscript database

Background: The purpose of this study was the development of multiplex fluorescence microsphere immunoassay (FMIA) for the detection of Rift Valley fever virus (RVFV) IgG and IgM antibodies by incorporation of antigens from whole cell lysates and purified virions from MP12. Methods and Findings: Vir...

Multiplexed detection of tumor markers with multicolor quantum dots based on fluorescence polarization immunoassay.

PubMed

Tian, Jianniao; Zhou, Liujin; Zhao, Yanchun; Wang, Yuan; Peng, Yan; Zhao, Shulin

2012-04-15

A multicolor quantum dot (QD)-based nanosensor for multiplex detection of two tumor markers in a homogeneous format based on fluorescence polarization immunoassay was proposed. QDs520 and QDs620 were labeled alpha-fetoprotein(α-AFP) and carcinoembryonic antigen (CEA), respectively. After separated and purified by ultrafiltration, they were used in fluorescence polarization immunoassay for the simultaneous detection of human serum alpha-fetoprotein and carcinoembryonic antigen. Under the optimal conditions, the multi-analyte immunosensor had a wide linear range (from 0.5 ng mL(-1) to 500 ng mL(-1)) for both two tumor markers and good correlation (0.996 for α-AFP and 0.993 for CEA). The detection limits (LOD) were 0.36 ng mL(-1) for CEA and 0.28 ng mL(-1) for α-AFP (S/N=3). The carcinoembryonic antigen and fetoprotein in clinical serum samples were simultaneously detected. The results from 28 serum samples had a good agreement with enzyme-linked immunosorbent assay (ELISA). The relative standard deviation and the recovery suggested that the precision and the accuracy of this analytical method were satisfactory. This strategy with high sensitivity, good specificity, easy procedures and short analysis time shows great promise for clinical diagnoses and basic discovery. The application of QDs with longer fluorescence lifetime and small fluorescence polarization can be used for the determination of high molecular-weight substances which cannot be analyzed using dye fluorescence polarization immunoassay. Copyright © 2012 Elsevier B.V. All rights reserved.

A space- and time-resolved single photon counting detector for fluorescence microscopy and spectroscopy

PubMed Central

Michalet, X.; Siegmund, O.H.W.; Vallerga, J.V.; Jelinsky, P.; Millaud, J.E.; Weiss, S.

2017-01-01

We have recently developed a wide-field photon-counting detector having high-temporal and high-spatial resolutions and capable of high-throughput (the H33D detector). Its design is based on a 25 mm diameter multi-alkali photocathode producing one photo electron per detected photon, which are then multiplied up to 107 times by a 3-microchannel plate stack. The resulting electron cloud is proximity focused on a cross delay line anode, which allows determining the incident photon position with high accuracy. The imaging and fluorescence lifetime measurement performances of the H33D detector installed on a standard epifluorescence microscope will be presented. We compare them to those of standard single-molecule detectors such as single-photon avalanche photodiode (SPAD) or electron-multiplying camera using model samples (fluorescent beads, quantum dots and live cells). Finally, we discuss the design and applications of future generation of H33D detectors for single-molecule imaging and high-throughput study of biomolecular interactions. PMID:29479130

[Multiplex real-time PCR method for rapid detection of Marburg virus and Ebola virus].

PubMed

Yang, Yu; Bai, Lin; Hu, Kong-Xin; Yang, Zhi-Hong; Hu, Jian-Ping; Wang, Jing

2012-08-01

Marburg virus and Ebola virus are acute infections with high case fatality rates. A rapid, sensitive detection method was established to detect Marburg virus and Ebola virus by multiplex real-time fluorescence quantitative PCR. Designing primers and Taqman probes from highly conserved sequences of Marburg virus and Ebola virus through whole genome sequences alignment, Taqman probes labeled by FAM and Texas Red, the sensitivity of the multiplex real-time quantitative PCR assay was optimized by evaluating the different concentrations of primers and Probes. We have developed a real-time PCR method with the sensitivity of 30.5 copies/microl for Marburg virus positive plasmid and 28.6 copies/microl for Ebola virus positive plasmids, Japanese encephalitis virus, Yellow fever virus, Dengue virus were using to examine the specificity. The Multiplex real-time PCR assays provide a sensitive, reliable and efficient method to detect Marburg virus and Ebola virus simultaneously.

Blocked impurity band hybrid infrared focal plane arrays for astronomy

NASA Technical Reports Server (NTRS)

Reynolds, D. B.; Seib, D. H.; Stetson, S. B.; Herter, T.; Rowlands, N.

1989-01-01

High-performance infrared hybrid focal plane arrays using 10- x 50-element Si:As blocked-impurity-band (BIB) detectors (cutoff wavelength = 28 microns) and matching switched MOSFET multiplexers have been developed and characterized for space astronomy. Use of impurity-band-conduction technology provides detectors which are nuclear-radiation-hard and free of the many anomalies associated with conventional silicon photoconductive detectors. Emphasis in the present work is on recent advances in detector material quality which have led to significantly improved detector and hybrid characteristics. Results demonstrating increased quantum efficiency (particularly at short-wavelength infrared), obtained by varying the BIB detector properties (infrared active layer thickness and arsenic doping profile), are summarized. Measured read noise and dark current for different temperatures are reported. The hybrid array performance achieved demonstrates that BIB detectors are well suited for use in astronomical instrumentation.

Mass cytometry: a highly multiplexed single-cell technology for advancing drug development.

PubMed

Atkuri, Kondala R; Stevens, Jeffrey C; Neubert, Hendrik

2015-02-01

Advanced single-cell analysis technologies (e.g., mass cytometry) that help in multiplexing cellular measurements in limited-volume primary samples are critical in bridging discovery efforts to successful drug approval. Mass cytometry is the state-of-the-art technology in multiparametric single-cell analysis. Mass cytometers (also known as cytometry by time-of-flight or CyTOF) combine the cellular analysis principles of traditional fluorescence-based flow cytometry with the selectivity and quantitative power of inductively coupled plasma-mass spectrometry. Standard flow cytometry is limited in the number of parameters that can be measured owing to the overlap in signal when detecting fluorescently labeled antibodies. Mass cytometry uses antibodies tagged to stable isotopes of rare earth metals, which requires minimal signal compensation between the different metal tags. This unique feature enables researchers to seamlessly multiplex up to 40 independent measurements on single cells. In this overview we first present an overview of mass cytometry and compare it with traditional flow cytometry. We then discuss the emerging and potential applications of CyTOF technology in the pharmaceutical industry, including quantitative and qualitative deep profiling of immune cells and their applications in assessing drug immunogenicity, extensive mapping of signaling networks in single cells, cell surface receptor quantification and multiplexed internalization kinetics, multiplexing sample analysis by barcoding, and establishing cell ontologies on the basis of phenotype and/or function. We end with a discussion of the anticipated impact of this technology on drug development lifecycle with special emphasis on the utility of mass cytometry in deciphering a drug's pharmacokinetics and pharmacodynamics relationship. Copyright © 2014 by The American Society for Pharmacology and Experimental Therapeutics.

High-throughput screening of hybridoma supernatants using multiplexed fluorescent cell barcoding on live cells.

PubMed

Lu, Mei; Chan, Brian M; Schow, Peter W; Chang, Wesley S; King, Chadwick T

2017-12-01

With current available assay formats using either immobilized protein (ELISA, enzyme-linked immunosorbent assay) or immunostaining of fixed cells for primary monoclonal antibody (mAb) screening, researchers often fail to identify and characterize antibodies that recognize the native conformation of cell-surface antigens. Therefore, screening using live cells has become an integral and important step contributing to the successful identification of therapeutic antibody candidates. Thus the need for developing high-throughput screening (HTS) technologies using live cells has become a major priority for therapeutic mAb discovery and development. We have developed a novel technique called Multiplexed Fluorescent Cell Barcoding (MFCB), a flow cytometry-based method based upon the Fluorescent Cell Barcoding (FCB) technique and the Luminex fluorescent bead array system, but is applicable to high-through mAb screens on live cells. Using this technique in our system, we can simultaneously identify or characterize the antibody-antigen binding of up to nine unique fluorescent labeled cell populations in the time that it would normally take to process a single population. This has significantly reduced the amount of time needed for the identification of potential lead candidates. This new technology enables investigators to conduct large-scale primary hybridoma screens using flow cytometry. This in turn has allowed us to screen antibodies more efficiently than before and streamline identification and characterization of lead molecules. Copyright © 2017 Elsevier B.V. All rights reserved.

Measuring fluorescence polarization with a dichrometer.

PubMed

Sutherland, John C

2017-09-01

A method for obtaining fluorescence polarization data from an instrument designed to measure circular and linear dichroism is compared with a previously reported approach. The new method places a polarizer between the sample and a detector mounted perpendicular to the direction of the incident beam and results in determination of the fluorescence polarization ratio, whereas the previous method does not use a polarizer and yields the fluorescence anisotropy. A similar analysis with the detector located axially with the excitation beam demonstrates that there is no frequency modulated signal due to fluorescence polarization in the absence of a polarizer. Copyright © 2017. Published by Elsevier Inc.

Development of a novel depth of interaction PET detector using highly multiplexed G-APD cross-strip encoding

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kolb, A., E-mail: armin.kolb@med.uni-tuebingen.de; Parl, C.; Liu, C. C.

Purpose: The aim of this study was to develop a prototype PET detector module for a combined small animal positron emission tomography and magnetic resonance imaging (PET/MRI) system. The most important factor for small animal imaging applications is the detection sensitivity of the PET camera, which can be optimized by utilizing longer scintillation crystals. At the same time, small animal PET systems must yield a high spatial resolution. The measured object is very close to the PET detector because the bore diameter of a high field animal MR scanner is limited. When used in combination with long scintillation crystals, thesemore » small-bore PET systems generate parallax errors that ultimately lead to a decreased spatial resolution. Thus, we developed a depth of interaction (DoI) encoding PET detector module that has a uniform spatial resolution across the whole field of view (FOV), high detection sensitivity, compactness, and insensitivity to magnetic fields. Methods: The approach was based on Geiger mode avalanche photodiode (G-APD) detectors with cross-strip encoding. The number of readout channels was reduced by a factor of 36 for the chosen block elements. Two 12 × 2 G-APD strip arrays (25μm cells) were placed perpendicular on each face of a 12 × 12 lutetium oxyorthosilicate crystal block with a crystal size of 1.55 × 1.55 × 20 mm. The strip arrays were multiplexed into two channels and used to calculate the x, y coordinates for each array and the deposited energy. The DoI was measured in step sizes of 1.8 mm by a collimated {sup 18}F source. The coincident resolved time (CRT) was analyzed at all DoI positions by acquiring the waveform for each event and applying a digital leading edge discriminator. Results: All 144 crystals were well resolved in the crystal flood map. The average full width half maximum (FWHM) energy resolution of the detector was 12.8% ± 1.5% with a FWHM CRT of 1.14 ± 0.02 ns. The average FWHM DoI resolution over 12 crystals was 2.90 ± 0.15 mm. Conclusions: The novel DoI PET detector, which is based on strip G-APD arrays, yielded a DoI resolution of 2.9 mm and excellent timing and energy resolution. Its high multiplexing factor reduces the number of electronic channels. Thus, this cross-strip approach enables low-cost, high-performance PET detectors for dedicated small animal PET and PET/MRI and potentially clinical PET/MRI systems.« less

Identification of nonclassical properties of light with multiplexing layouts

NASA Astrophysics Data System (ADS)

Sperling, J.; Eckstein, A.; Clements, W. R.; Moore, M.; Renema, J. J.; Kolthammer, W. S.; Nam, S. W.; Lita, A.; Gerrits, T.; Walmsley, I. A.; Agarwal, G. S.; Vogel, W.

2017-07-01

In Sperling et al. [Phys. Rev. Lett. 118, 163602 (2017), 10.1103/PhysRevLett.118.163602], we introduced and applied a detector-independent method to uncover nonclassicality. Here, we extend those techniques and give more details on the performed analysis. We derive a general theory of the positive-operator-valued measure that describes multiplexing layouts with arbitrary detectors. From the resulting quantum version of a multinomial statistics, we infer nonclassicality probes based on a matrix of normally ordered moments. We discuss these criteria and apply the theory to our data which are measured with superconducting transition-edge sensors. Our experiment produces heralded multiphoton states from a parametric down-conversion light source. We show that the known notions of sub-Poisson and sub-binomial light can be deduced from our general approach, and we establish the concept of sub-multinomial light, which is shown to outperform the former two concepts of nonclassicality for our data.

Identification of nonclassical properties of light with multiplexing layouts

PubMed Central

Sperling, J.; Eckstein, A.; Clements, W. R.; Moore, M.; Renema, J. J.; Kolthammer, W. S.; Nam, S. W.; Lita, A.; Gerrits, T.; Walmsley, I. A.; Agarwal, G. S.; Vogel, W.

2018-01-01

In Sperling et al. [Phys. Rev. Lett. 118, 163602 (2017)], we introduced and applied a detector-independent method to uncover nonclassicality. Here, we extend those techniques and give more details on the performed analysis. We derive a general theory of the positive-operator-valued measure that describes multiplexing layouts with arbitrary detectors. From the resulting quantum version of a multinomial statistics, we infer nonclassicality probes based on a matrix of normally ordered moments. We discuss these criteria and apply the theory to our data which are measured with superconducting transition-edge sensors. Our experiment produces heralded multiphoton states from a parametric down-conversion light source. We show that the known notions of sub-Poisson and sub-binomial light can be deduced from our general approach, and we establish the concept of sub-multinomial light, which is shown to outperform the former two concepts of nonclassicality for our data. PMID:29670949

Multiple-Locus Variable-Number Tandem-Repeats Analysis of Escherichia coli O157 using PCR multiplexing and multi-colored capillary electrophoresis.

PubMed

Lindstedt, Bjørn-Arne; Vardund, Traute; Kapperud, Georg

2004-08-01

The Multiple-Locus Variable-Number Tandem-Repeats Analysis (MLVA) method is currently being used as the primary typing tool for Shiga-toxin-producing Escherichia coli (STEC) O157 isolates in our laboratory. The initial assay was performed using a single fluorescent dye and the different patterns were assigned using a gel image. Here, we present a significantly improved assay using multiple dye colors and enhanced PCR multiplexing to increase speed, and ease the interpretation of the results. The different MLVA patterns are now based on allele sizes entered as character values, thus removing the uncertainties introduced when analyzing band patterns from the gel image. We additionally propose an easy numbering scheme for the identification of separate isolates that will facilitate exchange of typing data. Seventy-two human and animal strains of Shiga-toxin-producing E. coli O157 were used for the development of the improved MLVA assay. The method is based on capillary separation of multiplexed PCR products of VNTR loci in the E. coli O157 genome labeled with multiple fluorescent dyes. The different alleles at each locus were then assigned to allele numbers, which were used for strain comparison.

Pilot-multiplexed continuous-variable quantum key distribution with a real local oscillator

NASA Astrophysics Data System (ADS)

Wang, Tao; Huang, Peng; Zhou, Yingming; Liu, Weiqi; Zeng, Guihua

2018-01-01

We propose a pilot-multiplexed continuous-variable quantum key distribution (CVQKD) scheme based on a local local oscillator (LLO). Our scheme utilizes time-multiplexing and polarization-multiplexing techniques to dramatically isolate the quantum signal from the pilot, employs two heterodyne detectors to separately detect the signal and the pilot, and adopts a phase compensation method to almost eliminate the multifrequency phase jitter. In order to analyze the performance of our scheme, a general LLO noise model is constructed. Besides the phase noise and the modulation noise, the photon-leakage noise from the reference path and the quantization noise due to the analog-to-digital converter (ADC) are also considered, which are first analyzed in the LLO regime. Under such general noise model, our scheme has a higher key rate and longer secure distance compared with the preexisting LLO schemes. Moreover, we also conduct an experiment to verify our pilot-multiplexed scheme. Results show that it maintains a low level of the phase noise and is expected to obtain a 554-Kbps secure key rate within a 15-km distance under the finite-size effect.

Towards a disposable in vivo miniature implantable fluorescence detector

NASA Astrophysics Data System (ADS)

Bellis, Stephen; Jackson, J. Carlton; Mathewson, Alan

2006-02-01

In the field of fluorescent microscopy, neuronal activity, diabetes and drug treatment are a few of the wide ranging biomedical applications that can be monitored with the use of dye markers. Historically, in-vivo fluorescent detectors consist of implantable probes coupled by optical fibre to sophisticated bench-top instrumentation. These systems typically use laser light to excite the fluorescent marker dies and using sensors, such as the photo-multiplier tube (PMT) or charge coupled devices (CCD), detect the fluorescent light that is filtered from the total excitation. Such systems are large and expensive. In this paper we highlight the first steps toward a fully implantable in-vivo fluorescence detection system. The aim is to make the detector system small, low cost and disposable. The current prototype is a hybrid platform consisting of a vertical cavity surface emitting laser (VCSEL) to provide the excitation and a filtered solid state Geiger mode avalanche photo-diode (APD) to detect the emitted fluorescence. Fluorescence detection requires measurement of extremely low levels of light so the proposed APD detectors combine the ability to count individual photons with the added advantage of being small in size. At present the exciter and sensor are mounted on a hybrid PCB inside a 3mm diameter glass tube.This is wired to external electronics, which provide quenching, photon counting and a PC interface. In this configuration, the set-up can be used for in-vitro experimentation and in-vivo analysis conducted on animals such as mice.

Fluorescence of Picrosirius Red Multiplexed With Immunohistochemistry for the Quantitative Assessment of Collagen in Tissue Sections.

PubMed

Wegner, Kyle A; Keikhosravi, Adib; Eliceiri, Kevin W; Vezina, Chad M

2017-08-01

The low cost and simplicity of picrosirius red (PSR) staining have driven its popularity for collagen detection in tissue sections. We extended the versatility of this method by using fluorescent imaging to detect the PSR signal and applying automated quantification tools. We also developed the first PSR protocol that is fully compatible with multiplex immunostaining, making it possible to test whether collagen structure differs across immunohistochemically labeled regions of the tissue landscape. We compared our imaging method with two gold standards in collagen imaging, linear polarized light microscopy and second harmonic generation imaging, and found that it is at least as sensitive and robust to changes in sample orientation. As proof of principle, we used a genetic approach to overexpress beta catenin in a patchy subset of mouse prostate epithelial cells distinguished only by immunolabeling. We showed that collagen fiber length is significantly greater near beta catenin overexpressing cells than near control cells. Our fluorescent PSR imaging method is sensitive, reproducible, and offers a new way to guide region of interest selection for quantifying collagen in tissue sections.

Multiplex quantification of four DNA targets in one reaction with Bio-Rad droplet digital PCR system for GMO detection

NASA Astrophysics Data System (ADS)

Dobnik, David; Štebih, Dejan; Blejec, Andrej; Morisset, Dany; Žel, Jana

2016-10-01

The advantages of the digital PCR technology are already well documented until now. One way to achieve better cost efficiency of the technique is to use it in a multiplexing strategy. Droplet digital PCR platforms, which include two fluorescence filters, support at least duplex reactions and with some developments and optimization higher multiplexing is possible. The present study not only shows a development of multiplex assays in droplet digital PCR, but also presents a first thorough evaluation of several parameters in such multiplex digital PCR. Two 4-plex assays were developed for quantification of 8 different DNA targets (7 genetically modified maize events and maize endogene). Per assay, two of the targets were labelled with one fluorophore and two with another. As current analysis software does not support analysis of more than duplex, a new R- and Shiny-based web application analysis tool (http://bit.ly/ddPCRmulti) was developed that automates the analysis of 4-plex results. In conclusion, the two developed multiplex assays are suitable for quantification of GMO maize events and the same approach can be used in any other field with a need for accurate and reliable quantification of multiple DNA targets.

Multiplex quantification of four DNA targets in one reaction with Bio-Rad droplet digital PCR system for GMO detection.

PubMed

Dobnik, David; Štebih, Dejan; Blejec, Andrej; Morisset, Dany; Žel, Jana

2016-10-14

The advantages of the digital PCR technology are already well documented until now. One way to achieve better cost efficiency of the technique is to use it in a multiplexing strategy. Droplet digital PCR platforms, which include two fluorescence filters, support at least duplex reactions and with some developments and optimization higher multiplexing is possible. The present study not only shows a development of multiplex assays in droplet digital PCR, but also presents a first thorough evaluation of several parameters in such multiplex digital PCR. Two 4-plex assays were developed for quantification of 8 different DNA targets (7 genetically modified maize events and maize endogene). Per assay, two of the targets were labelled with one fluorophore and two with another. As current analysis software does not support analysis of more than duplex, a new R- and Shiny-based web application analysis tool (http://bit.ly/ddPCRmulti) was developed that automates the analysis of 4-plex results. In conclusion, the two developed multiplex assays are suitable for quantification of GMO maize events and the same approach can be used in any other field with a need for accurate and reliable quantification of multiple DNA targets.

Means and method for capillary zone electrophoresis with laser-induced indirect fluorescence detection

DOEpatents

Yeung, Edward S.; Kuhr, Werner G.

1996-02-20

A means and method for capillary zone electrphoresis with laser-induced indirect fluorescence detection. A detector is positioned on the capillary tube of a capillary zone electrophoresis system. The detector includes a laser which generates a laser beam which is imposed upon a small portion of the capillary tube. Fluorescence of the elutant electromigrating through the capillary tube is indirectly detected and recorded.

Means and method for capillary zone electrophoresis with laser-induced indirect fluorescence detection

DOEpatents

Yeung, Edwards; Kuhr, Werner G.

1991-04-09

A means and method for capillary zone electrphoresis with laser-induced indirect fluorescence detection. A detector is positioned on the capillary tube of a capillary zone electrophoresis system. The detector includes a laser which generates a laser beam which is imposed upon a small portion of the capillary tube. Fluorescence of the elutant electromigrating through the capillary tube is indirectly detected and recorded.

Feasibility Study of an Axially Extendable Multiplex Cylinder PET

NASA Astrophysics Data System (ADS)

Yoshida, Eiji; Hirano, Yoshiyuki; Tashima, Hideaki; Inadama, Naoko; Nishikido, Fumihiko; Murayama, Hideo; Ito, Hiroshi; Yamaya, Taiga

2013-10-01

Current clinical PET scanners have a 15-22 cm axial field-of-view (FOV). These scanners image the whole body using six or more bed positions. We designed an axially extendable multiplex cylinder (AEMC) PET scanner to provide high versatility for clinical and research studies using semiconductor photo-sensor based, depth-of-interaction (DOI) detectors. Since silicon-photomultipliers (Si-PMs) have high gain like conventional photomultiplier tubes and a compact design, the Si-PM-based detector is particularly expected to enable various new detector arrangements. The AEMC-PET scanner consists of multiple independent and laminated detector rings using the DOI detectors. The AEMC-PET scanner can extend the axial FOV as each stacked detector ring can be slid sideways. When this PET scanner is used for the four-layer DOI detector, its minimum axial FOV is 24 cm and its maximum crystal thickness is 3 cm. On the other hand, the axial FOV can be extended to 96 cm when laminated detector rings are slid sideways, but the crystal thickness must be 1/4 of 3 cm. In this work, we evaluated performance characteristics of the PET scanner with a variable axial FOV using Monte Carlo simulation. From the simulation of the 180-cm line source, the 96-cm axial FOV was found to have two-fold better sensitivity compared to the 24-cm axial FOV. For extension of the axial FOV, scatter and attenuation of oblique lines-of-response reduced the yield of true coincidences, but effects of scatter and attenuation were small. Conclusive results were obtained showing the 52.8-cm axial FOV yielded an increase in the noise equivalent count rate of approximately 30% relative to the 24-cm axial FOV. We expect the designed AEMC-PET scanner will provide high versatility in applications such as for measuring whole-body tracer uptakes while keeping the continuous axial FOV; as well, the scan time for static images will be reduced for a comparable number of detectors as conventional PET scanners.

Polymeric nanoparticles with sequential and multiple FRET cascade mechanisms for multicolor and multiplexed imaging.

PubMed

Wagh, Anil; Jyoti, Faidat; Mallik, Sanku; Qian, Steven; Leclerc, Estelle; Law, Benedict

2013-06-24

The ability to map multiple biomarkers at the same time has far-reaching biomedical and diagnostic applications. Here, a series of biocompatible poly(D,L-lactic-co-glycolic acid) and polyethylene glycol particles for multicolor and multiplexed imaging are reported. More than 30 particle formulations that exhibit distinct emission signatures (ranging from the visible to NIR wavelength region) are designed and synthesized. These particles are encapsulated with combinations of carbocyanine-based fluorophores DiO, Dil, DiD, and DiR, and are characterized as <100 nm in size and brighter than commercial quantum dots. A particle formulation is identified that simultaneously emits fluorescence at three different wavelengths upon a single excitation at 485 nm via sequential and multiple FRET cascade events for multicolor imaging. Three other particles that display maximum fluorescence intensities at 570, 672, or 777 nm for multiplexed imaging are also identified. These particles are individually conjugated with specific (Herceptin or IgG2A11 antibody) or nonspecific (heptaarginine) ligands for targeting and, thus, could be applied to differentiate different cancer cells from a cell mixture according to the expressions of cell-surface human epidermal growth factor receptor 2 and the receptor for advanced glycation endproducts. Using an animal model subcutaneously implanted with the particles, it is further demonstrated that the developed platform could be useful for in vivo multiplexed imaging. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Rapid, sensitive, and simultaneous detection of three foodborne pathogens using magnetic nanobead-based immunoseparation and quantum dot-based multiplex immunoassay.

PubMed

Wang, Hong; Li, Yanbin; Wang, Andrew; Slavik, Michael

2011-12-01

Losses caused by foodborne diseases are enormous in terms of human life, illness, medical costs, and food product recalls. Rapid detection of multiple bacterial pathogens in foods is extremely important to ensure food safety. The objective of this research was to develop a multiplex immunoassay by integrating magnetic nanobeads (MNBs) for immunoseparation with quantum dots (QDs) as fluorescent labels for rapid, sensitive, and simultaneous detection of three major pathogenic bacteria, Salmonella Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes, in food products. In this research, both streptavidin-conjugated MNBs (30- and 150-nm diameter) and QDs (530-, 580-, and 620-nm emission wavelength) were separately coated with biotinylated anti-Salmonella, anti-E. coli, and anti-Listeria antibodies. The immuno-MNBs were mixed with a food sample to capture the three target bacteria. After being magnetically separated from the sample, the MNB-cell conjugates were mixed with the immuno-QDs to form the MNB-cell-QD complexes, and unattached QDs were removed. The fluorescence intensity of the MNB-cell-QD complexes was measured at wavelengths of 530, 580, and 620 nm to determine the populations of Salmonella Typhimurium, E. coli O157:H7, and L. monocytogenes, respectively. This multiplex immunoassay simultaneously detected Salmonella Typhimurium, E. coli O157:H7, and L. monocytogenes at levels as low as 20 to 50 CFU/ml in food samples in less than 2 h without enrichment. The change in fluorescence intensity was linearly correlated (R(2) > 0.96) with the logarithmic value of bacterial level in the range of 10 to 10(3) CFU/ml. More than 85% of the three target pathogens could be simultaneously separated from food samples. The multiplex immunoassay could be expanded to detect more target pathogens, depending on the availability of specific antibodies and QDs with different emission wavelengths.

A BODIPY-Based Fluorescent Probe to Visually Detect Phosgene: Toward the Development of a Handheld Phosgene Detector.

PubMed

Sayar, Melike; Karakuş, Erman; Güner, Tuğrul; Yildiz, Busra; Yildiz, Umit Hakan; Emrullahoğlu, Mustafa

2018-03-02

A boron-dipyrromethene (BODIPY)-based fluorescent probe with a phosgene-specific reactive motif shows remarkable selectivity toward phosgene, in the presence of which the nonfluorescent dye rapidly transforms into a new structure and induces a fluorescent response clearly observable to the naked eye under ultraviolet light. Given that dynamic, a prototypical handheld phosgene detector with a promising sensing capability that expedites the detection of gaseous phosgene without sophisticated instrumentation was developed. The proposed method using the handheld detector involves a rapid response period suitable for issuing early warnings during emergency situations. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Fluorescently labelled multiplex lateral flow immunoassay based on cadmium-free quantum dots.

PubMed

Beloglazova, Natalia V; Sobolev, Aleksander M; Tessier, Mickael D; Hens, Zeger; Goryacheva, Irina Yu; De Saeger, Sarah

2017-03-01

A sensitive tool for simultaneous qualitative detection of two mycotoxins based on use of non-cadmium quantum dots (QDs) is presented for the first time. QDs have proven themselves as promising fluorescent labels for biolabeling and chemical analysis. With an increasing global tendency to regulate and limit the use of hazardous elements, indium phosphide (InP) QDs are highlighted as environmentally-friendly alternatives to the highly efficient and well-studied, but potentially toxic Cd- and Pb-based QDs. Here, we developed water-soluble InP QDs-based fluorescent nanostructures. They consisted of core/shell InP/ZnS QDs enrobed in a silica shell that allowed the water solubility (QD@SiO 2 ). Then we applied the QD@SiO 2 as novel, silica shell-encapsulated fluorescent labels in immunoassays for rapid multiplexed screening. Two mycotoxins, zearalenone and deoxynivalenol, were simultaneously detected in maize and wheat, since the two QD@SiO 2 labelled conjugates emit at two different, individually detectable wavelengths. The cutoff values for the simultaneous determination were 50 and 500μgkg -1 for zearalenone and deoxynivalenol, respectively, in both maize and wheat. Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was used to confirm the result. Copyright © 2017 Elsevier Inc. All rights reserved.

System and method for monitoring cellular activity

NASA Technical Reports Server (NTRS)

Bearman, Gregory H. (Inventor); Fraser, Scott E. (Inventor); Lansford, Russell D. (Inventor)

2002-01-01

A system and method for monitoring cellular activity in a cellular specimen. According to one embodiment, a plurality of excitable markers are applied to the specimen. A multi-photon laser microscope is provided to excite a region of the specimen and cause fluorescence to be radiated from the region. The radiating fluorescence is processed by a spectral analyzer to separate the fluorescence into respective wavelength bands. The respective bands of fluorescence are then collected by an array of detectors, with each detector receiving a corresponding one of the wavelength bands.

System and method for monitoring cellular activity

NASA Technical Reports Server (NTRS)

Bearman, Gregory H. (Inventor); Fraser, Scott E. (Inventor); Lansford, Russell D. (Inventor)

2004-01-01

A system and method for monitoring cellular activity in a cellular specimen. According to one embodiment, a plurality of excitable markers are applied to the specimen. A multi-photon laser microscope is provided to excite a region of the specimen and cause fluorescence to be radiated from the region. The radiating fluorescence is processed by a spectral analyzer to separate the fluorescence into respective wavelength bands. The respective bands of fluorescence are then collected by an array of detectors, with each detector receiving a corresponding one of the wavelength bands.

Measuring fluorescence polarization with a dichrometer

DOE PAGES

Sutherland, John C.

2017-04-06

In this article, a method for obtaining fluorescence polarization data from an instrument designed to measure circular and linear dichroism is compared with a previously reported approach. The new method places a polarizer between the sample and a detector mounted perpendicular to the direction of the incident beam and results in determination of the fluorescence polarization ratio, whereas the previous method does not use a polarizer and yields the fluorescence anisotropy. A similar analysis with the detector located axially with the excitation beam demonstrates that there is no frequency modulated signal due to fluorescence polarization in the absence of amore » polarizer.« less

Delay grid multiplexing: simple time-based multiplexing and readout method for silicon photomultipliers

NASA Astrophysics Data System (ADS)

Won, Jun Yeon; Ko, Guen Bae; Lee, Jae Sung

2016-10-01

In this paper, we propose a fully time-based multiplexing and readout method that uses the principle of the global positioning system. Time-based multiplexing allows simplifying the multiplexing circuits where the only innate traces that connect the signal pins of the silicon photomultiplier (SiPM) channels to the readout channels are used as the multiplexing circuit. Every SiPM channel is connected to the delay grid that consists of the traces on a printed circuit board, and the inherent transit times from each SiPM channel to the readout channels encode the position information uniquely. Thus, the position of each SiPM can be identified using the time difference of arrival (TDOA) measurements. The proposed multiplexing can also allow simplification of the readout circuit using the time-to-digital converter (TDC) implemented in a field-programmable gate array (FPGA), where the time-over-threshold (ToT) is used to extract the energy information after multiplexing. In order to verify the proposed multiplexing method, we built a positron emission tomography (PET) detector that consisted of an array of 4  ×  4 LGSO crystals, each with a dimension of 3  ×  3  ×  20 mm3, and one- to-one coupled SiPM channels. We first employed the waveform sampler as an initial study, and then replaced the waveform sampler with an FPGA-TDC to further simplify the readout circuits. The 16 crystals were clearly resolved using only the time information obtained from the four readout channels. The coincidence resolving times (CRTs) were 382 and 406 ps FWHM when using the waveform sampler and the FPGA-TDC, respectively. The proposed simple multiplexing and readout methods can be useful for time-of-flight (TOF) PET scanners.

Multiplexed analysis of protein-ligand interactions by fluorescence anisotropy in a microfluidic platform.

PubMed

Cheow, Lih Feng; Viswanathan, Ramya; Chin, Chee-Sing; Jennifer, Nancy; Jones, Robert C; Guccione, Ernesto; Quake, Stephen R; Burkholder, William F

2014-10-07

Homogeneous assay platforms for measuring protein-ligand interactions are highly valued due to their potential for high-throughput screening. However, the implementation of these multiplexed assays in conventional microplate formats is considerably expensive due to the large amounts of reagents required and the need for automation. We implemented a homogeneous fluorescence anisotropy-based binding assay in an automated microfluidic chip to simultaneously interrogate >2300 pairwise interactions. We demonstrated the utility of this platform in determining the binding affinities between chromatin-regulatory proteins and different post-translationally modified histone peptides. The microfluidic chip assay produces comparable results to conventional microtiter plate assays, yet requires 2 orders of magnitude less sample and an order of magnitude fewer pipetting steps. This approach enables one to use small samples for medium-scale screening and could ease the bottleneck of large-scale protein purification.

Optimizing detector geometry for trace element mapping by X-ray fluorescence.

PubMed

Sun, Yue; Gleber, Sophie-Charlotte; Jacobsen, Chris; Kirz, Janos; Vogt, Stefan

2015-05-01

Trace metals play critical roles in a variety of systems, ranging from cells to photovoltaics. X-Ray Fluorescence (XRF) microscopy using X-ray excitation provides one of the highest sensitivities available for imaging the distribution of trace metals at sub-100 nm resolution. With the growing availability and increasing performance of synchrotron light source based instruments and X-ray nanofocusing optics, and with improvements in energy-dispersive XRF detectors, what are the factors that limit trace element detectability? To address this question, we describe an analytical model for the total signal incident on XRF detectors with various geometries, including the spectral response of energy dispersive detectors. This model agrees well with experimentally recorded X-ray fluorescence spectra, and involves much shorter calculation times than with Monte Carlo simulations. With such a model, one can estimate the signal when a trace element is illuminated with an X-ray beam, and when just the surrounding non-fluorescent material is illuminated. From this signal difference, a contrast parameter can be calculated and this can in turn be used to calculate the signal-to-noise ratio (S/N) for detecting a certain elemental concentration. We apply this model to the detection of trace amounts of zinc in biological materials, and to the detection of small quantities of arsenic in semiconductors. We conclude that increased detector collection solid angle is (nearly) always advantageous even when considering the scattered signal. However, given the choice between a smaller detector at 90° to the beam versus a larger detector at 180° (in a backscatter-like geometry), the 90° detector is better for trace element detection in thick samples, while the larger detector in 180° geometry is better suited to trace element detection in thin samples. Copyright © 2015. Published by Elsevier B.V.

Optimizing detector geometry for trace element mapping by X-ray fluorescence

PubMed Central

Sun, Yue; Gleber, Sophie-Charlotte; Jacobsen, Chris; Kirz, Janos; Vogt, Stefan

2016-01-01

Trace metals play critical roles in a variety of systems, ranging from cells to photovoltaics. X-Ray Fluorescence (XRF) microscopy using X-ray excitation provides one of the highest sensitivities available for imaging the distribution of trace metals at sub-100 nm resolution. With the growing availability and increasing performance of synchrotron light source based instruments and X-ray nanofocusing optics, and with improvements in energy-dispersive XRF detectors, what are the factors that limit trace element detectability? To address this question, we describe an analytical model for the total signal incident on XRF detectors with various geometries, including the spectral response of energy dispersive detectors. This model agrees well with experimentally recorded X-ray fluorescence spectra, and involves much shorter calculation times than with Monte Carlo simulations. With such a model, one can estimate the signal when a trace element is illuminated with an X-ray beam, and when just the surrounding non-fluorescent material is illuminated. From this signal difference, a contrast parameter can be calculated and this can in turn be used to calculate the signal-to-noise ratio (S/N) for detecting a certain elemental concentration. We apply this model to the detection of trace amounts of zinc in biological materials, and to the detection of small quantities of arsenic in semiconductors. We conclude that increased detector collection solid angle is (nearly) always advantageous even when considering the scattered signal. However, given the choice between a smaller detector at 90° to the beam versus a larger detector at 180° (in a backscatter-like geometry), the 90° detector is better for trace element detection in thick samples, while the larger detector in 180° geometry is better suited to trace element detection in thin samples. PMID:25600825

Optimizing detector geometry for trace element mapping by X-ray fluorescence

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sun, Yue; Gleber, Sophie-Charlotte; Jacobsen, Chris

Trace metals play critical roles in a variety of systems, ranging from cells to photovoltaics. X-Ray Fluorescence (XRF) microscopy using X-ray excitation provides one of the highest sensitivities available for imaging the distribution of trace metals at sub-100 nm resolution. With the growing availability and increasing performance of synchrotron light source based instruments and X-ray nanofocusing optics, and with improvements in energy-dispersive XRF detectors, what are the factors that limit trace element detectability? To address this question, we describe an analytical model for the total signal incident on XRF detectors with various geometries, including the spectral response of energy dispersivemore » detectors. This model agrees well with experimentally recorded X-ray fluorescence spectra, and involves much shorter calculation times than with Monte Carlo simulations. With such a model, one can estimate the signal when a trace element is illuminated with an X-ray beam, and when just the surrounding non-fluorescent material is illuminated. From this signal difference, a contrast parameter can be calculated and this can in turn be used to calculate the signal-to-noise ratio (S/N) for detecting a certain elemental concentration. We apply this model to the detection of trace amounts of zinc in biological materials, and to the detection of small quantities of arsenic in semiconductors. We conclude that increased detector collection solid angle is (nearly) always advantageous even when considering the scattered signal. However, given the choice between a smaller detector at 90° to the beam versus a larger detector at 180° (in a backscatter-like geometry), the 90° detector is better for trace element detection in thick samples, while the larger detector in 180° geometry is better suited to trace element detection in thin samples.« less

MO-F-CAMPUS-J-03: Development of a Human Brain PET for On-Line Proton Beam-Range Verification

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shao, Yiping

Purpose: To develop a prototype PET for verifying proton beam-range before each fractionated therapy that will enable on-line re-planning proton therapy. Methods: Latest “edge-less” silicon photomultiplier arrays and customized ASIC readout electronics were used to develop PET detectors with depth-of-interaction (DOI) measurement capability. Each detector consists of one LYSO array with each end coupled to a SiPM array. Multiple detectors can be seamlessly tiled together to form a large detector panel. Detectors with 1.5×1.5 and 2.0×2.0 mm crystals at 20 or 30 mm lengths were studied. Readout of individual SiPM or signal multiplexing was used to transfer 3D interaction position-codedmore » analog signals through flexible-print-circuit cables or PCB board to dedicated ASIC front-end electronics to output digital timing pulses that encode interaction information. These digital pulses can be transferred to, through standard LVDS cables, and decoded by a FPGA-based data acquisition of coincidence events and data transfer. The modular detector and scalable electronics/data acquisition will enable flexible PET system configuration for different imaging geometry. Results: Initial detector performance measurement shows excellent crystal identification even with 30 mm long crystals, ∼18% and 2.8 ns energy and timing resolutions, and around 2–3 mm DOI resolution. A small prototype PET scanner with one detector ring has been built and evaluated, validating the technology and design. A large size detector panel has been fabricated by scaling up from modular detectors. Different designs of resistor and capacitor based signal multiplexing boards were tested and selected based on optimal crystal identification and timing performance. Stackable readout electronics boards and FPGA-based data acquisition boards were developed and tested. A brain PET is under construction. Conclusion: Technology of large-size DOI detector based on SiPM array and advanced readout has been developed. PET imaging performance and initial phantom studies of on-line proton beam-range measurement will be conducted and reported. NIH grant R21CA187717; Cancer Prevention and Research Institute of Texas grant RP120326.« less

First Tests of Prototype SCUBA-2 Superconducting Bolometer Array

NASA Astrophysics Data System (ADS)

Woodcraft, Adam L.; Ade, Peter A. R.; Bintley, Dan; Hunt, Cynthia L.; Sudiwala, Rashmi V.; Hilton, Gene C.; Irwin, Kent D.; Reintsema, Carl D.; Audley, Michael D.; Holland, Wayne S.; MacIntosh, Mike

2006-09-01

We present results of the first tests on a 1280 pixel superconducting bolometer array, a prototype for SCUBA-2, a sub-mm camera being built for the James Clerk Maxwell Telescope in Hawaii. The bolometers are TES (transition edge sensor) detectors; these take advantage of the large variation of resistance with temperature through the superconducting transition. To keep the number of wires reasonable, a multiplexed read-out is used. Each pixel is read out through an individual DC SQUID; room temperature electronics switch between rows in the array by biasing the appropriate SQUIDs in turn. Arrays of 100 SQUIDs in series for each column then amplify the output. Unlike previous TES arrays, the multiplexing elements are located beneath each pixel, making large arrays possible, but construction more challenging. The detectors are constructed from Mo/Cu bi-layers; this technique enables the transition temperature to be tuned using the proximity effect by choosing the thickness of the normal and superconducting materials. To achieve the required performance, the detectors are operated at a temperature of approximately 120 mK. We describe the results of a basic characterisation of the array, demonstrating that it is fully operational, and give the results of signal to noise measurements.

Genotyping of alpha-thalassemia deletions using multiplex polymerase chain reactions and gold nanoparticle-filled capillary electrophoresis.

PubMed

Chen, Yen-Ling; Shih, Chi-Jen; Ferrance, Jerome; Chang, Ya-Sian; Chang, Jan-Gowth; Wu, Shou-Mei

2009-02-13

A gold nanoparticle-filled capillary electrophoresis method combined with three multiplex polymerase chain reactions (PCRs) was established for simultaneous diagnosis of five common alpha-thalassemia deletions, including the -alpha(3.7) deletion, -alpha(4.2) deletion, Southeast Asian (--(SEA)), Filipino (--(FIL)) and Thai (--(THAI)) deletions. Gold nanoparticles (GNPs) were used as a pseudostationary phase to improve the resolution between DNA fragments in a low-viscosity polymer. To achieve the best CE separation, several parameters were evaluated for optimizing the separation conditions, including the capillary coating, the concentrations of polymer sieving matrix, the sizes and concentrations of GNPs, the buffer concentrations, and the pH. The final CE method for separating a 200-base pair (bp) DNA ladder and alpha-thalassemia deletions used a DB-17 capillary, 0.6% poly(ethylene oxide) (PEO) prepared in a mixture of GNP(32nm) solution and glycine buffer (25mM, pH 9.0) (80:20, v/v) as the sieving matrix with 1microM YO-PRO-1 for fluorescence detection; the applied voltage was -10kV (detector at anode side) and the separation temperature was 25 degrees C. Under these optimal conditions, 15 DNA fragments with sizes ranging from 0.2kb to 3.0kb were resolved within 11.5min. The RSDs of migration times were less than 2.81%. A total of 21 patients with alpha-thalassemia deletions were analyzed using this method, and all results showed good agreement with those obtained by gel electrophoresis.

Volatile chemical reagent detector

DOEpatents

Chen, Liaohai; McBranch, Duncan; Wang, Rong; Whitten, David

2004-08-24

A device for detecting volatile chemical reagents based on fluorescence quenching analysis that is capable of detecting neutral electron acceptor molecules. The device includes a fluorescent material, a contact region, a light source, and an optical detector. The fluorescent material includes at least one polymer-surfactant complex. The polymer-surfactant complex is formed by combining a fluorescent ionic conjugated polymer with an oppositely charged surfactant. The polymer-surfactant complex may be formed in a polar solvent and included in the fluorescent material as a solution. Alternatively, the complex may be included in the fluorescent material as a thin film. The use of a polymer-surfactant complex in the fluorescent material allows the device to detect both neutral and ionic acceptor molecules. The use of a polymer-surfactant complex film allows the device and the fluorescent material to be reusable after exposing the fluorescent material to a vacuum for limited time.

Multiplexed Oversampling Digitizer in 65 nm CMOS for Column-Parallel CCD Readout

DOE Office of Scientific and Technical Information (OSTI.GOV)

Grace, Carl; Walder, Jean-Pierre; von der Lippe, Henrik

2012-04-10

A digitizer designed to read out column-parallel charge-coupled devices (CCDs) used for high-speed X-ray imaging is presented. The digitizer is included as part of the High-Speed Image Preprocessor with Oversampling (HIPPO) integrated circuit. The digitizer module comprises a multiplexed, oversampling, 12-bit, 80 MS/s pipelined Analog-to-Digital Converter (ADC) and a bank of four fast-settling sample-and-hold amplifiers to instrument four analog channels. The ADC multiplexes and oversamples to reduce its area to allow integration that is pitch-matched to the columns of the CCD. Novel design techniques are used to enable oversampling and multiplexing with a reduced power penalty. The ADC exhibits 188more » ?V-rms noise which is less than 1 LSB at a 12-bit level. The prototype is implemented in a commercially available 65 nm CMOS process. The digitizer will lead to a proof-of-principle 2D 10 Gigapixel/s X-ray detector.« less

Fluorescence-based ion-sensing with colloidal particles.

PubMed

Ashraf, Sumaira; Carrillo-Carrion, Carolina; Zhang, Qian; Soliman, Mahmoud G; Hartmann, Raimo; Pelaz, Beatriz; Del Pino, Pablo; Parak, Wolfgang J

2014-10-01

Particle-based fluorescence sensors for the quantification of specific ions can be made by coupling ion-sensitive fluorophores to carrier particles, or by using intrinsically fluorescent particles whose fluorescence properties depend on the concentration of the ions. Despite the advantages of such particle-based sensors for the quantitative detection of ions, such as the possibility to tune the surface chemistry and thus entry portal of the sensor particles to cells, they have also some associated problems. Problems involve for example crosstalk of the ion-sensitive fluorescence read-out with pH, or spectral overlap of the emission spectra of different fluorescent particles in multiplexing formats. Here the benefits of using particle-based fluorescence sensors, their limitations and strategies to overcome these limitations will be described and exemplified with selected examples. Copyright © 2014 Elsevier Ltd. All rights reserved.

Photocleavable DNA Barcoding Antibodies for Multiplexed Protein Analysis in Single Cells.

PubMed

Ullal, Adeeti V; Weissleder, Ralph

2015-01-01

We describe a DNA-barcoded antibody sensing technique for single cell protein analysis in which the barcodes are photocleaved and digitally detected without amplification steps (Ullal et al., Sci Transl Med 6:219, 2014). After photocleaving the unique ~70 mer DNA barcodes we use a fluorescent hybridization technology for detection, similar to what is commonly done for nucleic acid readouts. This protocol offers a simple method for multiplexed protein detection using 100+ antibodies and can be performed on clinical samples as well as single cells.

Implementing a Multiplexed System of Detectors for Higher Photon Counting Rates

DTIC Science & Technology

2007-01-01

D1 D2 Fig. 3. (a) Setup for testing different arrangements of InGaAs SPAD assemblies; (b) three different InGaAs SPAD assemblies; ( c ) schematic of...presently available, either commercial or prototype, the deadtimes range from ≈50 ns for actively quenched single photon avalanche detectors ( SPADs ...to ≈10 µs for passively quenched SPADs , although even actively quenched SPADs sometimes employ µs deadtimes to avoid excessive afterpulsing rates. In

Laser-Based Measurements of OH, Temperature, and Water Vapor Concentration in a Hydrocarbon-Fueled Scramjet (POSTPRINT)

DTIC Science & Technology

2008-07-01

hours. The detector signals are post-processed with a software lock-in amplifier to recover the WMS-1f and WMS-2f signals. The TDLAS sensor utilizes...Figure 6. Schematic of TDLAS sensor for temperature and water vapor concentration. Fiber Diode lasers Grating Fiber Detectors Demultiplexer Multiplexer...within the combustor. Tunable diode laser- based absorption spectroscopy ( TDLAS ) is used to measure water vapor concentration and static temperature near

Heterojunction-Internal-Photoemission Infrared Detectors

NASA Technical Reports Server (NTRS)

Maserjian, Joseph

1991-01-01

New type of photodetector adds options for design of imaging devices. Heterojunction-internal-photoemission (HIP) infrared photodetectors proposed for incorporation into planar arrays in imaging devices required to function well at wavelengths from 8 to 17 micrometers and at temperatures above 65 K. Photoexcited electrons cross energy barrier at heterojunction and swept toward collection layer. Array of such detectors made by etching mesa structures. HIP layers stacked to increase quantum efficiency. Also built into integrated circuits including silicon multiplexer/readout circuits.

A Silicon SPECT System for Molecular Imaging of the Mouse Brain.

PubMed

Shokouhi, Sepideh; Fritz, Mark A; McDonald, Benjamin S; Durko, Heather L; Furenlid, Lars R; Wilson, Donald W; Peterson, Todd E

2007-01-01

We previously demonstrated the feasibility of using silicon double-sided strip detectors (DSSDs) for SPECT imaging of the activity distribution of iodine-125 using a 300-micrometer thick detector. Based on this experience, we now have developed fully customized silicon DSSDs and associated readout electronics with the intent of developing a multi-pinhole SPECT system. Each DSSD has a 60.4 mm × 60.4 mm active area and is 1 mm thick. The strip pitch is 59 micrometers, and the readout of the 1024 strips on each side gives rise to a detector with over one million pixels. Combining four high-resolution DSSDs into a SPECT system offers an unprecedented space-bandwidth product for the imaging of single-photon emitters. The system consists of two camera heads with two silicon detectors stacked one behind the other in each head. The collimator has a focused pinhole system with cylindrical-shaped pinholes that are laser-drilled in a 250 μm tungsten plate. The unique ability to collect projection data at two magnifications simultaneously allows for multiplexed data at high resolution to be combined with lower magnification data with little or no multiplexing. With the current multi-pinhole collimator design, our SPECT system will be capable of offering high spatial resolution, sensitivity and angular sampling for small field-of-view applications, such as molecular imaging of the mouse brain.

Operation and performance of new NIR detectors from SELEX

NASA Astrophysics Data System (ADS)

Atkinson, D.; Bezawada, N.; Hipwood, L. G.; Shorrocks, N.; Milne, H.

2012-07-01

The European Space Agency (ESA) has funded SELEX Galileo, Southampton, UK to develop large format near infrared (NIR) detectors for its future space and ground based programmes. The UKATC has worked in collaboration with SELEX Galileo to test and characterise the new detectors produced during phase-1 of the development. In order to demonstrate the detector material performance, the HgCdTe (MCT) detector diodes (grown on GaAs substrate through MOVPE process in small 320×256, 24μm pixel format) are hybridised to the existing SELEX Galileo SWALLOW CMOS readout chip. The substrate removed and MCT thinned detector arrays were then tested and evaluated at the UKATC following screening tests at SELEX. This paper briefly describes the test setup, the operational aspects of the readout multiplexer and presents the performance parameters of the detector arrays including: conversion gain, detector dark current, read noise, linearity, quantum efficiency and persistence for various detector temperatures between 80K and 140K.

Peptide library synthesis on spectrally encoded beads for multiplexed protein/peptide bioassays

NASA Astrophysics Data System (ADS)

Nguyen, Huy Q.; Brower, Kara; Harink, Björn; Baxter, Brian; Thorn, Kurt S.; Fordyce, Polly M.

2017-02-01

Protein-peptide interactions are essential for cellular responses. Despite their importance, these interactions remain largely uncharacterized due to experimental challenges associated with their measurement. Current techniques (e.g. surface plasmon resonance, fluorescence polarization, and isothermal calorimetry) either require large amounts of purified material or direct fluorescent labeling, making high-throughput measurements laborious and expensive. In this report, we present a new technology for measuring antibody-peptide interactions in vitro that leverages spectrally encoded beads for biological multiplexing. Specific peptide sequences are synthesized directly on encoded beads with a 1:1 relationship between peptide sequence and embedded code, thereby making it possible to track many peptide sequences throughout the course of an experiment within a single small volume. We demonstrate the potential of these bead-bound peptide libraries by: (1) creating a set of 46 peptides composed of 3 commonly used epitope tags (myc, FLAG, and HA) and single amino-acid scanning mutants; (2) incubating with a mixture of fluorescently-labeled antimyc, anti-FLAG, and anti-HA antibodies; and (3) imaging these bead-bound libraries to simultaneously identify the embedded spectral code (and thus the sequence of the associated peptide) and quantify the amount of each antibody bound. To our knowledge, these data demonstrate the first customized peptide library synthesized directly on spectrally encoded beads. While the implementation of the technology provided here is a high-affinity antibody/protein interaction with a small code space, we believe this platform can be broadly applicable to any range of peptide screening applications, with the capability to multiplex into libraries of hundreds to thousands of peptides in a single assay.

Parallel-multiplexed excitation light-sheet microscopy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Xu, Dongli; Zhou, Weibin; Peng, Leilei

2017-02-01

Laser scanning light-sheet imaging allows fast 3D image of live samples with minimal bleach and photo-toxicity. Existing light-sheet techniques have very limited capability in multi-label imaging. Hyper-spectral imaging is needed to unmix commonly used fluorescent proteins with large spectral overlaps. However, the challenge is how to perform hyper-spectral imaging without sacrificing the image speed, so that dynamic and complex events can be captured live. We report wavelength-encoded structured illumination light sheet imaging (λ-SIM light-sheet), a novel light-sheet technique that is capable of parallel multiplexing in multiple excitation-emission spectral channels. λ-SIM light-sheet captures images of all possible excitation-emission channels in true parallel. It does not require compromising the imaging speed and is capable of distinguish labels by both excitation and emission spectral properties, which facilitates unmixing fluorescent labels with overlapping spectral peaks and will allow more labels being used together. We build a hyper-spectral light-sheet microscope that combined λ-SIM with an extended field of view through Bessel beam illumination. The system has a 250-micron-wide field of view and confocal level resolution. The microscope, equipped with multiple laser lines and an unlimited number of spectral channels, can potentially image up to 6 commonly used fluorescent proteins from blue to red. Results from in vivo imaging of live zebrafish embryos expressing various genetic markers and sensors will be shown. Hyper-spectral images from λ-SIM light-sheet will allow multiplexed and dynamic functional imaging in live tissue and animals.

Quantitative performance measurements of bent crystal Laue analyzers for X-ray fluorescence spectroscopy.

PubMed

Karanfil, C; Bunker, G; Newville, M; Segre, C U; Chapman, D

2012-05-01

Third-generation synchrotron radiation sources pose difficult challenges for energy-dispersive detectors for XAFS because of their count rate limitations. One solution to this problem is the bent crystal Laue analyzer (BCLA), which removes most of the undesired scatter and fluorescence before it reaches the detector, effectively eliminating detector saturation due to background. In this paper experimental measurements of BCLA performance in conjunction with a 13-element germanium detector, and a quantitative analysis of the signal-to-noise improvement of BCLAs are presented. The performance of BCLAs are compared with filters and slits.

Parallel detecting super-resolution microscopy using correlation based image restoration

NASA Astrophysics Data System (ADS)

Yu, Zhongzhi; Liu, Shaocong; Zhu, Dazhao; Kuang, Cuifang; Liu, Xu

2017-12-01

A novel approach to achieve the image restoration is proposed in which each detector's relative position in the detector array is no longer a necessity. We can identify each detector's relative location by extracting a certain area from one of the detector's image and scanning it on other detectors' images. According to this location, we can generate the point spread functions (PSF) for each detector and perform deconvolution for image restoration. Equipped with this method, the microscope with discretionally designed detector array can be easily constructed without the concern of exact relative locations of detectors. The simulated results and experimental results show the total improvement in resolution with a factor of 1.7 compared to conventional confocal fluorescence microscopy. With the significant enhancement in resolution and easiness for application of this method, this novel method should have potential for a wide range of application in fluorescence microscopy based on parallel detecting.

Simultaneous detection of eight avian influenza A virus subtypes by multiplex reverse transcription-PCR using a GeXP analyser.

PubMed

Li, Meng; Xie, Zhixun; Xie, Zhiqin; Liu, Jiabo; Xie, Liji; Deng, Xianwen; Luo, Sisi; Fan, Qing; Huang, Li; Huang, Jiaoling; Zhang, Yanfang; Zeng, Tingting; Wang, Sheng

2018-04-18

Recent studies have demonstrated that at least eight subtypes of avian influenza virus (AIV) can infect humans, including H1, H2, H3, H5, H6, H7, H9 and H10. A GeXP analyser-based multiplex reverse transcription (RT)-PCR (GeXP-multiplex RT-PCR) assay was developed in our recent studies to simultaneously detect these eight AIV subtypes using the haemagglutinin (HA) gene. The assay consists of chimeric primer-based PCR amplification with fluorescent labelling and capillary electrophoresis separation. RNA was extracted from chick embryo allantoic fluid or liquid cultures of viral isolates. In addition, RNA synthesised via in vitro transcription was used to determine the specificity and sensitivity of the assay. After selecting the primer pairs, their concentrations and GeXP-multiplex RT-PCR conditions were optimised. The established GeXP-multiplex RT-PCR assay can detect as few as 100 copies of premixed RNA templates. In the present study, 120 clinical specimens collected from domestic poultry at live bird markets and from wild birds were used to evaluate the performance of the assay. The GeXP-multiplex RT-PCR assay specificity was the same as that of conventional RT-PCR. Thus, the GeXP-multiplex RT-PCR assay is a rapid and relatively high-throughput method for detecting and identifying eight AIV subtypes that may infect humans.

Multiplexing a high-throughput liability assay to leverage efficiencies.

PubMed

Herbst, John; Anthony, Monique; Stewart, Jeremy; Connors, David; Chen, Taosheng; Banks, Martyn; Petrillo, Edward W; Agler, Michele

2009-06-01

In order to identify potential cytochrome P-450 3A4 (drug-metabolizing enzyme) inducers at an early stage of the drug discovery process, a cell-based transactivation high-throughput luciferase reporter assay for the human pregnane X receptor (PXR) in HepG2 cells has been implemented and multiplexed with a viability end point for data interpretation, as part of a Lead Profiling portfolio of assays. As a routine part of Lead Profiling operations, assays are periodically evaluated for utility as well as for potential improvements in technology or process. We used a recent evaluation of our PXR-transactivation assay as a model for the application of Lean Thinking-based process analysis to lab-bench assay optimization and automation. This resulted in the development of a 384-well multiplexed homogeneous assay simultaneously detecting PXR transactivation and HepG2 cell cytotoxicity. In order to multiplex fluorescent and luminescent read-outs, modifications to each assay were necessary, which included optimization of multiple assay parameters such as cell density, plate type, and reagent concentrations. Subsequently, a set of compounds including known cytotoxic compounds and PXR inducers were used to validate the multiplexed assay. Results from the multiplexed assay correlate well with those from the singleplexed assay formats measuring PXR transactivation and viability separately. Implementation of the multiplexed assay for routine compound profiling provides improved data quality, sample conservation, cost savings, and resource efficiencies.

Measuring the electron neutrino mass with improved sensitivity: the HOLMES experiment

NASA Astrophysics Data System (ADS)

Giachero, A.; Alpert, B. K.; Becker, D. T.; Bennett, D. A.; Biasotti, M.; Brofferio, C.; Ceriale, V.; Ceruti, G.; Corsini, D.; Day, P. K.; De Gerone, M.; Dressler, R.; Faverzani, M.; Ferri, E.; Fowler, J. W.; Fumagalli, E.; Gallucci, G.; Gard, J. D.; Gatti, F.; Hays-Wehle, J. P.; Heinitz, S.; Hilton, G. C.; Köster, U.; Lusignoli, M.; Mates, J. A. B.; Nisi, S.; Nucciotti, A.; Orlando, A.; Parodi, L.; Pessina, G.; Pizzigoni, G.; Puiu, A.; Ragazzi, S.; Reintsema, C. D.; Ribeiro Gomes, M.; Schmidt, D. R.; Schumann, D.; Siccardi, F.; Sisti, M.; Swetz, D. S.; Terranova, F.; Ullom, J. N.; Vale, L. R.

2017-02-01

HOLMES is a new experiment aiming at directly measuring the neutrino mass with a sensitivity below 2 eV . HOLMES will perform a calorimetric measurement of the energy released in the decay of 163Ho. The calorimetric measurement eliminates systematic uncertainties arising from the use of external beta sources, as in experiments with spectrometers. This measurement was proposed in 1982 by A. De Rujula and M. Lusignoli, but only recently the detector technological progress has allowed to design a sensitive experiment. HOLMES will deploy a 1000 pixels array of low temperature microcalorimeters with implanted 163Ho nuclei. HOLMES, besides being an important step forward in the direct neutrino mass measurement with a calorimetric approach, will also establish the potential of this approach to extend the sensitivity down to 0.1 eV and lower. The detectors used for the HOLMES experiment will be Mo/Cu bilayers TESs (Transition Edge Sensors) on SiNx membrane with gold absorbers. Microwave multiplexed rf-SQUIDs are the best available technique to read out large array of such detectors. An extensive R&D activity is in progress in order to maximize the multiplexing factor while preserving the performances of the individual detectors. To embed the 163Ho into the gold absorbers a custom mass separator ion implanter is being developed. The current activities are focused on the the single detector performances optimization and on the 163Ho isotope production and embedding. A preliminary measurement of a sub-array of 4× 16 detectors is planned late in 2017. In this contribution we present the HOLMES project with its technical challenges, its status and perspectives.

A kilo-pixel imaging system for future space based far-infrared observatories using microwave kinetic inductance detectors

NASA Astrophysics Data System (ADS)

Baselmans, J. J. A.; Bueno, J.; Yates, S. J. C.; Yurduseven, O.; Llombart, N.; Karatsu, K.; Baryshev, A. M.; Ferrari, L.; Endo, A.; Thoen, D. J.; de Visser, P. J.; Janssen, R. M. J.; Murugesan, V.; Driessen, E. F. C.; Coiffard, G.; Martin-Pintado, J.; Hargrave, P.; Griffin, M.

2017-05-01

Aims: Future astrophysics and cosmic microwave background space missions operating in the far-infrared to millimetre part of the spectrum will require very large arrays of ultra-sensitive detectors in combination with high multiplexing factors and efficient low-noise and low-power readout systems. We have developed a demonstrator system suitable for such applications. Methods: The system combines a 961 pixel imaging array based upon Microwave Kinetic Inductance Detectors (MKIDs) with a readout system capable of reading out all pixels simultaneously with only one readout cable pair and a single cryogenic amplifier. We evaluate, in a representative environment, the system performance in terms of sensitivity, dynamic range, optical efficiency, cosmic ray rejection, pixel-pixel crosstalk and overall yield at an observation centre frequency of 850 GHz and 20% fractional bandwidth. Results: The overall system has an excellent sensitivity, with an average detector sensitivity < NEPdet> =3×10-19 WHz measured using a thermal calibration source. At a loading power per pixel of 50 fW we demonstrate white, photon noise limited detector noise down to 300 mHz. The dynamic range would allow the detection of 1 Jy bright sources within the field of view without tuning the readout of the detectors. The expected dead time due to cosmic ray interactions, when operated in an L2 or a similar far-Earth orbit, is found to be <4%. Additionally, the achieved pixel yield is 83% and the crosstalk between the pixels is <-30 dB. Conclusions: This demonstrates that MKID technology can provide multiplexing ratios on the order of a 1000 with state-of-the-art single pixel performance, and that the technology is now mature enough to be considered for future space based observatories and experiments.

Experimental study of a depth-encoding PET detector inserting horizontal-striped glass between crystal layers

NASA Astrophysics Data System (ADS)

Yang, J.; Kim, K. B.; Choi, Y.; Kang, J.

2018-04-01

A depth-encoding positron emission tomography (PET) detector inserting a horizontal-striped glass between pixilated scintillation crystal layers was developed and experimentally evaluated. The detector consists of 2-layers of 4×4 LYSO array arranged with a 3.37 mm pitch. Horizontal-striped glasses with 1×4 array with different thickness of 3, 4 and 5 mm were inserted between top- and bottom-crystal layers. Bottom surface of bottom-layer was optically coupled to a 4×4 GAPD array. Sixteen output signals from DOI-PET detector were multiplexed by modified resistive charge division (RCD) networks and multiplexed signals were fed into custom-made charge-sensitive preamplifiers. The four amplified signals were digitized and recorded by the custom-made DAQ system based on FPGA. The four digitized outputs were post-processed and converted to flood histograms for each interaction event. Experimental results revealed that all crystal pixels were clearly identified on the 2D flood histogram without overlapping. Patterns of the 2D flood histogram were constituted with arrangements of [bottom–top–bottom–top–\\ldots–top–bottom–top–bottom] crystal responses in X-direction. These could be achieved by employing horizontal-striped glass that controlled the extent of light dispersion towards the X-direction in crystal layers for generation of a different position mapping for each layer and the modified RCD network that controls degree of charge sharing in readout electronics for reduction of identification error. This study demonstrated the proposed DOI-PET detector can extract the 3D γ-ray interaction position without considerable performance degradation of PET detector from the 2D flood histogram.

Holmes

NASA Astrophysics Data System (ADS)

Faverzani, M.; HOLMES Collaboration

2017-09-01

The experiment HOLMES, founded by the European Research Council, will perform a calorimetric measurement of the energy released in the electron capture of 163Ho to directly measure the neutrino mass with a sensitivity of ˜ 1 eV. This approach allows to eliminate the problematics connected to the use of external sources and the systematic uncertainties arising from decays on excited states. Such measurement will be performed with low temperature thermal detectors, where the decay energy is converted into a temperature signal measured by sensitive thermometers. HOLMES, besides of being an important step forward in the direct neutrino mass measurement with a calorimetric approach, will also establish the potential of this approach to extend the sensitivity down to 0.1 eV and lower. The best configuration has been defined with Monte Carlo simulations: HOLMES will collect about 3 × 1013 decays with 1000 detectors characterized by an instrumental energy resolution of the order of the eV and a time resolution of few microseconds. For a measuring time of 3 years, this translates in a total required 163Ho activity of about 300 kBq, equivalent to about 6.5 × 1016 163Ho nuclei, or 18 µg. The HOLMES detectors will have 163Ho implanted into Gold absorber coupled to Transition Edge Sensors, which will be read using microwave multiplexed rf-SQUIDs in combination with a ROACH2 based acquisition system. An extensive R&D activity is in progress in order to maximize the multiplexing factor while preserving the performances of the individual detectors. R&D activities aimed at optimizing the single detector performances, the 163Ho isotope production and embedding are in progress and will converge in a preliminary measurement of an array of 16 detectors planned by the end of 2016. We outline here the HOLMES project with its technical challenges, its status and perspectives.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sutherland, John C.

In this article, a method for obtaining fluorescence polarization data from an instrument designed to measure circular and linear dichroism is compared with a previously reported approach. The new method places a polarizer between the sample and a detector mounted perpendicular to the direction of the incident beam and results in determination of the fluorescence polarization ratio, whereas the previous method does not use a polarizer and yields the fluorescence anisotropy. A similar analysis with the detector located axially with the excitation beam demonstrates that there is no frequency modulated signal due to fluorescence polarization in the absence of amore » polarizer.« less

Metal Stable Isotope Tagging: Renaissance of Radioimmunoassay for Multiplex and Absolute Quantification of Biomolecules.

PubMed

Liu, Rui; Zhang, Shixi; Wei, Chao; Xing, Zhi; Zhang, Sichun; Zhang, Xinrong

2016-05-17

The unambiguous quantification of biomolecules is of great significance in fundamental biological research as well as practical clinical diagnosis. Due to the lack of a detectable moiety, the direct and highly sensitive quantification of biomolecules is often a "mission impossible". Consequently, tagging strategies to introduce detectable moieties for labeling target biomolecules were invented, which had a long and significant impact on studies of biomolecules in the past decades. For instance, immunoassays have been developed with radioisotope tagging by Yalow and Berson in the late 1950s. The later languishment of this technology can be almost exclusively ascribed to the use of radioactive isotopes, which led to the development of nonradioactive tagging strategy-based assays such as enzyme-linked immunosorbent assay, fluorescent immunoassay, and chemiluminescent and electrochemiluminescent immunoassay. Despite great success, these strategies suffered from drawbacks such as limited spectral window capacity for multiplex detection and inability to provide absolute quantification of biomolecules. After recalling the sequences of tagging strategies, an apparent question is why not use stable isotopes from the start? A reasonable explanation is the lack of reliable means for accurate and precise quantification of stable isotopes at that time. The situation has changed greatly at present, since several atomic mass spectrometric measures for metal stable isotopes have been developed. Among the newly developed techniques, inductively coupled plasma mass spectrometry is an ideal technique to determine metal stable isotope-tagged biomolecules, for its high sensitivity, wide dynamic linear range, and more importantly multiplex and absolute quantification ability. Since the first published report by our group, metal stable isotope tagging has become a revolutionary technique and gained great success in biomolecule quantification. An exciting research highlight in this area is the development and application of the mass cytometer, which fully exploited the multiplexing potential of metal stable isotope tagging. It realized the simultaneous detection of dozens of parameters in single cells, accurate immunophenotyping in cell populations, through modeling of intracellular signaling network and undoubted discrimination of function and connection of cell subsets. Metal stable isotope tagging has great potential applications in hematopoiesis, immunology, stem cells, cancer, and drug screening related research and opened a post-fluorescence era of cytometry. Herein, we review the development of biomolecule quantification using metal stable isotope tagging. Particularly, the power of multiplex and absolute quantification is demonstrated. We address the advantages, applicable situations, and limitations of metal stable isotope tagging strategies and propose suggestions for future developments. The transfer of enzymatic or fluorescent tagging to metal stable isotope tagging may occur in many aspects of biological and clinical practices in the near future, just as the revolution from radioactive isotope tagging to fluorescent tagging happened in the past.

Multiplex quantification of four DNA targets in one reaction with Bio-Rad droplet digital PCR system for GMO detection

PubMed Central

Dobnik, David; Štebih, Dejan; Blejec, Andrej; Morisset, Dany; Žel, Jana

2016-01-01

The advantages of the digital PCR technology are already well documented until now. One way to achieve better cost efficiency of the technique is to use it in a multiplexing strategy. Droplet digital PCR platforms, which include two fluorescence filters, support at least duplex reactions and with some developments and optimization higher multiplexing is possible. The present study not only shows a development of multiplex assays in droplet digital PCR, but also presents a first thorough evaluation of several parameters in such multiplex digital PCR. Two 4-plex assays were developed for quantification of 8 different DNA targets (7 genetically modified maize events and maize endogene). Per assay, two of the targets were labelled with one fluorophore and two with another. As current analysis software does not support analysis of more than duplex, a new R- and Shiny-based web application analysis tool (http://bit.ly/ddPCRmulti) was developed that automates the analysis of 4-plex results. In conclusion, the two developed multiplex assays are suitable for quantification of GMO maize events and the same approach can be used in any other field with a need for accurate and reliable quantification of multiple DNA targets. PMID:27739510

Large-area, low-noise, high-speed, photodiode-based fluorescence detectors with fast overdrive recovery

NASA Astrophysics Data System (ADS)

Bickman, S.; DeMille, D.

2005-11-01

Two large-area, low-noise, high-speed fluorescence detectors have been built. One detector consists of a photodiode with an area of 28mm×28mm and a low-noise transimpedance amplifier. This detector has a input light-equivalent spectral noise density of less than 3pW/√Hz , can recover from a large scattered light pulse within 10μs, and has a bandwidth of at least 900 kHz. The second detector consists of a 16-mm-diam avalanche photodiode and a low-noise transimpedance amplifier. This detector has an input light-equivalent spectral noise density of 0.08pW/√Hz , also can recover from a large scattered light pulse within 10μs, and has a bandwidth of 1 MHz.

Characterization of Geiger mode avalanche photodiodes for fluorescence decay measurements

NASA Astrophysics Data System (ADS)

Jackson, John C.; Phelan, Don; Morrison, Alan P.; Redfern, R. Michael; Mathewson, Alan

2002-05-01

Geiger mode avalanche photodiodes (APD) can be biased above the breakdown voltage to allow detection of single photons. Because of the increase in quantum efficiency, magnetic field immunity, robustness, longer operating lifetime and reduction in costs, solid-state detectors capable of operating at non-cryogenic temperatures and providing single photon detection capabilities provide attractive alternatives to the photomultiplier tube (PMT). Shallow junction Geiger mode APD detectors provide the ability to manufacture photon detectors and detector arrays with CMOS compatible processing steps and allows the use of novel Silicon-on-Insulator(SoI) technology to provide future integrated sensing solutions. Previous work on Geiger mode APD detectors has focused on increasing the active area of the detector to make it more PMT like, easing the integration of discrete reaction, detection and signal processing into laboratory experimental systems. This discrete model for single photon detection works well for laboratory sized test and measurement equipment, however the move towards microfluidics and systems on a chip requires integrated sensing solutions. As we move towards providing integrated functionality of increasingly nanoscopic sized emissions, small area detectors and detector arrays that can be easily integrated into marketable systems, with sensitive small area single photon counting detectors will be needed. This paper will demonstrate the 2-dimensional and 3-dimensional simulation of optical coupling that occurs in Geiger mode APDs. Fabricated Geiger mode APD detectors optimized for fluorescence decay measurements were characterized and preliminary results show excellent results for their integration into fluorescence decay measurement systems.

Simultaneous two-dimensional laser-induced-fluorescence measurements of argon ions.

PubMed

Hansen, A K; Galante, Matthew; McCarren, Dustin; Sears, Stephanie; Scime, E E

2010-10-01

Recent laser upgrades on the Hot Helicon Experiment at West Virginia University have enabled multiplexed simultaneous measurements of the ion velocity distribution function at a single location, expanding our capabilities in laser-induced fluorescence diagnostics. The laser output is split into two beams, each modulated with an optical chopper and injected perpendicular and parallel to the magnetic field. Light from the crossing point of the beams is transported to a narrow-band photomultiplier tube filtered at the fluorescence wavelength and monitored by two lock-in amplifiers, each referenced to one of the two chopper frequencies.

Developing a new nonbinary SNP fluorescent multiplex detection system for forensic application in China.

PubMed

Liu, Yanfang; Liao, Huidan; Liu, Ying; Guo, Juanjuan; Sun, Yi; Fu, Xiaoliang; Xiao, Ding; Cai, Jifeng; Lan, Lingmei; Xie, Pingli; Zha, Lagabaiyila

2017-04-01

Nonbinary single-nucleotide polymorphisms (SNPs) are potential forensic genetic markers because their discrimination power is greater than that of normal binary SNPs, and that they can detect highly degraded samples. We previously developed a nonbinary SNP multiplex typing assay. In this study, we selected additional 20 nonbinary SNPs from the NCBI SNP database and verified them through pyrosequencing. These 20 nonbinary SNPs were analyzed using the fluorescent-labeled SNaPshot multiplex SNP typing method. The allele frequencies and genetic parameters of these 20 nonbinary SNPs were determined among 314 unrelated individuals from Han populations from China. The total power of discrimination was 0.9999999999994, and the cumulative probability of exclusion was 0.9986. Moreover, the result of the combination of this 20 nonbinary SNP assay with the 20 nonbinary SNP assay we previously developed demonstrated that the cumulative probability of exclusion of the 40 nonbinary SNPs was 0.999991 and that no significant linkage disequilibrium was observed in all 40 nonbinary SNPs. Thus, we concluded that this new system consisting of new 20 nonbinary SNPs could provide highly informative polymorphic data which would be further used in forensic application and would serve as a potentially valuable supplement to forensic DNA analysis. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Functional characterization of Gram-negative bacteria from different genera as multiplex cadmium biosensors.

PubMed

Bereza-Malcolm, Lara; Aracic, Sanja; Kannan, Ruban; Mann, Gülay; Franks, Ashley E

2017-08-15

Widespread presence of cadmium in soil and water systems is a consequence of industrial and agricultural processes. Subsequent accumulation of cadmium in food and drinking water can result in accidental consumption of dangerous concentrations. As such, cadmium environmental contamination poses a significant threat to human health. Development of microbial biosensors, as a novel alternative method for in situ cadmium detection, may reduce human exposure by complementing traditional analytical methods. In this study, a multiplex cadmium biosensing construct was assembled by cloning a single-output cadmium biosensor element, cadRgfp, and a constitutively expressed mrfp1 onto a broad-host range vector. Incorporation of the duplex fluorescent output [green and red fluorescence proteins] allowed measurement of biosensor functionality and viability. The biosensor construct was tested in several Gram-negative bacteria including Pseudomonas, Shewanella and Enterobacter. The multiplex cadmium biosensors were responsive to cadmium concentrations ranging from 0.01 to 10µgml -1 , as well as several other heavy metals, including arsenic, mercury and lead at similar concentrations. The biosensors were also responsive within 20-40min following exposure to 3µgml -1 cadmium. This study highlights the importance of testing biosensor constructs, developed using synthetic biology principles, in different bacterial genera. Copyright © 2017 Elsevier B.V. All rights reserved.

Multiplexed protein detection using antibody-conjugated microbead arrays in a microfabricated electrophoretic device

PubMed Central

Barbee, Kristopher D.; Hsiao, Alexander P.; Roller, Eric E.; Huang, Xiaohua

2011-01-01

We report the development of a microfabricated electrophoretic device for assembling high-density arrays of antibody-conjugated microbeads for chip-based protein detection. The device consists of a flow cell formed between a gold-coated silicon chip with an array of microwells etched in a silicon dioxide film and a glass coverslip with a series of thin gold counter electrode lines. We have demonstrated that 0.4 and 1 μm beads conjugated with antibodies can be rapidly assembled into the microwells by applying a pulsed electric field across the chamber. By assembling step-wise a mixture of fluorescently labeled antibody-conjugated microbeads, we incorporated both spatial and fluorescence encoding strategies to demonstrate significant multiplexing capabilities. We have shown that these antibody-conjugated microbead arrays can be used to perform on-chip sandwich immunoassays to detect test antigens at concentrations as low as 40 pM (6 ng/mL). A finite element model was also developed to examine the electric field distribution within the device for different counter electrode configurations over a range of line pitches and chamber heights. This device will be useful for assembling high-density, encoded antibody arrays for multiplexed detection of proteins and other types of protein-conjugated microbeads for applications such as the analysis of protein-protein interactions. PMID:20820631

Single transmission-line readout method for silicon photomultiplier based time-of-flight and depth-of-interaction PET

NASA Astrophysics Data System (ADS)

Ko, Guen Bae; Lee, Jae Sung

2017-03-01

We propose a novel single transmission-line readout method for whole-body time-of-flight positron emission tomography applications, without compromising on performance. The basic idea of the proposed multiplexing method is the addition of a specially prepared tag signal ahead of the scintillation pulse. The tag signal is a square pulse that encodes photon arrival time and channel information. The 2D position of a silicon photomultiplier (SiPM) array is encoded by the specific width and height of the tag signal. A summing amplifier merges the tag and scintillation signals of each channel, and the final output signal can be acquired with a one-channel digitizer. The feasibility and performance of the proposed method were evaluated using a 1:1 coupled detector consisting of 4  ×  4 array of LGSO crystals and 16 channel SiPM. The sixteen 3 mm LGSO crystals were clearly separated in the crystal-positioning map with high reliability. The average energy resolution and coincidence resolving time were 11.31  ±  0.55% and 264.7  ±  10.7 ps, respectively. We also proved that the proposed method does not degrade timing performance with increasing multiplexing ratio. The two types of LGSO crystals (L0.95GSO and L0.20GSO) in phoswich detector were also clearly identified with the high-reliability using pulse shape discrimination, thanks to the well-preserved pulse shape information. In conclusion, the proposed multiplexing method allows decoding of the 3D interaction position of gamma rays in the scintillation detector with single-line readout.

Single transmission-line readout method for silicon photomultiplier based time-of-flight and depth-of-interaction PET.

PubMed

Ko, Guen Bae; Lee, Jae Sung

2017-03-21

We propose a novel single transmission-line readout method for whole-body time-of-flight positron emission tomography applications, without compromising on performance. The basic idea of the proposed multiplexing method is the addition of a specially prepared tag signal ahead of the scintillation pulse. The tag signal is a square pulse that encodes photon arrival time and channel information. The 2D position of a silicon photomultiplier (SiPM) array is encoded by the specific width and height of the tag signal. A summing amplifier merges the tag and scintillation signals of each channel, and the final output signal can be acquired with a one-channel digitizer. The feasibility and performance of the proposed method were evaluated using a 1:1 coupled detector consisting of 4  ×  4 array of LGSO crystals and 16 channel SiPM. The sixteen 3 mm LGSO crystals were clearly separated in the crystal-positioning map with high reliability. The average energy resolution and coincidence resolving time were 11.31  ±  0.55% and 264.7  ±  10.7 ps, respectively. We also proved that the proposed method does not degrade timing performance with increasing multiplexing ratio. The two types of LGSO crystals (L 0.95 GSO and L 0.20 GSO) in phoswich detector were also clearly identified with the high-reliability using pulse shape discrimination, thanks to the well-preserved pulse shape information. In conclusion, the proposed multiplexing method allows decoding of the 3D interaction position of gamma rays in the scintillation detector with single-line readout.

Fabrication of the pinhole aperture for AdaptiSPECT

PubMed Central

Kovalsky, Stephen; Kupinski, Matthew A.; Barrett, Harrison H.; Furenlid, Lars R.

2015-01-01

AdaptiSPECT is a pre-clinical pinhole SPECT imaging system under final construction at the Center for Gamma-Ray Imaging. The system is designed to be able to autonomously change its imaging configuration. The system comprises 16 detectors mounted on translational stages to move radially away and towards the center of the field-of-view. The system also possesses an adaptive pinhole aperture with multiple collimator diameters and pinhole sizes, as well as the possibility to switch between multiplexed and non-multiplexed imaging configurations. In this paper, we describe the fabrication of the AdaptiSPECT pinhole aperture and its controllers. PMID:26146443

Analysis of synthetic derivatives of peptide hormones by capillary zone electrophoresis and micellar electrokinetic chromatography with ultraviolet-absorption and laser-induced fluorescence detection.

PubMed

Solínová, Veronika; Kasicka, Václav; Koval, Dusan; Barth, Tomislav; Ciencialová, Alice; Záková, Lenka

2004-08-25

Capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC) were used for the analysis of new synthetic derivatives of hypophysis neurohormones--vasopressin and oxytocin, and pancreatic hormone--human insulin (HI) and its octapeptide fragment, derivatized by fluorescent probe, 4-chloro-7-nitrobenzo[1,2,5]oxadiazol (NBD). The suitable composition of background electrolytes (BGEs) was selected on the basis of calculated pH dependence of effective charge of analyzed peptides. Basic ionogenic peptides were analyzed by CZE in the acidic BGE composed of 100 mM H3PO4, 50 mM Tris, pH 2.25. The ionogenic peptides with fluorescent label, NBD, were analyzed in 0.5 M acetic acid, pH 2.5. The best MEKC separation of non-ionogenic peptides was achieved in alkaline BGE, 20 mM Tris, 5 mM H3PO4, with micellar pseudophase formed by 50 mM sodium dodecylsulfate (SDS), pH 8.8. Selected characteristics (noise, detectability of substance, sensitivity of detector) of the UV-absorption detectors (single wavelength detector, multiple-wavelength photodiode array detector (PDA), both of them operating at constant wavelength 206 nm) and laser-induced fluorescence (LIF) detector (excitation/emission wavelength 488/520 nm) were determined. The detectability of peptides in the single wavelength detector was 1.3-6.0 micromol dm(-3) and in the PDA detector 1.6-3.1 micromol dm(-3). The LIF detection was more sensitive, the applied concentration of NBD derivative of insulin fragment in CZE analysis with LIF detection was three orders lower than in CZE with UV-absorption detector, and the detectability of this peptide was improved to 15.8 nmol dm(-3).

Effect of time discretization of the imaging process on the accuracy of trajectory estimation in fluorescence microscopy

PubMed Central

Wong, Yau; Chao, Jerry; Lin, Zhiping; Ober, Raimund J.

2014-01-01

In fluorescence microscopy, high-speed imaging is often necessary for the proper visualization and analysis of fast subcellular dynamics. Here, we examine how the speed of image acquisition affects the accuracy with which parameters such as the starting position and speed of a microscopic non-stationary fluorescent object can be estimated from the resulting image sequence. Specifically, we use a Fisher information-based performance bound to investigate the detector-dependent effect of frame rate on the accuracy of parameter estimation. We demonstrate that when a charge-coupled device detector is used, the estimation accuracy deteriorates as the frame rate increases beyond a point where the detector’s readout noise begins to overwhelm the low number of photons detected in each frame. In contrast, we show that when an electron-multiplying charge-coupled device (EMCCD) detector is used, the estimation accuracy improves with increasing frame rate. In fact, at high frame rates where the low number of photons detected in each frame renders the fluorescent object difficult to detect visually, imaging with an EMCCD detector represents a natural implementation of the Ultrahigh Accuracy Imaging Modality, and enables estimation with an accuracy approaching that which is attainable only when a hypothetical noiseless detector is used. PMID:25321248

Detector Having A Transmission Grating Beam Splitter For Multi-Wavelength Sample Analysis.

DOEpatents

Liu, Changsheng; Li, Qingbo

2000-09-12

A detector for DNA sample identification is provided with a transmission grating beam splitter (TGBS). The TGBS split fluoresced light from a tagged DNA sample into 0th order and a 1st order components, both of which are detected on a two-dimensional detector array of a CCD camera. The 0th and 1st order components are detected along a column of pixels in the detector array, and are spaced apart from one another. The DNA samples are tagged with four fluorescent dyes, one dye specific for each nucleotide, and all four dyes responding in slightly different manner to the same monochromatic excitation signal. The TGBS splits fluoresced incoming light into 0th and 1st order components, which are then spread out among a number of pixels in the detector array. The 1st component of this light is received by pixels whose position relative to the 0th order component depends on the frequency of fluorescence. Thus, the position at which signal energy is detected on the array is indicative of the particular dye, and therefore, the corresponding nucleotide tagged by that dye. Monitoring signal energy at the 0th order pixel and selected 1st order pixels, provides a set of data from which one may then identify the particular nucleotide.

Detector Having A Transmission Grating Beam Splitter For Multi-Wavelength.

DOEpatents

Liu, Changsheng; Li, Qingbo (State College, PA

1999-12-07

A detector for DNA sample identification is provided with a transmission grating beam splitter (TGBS). The TGBS split fluoresced light from a tagged DNA sample into 0th order and a 1st order components, both of which are detected on a two-dimensional detector array of a CCD camera. The 0th and 1st order components are detected along a column of pixels in the detector array, and are spaced apart from one another. The DNA samples are tagged with four fluorescent dyes, one dye specific for each nucleotide, and all four dyes responding in slightly different manner to the same monochromatic excitation signal. The TGBS splits fluoresced incoming light into 0th and 1st order components, which are then spread out among a number of pixels in the detector array. The 1st component of this light is received by pixels whose position relative to the 0th order component depends on the frequency of fluorescence. Thus, the position at which signal energy is detected on the array is indicative of the particular dye, and therefore, the corresponding nucleotide tagged by that dye. Monitoring signal energy at the 0th order pixel and selected 1st order pixels, provides a set of data from which one may then identify the particular nucleotide.

Molecular identification of common Salmonella serovars using multiplex DNA sensor-based suspension array.

PubMed

Aydin, Muhsin; Carter-Conger, Jacqueline; Gao, Ning; Gilmore, David F; Ricke, Steven C; Ahn, Soohyoun

2018-04-01

Salmonella is one of major foodborne pathogens and the leading cause of foodborne illness-related hospitalizations and deaths. It is critical to develop a sensitive and rapid detection assay that can identify Salmonella to ensure food safety. In this study, a DNA sensor-based suspension array system of high multiplexing ability was developed to identify eight Salmonella serovars commonly associated with foodborne outbreaks to the serotype level. Each DNA sensor was prepared by activating pre-encoded microspheres with oligonucleotide probes that are targeting virulence genes and serovar-specific regions. The mixture of 12 different types of DNA sensors were loaded into a 96-well microplate and used as a 12-plex DNA sensor array platform. DNA isolated from Salmonella was amplified by multiplex polymerase chain reaction (mPCR), and the presence of Salmonella was determined by reading fluorescent signals from hybridization between probes on DNA sensors and fluorescently labeled target DNA using the Bio-Plex® system. The developed multiplex array was able to detect synthetic DNA at the concentration as low as 100 fM and various Salmonella serovars as low as 100 CFU/mL within 1 h post-PCR. Sensitivity of this assay was further improved to 1 CFU/mL with 6-h enrichment. The array system also correctly and specifically identified serotype of tested Salmonella strains without any cross-reactivity with other common foodborne pathogens. Our results indicate the developed DNA sensor suspension array can be a rapid and reliable high-throughput method for simultaneous detection and molecular identification of common Salmonella serotypes.

Charging YOYO-1 on Capillary Wall for Online DNA Intercalation and Integrating This Approach with Multiplex PCR and Bare Narrow Capillary–Hydrodynamic Chromatography for Online DNA Analysis

PubMed Central

2016-01-01

Multiplex polymerase chain reaction (PCR) has been widely utilized for high-throughput pathogen identification. Often, a dye is used to intercalate the amplified DNA fragments, and identifications of the pathogens are carried out by DNA melting curve analysis or gel electrophoresis. Integrating DNA amplification and identification is a logic path toward maximizing the benefit of multiplex PCR. Although PCR and gel electrophoresis have been integrated, replenishing the gels after each run is tedious and time-consuming. In this technical note, we develop an approach to address this issue. We perform multiplex PCR inside a capillary, transfer the amplified fragments to a bare narrow capillary, and measure their lengths online using bare narrow capillary–hydrodynamic chromatography (BaNC-HDC), a new technique recently developed in our laboratory for free-solution DNA separation. To intercalate the DNA with YOYO-1 (a fluorescent dye) for BaNC-HDC, we flush the capillary column with a YOYO-1 solution; positively charged YOYO-1 is adsorbed (or charged) onto the negatively charged capillary wall. As DNA molecules are driven down the column for separation, they react with the YOYO-1 stored on the capillary wall and are online-intercalated with the dye. With a single YOYO-1 charging, the column can be used for more than 40 runs, although the fluorescence signal intensities of the DNA peaks decrease gradually. Although the dye-DNA intercalation occurs during the separation, it does not affect the retention times, separation efficiencies, or resolutions. PMID:25555111

Quantitative and multiplexed detection for blood typing based on quantum dot-magnetic bead assay.

PubMed

Xu, Ting; Zhang, Qiang; Fan, Ya-Han; Li, Ru-Qing; Lu, Hua; Zhao, Shu-Ming; Jiang, Tian-Lun

2017-01-01

Accurate and reliable blood grouping is essential for safe blood transfusion. However, conventional methods are qualitative and use only single-antigen detection. We overcame these limitations by developing a simple, quantitative, and multiplexed detection method for blood grouping using quantum dots (QDs) and magnetic beads. In the QD fluorescence assay (QFA), blood group A and B antigens were quantified using QD labeling and magnetic beads, and the blood groups were identified according to the R value (the value was calculated with the fluorescence intensity from dual QD labeling) of A and B antigens. The optimized performance of QFA was established by blood typing 791 clinical samples. Quantitative and multiplexed detection for blood group antigens can be completed within 35 min with more than 10 5 red blood cells. When conditions are optimized, the assay performance is satisfactory for weak samples. The coefficients of variation between and within days were less than 10% and the reproducibility was good. The ABO blood groups of 791 clinical samples were identified by QFA, and the accuracy obtained was 100% compared with the tube test. Receiver-operating characteristic curves revealed that the QFA has high sensitivity and specificity toward clinical samples, and the cutoff points of the R value of A and B antigens were 1.483 and 1.576, respectively. In this study, we reported a novel quantitative and multiplexed method for the identification of ABO blood groups and presented an effective alternative for quantitative blood typing. This method can be used as an effective tool to improve blood typing and further guarantee clinical transfusion safety.

Development of an integrated endoscopic device for multiplexed low coherence interferometry measurements of microbicide gel coating thickness

NASA Astrophysics Data System (ADS)

Drake, Tyler K.; Robles, Francisco E.; DeSoto, Michael; Henderson, Marcus H.; Katz, David F.; Wax, Adam P.

2009-02-01

Microbicide gels are topical products that have recently been developed to combat sexually transmitted diseases including HIV/AIDS. The extent of gel coverage, thickness, and structure are crucial factors in gel effectiveness. It is necessary to be able to monitor gel distribution and behavior under various circumstances, such as coatis, and over an extended time scale in vivo. We have developed a multiplexed, Fourier-domain low coherence interferometry (LCI) system as a practical method of measuring microbicide gel distribution, with precision and accuracy comparable to currently used fluorometric techniques techniques. The multiplexed system achieved a broad scanning area without the need for a mechanical scanning device, typical of OCT systems, by utilizing six parallel channels with simultaneous data collection. We now propose an imaging module which will allow the integration of the multiplexed LCI system into the current fluorescence system in conjunction with an endoscope. The LCI imaging module will meet several key criteria in order to be compatible with the current system. The fluorescent system features a 4-mm diameter rigid endsoscope enclosed in a 27-mm diameter polycarbonate tube, with a water immersion tip. Therefore, the LCI module must be low-profile as well as water-resistant to fit inside the current design. It also must fulfill its primary function of delivering light from each of the six channels to the gel and collecting backscattered light. The performance of the imaging module will be characterized by scanning a calibration socket which contains grooves of known depths, and comparing these measurements to the fluorometric results.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, R.; Lu, R.; Gong, S.

We demonstrate a room-temperature semiconductor-based photodetector where readout is achieved using a resonant radio-frequency (RF) circuit consisting of a microstrip split-ring resonator coupled to a microstrip busline, fabricated on a semiconductor substrate. The RF resonant circuits are characterized at RF frequencies as function of resonator geometry, as well as for their response to incident IR radiation. The detectors are modeled analytically and using commercial simulation software, with good agreement to our experimental results. Though the detector sensitivity is weak, the detector architecture offers the potential for multiplexing arrays of detectors on a single read-out line, in addition to high speedmore » response for either direct coupling of optical signals to RF circuitry, or alternatively, carrier dynamics characterization of semiconductor, or other, material systems.« less

Saturated virtual fluorescence emission difference microscopy based on detector array

NASA Astrophysics Data System (ADS)

Liu, Shaocong; Sun, Shiyi; Kuang, Cuifang; Ge, Baoliang; Wang, Wensheng; Liu, Xu

2017-07-01

Virtual fluorescence emission difference microscopy (vFED) has been proposed recently to enhance the lateral resolution of confocal microscopy with a detector array, implemented by scanning a doughnut-shaped pattern. Theoretically, the resolution can be enhanced by around 1.3-fold compared with that in confocal microscopy. For further improvement of the resolving ability of vFED, a novel method is presented utilizing fluorescence saturation for super-resolution imaging, which we called saturated virtual fluorescence emission difference microscopy (svFED). With a point detector array, matched solid and hollow point spread functions (PSF) can be obtained by photon reassignment, and the difference results between them can be used to boost the transverse resolution. Results show that the diffraction barrier can be surpassed by at least 34% compared with that in vFED and the resolution is around 2-fold higher than that in confocal microscopy.

CALDER: Cryogenic light detectors for background-free searches

NASA Astrophysics Data System (ADS)

Di Domizio, S.; Bellini, F.; Cardani, L.; Casali, N.; Castellano, M. G.; Colantoni, I.; Cosmelli, C.; Cruciani, A.; D'Addabbo, A.; Martinez, M.; Minutolo, L.; Tomei, C.; Vignati, M.

2018-01-01

CALDER is a R&D project for the development of cryogenic light detectors with an active surface of 5x5cm2 and an energy resolution of 20 eV RMS for visible and UV photons. These devices can enhance the sensitivity of next generation large mass bolometric detectors for rare event searches, providing an active background rejection method based on particle discrimination. A CALDER detector is composed by a large area Si absorber substrate with superconducting kinetic inductance detectors (KIDs) deposited on it. The substrate converts the incoming light into athermal phonons, that are then sensed by the KIDs. KID technology combine fabrication simplicity with natural attitude to frequency-domain multiplexing, making it an ideal candidate for a large scale bolometric experiments. We will give an overview of the CALDER project and show the performances obtained with prototype detectors both in terms of energy resolution and efficiency.

Development of a Crosstalk Suppression Algorithm for KID Readout

NASA Astrophysics Data System (ADS)

Lee, Kyungmin; Ishitsuka, H.; Oguri, S.; Suzuki, J.; Tajima, O.; Tomita, N.; Won, Eunil; Yoshida, M.

2018-06-01

The GroundBIRD telescope aims to detect B-mode polarization of the cosmic microwave background radiation using the kinetic inductance detector array as a polarimeter. For the readout of the signal from detector array, we have developed a frequency division multiplexing readout system based on a digital down converter method. These techniques in general have the leakage problems caused by the crosstalks. The window function was applied in the field programmable gate arrays to mitigate the effect of these problems and tested it in algorithm level.

Sensors with centroid-based common sensing scheme and their multiplexing

NASA Astrophysics Data System (ADS)

Berkcan, Ertugrul; Tiemann, Jerome J.; Brooksby, Glen W.

1993-03-01

The ability to multiplex sensors with different measurands but with a common sensing scheme is of importance in aircraft and aircraft engine applications; this unification of the sensors into a common interface has major implications for weight, cost, and reliability. A new class of sensors based on a common sensing scheme and their E/O Interface has been developed. The approach detects the location of the centroid of a beam of light; the set of fiber optic sensors with this sensing scheme include linear and rotary position, temperature, pressure, as well as duct Mach number. The sensing scheme provides immunity to intensity variations of the source or due to environmental effects on the fiber. A detector spatially multiplexed common electro-optic interface for the sensors has been demonstrated with a position and a temperature sensor.

Fiber optical assembly for fluorescence spectrometry

DOEpatents

Carpenter, II, Robert W.; Rubenstein, Richard; Piltch, Martin; Gray, Perry

2010-12-07

A system for analyzing a sample for the presence of an analyte in a sample. The system includes a sample holder for containing the sample; an excitation source, such as a laser, and at least one linear array radially disposed about the sample holder. Radiation from the excitation source is directed to the sample, and the radiation induces fluorescent light in the sample. Each linear array includes a plurality of fused silica optical fibers that receive the fluorescent light and transmits a fluorescent light signal from the first end to an optical end port of the linear array. An end port assembly having a photo-detector is optically coupled to the optical end port. The photo-detector detects the fluorescent light signal and converts the fluorescent light signal into an electrical signal.

Multi-pinhole SPECT Imaging with Silicon Strip Detectors

PubMed Central

Peterson, Todd E.; Shokouhi, Sepideh; Furenlid, Lars R.; Wilson, Donald W.

2010-01-01

Silicon double-sided strip detectors offer outstanding instrinsic spatial resolution with reasonable detection efficiency for iodine-125 emissions. This spatial resolution allows for multiple-pinhole imaging at low magnification, minimizing the problem of multiplexing. We have conducted imaging studies using a prototype system that utilizes a detector of 300-micrometer thickness and 50-micrometer strip pitch together with a 23-pinhole collimator. These studies include an investigation of the synthetic-collimator imaging approach, which combines multiple-pinhole projections acquired at multiple magnifications to obtain tomographic reconstructions from limited-angle data using the ML-EM algorithm. Sub-millimeter spatial resolution was obtained, demonstrating the basic validity of this approach. PMID:20953300

Assessment of illumination conditions in a single-pixel imaging configuration

NASA Astrophysics Data System (ADS)

Garoi, Florin; Udrea, Cristian; Damian, Cristian; Logofǎtu, Petre C.; Colţuc, Daniela

2016-12-01

Single-pixel imaging based on multiplexing is a promising technique, especially in applications where 2D detectors or raster scanning imaging are not readily applicable. With this method, Hadamard masks are projected on a spatial light modulator to encode an incident scene and a signal is recorded at the photodiode detector for each of these masks. Ultimately, the image is reconstructed on the computer by applying the inverse transform matrix. Thus, various algorithms were optimized and several spatial light modulators already characterized for such a task. This work analyses the imaging quality of such a single-pixel arrangement, when various illumination conditions are used. More precisely, the main comparison is made between coherent and incoherent ("white light") illumination and between two multiplexing methods, namely Hadamard and Scanning. The quality of the images is assessed by calculating their SNR, using two relations. The results show better images are obtained with "white light" illumination for the first method and coherent one for the second.

Quantitative in vivo optical tomography of cancer progression & vasculature development in adult zebrafish

PubMed Central

Kumar, Sunil; Lockwood, Nicola; Ramel, Marie-Christine; Correia, Teresa; Ellis, Matthew; Alexandrov, Yuriy; Andrews, Natalie; Patel, Rachel; Bugeon, Laurence; Dallman, Margaret J.; Brandner, Sebastian; Arridge, Simon; Katan, Matilda; McGinty, James; Frankel, Paul; French, Paul M.W.

2016-01-01

We describe a novel approach to study tumour progression and vasculature development in vivo via global 3-D fluorescence imaging of live non-pigmented adult zebrafish utilising angularly multiplexed optical projection tomography with compressive sensing (CS-OPT). This “mesoscopic” imaging method bridges a gap between established ~μm resolution 3-D fluorescence microscopy techniques and ~mm-resolved whole body planar imaging and diffuse tomography. Implementing angular multiplexing with CS-OPT, we demonstrate the in vivo global imaging of an inducible fluorescently labelled genetic model of liver cancer in adult non-pigmented zebrafish that also present fluorescently labelled vasculature. In this disease model, addition of a chemical inducer (doxycycline) drives expression of eGFP tagged oncogenic K-RASV12 in the liver of immune competent animals. We show that our novel in vivo global imaging methodology enables non-invasive quantitative imaging of the development of tumour and vasculature throughout the progression of the disease, which we have validated against established methods of pathology including immunohistochemistry. We have also demonstrated its potential for longitudinal imaging through a study of vascular development in the same zebrafish from early embryo to adulthood. We believe that this instrument, together with its associated analysis and data management tools, constitute a new platform for in vivo cancer studies and drug discovery in zebrafish disease models. PMID:27259259

Multicolor fluorescent biosensor for multiplexed detection of DNA.

PubMed

Hu, Rong; Liu, Tao; Zhang, Xiao-Bing; Huan, Shuang-Yan; Wu, Cuichen; Fu, Ting; Tan, Weihong

2014-05-20

Development of efficient methods for highly sensitive and rapid screening of specific oligonucleotide sequences is essential to the early diagnosis of serious diseases. In this work, an aggregated cationic perylene diimide (PDI) derivative was found to efficiently quench the fluorescence emission of a variety of anionic oligonucleotide-labeled fluorophores that emit at wavelengths from the visible to NIR region. This broad-spectrum quencher was then adopted to develop a multicolor biosensor via a label-free approach for multiplexed fluorescent detection of DNA. The aggregated perylene derivative exhibits a very high quenching efficiency on all ssDNA-labeled dyes associated with biosensor detection, having efficiency values of 98.3 ± 0.9%, 97 ± 1.1%, and 98.2 ± 0.6% for FAM, TAMRA, and Cy5, respectively. An exonuclease-assisted autocatalytic target recycling amplification was also integrated into the sensing system. High quenching efficiency combined with autocatalytic target recycling amplification afforded the biosensor with high sensitivity toward target DNA, resulting in a detection limit of 20 pM, which is about 50-fold lower than that of traditional unamplified homogeneous fluorescent assay methods. The quencher did not interfere with the catalytic activity of nuclease, and the biosensor could be manipulated in either preaddition or postaddition manner with similar sensitivity. Moreover, the proposed sensing system allows for simultaneous and multicolor analysis of several oligonucleotides in homogeneous solution, demonstrating its potential application in the rapid screening of multiple biotargets.

Frequency division multiplexed multi-color fluorescence microscope system

NASA Astrophysics Data System (ADS)

Le, Vu Nam; Yang, Huai Dong; Zhang, Si Chun; Zhang, Xin Rong; Jin, Guo Fan

2017-10-01

Grayscale camera can only obtain gray scale image of object, while the multicolor imaging technology can obtain the color information to distinguish the sample structures which have the same shapes but in different colors. In fluorescence microscopy, the current method of multicolor imaging are flawed. Problem of these method is affecting the efficiency of fluorescence imaging, reducing the sampling rate of CCD etc. In this paper, we propose a novel multiple color fluorescence microscopy imaging method which based on the Frequency division multiplexing (FDM) technology, by modulating the excitation lights and demodulating the fluorescence signal in frequency domain. This method uses periodic functions with different frequency to modulate amplitude of each excitation lights, and then combine these beams for illumination in a fluorescence microscopy imaging system. The imaging system will detect a multicolor fluorescence image by a grayscale camera. During the data processing, the signal obtained by each pixel of the camera will be processed with discrete Fourier transform, decomposed by color in the frequency domain and then used inverse discrete Fourier transform. After using this process for signals from all of the pixels, monochrome images of each color on the image plane can be obtained and multicolor image is also acquired. Based on this method, this paper has constructed and set up a two-color fluorescence microscope system with two excitation wavelengths of 488 nm and 639 nm. By using this system to observe the linearly movement of two kinds of fluorescent microspheres, after the data processing, we obtain a two-color fluorescence dynamic video which is consistent with the original image. This experiment shows that the dynamic phenomenon of multicolor fluorescent biological samples can be generally observed by this method. Compared with the current methods, this method can obtain the image signals of each color at the same time, and the color video's frame rate is consistent with the frame rate of the camera. The optical system is simpler and does not need extra color separation element. In addition, this method has a good filtering effect on the ambient light or other light signals which are not affected by the modulation process.

Modular optical detector system

DOEpatents

Horn, Brent A [Livermore, CA; Renzi, Ronald F [Tracy, CA

2006-02-14

A modular optical detector system. The detector system is designed to detect the presence of molecules or molecular species by inducing fluorescence with exciting radiation and detecting the emitted fluorescence. Because the system is capable of accurately detecting and measuring picomolar concentrations it is ideally suited for use with microchemical analysis systems generally and capillary chromatographic systems in particular. By employing a modular design, the detector system provides both the ability to replace various elements of the detector system without requiring extensive realignment or recalibration of the components as well as minimal user interaction with the system. In addition, the modular concept provides for the use and addition of a wide variety of components, including optical elements (lenses and filters), light sources, and detection means, to fit particular needs.

Simple, Sensitive and Accurate Multiplex Detection of Clinically Important Melanoma DNA Mutations in Circulating Tumour DNA with SERS Nanotags

PubMed Central

Wee, Eugene J.H.; Wang, Yuling; Tsao, Simon Chang-Hao; Trau, Matt

2016-01-01

Sensitive and accurate identification of specific DNA mutations can influence clinical decisions. However accurate diagnosis from limiting samples such as circulating tumour DNA (ctDNA) is challenging. Current approaches based on fluorescence such as quantitative PCR (qPCR) and more recently, droplet digital PCR (ddPCR) have limitations in multiplex detection, sensitivity and the need for expensive specialized equipment. Herein we describe an assay capitalizing on the multiplexing and sensitivity benefits of surface-enhanced Raman spectroscopy (SERS) with the simplicity of standard PCR to address the limitations of current approaches. This proof-of-concept method could reproducibly detect as few as 0.1% (10 copies, CV < 9%) of target sequences thus demonstrating the high sensitivity of the method. The method was then applied to specifically detect three important melanoma mutations in multiplex. Finally, the PCR/SERS assay was used to genotype cell lines and ctDNA from serum samples where results subsequently validated with ddPCR. With ddPCR-like sensitivity and accuracy yet at the convenience of standard PCR, we believe this multiplex PCR/SERS method could find wide applications in both diagnostics and research. PMID:27446486

Simple, Sensitive and Accurate Multiplex Detection of Clinically Important Melanoma DNA Mutations in Circulating Tumour DNA with SERS Nanotags.

PubMed

Wee, Eugene J H; Wang, Yuling; Tsao, Simon Chang-Hao; Trau, Matt

2016-01-01

Sensitive and accurate identification of specific DNA mutations can influence clinical decisions. However accurate diagnosis from limiting samples such as circulating tumour DNA (ctDNA) is challenging. Current approaches based on fluorescence such as quantitative PCR (qPCR) and more recently, droplet digital PCR (ddPCR) have limitations in multiplex detection, sensitivity and the need for expensive specialized equipment. Herein we describe an assay capitalizing on the multiplexing and sensitivity benefits of surface-enhanced Raman spectroscopy (SERS) with the simplicity of standard PCR to address the limitations of current approaches. This proof-of-concept method could reproducibly detect as few as 0.1% (10 copies, CV < 9%) of target sequences thus demonstrating the high sensitivity of the method. The method was then applied to specifically detect three important melanoma mutations in multiplex. Finally, the PCR/SERS assay was used to genotype cell lines and ctDNA from serum samples where results subsequently validated with ddPCR. With ddPCR-like sensitivity and accuracy yet at the convenience of standard PCR, we believe this multiplex PCR/SERS method could find wide applications in both diagnostics and research.

Laser-induced fluorescence microscopic system using an optical parametric oscillator for tunable detection in microchip analysis.

PubMed

Kumemura, Momoko; Odake, Tamao; Korenaga, Takashi

2005-06-01

A laser-induced fluorescence microscopic system based on optical parametric oscillation has been constructed as a tunable detector for microchip analysis. The detection limit of sulforhodamine B (Ex. 520 nm, Em. 570 nm) was 0.2 mumol, which was approximately eight orders of magnitude better than with a conventional fluorophotometer. The system was applied to the determination of fluorescence-labeled DNA (Ex. 494 nm, Em. 519 nm) in a microchannel and the detection limit reached a single molecule. These results showed the feasibility of this system as a highly sensitive and tunable fluorescence detector for microchip analysis.

Theoretical considerations on the optogalvanic detection of laser induced fluorescence in atmospheric pressure atomizers

NASA Astrophysics Data System (ADS)

Omenetto, N.; Smith, B. W.; Winefordner, J. D.

1989-01-01

Several theoretical considerations are given on the potential and practical capabilities of a detector of fluorescence radiation whose operating principle is based on a multi-step excitation-ionization scheme involving the fluorescence photons as the first excitation step. This detection technique, which was first proposed by MATVEEVet al. [ Zh. Anal Khim.34, 846 (1979)], combines two independent atomizers, one analytical cell for the excitation of the sample fluorescence and one cell, filled with pure analyte atomic vapor, acting as the ionization detector. One laser beam excites the analyte fluorescence in the analytical cell and one (or two) laser beams are used to ionize the excited atoms in the detector. Several different causes of signal and noise are evaluated, together with a discussion on possible analytical atom reservoirs (flames, furnaces) and laser sources which could be used with this approach. For properly devised conditions, i.e. optical saturation of the fluorescence and unity ionization efficiency, detection limits well below pg/ml in solution and well below femtograms as absolute amounts in furnaces can be predicted. However, scattering problems, which are absent in a conventional laser-enhanced ionization set-up, may be important in this approach.

Multiplex quantification of 12 European Union authorized genetically modified maize lines with droplet digital polymerase chain reaction.

PubMed

Dobnik, David; Spilsberg, Bjørn; Bogožalec Košir, Alexandra; Holst-Jensen, Arne; Žel, Jana

2015-08-18

Presence of genetically modified organisms (GMO) in food and feed products is regulated in many countries. The European Union (EU) has implemented a threshold for labeling of products containing more than 0.9% of authorized GMOs per ingredient. As the number of GMOs has increased over time, standard-curve based simplex quantitative polymerase chain reaction (qPCR) analyses are no longer sufficiently cost-effective, despite widespread use of initial PCR based screenings. Newly developed GMO detection methods, also multiplex methods, are mostly focused on screening and detection but not quantification. On the basis of droplet digital PCR (ddPCR) technology, multiplex assays for quantification of all 12 EU authorized GM maize lines (per April first 2015) were developed. Because of high sequence similarity of some of the 12 GM targets, two separate multiplex assays were needed. In both assays (4-plex and 10-plex), the transgenes were labeled with one fluorescence reporter and the endogene with another (GMO concentration = transgene/endogene ratio). It was shown that both multiplex assays produce specific results and that performance parameters such as limit of quantification, repeatability, and trueness comply with international recommendations for GMO quantification methods. Moreover, for samples containing GMOs, the throughput and cost-effectiveness is significantly improved compared to qPCR. Thus, it was concluded that the multiplex ddPCR assays could be applied for routine quantification of 12 EU authorized GM maize lines. In case of new authorizations, the events can easily be added to the existing multiplex assays. The presented principle of quantitative multiplexing can be applied to any other domain.

Large-area, low-noise, high-speed, photodiode-based fluorescence detectors with fast overdrive recovery

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bickman, S.; DeMille, D.

2005-11-15

Two large-area, low-noise, high-speed fluorescence detectors have been built. One detector consists of a photodiode with an area of 28 mmx28 mm and a low-noise transimpedance amplifier. This detector has a input light-equivalent spectral noise density of less than 3 pW/{radical}(Hz), can recover from a large scattered light pulse within 10 {mu}s, and has a bandwidth of at least 900 kHz. The second detector consists of a 16-mm-diam avalanche photodiode and a low-noise transimpedance amplifier. This detector has an input light-equivalent spectral noise density of 0.08 pW/{radical}(Hz), also can recover from a large scattered light pulse within 10 {mu}s, andmore » has a bandwidth of 1 MHz.« less

21 CFR 872.1745 - Laser fluorescence caries detection device.

Code of Federal Regulations, 2011 CFR

2011-04-01

... SERVICES (CONTINUED) MEDICAL DEVICES DENTAL DEVICES Diagnostic Devices § 872.1745 Laser fluorescence caries... fluorescence detector housed in a dental handpiece, and a control console that performs device calibration, as...

21 CFR 872.1745 - Laser fluorescence caries detection device.

Code of Federal Regulations, 2014 CFR

2014-04-01

... SERVICES (CONTINUED) MEDICAL DEVICES DENTAL DEVICES Diagnostic Devices § 872.1745 Laser fluorescence caries... fluorescence detector housed in a dental handpiece, and a control console that performs device calibration, as...

Multiplex-Ready Technology for mid-throughput genotyping of molecular markers.

PubMed

Bonneau, Julien; Hayden, Matthew

2014-01-01

Screening molecular markers across large populations in breeding programs is generally time consuming and expensive. The Multiplex-Ready Technology (MRT) (Hayden et al., BMC genomics 9:80, 2008) was created to optimize polymorphism screening and genotyping using standardized PCR reaction conditions. The flexibility of this method maximizes the number of markers (up to 24 markers SSR or SNP, ideally small PCR product <500 bp and highly polymorphic) by using fluorescent dye (VIC, FAM, NED, and PET) and a semiautomated DNA fragment analyzer (ABI3730) capillary electrophoresis for large numbers of DNA samples (96 or 384 samples).

Optimizing detector geometry for trace element mapping by X-ray fluorescence

DOE PAGES

Sun, Yue; Gleber, Sophie -Charlotte; Jacobsen, Chris; ...

2015-01-01

We report that trace metals play critical roles in a variety of systems, ranging from cells to photovoltaics. X-Ray Fluorescence (XRF) microscopy using X-ray excitation provides one of the highest sensitivities available for imaging the distribution of trace metals at sub-100 nm resolution. With the growing availability and increasing performance of synchrotron light source based instruments and X-ray nanofocusing optics, and with improvements in energy-dispersive XRF detectors, what are the factors that limit trace element detectability? To address this question, we describe an analytical model for the total signal incident on XRF detectors with various geometries, including the spectral responsemore » of energy dispersive detectors. This model agrees well with experimentally recorded X-ray fluorescence spectra, and involves much shorter calculation times than with Monte Carlo simulations. With such a model, one can estimate the signal when a trace element is illuminated with an X-ray beam, and when just the surrounding non-fluorescent material is illuminated. From this signal difference, a contrast parameter can be calculated and this can in turn be used to calculate the signal-to-noise ratio (S/N) for detecting a certain elemental concentration. We apply this model to the detection of trace amounts of zinc in biological materials, and to the detection of small quantities of arsenic in semiconductors. In conclusion, we conclude that increased detector collection solid angle is (nearly) always advantageous even when considering the scattered signal. However, given the choice between a smaller detector at 90° to the beam versus a larger detector at 180° (in a backscatter-like geometry), the 90° detector is better for trace element detection in thick samples, while the larger detector in 180° geometry is better suited to trace element detection in thin samples.« less

Optimizing detector geometry for trace element mapping by X-ray fluorescence

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sun, Yue; Gleber, Sophie -Charlotte; Jacobsen, Chris

We report that trace metals play critical roles in a variety of systems, ranging from cells to photovoltaics. X-Ray Fluorescence (XRF) microscopy using X-ray excitation provides one of the highest sensitivities available for imaging the distribution of trace metals at sub-100 nm resolution. With the growing availability and increasing performance of synchrotron light source based instruments and X-ray nanofocusing optics, and with improvements in energy-dispersive XRF detectors, what are the factors that limit trace element detectability? To address this question, we describe an analytical model for the total signal incident on XRF detectors with various geometries, including the spectral responsemore » of energy dispersive detectors. This model agrees well with experimentally recorded X-ray fluorescence spectra, and involves much shorter calculation times than with Monte Carlo simulations. With such a model, one can estimate the signal when a trace element is illuminated with an X-ray beam, and when just the surrounding non-fluorescent material is illuminated. From this signal difference, a contrast parameter can be calculated and this can in turn be used to calculate the signal-to-noise ratio (S/N) for detecting a certain elemental concentration. We apply this model to the detection of trace amounts of zinc in biological materials, and to the detection of small quantities of arsenic in semiconductors. In conclusion, we conclude that increased detector collection solid angle is (nearly) always advantageous even when considering the scattered signal. However, given the choice between a smaller detector at 90° to the beam versus a larger detector at 180° (in a backscatter-like geometry), the 90° detector is better for trace element detection in thick samples, while the larger detector in 180° geometry is better suited to trace element detection in thin samples.« less

Quantitative multiplex detection of biomarkers on a waveguide-based biosensor using quantum dots

DOE Office of Scientific and Technical Information (OSTI.GOV)

Xie, Hongzhi; Mukundan, Harshini; Martinez, Jennifer S

2009-01-01

The quantitative, simultaneous detection of multiple biomarkers with high sensitivity and specificity is critical for biomedical diagnostics, drug discovery and biomarker characterization [Wilson 2006, Tok 2006, Straub 2005, Joos 2002, Jani 2000]. Detection systems relying on optical signal transduction are, in general, advantageous because they are fast, portable, inexpensive, sensitive, and have the potential for multiplex detection of analytes of interest. However, conventional immunoassays for the detection of biomarkers, such as the Enzyme Linked Immunosorbant Assays (ELISAs) are semi-quantitative, time consuming and insensitive. ELISA assays are also limited by high non-specific binding, especially when used with complex biological samples suchmore » as serum and urine (REF). Organic fluorophores that are commonly used in such applications lack photostability and possess a narrow Stoke's shift that makes simultaneous detection of multiple fluorophores with a single excitation source difficult, thereby restricting their use in multiplex assays. The above limitations with traditional assay platforms have resulted in the increased use of nanotechnology-based tools and techniques in the fields of medical imaging [ref], targeted drug delivery [Caruthers 2007, Liu 2007], and sensing [ref]. One such area of increasing interest is the use of semiconductor quantum dots (QDs) for biomedical research and diagnostics [Gao and Cui 2004, Voura 2004, Michalet 2005, Chan 2002, Jaiswal 2004, Gao 2005, Medintz 2005, So 2006 2006, Wu 2003]. Compared to organic dyes, QDs provide several advantages for use in immunoassay platforms, including broad absorption bands with high extinction coefficients, narrow and symmetric emission bands with high quantum yields, high photostablility, and a large Stokes shift [Michalet 2005, Gu 2002]. These features prompted the use of QDs as probes in biodetection [Michalet 2005, Medintz 2005]. For example, Jaiswal et al. reported long term multiple color imaging of live cells using QD-bioconjugates [Jaiswal 2003]. Gao [Gao 2004] and So [So 2006] have used QDs as probes for in-vivo cancer targeting and imaging. Medintz et al. reported self-assembled QD-based biosensors for detection of analytes based on energy transfer [Medintz 2003]. Others have developed an approach for multiplex optical encoding of biomolecules using QDs [Han 2001]. Immunoassays have also benefited from the advantages of QDs. Recently, dihydrolipoic acid (DHLA) capped-QDs have been attached to antibodies and used as fluorescence reporters in plate-based multiplex immunoassays [Goodman 2004]. However, DHLA-QDs are associated with low quantum efficiency and are unstable at neutral pH. These problems limit the application of this technology to the sensitive detection of biomolecules, especially in complex biological samples. Thus, the development of a rapid, sensitive, quantitative, and specific multiplex platform for the detection of biomarkers in difficult samples remains an elusive target. The goal stated above has applications in many fields including medical diagnostics, biological research, and threat reduction. The current decade alone has seen the development of a need to rapidly and accurately detect potential biological warfare agents. For example, current methods for the detection of anthrax are grossly inadequate for a variety of reasons including long incubation time (5 days from time of exposure to onset of symptoms) and non-specific ('flu-like') symptoms. When five employees of the United State Senate were exposed to B. anthracis in the mail (2001), only one patient had a confirmed diagnosis before death. Since then, sandwich immunoassays using both colorimetric and fluorescence detectors have been developed for key components of the anthrax lethal toxin, namely protective antigen (PA), lethal factor (LF), and the edema factor [Mourez 2001]. While these platforms were successful in assays against anthrax toxins, the sensitivity was poor. Furthermore, no single platform exists for the simultaneous and quantitative detection of multiple components of the B. anthracis toxin. Addressing multiple biomarkers at the same time will increase confidence in a positive result, and may lead to application in the simultaneous detection of anthrax and other biowarfare agents.« less

On-chip wavelength multiplexed detection of cancer DNA biomarkers in blood

PubMed Central

Cai, H.; Stott, M. A.; Ozcelik, D.; Parks, J. W.; Hawkins, A. R.; Schmidt, H.

2016-01-01

We have developed an optofluidic analysis system that processes biomolecular samples starting from whole blood and then analyzes and identifies multiple targets on a silicon-based molecular detection platform. We demonstrate blood filtration, sample extraction, target enrichment, and fluorescent labeling using programmable microfluidic circuits. We detect and identify multiple targets using a spectral multiplexing technique based on wavelength-dependent multi-spot excitation on an antiresonant reflecting optical waveguide chip. Specifically, we extract two types of melanoma biomarkers, mutated cell-free nucleic acids —BRAFV600E and NRAS, from whole blood. We detect and identify these two targets simultaneously using the spectral multiplexing approach with up to a 96% success rate. These results point the way toward a full front-to-back chip-based optofluidic compact system for high-performance analysis of complex biological samples. PMID:28058082

Multiplexed quantitation of protein expression and phosphorylation based on functionalized soluble nanopolymers

PubMed Central

Pan, Li; Iliuk, Anton; Yu, Shuai; Geahlen, Robert L.; Tao, W. Andy

2012-01-01

We report here for the first time the multiplexed quantitation of phosphorylation and protein expression based on a functionalized soluble nanopolymer. The soluble nanopolymer, pIMAGO, is functionalized with Ti (IV) ions for chelating phosphoproteins in high specificity, and with infrared fluorescent tags for direct, multiplexed assays. The nanopolymer allows for direct competition for epitopes on proteins of interest, thus facilitating simultaneous detection of phosphorylation by pIMAGO and total protein amount by protein antibody in the same well of microplates. The new strategy has a great potential to measure cell signaling events by clearly distinguishing actual phosphorylation signals from protein expression changes, thus providing a powerful tool to accurately profile cellular signal transduction in healthy and disease cells. We anticipate broad applications of this new strategy in monitoring cellular signaling pathways and discovering new signaling events. PMID:23088311

Simplified Real-Time Multiplex Detection of Loop-Mediated Isothermal Amplification Using Novel Mediator Displacement Probes with Universal Reporters.

PubMed

Becherer, Lisa; Bakheit, Mohammed; Frischmann, Sieghard; Stinco, Silvina; Borst, Nadine; Zengerle, Roland; von Stetten, Felix

2018-04-03

A variety of real-time detection techniques for loop-mediated isothermal amplification (LAMP) based on the change in fluorescence intensity during DNA amplification enable simultaneous detection of multiple targets. However, these techniques depend on fluorogenic probes containing target-specific sequences. That complicates the adaption to different targets leading to time-consuming assay optimization. Here, we present the first universal real-time detection technique for multiplex LAMP. The novel approach allows simple assay design and is easy to implement for various targets. The innovation features a mediator displacement probe and a universal reporter. During amplification of target DNA the mediator is displaced from the mediator displacement probe. Then it hybridizes to the reporter generating a fluorescence signal. The novel mediator displacement (MD) detection was validated against state-of-the-art molecular beacon (MB) detection by means of a HIV-1 RT-LAMP: MD surpassed MB detection by accelerated probe design (MD: 10 min, MB: 3-4 h), shorter times to positive (MD 4.1 ± 0.1 min shorter than MB, n = 36), improved signal-to-noise fluorescence ratio (MD: 5.9 ± 0.4, MB: 2.7 ± 0.4; n = 15), and showed equally good or better analytical performance parameters. The usability of one universal mediator-reporter set in different multiplex assays was successfully demonstrated for a biplex RT-LAMP of HIV-1 and HTLV-1 and a biplex LAMP of Haemophilus ducreyi and Treponema pallidum, both showing good correlation between target concentration and time to positive. Due to its simple implementation it is suggested to extend the use of the universal mediator-reporter sets to the detection of various other diagnostic panels.

Energy resolution and efficiency of phonon-mediated kinetic inductance detectors for light detection

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cardani, L., E-mail: laura.cardani@roma1.infn.it; Physics Department, Princeton University, Washington Road, 08544, Princeton, New Jersey; Colantoni, I.

The development of sensitive cryogenic light detectors is of primary interest for bolometric experiments searching for rare events like dark matter interactions or neutrino-less double beta decay. Thanks to their good energy resolution and the natural multiplexed read-out, Kinetic Inductance Detectors (KIDs) are particularly suitable for this purpose. To efficiently couple KIDs-based light detectors to the large crystals used by the most advanced bolometric detectors, active surfaces of several cm{sup 2} are needed. For this reason, we are developing phonon-mediated detectors. In this paper, we present the results obtained with a prototype consisting of four 40 nm thick aluminum resonators patternedmore » on a 2 × 2 cm{sup 2} silicon chip, and calibrated with optical pulses and X-rays. The detector features a noise resolution σ{sub E} = 154 ± 7 eV and an (18 ± 2)% efficiency.« less

A multiplexed TOF and DOI capable PET detector using a binary position sensitive network.

PubMed

Bieniosek, M F; Cates, J W; Levin, C S

2016-11-07

Time of flight (TOF) and depth of interaction (DOI) capabilities can significantly enhance the quality and uniformity of positron emission tomography (PET) images. Many proposed TOF/DOI PET detectors require complex readout systems using additional photosensors, active cooling, or waveform sampling. This work describes a high performance, low complexity, room temperature TOF/DOI PET module. The module uses multiplexed timing channels to significantly reduce the electronic readout complexity of the PET detector while maintaining excellent timing, energy, and position resolution. DOI was determined using a two layer light sharing scintillation crystal array with a novel binary position sensitive network. A 20 mm effective thickness LYSO crystal array with four 3 mm  ×  3 mm silicon photomultipliers (SiPM) read out by a single timing channel, one energy channel and two position channels achieved a full width half maximum (FWHM) coincidence time resolution of 180  ±  2 ps with 10 mm of DOI resolution and 11% energy resolution. With sixteen 3 mm  ×  3 mm SiPMs read out by a single timing channel, one energy channel and four position channels a coincidence time resolution 204  ±  1 ps was achieved with 10 mm of DOI resolution and 15% energy resolution. The methods presented here could significantly simplify the construction of high performance TOF/DOI PET detectors.

Relevancies of multiple-interaction events and signal-to-noise ratio for Anger-logic based PET detector designs

NASA Astrophysics Data System (ADS)

Peng, Hao

2015-10-01

A fundamental challenge for PET block detector designs is to deploy finer crystal elements while limiting the number of readout channels. The standard Anger-logic scheme including light sharing (an 8 by 8 crystal array coupled to a 2×2 photodetector array with an optical diffuser, multiplexing ratio: 16:1) has been widely used to address such a challenge. Our work proposes a generalized model to study the impacts of two critical parameters on spatial resolution performance of a PET block detector: multiple interaction events and signal-to-noise ratio (SNR). The study consists of the following three parts: (1) studying light output profile and multiple interactions of 511 keV photons within crystal arrays of different crystal widths (from 4 mm down to 1 mm, constant height: 20 mm); (2) applying the Anger-logic positioning algorithm to investigate positioning/decoding uncertainties (i.e., "block effect") in terms of peak-to-valley ratio (PVR), with light sharing, multiple interactions and photodetector SNR taken into account; and (3) studying the dependency of spatial resolution on SNR in the context of modulation transfer function (MTF). The proposed model can be used to guide the development and evaluation of a standard Anger-logic based PET block detector including: (1) selecting/optimizing the configuration of crystal elements for a given photodetector SNR; and (2) predicting to what extent additional electronic multiplexing may be implemented to further reduce the number of readout channels.

Multiplexing 32,000 spectra onto 8 detectors: the HARMONI field splitting, image slicing, and wavelength selecting optics

NASA Astrophysics Data System (ADS)

Tecza, Matthias; Thatte, Niranjan; Clarke, Fraser; Freeman, David; Kosmalski, Johan

2012-09-01

HARMONI, the High Angular Resolution Monolithic Optical & Near-infrared Integral field spectrograph is one of two first-light instruments for the European Extremely Large Telescope. Over a 256x128 pixel field-of-view HARMONI will simultaneously measure approximately 32,000 spectra. Each spectrum is about 4000 spectral pixels long, and covers a selectable part of the 0.47-2.45 μm wavelength range at resolving powers of either R≍4000, 10000, or 20000. All 32,000 spectra are imaged onto eight HAWAII4RG detectors using a multiplexing scheme that divides the input field into four sub-fields, each imaged onto one image slicer that in turn re-arranges a single sub-field into two long exit slits feeding one spectrograph each. In total we require eight spectrographs, each with one HAWAII4RG detector. A system of articulated and exchangeable fold-mirrors and VPH gratings allows one to select different spectral resolving powers and wavelength ranges of interest while keeping a fixed geometry between the spectrograph collimator and camera avoiding the need for an articulated grating and camera. In this paper we describe both the field splitting and image slicing optics as well as the optics that will be used to select both spectral resolving power and wavelength range.

The background in a balloon-borne fluorescence-gated proportional counter

NASA Technical Reports Server (NTRS)

Ramsey, B. D.; Bower, C. R.; Dietz, K. L.; Weisskopf, M. C.

1990-01-01

The results of an analysis of the background in a fluorescence-gated proportional counter operating over the energy range 3-150 keV are presented. It is found that the dominant background component is that produced by high energy qamma-rays that penetrate the shields and undergo multiple scattering in the detector body, resulting in photoelectric absorption in the detector gas. A careful choice of materials and thickness can move the peak of this emission outside of the detector sensitive range, thereby dramatically reducing the residual background.

First Astronomical Use of Multiplexed Transition Edge Sensor Bolometers

NASA Technical Reports Server (NTRS)

Staguhn, J. G.; Ames, T. A.; Benford, D. J.; Chervenak, J. A.; Grossman, E. N.; Irwin, K. D.; Khan, S. A.; Maffei, B.; Moseley, S. H.; Pajot, F.

2004-01-01

We present performance results based on the first astronomical use of multiplexed superconducting bolometers. The Fabry-Perot Interferometer Bolometer Research Experiment (FIBRE) is a broadband submillimeter spectrometer that achieved first light in June 2001 at the Caltech Submillimeter Observatory (CSO). FIBRE's detectors are superconducting transition edge sensor (TES) bolometers read out by a SQUID multiplexer. The Fabry-Perot uses a low resolution grating to order sort the incoming light. A linear bolometer array consisting of 16 elements detects this dispersed light, capturing 5 orders simultaneously from one position on the sky. With tuning of the Fabry-Perot over one free spectral range, a spectrum covering Delta lambda/lambda = 1/7 at a resolution of delta lambda/lambda approx. 1/1200 can be acquired. This spectral resolution is sufficient to resolve Doppler-broadened line emission from external galaxies. FIBRE operates in the 350 m and 450 m bands. These bands cover line emission from the important star formation tracers neutral carbon (CI) and carbon monoxide (CO). We have verified that the multiplexed bolometers are photon noise limited even with the low power present in moderate resolution spectrometry.

First Astronomical Use Of Multiplexed Transition Edge Bolometers

NASA Technical Reports Server (NTRS)

Benford, D. J.; Chervenak, J. A.; Grossman, E. N.; Irwin, K. D.; DeKotwara, S. A.; Maffei, B.; Moseley, S. H.; Pajot, F.; Phillips, T. G.; Reintsema, C. D.

2001-01-01

We present performance results based on the first astronomical use of multiplexed superconducting bolometers. The Fabry-Perot Interferometer Bolometer Research Experiment (FIBRE) is a broadband submillimeter spectrometer that achieved first light in June 2001 at the Caltech Submillimeter Observatory (CSO). FIBRE's detectors are superconducting transition edge sensor (TES) bolometers read out by a SQUID multiplexer. The Fabry-Perot uses a low resolution grating to order sort the incoming light. A linear bolometer array consisting of 16 elements detects this dispersed light, capturing five orders simultaneously from one position on the sky. With tuning of the Fabry-Perot over one free spectral range, a spectrum covering delta-lamda/lamda = 1/7 at a resolution of delta-lamda/lamda = 1/1200 can be acquired. This spectral resolution is sufficient to resolve doppler broadened line emission from external galaxies. FIBRE operates in the 350 micrometer and 450 micrometer bands. These bands cover line emission from the important PDR tracers neutral carbon [CI] and carbon monoxide (CO). We have verified that the multiplexed bolometers are photon noise limited even with the low power present in moderate resolution spectrometry.

Fluorescence decay time imaging using an imaging photon detector with a radio frequency photon correlation system

NASA Astrophysics Data System (ADS)

Morgan, Christopher G.; Mitchell, A. C.; Murray, J. G.

1990-05-01

An imaging photon detector has been modified to incorporate fast timing electronics coupled to a custom built photon correlator interfaced to a RISC computer. Using excitation with intensity- muodulated light, fluorescence images can be readily obtained where contrast is determined by the decay time of emission, rather than by intensity. This technology is readily extended to multifrequency phase/demodulation fluorescence imaging or to differential polarised phase fluorometry. The potential use of the correlator for confocal imaging with a laser scanner is also briefly discussed.

LAMP2GO

DOE Office of Scientific and Technical Information (OSTI.GOV)

Priye, Aashish

2017-06-08

The app is a unique image analysis software and acts as a fluorescence reader for multiplexed nucleic acid amplification reactions. If the reaction is positive, a bright fluorescent signal is emitted from the solution depending on the choice of fluorophore molecule. The app reads the Red, blue and green (RGB) channels of the emitted signal and transforms it to yield the chromaticity (x and y) and luminance of the signal. This new representation of signal is far superior to the RGB system in determining the signal color and intensity.

Visualization and tracking of tumour extracellular vesicle delivery and RNA translation using multiplexed reporters

PubMed Central

Lai, Charles P.; Kim, Edward Y.; Badr, Christian E.; Weissleder, Ralph; Mempel, Thorsten R.; Tannous, Bakhos A.; Breakefield, Xandra O.

2015-01-01

Accurate spatiotemporal assessment of extracellular vesicle (EV) delivery and cargo RNA translation requires specific and robust live-cell imaging technologies. Here we engineer optical reporters to label multiple EV populations for visualization and tracking of tumour EV release, uptake and exchange between cell populations both in culture and in vivo. Enhanced green fluorescence protein (EGFP) and tandem dimer Tomato (tdTomato) were fused at NH2-termini with a palmitoylation signal (PalmGFP, PalmtdTomato) for EV membrane labelling. To monitor EV-RNA cargo, transcripts encoding PalmtdTomato were tagged with MS2 RNA binding sequences and detected by co-expression of bacteriophage MS2 coat protein fused with EGFP. By multiplexing fluorescent and bioluminescent EV membrane reporters, we reveal the rapid dynamics of both EV uptake and translation of EV-delivered cargo mRNAs in cancer cells that occurred within 1-hour post-horizontal transfer between cells. These studies confirm that EV-mediated communication is dynamic and multidirectional between cells with delivery of functional mRNA. PMID:25967391

New photon-counting detectors for single-molecule fluorescence spectroscopy and imaging

PubMed Central

Michalet, X.; Colyer, R. A.; Scalia, G.; Weiss, S.; Siegmund, Oswald H. W.; Tremsin, Anton S.; Vallerga, John V.; Villa, F.; Guerrieri, F.; Rech, I.; Gulinatti, A.; Tisa, S.; Zappa, F.; Ghioni, M.; Cova, S.

2013-01-01

Solution-based single-molecule fluorescence spectroscopy is a powerful new experimental approach with applications in all fields of natural sciences. Two typical geometries can be used for these experiments: point-like and widefield excitation and detection. In point-like geometries, the basic concept is to excite and collect light from a very small volume (typically femtoliter) and work in a concentration regime resulting in rare burst-like events corresponding to the transit of a single-molecule. Those events are accumulated over time to achieve proper statistical accuracy. Therefore the advantage of extreme sensitivity is somewhat counterbalanced by a very long acquisition time. One way to speed up data acquisition is parallelization. Here we will discuss a general approach to address this issue, using a multispot excitation and detection geometry that can accommodate different types of novel highly-parallel detector arrays. We will illustrate the potential of this approach with fluorescence correlation spectroscopy (FCS) and single-molecule fluorescence measurements. In widefield geometries, the same issues of background reduction and single-molecule concentration apply, but the duration of the experiment is fixed by the time scale of the process studied and the survival time of the fluorescent probe. Temporal resolution on the other hand, is limited by signal-to-noise and/or detector resolution, which calls for new detector concepts. We will briefly present our recent results in this domain. PMID:24729836

New photon-counting detectors for single-molecule fluorescence spectroscopy and imaging.

PubMed

Michalet, X; Colyer, R A; Scalia, G; Weiss, S; Siegmund, Oswald H W; Tremsin, Anton S; Vallerga, John V; Villa, F; Guerrieri, F; Rech, I; Gulinatti, A; Tisa, S; Zappa, F; Ghioni, M; Cova, S

2011-05-13

Solution-based single-molecule fluorescence spectroscopy is a powerful new experimental approach with applications in all fields of natural sciences. Two typical geometries can be used for these experiments: point-like and widefield excitation and detection. In point-like geometries, the basic concept is to excite and collect light from a very small volume (typically femtoliter) and work in a concentration regime resulting in rare burst-like events corresponding to the transit of a single-molecule. Those events are accumulated over time to achieve proper statistical accuracy. Therefore the advantage of extreme sensitivity is somewhat counterbalanced by a very long acquisition time. One way to speed up data acquisition is parallelization. Here we will discuss a general approach to address this issue, using a multispot excitation and detection geometry that can accommodate different types of novel highly-parallel detector arrays. We will illustrate the potential of this approach with fluorescence correlation spectroscopy (FCS) and single-molecule fluorescence measurements. In widefield geometries, the same issues of background reduction and single-molecule concentration apply, but the duration of the experiment is fixed by the time scale of the process studied and the survival time of the fluorescent probe. Temporal resolution on the other hand, is limited by signal-to-noise and/or detector resolution, which calls for new detector concepts. We will briefly present our recent results in this domain.

Toward Precision Medicine: A Cancer Molecular Subtyping Nano-Strategy for RNA Biomarkers in Tumor and Urine.

PubMed

Koo, Kevin M; Wee, Eugene J H; Mainwaring, Paul N; Wang, Yuling; Trau, Matt

2016-12-01

Cancer is a heterogeneous disease which manifests as different molecular subtypes due to the complex nature of tumor initiation, progression, and metastasis. The concept of precision medicine aims to exploit this cancer heterogeneity by incorporating diagnostic technology to characterize each cancer patient's molecular subtype for tailored treatments. To characterize cancer molecular subtypes accurately, a suite of multiplexed bioassays have currently been developed to detect multiple oncogenic biomarkers. Despite the reliability of current multiplexed detection techniques, novel strategies are still needed to resolve limitations such as long assay time, complex protocols, and difficulty in interpreting broad overlapping spectral peaks of conventional fluorescence readouts. Herein a rapid (80 min) multiplexed platform strategy for subtyping prostate cancer tumor and urine samples based on their RNA biomarker profiles is presented. This is achieved by combining rapid multiplexed isothermal reverse transcription-recombinase polymerase amplification (RT-RPA) of target RNA biomarkers with surface-enhanced Raman spectroscopy (SERS) nanotags for "one-pot" readout. This is the first translational application of a RT-RPA/SERS-based platform for multiplexed cancer biomarker detection to address a clinical need. With excellent sensitivity of 200 zmol (100 copies) and specificity, we believed that this platform methodology could be a useful tool for rapid multiplexed subtyping of cancers. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Single Fluorescence Channel-based Multiplex Detection of Avian Influenza Virus by Quantitative PCR with Intercalating Dye

PubMed Central

Ahberg, Christian D.; Manz, Andreas; Neuzil, Pavel

2015-01-01

Since its invention in 1985 the polymerase chain reaction (PCR) has become a well-established method for amplification and detection of segments of double-stranded DNA. Incorporation of fluorogenic probe or DNA intercalating dyes (such as SYBR Green) into the PCR mixture allowed real-time reaction monitoring and extraction of quantitative information (qPCR). Probes with different excitation spectra enable multiplex qPCR of several DNA segments using multi-channel optical detection systems. Here we show multiplex qPCR using an economical EvaGreen-based system with single optical channel detection. Previously reported non quantitative multiplex real-time PCR techniques based on intercalating dyes were conducted once the PCR is completed by performing melting curve analysis (MCA). The technique presented in this paper is both qualitative and quantitative as it provides information about the presence of multiple DNA strands as well as the number of starting copies in the tested sample. Besides important internal control, multiplex qPCR also allows detecting concentrations of more than one DNA strand within the same sample. Detection of the avian influenza virus H7N9 by PCR is a well established method. Multiplex qPCR greatly enhances its specificity as it is capable of distinguishing both haemagglutinin (HA) and neuraminidase (NA) genes as well as their ratio. PMID:26088868

Gaseous detectors for energy dispersive X-ray fluorescence analysis

NASA Astrophysics Data System (ADS)

Veloso, J. F. C. A.; Silva, A. L. M.

2018-01-01

The energy resolution capability of gaseous detectors is being used in the last years to perform studies on the detection of characteristic X-ray lines emitted by elements when excited by external radiation sources. One of the most successful techniques is the Energy Dispersive X-ray Fluorescence (EDXRF) analysis. Recent developments in the new generation of micropatterned gaseous detectors (MPGDs), triggered the possibility not only of recording the photon energy, but also of providing position information, extending their application to EDXRF imaging. The relevant features and strategies to be applied in gaseous detectors in order to better fit the requirements for EDXRF imaging will be reviewed and discussed, and some application examples will be presented.

Spectroscopic optical coherence tomography based on wavelength de-multiplexing and smart pixel array detection

NASA Astrophysics Data System (ADS)

Laubscher, Markus; Bourquin, Stéphane; Froehly, Luc; Karamata, Boris; Lasser, Theo

2004-07-01

Current spectroscopic optical coherence tomography (OCT) methods rely on a posteriori numerical calculation. We present an experimental alternative for accessing spectroscopic information in OCT without post-processing based on wavelength de-multiplexing and parallel detection using a diffraction grating and a smart pixel detector array. Both a conventional A-scan with high axial resolution and the spectrally resolved measurement are acquired simultaneously. A proof-of-principle demonstration is given on a dynamically changing absorbing sample. The method's potential for fast spectroscopic OCT imaging is discussed. The spectral measurements obtained with this approach are insensitive to scan non-linearities or sample movements.

The LiteBIRD Satellite Mission: Sub-Kelvin Instrument

NASA Astrophysics Data System (ADS)

Suzuki, A.; Ade, P. A. R.; Akiba, Y.; Alonso, D.; Arnold, K.; Aumont, J.; Baccigalupi, C.; Barron, D.; Basak, S.; Beckman, S.; Borrill, J.; Boulanger, F.; Bucher, M.; Calabrese, E.; Chinone, Y.; Cho, S.; Crill, B.; Cukierman, A.; Curtis, D. W.; de Haan, T.; Dobbs, M.; Dominjon, A.; Dotani, T.; Duband, L.; Ducout, A.; Dunkley, J.; Duval, J. M.; Elleflot, T.; Eriksen, H. K.; Errard, J.; Fischer, J.; Fujino, T.; Funaki, T.; Fuskeland, U.; Ganga, K.; Goeckner-Wald, N.; Grain, J.; Halverson, N. W.; Hamada, T.; Hasebe, T.; Hasegawa, M.; Hattori, K.; Hattori, M.; Hayes, L.; Hazumi, M.; Hidehira, N.; Hill, C. A.; Hilton, G.; Hubmayr, J.; Ichiki, K.; Iida, T.; Imada, H.; Inoue, M.; Inoue, Y.; Irwin, K. D.; Ishino, H.; Jeong, O.; Kanai, H.; Kaneko, D.; Kashima, S.; Katayama, N.; Kawasaki, T.; Kernasovskiy, S. A.; Keskitalo, R.; Kibayashi, A.; Kida, Y.; Kimura, K.; Kisner, T.; Kohri, K.; Komatsu, E.; Komatsu, K.; Kuo, C. L.; Kurinsky, N. A.; Kusaka, A.; Lazarian, A.; Lee, A. T.; Li, D.; Linder, E.; Maffei, B.; Mangilli, A.; Maki, M.; Matsumura, T.; Matsuura, S.; Meilhan, D.; Mima, S.; Minami, Y.; Mitsuda, K.; Montier, L.; Nagai, M.; Nagasaki, T.; Nagata, R.; Nakajima, M.; Nakamura, S.; Namikawa, T.; Naruse, M.; Nishino, H.; Nitta, T.; Noguchi, T.; Ogawa, H.; Oguri, S.; Okada, N.; Okamoto, A.; Okamura, T.; Otani, C.; Patanchon, G.; Pisano, G.; Rebeiz, G.; Remazeilles, M.; Richards, P. L.; Sakai, S.; Sakurai, Y.; Sato, Y.; Sato, N.; Sawada, M.; Segawa, Y.; Sekimoto, Y.; Seljak, U.; Sherwin, B. D.; Shimizu, T.; Shinozaki, K.; Stompor, R.; Sugai, H.; Sugita, H.; Suzuki, J.; Tajima, O.; Takada, S.; Takaku, R.; Takakura, S.; Takatori, S.; Tanabe, D.; Taylor, E.; Thompson, K. L.; Thorne, B.; Tomaru, T.; Tomida, T.; Tomita, N.; Tristram, M.; Tucker, C.; Turin, P.; Tsujimoto, M.; Uozumi, S.; Utsunomiya, S.; Uzawa, Y.; Vansyngel, F.; Wehus, I. K.; Westbrook, B.; Willer, M.; Whitehorn, N.; Yamada, Y.; Yamamoto, R.; Yamasaki, N.; Yamashita, T.; Yoshida, M.

2018-05-01

Inflation is the leading theory of the first instant of the universe. Inflation, which postulates that the universe underwent a period of rapid expansion an instant after its birth, provides convincing explanation for cosmological observations. Recent advancements in detector technology have opened opportunities to explore primordial gravitational waves generated by the inflation through "B-mode" (divergent-free) polarization pattern embedded in the cosmic microwave background anisotropies. If detected, these signals would provide strong evidence for inflation, point to the correct model for inflation, and open a window to physics at ultra-high energies. LiteBIRD is a satellite mission with a goal of detecting degree-and-larger-angular-scale B-mode polarization. LiteBIRD will observe at the second Lagrange point with a 400 mm diameter telescope and 2622 detectors. It will survey the entire sky with 15 frequency bands from 40 to 400 GHz to measure and subtract foregrounds. The US LiteBIRD team is proposing to deliver sub-Kelvin instruments that include detectors and readout electronics. A lenslet-coupled sinuous antenna array will cover low-frequency bands (40-235 GHz) with four frequency arrangements of trichroic pixels. An orthomode-transducer-coupled corrugated horn array will cover high-frequency bands (280-402 GHz) with three types of single frequency detectors. The detectors will be made with transition edge sensor (TES) bolometers cooled to a 100 milli-Kelvin base temperature by an adiabatic demagnetization refrigerator. The TES bolometers will be read out using digital frequency multiplexing with Superconducting QUantum Interference Device (SQUID) amplifiers. Up to 78 bolometers will be multiplexed with a single SQUID amplifier. We report on the sub-Kelvin instrument design and ongoing developments for the LiteBIRD mission.

Kinetic inductance detectors for far-infrared spectroscopy

NASA Astrophysics Data System (ADS)

Barlis, Alyssa; Aguirre, James; Stevenson, Thomas

2016-07-01

The star formation mechanisms at work in the early universe remain one of the major unsolved problems of modern astrophysics. Many of the luminous galaxies present during the period of peak star formation (between redshifts 1 and 3) were heavily enshrouded in dust, which makes observing their properties difficult at optical wavelengths. However, many spectral lines exist at far-infrared wavelengths that serve as tracers of star formation during that period, in particular fine structure lines of nitrogen, carbon, and oxygen, as well as the carbon monoxide molecule. Using an observation technique known as intensity mapping, it would be possible to observe the total line intensity for a given redshift range even without detecting individual sources. Here, we describe a detector system suitable for a balloonborne spectroscopic intensity mapping experiment at far-infrared wavelengths. The experiment requires an "integralfield" type spectrograph, with modest spectral resolution (R 100) for each of a number of spatial pixels spanning several octaves in wavelength. The detector system uses lumped-element kinetic inductance detectors (LEKIDs), which have the potential to achieve the high sensitivity, low noise, and high multiplexing factor required for this experiment. We detail the design requirements and considerations, and the fabrication process for a prototype LEKID array of 1600 pixels. The pixel design is driven by the need for high responsivity, which requires a small physical volume for the LEKID inductor. In order to minimize two-level system noise, the resonators include large-area interdigitated capacitors. High quality factor resonances are required for a large frequency multiplexing factor. Detectors were fabricated using both trilayer TiN/Ti/TiN recipes and thin-film Al, and are operated at base temperatures near 250 mK.

Differential phase contrast with a segmented detector in a scanning X-ray microprobe

PubMed Central

Hornberger, B.; de Jonge, M. D.; Feser, M.; Holl, P.; Holzner, C.; Jacobsen, C.; Legnini, D.; Paterson, D.; Rehak, P.; Strüder, L.; Vogt, S.

2008-01-01

Scanning X-ray microprobes are unique tools for the nanoscale investigation of specimens from the life, environmental, materials and other fields of sciences. Typically they utilize absorption and fluorescence as contrast mechanisms. Phase contrast is a complementary technique that can provide strong contrast with reduced radiation dose for weakly absorbing structures in the multi-keV range. In this paper the development of a segmented charge-integrating silicon detector which provides simultaneous absorption and differential phase contrast is reported. The detector can be used together with a fluorescence detector for the simultaneous acquisition of transmission and fluorescence data. It can be used over a wide range of photon energies, photon rates and exposure times at third-generation synchrotron radiation sources, and is currently operating at two beamlines at the Advanced Photon Source. Images obtained at around 2 keV and 10 keV demonstrate the superiority of phase contrast over absorption for specimens composed of light elements. PMID:18552427

Ultrasensitive Single Fluorescence-Labeled Probe-Mediated Single Universal Primer-Multiplex-Droplet Digital Polymerase Chain Reaction for High-Throughput Genetically Modified Organism Screening.

PubMed

Niu, Chenqi; Xu, Yuancong; Zhang, Chao; Zhu, Pengyu; Huang, Kunlun; Luo, Yunbo; Xu, Wentao

2018-05-01

As genetically modified (GM) technology develops and genetically modified organisms (GMOs) become more available, GMOs face increasing regulations and pressure to adhere to strict labeling guidelines. A singleplex detection method cannot perform the high-throughput analysis necessary for optimal GMO detection. Combining the advantages of multiplex detection and droplet digital polymerase chain reaction (ddPCR), a single universal primer-multiplex-ddPCR (SUP-M-ddPCR) strategy was proposed for accurate broad-spectrum screening and quantification. The SUP increases efficiency of the primers in PCR and plays an important role in establishing a high-throughput, multiplex detection method. Emerging ddPCR technology has been used for accurate quantification of nucleic acid molecules without a standard curve. Using maize as a reference point, four heterologous sequences ( 35S, NOS, NPTII, and PAT) were selected to evaluate the feasibility and applicability of this strategy. Surprisingly, these four genes cover more than 93% of the transgenic maize lines and serve as preliminary screening sequences. All screening probes were labeled with FAM fluorescence, which allows the signals from the samples with GMO content and those without to be easily differentiated. This fiveplex screening method is a new development in GMO screening. Utilizing an optimal amplification assay, the specificity, limit of detection (LOD), and limit of quantitation (LOQ) were validated. The LOD and LOQ of this GMO screening method were 0.1% and 0.01%, respectively, with a relative standard deviation (RSD) < 25%. This method could serve as an important tool for the detection of GM maize from different processed, commercially available products. Further, this screening method could be applied to other fields that require reliable and sensitive detection of DNA targets.

Multiplex digital PCR: breaking the one target per color barrier of quantitative PCR.

PubMed

Zhong, Qun; Bhattacharya, Smiti; Kotsopoulos, Steven; Olson, Jeff; Taly, Valérie; Griffiths, Andrew D; Link, Darren R; Larson, Jonathan W

2011-07-07

Quantitative polymerase chain reactions (qPCR) based on real-time PCR constitute a powerful and sensitive method for the analysis of nucleic acids. However, in qPCR, the ability to multiplex targets using differently colored fluorescent probes is typically limited to 4-fold by the spectral overlap of the fluorophores. Furthermore, multiplexing qPCR assays requires expensive instrumentation and most often lengthy assay development cycles. Digital PCR (dPCR), which is based on the amplification of single target DNA molecules in many separate reactions, is an attractive alternative to qPCR. Here we report a novel and easy method for multiplexing dPCR in picolitre droplets within emulsions-generated and read out in microfluidic devices-that takes advantage of both the very high numbers of reactions possible within emulsions (>10(6)) as well as the high likelihood that the amplification of only a single target DNA molecule will initiate within each droplet. By varying the concentration of different fluorogenic probes of the same color, it is possible to identify the different probes on the basis of fluorescence intensity. Adding multiple colors increases the number of possible reactions geometrically, rather than linearly as with qPCR. Accurate and precise copy numbers of up to sixteen per cell were measured using a model system. A 5-plex assay for spinal muscular atrophy was demonstrated with just two fluorophores to simultaneously measure the copy number of two genes (SMN1 and SMN2) and to genotype a single nucleotide polymorphism (c.815A>G, SMN1). Results of a pilot study with SMA patients are presented. This journal is © The Royal Society of Chemistry 2011

Monochrome Multiplexing in Polymerase Chain Reaction by Photobleaching of Fluorogenic Hydrolysis Probes.

PubMed

Schuler, Friedrich; Trotter, Martin; Zengerle, Roland; von Stetten, Felix

2016-03-01

Multiplexing in polymerase chain reaction (PCR) is a technique widely used to save cost and sample material and to increase sensitivity compared to distributing a sample to several singleplex reactions. One of the most common methods to detect the different amplification products is the use of fluorogenic probes that emit at different wavelengths (colors). To reduce the number of detection channels, several methods for monochrome multiplexing have been suggested. However, they pose restrictions to the amplifiable target length, the sequence, or the melting temperature. To circumvent these limitations, we suggest a novel approach that uses different fluorophores with the same emission maximum. Discrimination is achieved by their different fluorescence stability during photobleaching. Atto488 (emitting at the same wavelength as 6-carboxyfluorescein, FAM) and Atto467N (emitting at the same wavelength as cyanine 5, Cy5) were found to bleach significantly less than FAM and Cy5; i.e., the final fluorescence of Atto dyes was more than tripled compared to FAM and Cy5. We successfully applied this method by performing a 4-plex PCR targeting antibiotic resistance genes in S. aureus using only 2 color channels. Confidence of discrimination between the targets was >99.9% at high copy initial copy numbers of 100 000 copies. Cases where both targets were present could be discriminated with equal confidence for Cy5 channel and reduced levels of confidence (>68%) for FAM channel. Moreover, a 2-plex digital PCR reaction in 1 color channel was shown. In the future, the degree of multiplexing may be increased by adding fluorogenic probe pairs with other emission wavelengths. The method may also be applied to other probe and assay formats, such as Förster resonance energy transfer (FRET) probes and immunoassays.

Code-division-multiplexed readout of large arrays of TES microcalorimeters

NASA Astrophysics Data System (ADS)

Morgan, K. M.; Alpert, B. K.; Bennett, D. A.; Denison, E. V.; Doriese, W. B.; Fowler, J. W.; Gard, J. D.; Hilton, G. C.; Irwin, K. D.; Joe, Y. I.; O'Neil, G. C.; Reintsema, C. D.; Schmidt, D. R.; Ullom, J. N.; Swetz, D. S.

2016-09-01

Code-division multiplexing (CDM) offers a path to reading out large arrays of transition edge sensor (TES) X-ray microcalorimeters with excellent energy and timing resolution. We demonstrate the readout of X-ray TESs with a 32-channel flux-summed code-division multiplexing circuit based on superconducting quantum interference device (SQUID) amplifiers. The best detector has energy resolution of 2.28 ± 0.12 eV FWHM at 5.9 keV and the array has mean energy resolution of 2.77 ± 0.02 eV over 30 working sensors. The readout channels are sampled sequentially at 160 ns/row, for an effective sampling rate of 5.12 μs/channel. The SQUID amplifiers have a measured flux noise of 0.17 μΦ0/√Hz (non-multiplexed, referred to the first stage SQUID). The multiplexed noise level and signal slew rate are sufficient to allow readout of more than 40 pixels per column, making CDM compatible with requirements outlined for future space missions. Additionally, because the modulated data from the 32 SQUID readout channels provide information on each X-ray event at the row rate, our CDM architecture allows determination of the arrival time of an X-ray event to within 275 ns FWHM with potential benefits in experiments that require detection of near-coincident events.

Fluorescence lifetime imaging with near-infrared dyes

NASA Astrophysics Data System (ADS)

Becker, Wolfgang; Shcheslavskiy, Vladislav

2013-02-01

Near-infrared (NIR) dyes are used as fluorescence markers in small-animal imaging and in diffuse optical tomography of the human brain. In these applications it is important to know whether the dyes bind to proteins or other tissue constituents, and whether their fluorescence lifetimes depend on the targets they are bound to. Unfortunately, neither the lasers nor the detectors of commonly used confocal and multiphoton laser scanning microscopes allow for excitation and detection of NIR fluorescence. We therefore upgraded existing confocal TCSPC FLIM systems with NIR lasers and NIR sensitive detectors. In multiphoton systems we used the Ti:Sa laser as a one-photon excitation source in combination with an NIR-sensitive detector in the confocal beam path. We tested a number of NIR dyes in biological tissue. Some of them showed clear lifetime changes depending on the tissue structures they are bound to. We therefore believe that NIR FLIM can deliver supplementary information on the tissue constitution and on local biochemical parameters.

Novel Multiplexing Technique for Detector and Mixer Arrays

NASA Technical Reports Server (NTRS)

Karasik, Boris S.; McGrath, William R.

2001-01-01

Future submillimeter and far-infrared space telescopes will require large-format (many 1000's of elements) imaging detector arrays to perform state-of-the-art astronomical observations. A crucial issue related to a focal plane array is a readout scheme which is compatible with large numbers of cryogenically-cooled (typically < 1 K) detectors elements. When the number of elements becomes of the order of thousands, the physical layout for individual readout amplifiers becomes nearly impossible to realize for practical systems. Another important concern is the large number of wires leading to a 0.1-0.3 K platform. In the case of superconducting transition edge sensors (TES), a scheme for time-division multiplexing of SQUID read-out amplifiers has been recently demonstrated. In this scheme the number of SQUIDs is equal to the number (N) of the detectors, but only one SQUID is turned on at a time. The SQUIDs are connected in series in each column of the array, so the number of wires leading to the amplifiers can be reduced, but it is still of the order of N. Another approach uses a frequency domain multiplexing scheme of the bolometer array. The bolometers are biased with ac currents whose frequencies are individual for each element and are much higher than the bolometer bandwidth. The output signals are connected in series in a summing loop which is coupled to a single SQUID amplifier. The total number of channels depends on the ratio between the SQUID bandwidth and the bolometer bandwidth and can be at least 100 according to the authors. An important concern about this technique is a contribution of the out-of-band Johnson noise which multiplies by factor N(exp 1/2) for each frequency channel. We propose a novel solution for large format arrays based on the Hadamard transform coding technique which requires only one amplifier to read out the entire array of potentially many 1000's of elements and uses approximately 10 wires between the cold stage and room temperature electronics. This can significantly reduce the complexity of the readout circuits.

Simultaneous dual-color fluorescence microscope: a characterization study.

PubMed

Li, Zheng; Chen, Xiaodong; Ren, Liqiang; Song, Jie; Li, Yuhua; Zheng, Bin; Liu, Hong

2013-01-01

High spatial resolution and geometric accuracy is crucial for chromosomal analysis of clinical cytogenetic applications. High resolution and rapid simultaneous acquisition of multiple fluorescent wavelengths can be achieved by utilizing concurrent imaging with multiple detectors. However, such class of microscopic systems functions differently from traditional fluorescence microscopes. To develop a practical characterization framework to assess and optimize the performance of a high resolution and dual-color fluorescence microscope designed for clinical chromosomal analysis. A dual-band microscopic imaging system utilizes a dichroic mirror, two sets of specially selected optical filters, and two detectors to simultaneously acquire two fluorescent wavelengths. The system's geometric distortion, linearity, the modulation transfer function, and the dual detectors' alignment were characterized. Experiment results show that the geometric distortion at lens periphery is less than 1%. Both fluorescent channels show linear signal responses, but there exists discrepancy between the two due to the detectors' non-uniform response ratio to different wavelengths. In terms of the spatial resolution, the two contrast transfer function curves trend agreeably with the spatial frequency. The alignment measurement allows quantitatively assessing the cameras' alignment. A result image of adjusted alignment is demonstrated to show the reduced discrepancy by using the alignment measurement method. In this paper, we present a system characterization study and its methods for a specially designed imaging system for clinical cytogenetic applications. The presented characterization methods are not only unique to this dual-color imaging system but also applicable to evaluation and optimization of other similar multi-color microscopic image systems for improving their clinical utilities for future cytogenetic applications.

Large-Format AlGaN PIN Photodiode Arrays for UV Images

NASA Technical Reports Server (NTRS)

Aslam, Shahid; Franz, David

2010-01-01

A large-format hybridized AlGaN photodiode array with an adjustable bandwidth features stray-light control, ultralow dark-current noise to reduce cooling requirements, and much higher radiation tolerance than previous technologies. This technology reduces the size, mass, power, and cost of future ultraviolet (UV) detection instruments by using lightweight, low-voltage AlGaN detectors in a hybrid detector/multiplexer configuration. The solar-blind feature eliminates the need for additional visible light rejection and reduces the sensitivity of the system to stray light that can contaminate observations.

[Determination of fluorescent whitening agents in plastic food contact materials by high performance liquid chromatography with fluorescence detector].

PubMed

Jiao, Yanna; Ding, Li; Zhu, Shaohua; Fu, Shanliang; Gong, Qiang; Li, Hui; Wang, Libing

2013-01-01

A method for the determination of fluorescent whitening agents in plastic food contact materials by high performance liquid chromatography (HPLC) with fluorescence detector was developed. The samples were extracted with trichloromethane by sonication for 30 min at 40 degrees C. The HPLC method was performed on a column of Eclipse XDB-C18 (250 mm x 4.6 mm, 5 microm) by gradient elution using 5 mmol/L ammonium acetate and acetonitrile as the mobile phases, and detected by the fluorescence detector at an excitation wavelength of 350 nm and an emission wavelength of 430 nm. The experimental results indicated that the four fluorescent whitening agents were separated well. The limits of detection (LOD) (S/N = 3) were 0.3, 0.1, 0.05, 0.14 mg/L, and the limits of quantification (LOQ) (S/N = 10) were 1.0, 0.4, 0.2, 0.5 mg/L for 1,4-bis (4-cyanostyryl) benzene (C. I. 199), 1,4-bis (2-benzoxazolyl) naphthalene (C. I. 367), 4,4'-bis(2-methoxystyryl) biphenyl (C. I. 378) and 2,5-thiophenediylbis (5-tert-butyl-1,3-benzoxazole) (C. I. 184), respectively. Good linearities with correlation coefficients (r2) not less than 0.991 were obtained. The proposed method is simple, accurate, sensitive and can meet the requirements of the routine determination of fluorescent whitening agents in entry-exit products.

The small contribution of molecular Bremsstrahlung radiation to the air-fluorescence yield of cosmic ray shower particles

NASA Astrophysics Data System (ADS)

Al Samarai, Imen; Deligny, Olivier; Rosado, Jaime

2016-10-01

A small contribution of molecular Bremsstrahlung radiation to the air-fluorescence yield in the UV range is estimated based on an approach previously developed in the framework of the radio-detection of showers in the gigahertz frequency range. First, this approach is shown to provide an estimate of the main contribution of the fluorescence yield due to the de-excitation of the C 3Πu electronic level of nitrogen molecules to the B 3Πg one amounting to Y[ 337 ] =(6.05 ± 1.50) MeV-1 at 800 hPa pressure and 293 K temperature conditions, which compares well to previous dedicated works and to experimental results. Then, under the same pressure and temperature conditions, the fluorescence yield induced by molecular Bremsstrahlung radiation is found to be Y[330-400]MBR = 0.10 MeV-1 in the wavelength range of interest for the air-fluorescence detectors used to detect extensive air showers induced in the atmosphere by ultra-high energy cosmic rays. This means that out of ≃175 photons with wavelength between 330 and 400 nm detected by fluorescence detectors, one of them has been produced by molecular Bremsstrahlung radiation. Although small, this contribution is not negligible in regards to the total budget of systematic uncertainties when considering the absolute energy scale of fluorescence detectors.

Choice of Illumination System & Fluorophore for Multiplex Immunofluorescence on FFPE Tissue Sections

PubMed Central

Kishen, Ria E. B.; Kluth, David C.; Bellamy, Christopher O. C.

2016-01-01

The recent availability of novel dyes and alternative light sources to facilitate complex tissue immunofluorescence studies such as multiplex labelling has not been matched by reports critically evaluating the considerations and relative benefits of these new tools, particularly in combination. Product information is often limited to wavelengths used for older fluorophores (FITC, TRITC & corresponding Alexa dyes family). Consequently, novel agents such as Quantum dots are not widely appreciated or used, despite highly favourable properties including extremely bright emission, stability and potentially reduced tissue autofluorescence at the excitation wavelength. Using spectral analysis, we report here a detailed critical appraisal and comparative evaluation of different light sources and fluorophores in multiplex immunofluorescence of clinical biopsy sections. The comparison includes mercury light, metal halide and 3 different LED-based systems, using 7 Qdots (525, 565, 585, 605, 625, 705), Cy3 and Cy5. We discuss the considerations relevant to achieving the best combination of light source and fluorophore for accurate multiplex fluorescence quantitation. We highlight practical limitations and confounders to quantitation with filter-based approaches. PMID:27632367

Mosaic-Detector-Based Fluorescence Spectral Imager

NASA Technical Reports Server (NTRS)

Son, Kyung-Ah; Moon, Jeong

2007-01-01

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.

Development and Application of Chemical Probes for Vibrational Imaging by Stimulated Raman Scattering

NASA Astrophysics Data System (ADS)

Hu, Fanghao

During the last decade, Raman microscopy is experiencing rapid development and increasingly applied in biological and medical systems. Especially, stimulated Raman scattering (SRS) microscopy, which significantly improves the sensitivity of Raman scattering through stimulated emission, has allowed direct visualization of many species that are previously challenging with conventional fluorescence imaging. Compared to fluorescence, SRS imaging requires no label or small label on the target molecule, thus with minimal perturbation to the molecule of interest. Moreover, Raman scattering is free from complicated photophysical and photochemical processes such as photobleaching, and has intrinsically narrower linewidth than fluorescence emission. This allows multiplexed Raman imaging with minimal spectral crosstalk and excellent photo-stability. To achieve the full potential of Raman microscopy, vibrational probes have been developed for Raman imaging. Multiple Raman probes with a few atoms in size are applied in Raman imaging with high sensitivity and specificity. An overview of both fluorescence and Raman microscopy and their imaging probes is given in Chapter 1 with a brief discussion on the SRS theory. Built on the current progress of Raman microscopy and vibrational probes, I write on my research in the development of carbon-deuterium, alkyne and nitrile probes for visualizing choline metabolism (Chapter 2), glucose uptake activity (Chapter 3), complex brain metabolism (Chapter 4) and polymeric nanoparticles (Chapter 5) in live cells and tissues, as well as the development of polyyne-based vibrational probes for super-multiplexed imaging, barcoding and analysis (Chapter 6).

Quantum Dot Enabled Molecular Sensing and Diagnostics

PubMed Central

Zhang, Yi; Wang, Tza-Huei

2012-01-01

Since its emergence, semiconductor nanoparticles known as quantum dots (QDs) have drawn considerable attention and have quickly extended their applicability to numerous fields within the life sciences. This is largely due to their unique optical properties such as high brightness and narrow emission band as well as other advantages over traditional organic fluorophores. New molecular sensing strategies based on QDs have been developed in pursuit of high sensitivity, high throughput, and multiplexing capabilities. For traditional biological applications, QDs have already begun to replace traditional organic fluorophores to serve as simple fluorescent reporters in immunoassays, microarrays, fluorescent imaging applications, and other assay platforms. In addition, smarter, more advanced QD probes such as quantum dot fluorescence resonance energy transfer (QD-FRET) sensors, quenching sensors, and barcoding systems are paving the way for highly-sensitive genetic and epigenetic detection of diseases, multiplexed identification of infectious pathogens, and tracking of intracellular drug and gene delivery. When combined with microfluidics and confocal fluorescence spectroscopy, the detection limit is further enhanced to single molecule level. Recently, investigations have revealed that QDs participate in series of new phenomena and exhibit interesting non-photoluminescent properties. Some of these new findings are now being incorporated into novel assays for gene copy number variation (CNV) studies and DNA methylation analysis with improved quantification resolution. Herein, we provide a comprehensive review on the latest developments of QD based molecular diagnostic platforms in which QD plays a versatile and essential role. PMID:22916072

Multiplex Detection of IgG and IgM to Rift Valley Fever Virus Nucleoprotein, Nonstructural Proteins, and Glycoprotein in Ovine and Bovine.

PubMed

Hossain, Mohammad M; Wilson, William C; Faburay, Bonto; Richt, Jürgen; McVey, David S; Rowland, Raymond R

2016-08-01

A multiplex fluorescence microsphere immunoassay (FMIA) was used to detect bovine and ovine IgM and IgG antibodies to several Rift Valley fever virus (RVFV) proteins, including the major surface glycoprotein, Gn; the nonstructural proteins, NSs and NSm; and the nucleoprotein, N. Target antigens were assembled into a multiplex and tested in serum samples from infected wild-type RVFV or MP12, a modified live virus vaccine. As expected, the N protein was immunodominant and the best target for early detection of infection. Antibody activity against the other targets was also detected. The experimental results demonstrate the capabilities of FMIA for the detection of antibodies to RVFV structural and nonstructural proteins, which can be applied to future development and validation of diagnostic tests that can be used to differentiate vaccinated from infected animals.

A novel multiplexed fluorescence polarisation immunoassay based on a recombinant bi-specific single-chain diabody for simultaneous detection of fluoroquinolones and sulfonamides in milk.

PubMed

Chen, Min; Wen, Kai; Tao, Xiaoqi; Ding, Shuangyang; Xie, Jie; Yu, Xuezhi; Li, Jiancheng; Xia, Xi; Wang, Yang; Xie, Sanlei; Jiang, Haiyang

2014-01-01

Major research efforts are focusing on the development of simultaneous multiplexed immunoassays. In this study, a novel dual-binding fluorescence polarisation immunoassay (DB-FPIA) using a broad-specificity bi-specific single-chain diabody (scDb) and two fluorescent-labelled tracers (sulfamethoxypyridazine-fluorescein isothiocyanate (SMP-FITC) and sarafloxacin-Texas Red (SAR-TR)) with different excitation and emission wavelengths was developed for simultaneous and high-throughput detection of 19 fluoroquinolones (FQs) and 13 sulfonamides (SAs) at the maximum residue limits in milk samples. Recoveries for spiked milk samples were from 76.4% to 128.4%, with a relative standard deviation lower than 13.9%. The developed DB-FPIA was then applied to field samples, followed by confirmation by LC-MS/MS. All three instances in which FQs and SAs were present at concentrations near or above the assay limit of detection were identified as positive by the developed DB-FPIA, demonstrating that the method is suitable for rapid screening of FQs and SAs contamination. The novel methodology combines the advantage of the FPIA and the broad sensitivity of scDb and shows great promise for fast multi-analyte screening of low-molecular weight chemical residues in food samples.

A sub-millimeter resolution PET detector module using a multi-pixel photon counter array

NASA Astrophysics Data System (ADS)

Song, Tae Yong; Wu, Heyu; Komarov, Sergey; Siegel, Stefan B.; Tai, Yuan-Chuan

2010-05-01

A PET block detector module using an array of sub-millimeter lutetium oxyorthosilicate (LSO) crystals read out by an array of surface-mount, semiconductor photosensors has been developed. The detector consists of a LSO array, a custom acrylic light guide, a 3 × 3 multi-pixel photon counter (MPPC) array (S10362-11-050P, Hamamatsu Photonics, Japan) and a readout board with a charge division resistor network. The LSO array consists of 100 crystals, each measuring 0.8 × 0.8 × 3 mm3 and arranged in 0.86 mm pitches. A Monte Carlo simulation was used to aid the design and fabrication of a custom light guide to control distribution of scintillation light over the surface of the MPPC array. The output signals of the nine MPPC are multiplexed by a charge division resistor network to generate four position-encoded analog outputs. Flood image, energy resolution and timing resolution measurements were performed using standard NIM electronics. The linearity of the detector response was investigated using gamma-ray sources of different energies. The 10 × 10 array of 0.8 mm LSO crystals was clearly resolved in the flood image. The average energy resolution and standard deviation were 20.0% full-width at half-maximum (FWHM) and ±5.0%, respectively, at 511 keV. The timing resolution of a single MPPC coupled to a LSO crystal was found to be 857 ps FWHM, and the value for the central region of detector module was 1182 ps FWHM when ±10% energy window was applied. The nonlinear response of a single MPPC when used to read out a single LSO was observed among the corner crystals of the proposed detector module. However, the central region of the detector module exhibits significantly less nonlinearity (6.5% for 511 keV). These results demonstrate that (1) a charge-sharing resistor network can effectively multiplex MPPC signals and reduce the number of output signals without significantly degrading the performance of a PET detector and (2) a custom light guide to permit light sharing among multiple MPPC and to diffuse and direct scintillation light can reduce the nonlinearity of the detector response within the limited dynamic range of a typical MPPC. As a result, the proposed PET detector module has the potential to be refined for use in high-resolution PET insert applications.

A sub-millimeter resolution PET detector module using a multi-pixel photon counter array.

PubMed

Song, Tae Yong; Wu, Heyu; Komarov, Sergey; Siegel, Stefan B; Tai, Yuan-Chuan

2010-05-07

A PET block detector module using an array of sub-millimeter lutetium oxyorthosilicate (LSO) crystals read out by an array of surface-mount, semiconductor photosensors has been developed. The detector consists of a LSO array, a custom acrylic light guide, a 3 x 3 multi-pixel photon counter (MPPC) array (S10362-11-050P, Hamamatsu Photonics, Japan) and a readout board with a charge division resistor network. The LSO array consists of 100 crystals, each measuring 0.8 x 0.8 x 3 mm(3) and arranged in 0.86 mm pitches. A Monte Carlo simulation was used to aid the design and fabrication of a custom light guide to control distribution of scintillation light over the surface of the MPPC array. The output signals of the nine MPPC are multiplexed by a charge division resistor network to generate four position-encoded analog outputs. Flood image, energy resolution and timing resolution measurements were performed using standard NIM electronics. The linearity of the detector response was investigated using gamma-ray sources of different energies. The 10 x 10 array of 0.8 mm LSO crystals was clearly resolved in the flood image. The average energy resolution and standard deviation were 20.0% full-width at half-maximum (FWHM) and +/-5.0%, respectively, at 511 keV. The timing resolution of a single MPPC coupled to a LSO crystal was found to be 857 ps FWHM, and the value for the central region of detector module was 1182 ps FWHM when +/-10% energy window was applied. The nonlinear response of a single MPPC when used to read out a single LSO was observed among the corner crystals of the proposed detector module. However, the central region of the detector module exhibits significantly less nonlinearity (6.5% for 511 keV). These results demonstrate that (1) a charge-sharing resistor network can effectively multiplex MPPC signals and reduce the number of output signals without significantly degrading the performance of a PET detector and (2) a custom light guide to permit light sharing among multiple MPPC and to diffuse and direct scintillation light can reduce the nonlinearity of the detector response within the limited dynamic range of a typical MPPC. As a result, the proposed PET detector module has the potential to be refined for use in high-resolution PET insert applications.

A sub-millimeter resolution PET detector module using a multi-pixel photon counter array

PubMed Central

Song, Tae Yong; Wu, Heyu; Komarov, Sergey; Siegel, Stefan B; Tai, Yuan-Chuan

2010-01-01

A PET block detector module using an array of sub-millimeter lutetium oxyorthosilicate (LSO) crystals read out by an array of surface-mount, semiconductor photosensors has been developed. The detector consists of a LSO array, a custom acrylic light guide, a 3 × 3 multi-pixel photon counter (MPPC) array (S10362-11-050P, Hamamatsu Photonics, Japan) and a readout board with a charge division resistor network. The LSO array consists of 100 crystals, each measuring 0.8 × 0.8 × 3 mm3 and arranged in 0.86 mm pitches. A Monte Carlo simulation was used to aid the design and fabrication of a custom light guide to control distribution of scintillation light over the surface of the MPPC array. The output signals of the nine MPPC are multiplexed by a charge division resistor network to generate four position-encoded analog outputs. Flood image, energy resolution and timing resolution measurements were performed using standard NIM electronics. The linearity of the detector response was investigated using gamma-ray sources of different energies. The 10 × 10 array of 0.8 mm LSO crystals was clearly resolved in the flood image. The average energy resolution and standard deviation were 20.0% full-width at half-maximum (FWHM) and ±5.0%, respectively, at 511 keV. The timing resolution of a single MPPC coupled to a LSO crystal was found to be 857 ps FWHM, and the value for the central region of detector module was 1182 ps FWHM when ±10% energy window was applied. The nonlinear response of a single MPPC when used to read out a single LSO was observed among the corner crystals of the proposed detector module. However, the central region of the detector module exhibits significantly less nonlinearity (6.5% for 511 keV). These results demonstrate that (1) a charge-sharing resistor network can effectively multiplex MPPC signals and reduce the number of output signals without significantly degrading the performance of a PET detector and (2) a custom light guide to permit light sharing among multiple MPPC and to diffuse and direct scintillation light can reduce the nonlinearity of the detector response within the limited dynamic range of a typical MPPC. As a result, the proposed PET detector module has the potential to be refined for use in high-resolution PET insert applications. PMID:20393236

Bolometric kinetic inductance detector technology for sub-millimeter radiometric imaging

NASA Astrophysics Data System (ADS)

Hassel, Juha; Timofeev, Andrey V.; Vesterinen, Visa; Sipola, Hannu; Helistö, Panu; Aikio, Mika; Mäyrä, Aki; Grönberg, Leif; Luukanen, Arttu

2015-10-01

Radiometric sub-millimeter imaging is a candidate technology especially in security screening applications utilizing the property of radiation in the band of 0.2 - 1.0 THz to penetrate through dielectric substances such as clothing. The challenge of the passive technology is the fact that the irradiance corresponding to the blackbody radiation is very weak in this spectral band: about two orders of magnitude below that of the infrared band. Therefore the role of the detector technology is of ultimate importance to achieve sufficient sensitivity. In this paper we present results related to our technology relying on superconducting kinetic inductance detectors operating in a thermal (bolometric) mode. The detector technology is motivated by the fact that it is naturally suitable for scalable multiplexed readout systems, and operates with relatively simple cryogenics. We will review the basic concepts of the detectors, and provide experimental figures of merit. Furthermore, we will discuss the issues related to the scale-up of our detector technology into large 2D focal plane arrays.

Laser-excited fluorescence for measuring atmospheric pollution

NASA Technical Reports Server (NTRS)

Menzies, R. T.

1975-01-01

System measures amount of given pollutant at specific location. Infrared laser aimed at location has wavelength that will cause molecules of pollutant to fluoresce. Detector separates fluorescence from other radiation and measures its intensity to indicate concentration of pollutant.

Generating multiplex gradients of biomolecules for controlling cellular adhesion in parallel microfluidic channels.

PubMed

Didar, Tohid Fatanat; Tabrizian, Maryam

2012-11-07

Here we present a microfluidic platform to generate multiplex gradients of biomolecules within parallel microfluidic channels, in which a range of multiplex concentration gradients with different profile shapes are simultaneously produced. Nonlinear polynomial gradients were also generated using this device. The gradient generation principle is based on implementing parrallel channels with each providing a different hydrodynamic resistance. The generated biomolecule gradients were then covalently functionalized onto the microchannel surfaces. Surface gradients along the channel width were a result of covalent attachments of biomolecules to the surface, which remained functional under high shear stresses (50 dyn/cm(2)). An IgG antibody conjugated to three different fluorescence dyes (FITC, Cy5 and Cy3) was used to demonstrate the resulting multiplex concentration gradients of biomolecules. The device enabled generation of gradients with up to three different biomolecules in each channel with varying concentration profiles. We were also able to produce 2-dimensional gradients in which biomolecules were distributed along the length and width of the channel. To demonstrate the applicability of the developed design, three different multiplex concentration gradients of REDV and KRSR peptides were patterned along the width of three parallel channels and adhesion of primary human umbilical vein endothelial cell (HUVEC) in each channel was subsequently investigated using a single chip.

Multiplexed SNP genotyping using the Qbead™ system: a quantum dot-encoded microsphere-based assay

PubMed Central

Xu, Hongxia; Sha, Michael Y.; Wong, Edith Y.; Uphoff, Janet; Xu, Yanzhang; Treadway, Joseph A.; Truong, Anh; O’Brien, Eamonn; Asquith, Steven; Stubbins, Michael; Spurr, Nigel K.; Lai, Eric H.; Mahoney, Walt

2003-01-01

We have developed a new method using the Qbead™ system for high-throughput genotyping of single nucleotide polymorphisms (SNPs). The Qbead system employs fluorescent Qdot™ semiconductor nanocrystals, also known as quantum dots, to encode microspheres that subsequently can be used as a platform for multiplexed assays. By combining mixtures of quantum dots with distinct emission wavelengths and intensities, unique spectral ‘barcodes’ are created that enable the high levels of multiplexing required for complex genetic analyses. Here, we applied the Qbead system to SNP genotyping by encoding microspheres conjugated to allele-specific oligonucleotides. After hybridization of oligonucleotides to amplicons produced by multiplexed PCR of genomic DNA, individual microspheres are analyzed by flow cytometry and each SNP is distinguished by its unique spectral barcode. Using 10 model SNPs, we validated the Qbead system as an accurate and reliable technique for multiplexed SNP genotyping. By modifying the types of probes conjugated to microspheres, the Qbead system can easily be adapted to other assay chemistries for SNP genotyping as well as to other applications such as analysis of gene expression and protein–protein interactions. With its capability for high-throughput automation, the Qbead system has the potential to be a robust and cost-effective platform for a number of applications. PMID:12682378

Detection of inflammatory cytokines using a fiber optic microsphere immunoassay array

NASA Astrophysics Data System (ADS)

Blicharz, Timothy M.; Walt, David R.

2006-10-01

A multiplexed fiber optic microsphere-based immunoassay array capable of simultaneously measuring five inflammatory cytokines has been developed. Five groups of amine-functionalized 3.1 micron microspheres were internally encoded with five distinct concentrations of a europium dye and converted to cytokine probes by covalently coupling monoclonal capture antibodies specific for human VEGF, IFN-gamma, RANTES, IP-10, and Eotaxin-3 to the microspheres via glutaraldehyde chemistry. The microspheres were pooled and loaded into a 1 mm diameter fiber optic bundle containing ~50,000 individual etched microwells, producing the multiplexed cytokine immunoassay array. Multiple arrays can be created from a single microsphere pool for high throughput sample analysis. Sandwich fluoroimmunoassays were performed by incubating the probe array in a sample, followed by incubation in a mixture of biotin-labeled detection antibodies that are complementary to the five cytokines. Finally, universal detection of each protein was performed using a fluorescence imaging system after briefly immersing the array in a solution of fluorophore-labeled streptavidin. The multiplexed cytokine array has been shown to respond selectively to VEGF, IFNgamma, RANTES, IP-10, and Eotaxin-3, permitting multiplexed quantitative analysis. Ultimately, the multiplexed cytokine array will be utilized to evaluate the potential of using saliva as a noninvasive diagnostic fluid for pulmonary inflammatory diseases such as asthma.

Parallel separations using capillary electrophoresis on a multilane microchip with multiplexed laser-induced fluorescence detection.

PubMed

Nikcevic, Irena; Piruska, Aigars; Wehmeyer, Kenneth R; Seliskar, Carl J; Limbach, Patrick A; Heineman, William R

2010-08-01

Parallel separations using CE on a multilane microchip with multiplexed LIF detection is demonstrated. The detection system was developed to simultaneously record data on all channels using an expanded laser beam for excitation, a camera lens to capture emission, and a CCD camera for detection. The detection system enables monitoring of each channel continuously and distinguishing individual lanes without significant crosstalk between adjacent lanes. Multiple analytes can be determined in parallel lanes within a single microchip in a single run, leading to increased sample throughput. The pK(a) determination of small molecule analytes is demonstrated with the multilane microchip.

Muon Detector R&D in Telescope Array Experiment

NASA Astrophysics Data System (ADS)

Nonaka, T.; Takamura, M.; Honda, K.; Matthews, J. N.; Ogio, S.; Sakurai, N.; Sagawa, H.; Stokes, B. T.; Tsujimoto, M.; Yashiro, K.

The Telescope Array (TA) experiment, located in the western desert of Utah, U.S.A., at 39.38° north and 112.9° west, is collecting data of ultra high energy cosmic rays in the energy range 1018-1020 eV. The experiment has a Surface Detector (SD) array surrounded by three Fluorescence Detector (FD) stations to enable simultaneous detection of shower particles and fluorescence photons generated by the extensive air shower. Measurement of shower particles at the ground level, with different absorber thickness, enables a more detailed studies of the experiment's energy scale and of hadron interaction models. In this report, we present a design and the first observation result of a surface muon detector using lead plates and concrete as absorbers.

Data transformation methods for multiplexed assays

DOEpatents

Tammero, Lance F. Bentley; Dzenitis, John M; Hindson, Benjamin J

2013-07-23

Methods to improve the performance of an array assay are described. A correlation between fluorescence intensity-related parameters and negative control values of the assay is determined. The parameters are then adjusted as a function of the correlation. As a result, sensitivity of the assay is improved without changes in its specificity.

Definition of a near real time microbiological monitor for space vehicles

NASA Technical Reports Server (NTRS)

Kilgore, Melvin V., Jr.; Zahorchak, Robert J.; Arendale, William F.

1989-01-01

Efforts to identify the ideal candidate to serve as the biological monitor on the space station Freedom are discussed. The literature review, the evaluation scheme, descriptions of candidate monitors, experimental studies, test beds, and culture techniques are discussed. Particular attention is given to descriptions of five candidate monitors or monitoring techniques: laser light scattering, primary fluorescence, secondary fluorescence, the volatile product detector, and the surface acoustic wave detector.

Development of new photon-counting detectors for single-molecule fluorescence microscopy.

PubMed

Michalet, X; Colyer, R A; Scalia, G; Ingargiola, A; Lin, R; Millaud, J E; Weiss, S; Siegmund, Oswald H W; Tremsin, Anton S; Vallerga, John V; Cheng, A; Levi, M; Aharoni, D; Arisaka, K; Villa, F; Guerrieri, F; Panzeri, F; Rech, I; Gulinatti, A; Zappa, F; Ghioni, M; Cova, S

2013-02-05

Two optical configurations are commonly used in single-molecule fluorescence microscopy: point-like excitation and detection to study freely diffusing molecules, and wide field illumination and detection to study surface immobilized or slowly diffusing molecules. Both approaches have common features, but also differ in significant aspects. In particular, they use different detectors, which share some requirements but also have major technical differences. Currently, two types of detectors best fulfil the needs of each approach: single-photon-counting avalanche diodes (SPADs) for point-like detection, and electron-multiplying charge-coupled devices (EMCCDs) for wide field detection. However, there is room for improvements in both cases. The first configuration suffers from low throughput owing to the analysis of data from a single location. The second, on the other hand, is limited to relatively low frame rates and loses the benefit of single-photon-counting approaches. During the past few years, new developments in point-like and wide field detectors have started addressing some of these issues. Here, we describe our recent progresses towards increasing the throughput of single-molecule fluorescence spectroscopy in solution using parallel arrays of SPADs. We also discuss our development of large area photon-counting cameras achieving subnanosecond resolution for fluorescence lifetime imaging applications at the single-molecule level.

Development of new photon-counting detectors for single-molecule fluorescence microscopy

PubMed Central

Michalet, X.; Colyer, R. A.; Scalia, G.; Ingargiola, A.; Lin, R.; Millaud, J. E.; Weiss, S.; Siegmund, Oswald H. W.; Tremsin, Anton S.; Vallerga, John V.; Cheng, A.; Levi, M.; Aharoni, D.; Arisaka, K.; Villa, F.; Guerrieri, F.; Panzeri, F.; Rech, I.; Gulinatti, A.; Zappa, F.; Ghioni, M.; Cova, S.

2013-01-01

Two optical configurations are commonly used in single-molecule fluorescence microscopy: point-like excitation and detection to study freely diffusing molecules, and wide field illumination and detection to study surface immobilized or slowly diffusing molecules. Both approaches have common features, but also differ in significant aspects. In particular, they use different detectors, which share some requirements but also have major technical differences. Currently, two types of detectors best fulfil the needs of each approach: single-photon-counting avalanche diodes (SPADs) for point-like detection, and electron-multiplying charge-coupled devices (EMCCDs) for wide field detection. However, there is room for improvements in both cases. The first configuration suffers from low throughput owing to the analysis of data from a single location. The second, on the other hand, is limited to relatively low frame rates and loses the benefit of single-photon-counting approaches. During the past few years, new developments in point-like and wide field detectors have started addressing some of these issues. Here, we describe our recent progresses towards increasing the throughput of single-molecule fluorescence spectroscopy in solution using parallel arrays of SPADs. We also discuss our development of large area photon-counting cameras achieving subnanosecond resolution for fluorescence lifetime imaging applications at the single-molecule level. PMID:23267185

MOD: An Organic Detector for the Future Exploration of Mars

NASA Technical Reports Server (NTRS)

Kminek, G.; Bada, J. L.; Botta, O.; Grunthaner, F.; Glavin, D. P.

1999-01-01

The Mars Organic Detector (MOD) is designed to assess whether organic compounds, possibly associated with life, are present in Martian rock and soil samples. MOD has a detection limit that is at least two orders of magnitude more sensitive than the Viking GCMS. MOD is focused on detecting amino acids, amines and PAH (polycyclic aromatic hydrocarbons). Amino acids play an essential role in biochemistry on Earth and PAH are widespread throughout the universe and can provide an indication of the delivery of meteoritic organic material to Mars. The advantage of MOD is the absence of wet chemistry and its simple and robust design. The sample will be extracted from the mineral matrix (0.1 - 1 g of rock-powder) using sublimation and analyzed with a fluorescence detector. The isolation method is based on the fact that amino acids and PAH are volatile at temperatures greater than 150C. The fluorescence detection scheme is based on UV excitation with LED's, optical filters, PrN diode photon detector and a sample calibration reservoir. Fluorescamine is used as a fluorescing reagent for amino acids and amines, while PAH are naturally fluorescent. There is no sample preparation required and the turnaround time for a single analysis is on the order of minutes.

A new large solid angle multi-element silicon drift detector system for low energy X-ray fluorescence spectroscopy

NASA Astrophysics Data System (ADS)

Bufon, J.; Schillani, S.; Altissimo, M.; Bellutti, P.; Bertuccio, G.; Billè, F.; Borghes, R.; Borghi, G.; Cautero, G.; Cirrincione, D.; Fabiani, S.; Ficorella, F.; Gandola, M.; Gianoncelli, A.; Giuressi, D.; Kourousias, G.; Mele, F.; Menk, R. H.; Picciotto, A.; Rachevski, A.; Rashevskaya, I.; Sammartini, M.; Stolfa, A.; Zampa, G.; Zampa, N.; Zorzi, N.; Vacchi, A.

2018-03-01

Low-energy X-ray fluorescence (LEXRF) is an essential tool for bio-related research of organic samples, whose composition is dominated by light elements. Working at energies below 2 keV and being able to detect fluorescence photons of lightweight elements such as carbon (277 eV) is still a challenge, since it requires in-vacuum operations to avoid in-air photon absorption. Moreover, the detectors must have a thin entrance window and collect photons at an angle of incidence near 90 degrees to minimize the absorption by the protective coating. Considering the low fluorescence yield of light elements, it is important to cover a substantial part of the solid angle detecting ideally all emitted X-ray fluorescence (XRF) photons. Furthermore, the energy resolution of the detection system should be close to the Fano limit in order to discriminate elements whose XRF emission lines are often very close within the energy spectra. To ensure all these features, a system consisting of four monolithic multi-element silicon drift detectors was developed. The use of four separate detector units allows optimizing the incidence angle on all the sensor elements. The multi-element approach in turn provides a lower leakage current on each anode, which, in combination with ultra-low noise preamplifiers, is necessary to achieve an energy resolution close to the Fano limit. The potential of the new detection system and its applicability for typical LEXRF applications has been proved on the Elettra TwinMic beamline.

Application of CORSIKA Simulation Code to Study Lateral and Longitudinal Distribution of Fluorescence Light in Cosmic Ray Extensive Air Showers

NASA Astrophysics Data System (ADS)

Bagheri, Zahra; Davoudifar, Pantea; Rastegarzadeh, Gohar; Shayan, Milad

2017-03-01

In this paper, we used CORSIKA code to understand the characteristics of cosmic ray induced showers at extremely high energy as a function of energy, detector distance to shower axis, number, and density of secondary charged particles and the nature particle producing the shower. Based on the standard properties of the atmosphere, lateral and longitudinal development of the shower for photons and electrons has been investigated. Fluorescent light has been collected by the detector for protons, helium, oxygen, silicon, calcium and iron primary cosmic rays in different energies. So we have obtained a number of electrons per unit area, distance to the shower axis, shape function of particles density, percentage of fluorescent light, lateral distribution of energy dissipated in the atmosphere and visual field angle of detector as well as size of the shower image. We have also shown that location of highest percentage of fluorescence light is directly proportional to atomic number of elements. Also we have shown when the distance from shower axis increases and the shape function of particles density decreases severely. At the first stages of development, shower axis distance from detector is high and visual field angle is small; then with shower moving toward the Earth, angle increases. Overall, in higher energies, the fluorescent light method has more efficiency. The paper provides standard calibration lines for high energy showers which can be used to determine the nature of the particles.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Allman, M. S., E-mail: shane.allman@boulder.nist.gov; Verma, V. B.; Stevens, M.

We demonstrate a 64-pixel free-space-coupled array of superconducting nanowire single photon detectors optimized for high detection efficiency in the near-infrared range. An integrated, readily scalable, multiplexed readout scheme is employed to reduce the number of readout lines to 16. The cryogenic, optical, and electronic packaging to read out the array as well as characterization measurements are discussed.

HgCdTe 256x256 NWIR FPA

NASA Technical Reports Server (NTRS)

Vural, Kadri; Blessinger, Michael; Chen, Jenkon; Kleinhans, William

1989-01-01

Researchers developed a HgCdTe 256x256 focal plane array (FPA) which operates in the 1 to 5 micron band. This is presently the largest demonstrated HgCdTe FPA. The detector material is HgCdTe on sapphire (PACE-1 technology) which has a low thermal expansion mismatch with silicon. The multiplexer is a CMOS FET-switch device processed through a commercial silicon foundry. The multiplexer input is direct injection and the charge capacity is about 2 times 10 to the 7th power electrons. The kTC limited read noise is 400 electrons. Researchers demonstrated high background imaging using the device. The broadband quantum efficiency is measured to be 59 percent. Dark currents less than 0.1 pA were measured at 77 K for detectors processed on PACE-1 material with 4.9 microns cutoff. The dark currents decrease as the temperature is lowered, and researchers are presently studying the T less than 77 K characteristics. The interconnect yield is greater than 95 percent. The devices are available for astronomical applications.

Advanced ACTPol Cryogenic Detector Arrays and Readout

NASA Astrophysics Data System (ADS)

Henderson, S. W.; Allison, R.; Austermann, J.; Baildon, T.; Battaglia, N.; Beall, J. A.; Becker, D.; De Bernardis, F.; Bond, J. R.; Calabrese, E.; Choi, S. K.; Coughlin, K. P.; Crowley, K. T.; Datta, R.; Devlin, M. J.; Duff, S. M.; Dunkley, J.; Dünner, R.; van Engelen, A.; Gallardo, P. A.; Grace, E.; Hasselfield, M.; Hills, F.; Hilton, G. C.; Hincks, A. D.; Hloẑek, R.; Ho, S. P.; Hubmayr, J.; Huffenberger, K.; Hughes, J. P.; Irwin, K. D.; Koopman, B. J.; Kosowsky, A. B.; Li, D.; McMahon, J.; Munson, C.; Nati, F.; Newburgh, L.; Niemack, M. D.; Niraula, P.; Page, L. A.; Pappas, C. G.; Salatino, M.; Schillaci, A.; Schmitt, B. L.; Sehgal, N.; Sherwin, B. D.; Sievers, J. L.; Simon, S. M.; Spergel, D. N.; Staggs, S. T.; Stevens, J. R.; Thornton, R.; Van Lanen, J.; Vavagiakis, E. M.; Ward, J. T.; Wollack, E. J.

2016-08-01

Advanced ACTPol is a polarization-sensitive upgrade for the 6 m aperture Atacama Cosmology Telescope, adding new frequencies and increasing sensitivity over the previous ACTPol receiver. In 2016, Advanced ACTPol will begin to map approximately half the sky in five frequency bands (28-230 GHz). Its maps of primary and secondary cosmic microwave background anisotropies—imaged in intensity and polarization at few arcminute-scale resolution—will enable precision cosmological constraints and also a wide array of cross-correlation science that probes the expansion history of the universe and the growth of structure via gravitational collapse. To accomplish these scientific goals, the Advanced ACTPol receiver will be a significant upgrade to the ACTPol receiver, including four new multichroic arrays of cryogenic, feedhorn-coupled AlMn transition edge sensor polarimeters (fabricated on 150 mm diameter wafers); a system of continuously rotating meta-material silicon half-wave plates; and a new multiplexing readout architecture which uses superconducting quantum interference devices and time division to achieve a 64-row multiplexing factor. Here we present the status and scientific goals of the Advanced ACTPol instrument, emphasizing the design and implementation of the Advanced ACTPol cryogenic detector arrays.

Advanced ACTPol Cryogenic Detector Arrays and Readout

NASA Technical Reports Server (NTRS)

Henderson, S.W.; Allison, R.; Austermann, J.; Baildon, T.; Battaglia, N.; Beall, J. A.; Becker, D.; De Bernardis, F.; Bond, J. R.; Wollack, E. J.

2016-01-01

Advanced ACTPol is a polarization-sensitive upgrade for the 6 m aperture Atacama Cosmology Telescope, adding new frequencies and increasing sensitivity over the previous ACTPol receiver. In 2016, Advanced ACTPol will begin to map approximately half the sky in five frequency bands (28-230 GHz). Its maps of primary and secondary cosmic microwave background anisotropies-imaged in intensity and polarization at few arcminute-scale resolution-will enable precision cosmological constraints and also awide array of cross-correlation science that probes the expansion history of the universe and the growth of structure via gravitational collapse. To accomplish these scientific goals, the AdvancedACTPol receiver will be a significant upgrade to the ACTPol receiver, including four new multichroic arrays of cryogenic, feedhorn-coupled AlMn transition edge sensor polarimeters (fabricated on 150 mm diameter wafers); a system of continuously rotating meta-material silicon half-wave plates; and a new multiplexing readout architecture which uses superconducting quantum interference devices and time division to achieve a 64-row multiplexing factor. Here we present the status and scientific goals of the Advanced ACTPol instrument, emphasizing the design and implementation of the AdvancedACTPol cryogenic detector arrays.

Development status of the life marker chip instrument for ExoMars

NASA Astrophysics Data System (ADS)

Sims, Mark R.; Cullen, David C.; Rix, Catherine S.; Buckley, Alan; Derveni, Mariliza; Evans, Daniel; Miguel García-Con, Luis; Rhodes, Andrew; Rato, Carla C.; Stefinovic, Marijan; Sephton, Mark A.; Court, Richard W.; Bulloch, Christopher; Kitchingman, Ian; Ali, Zeshan; Pullan, Derek; Holt, John; Blake, Oliver; Sykes, Jonathan; Samara-Ratna, Piyal; Canali, Massimiliano; Borst, Guus; Leeuwis, Henk; Prak, Albert; Norfini, Aleandro; Geraci, Ennio; Tavanti, Marco; Brucato, John; Holm, Nils

2012-11-01

The Life Marker Chip (LMC) is one of the instruments being developed for possible flight on the 2018 ExoMars mission. The instrument uses solvents to extract organic compounds from samples of martian regolith and to transfer the extracts to dedicated detectors based around the use of antibodies. The scientific aims of the instrument are to detect organics in the form of biomarkers that might be associated with extinct life, extant life or abiotic sources of organics. The instrument relies on a novel surfactant-based solvent system and bespoke, commercial and research-developed antibodies against a number of distinct biomarkers or molecular types. The LMC comprises of a number of subsystems designed to accept up to four discrete samples of martian regolith or crushed rock, implement the solvent extraction, perform microfluidic-based multiplexed antibody-assays for biomarkers and other targets, optically detect the fluorescent output of the assays, control the internal instrument pressure and temperature, in addition to the associated instrument control electronics and software. The principle of operation, the design and the instrument development status as of December 2011 are reported here. The instrument principle can be extended to other configurations and missions as needed.

A transmission imaging spectrograph and microfabricated channel system for DNA analysis.

PubMed

Simpson, J W; Ruiz-Martinez, M C; Mulhern, G T; Berka, J; Latimer, D R; Ball, J A; Rothberg, J M; Went, G T

2000-01-01

In this paper we present the development of a DNA analysis system using a microfabricated channel device and a novel transmission imaging spectrograph which can be efficiently incorporated into a high throughput genomics facility for both sizing and sequencing of DNA fragments. The device contains 48 channels etched on a glass substrate. The channels are sealed with a flat glass plate which also provides a series of apertures for sample loading and contact with buffer reservoirs. Samples can be easily loaded in volumes up to 640 nL without band broadening because of an efficient electrokinetic stacking at the electrophoresis channel entrance. The system uses a dual laser excitation source and a highly sensitive charge-coupled device (CCD) detector allowing for simultaneous detection of many fluorescent dyes. The sieving matrices for the separation of single-stranded DNA fragments are polymerized in situ in denaturing buffer systems. Examples of separation of single-stranded DNA fragments up to 500 bases in length are shown, including accurate sizing of GeneCalling fragments, and sequencing samples prepared with a reduced amount of dye terminators. An increase in sample throughput has been achieved by color multiplexing.

Demonstrating Enabling Technologies for the High-Resolution Imaging Spectrometer of the Next NASA X-ray Astronomy Mission

NASA Astrophysics Data System (ADS)

Kilbourne, Caroline; Adams, J. S.; Bandler, S.; Chervenak, J.; Chiao, M.; Doriese, R.; Eckart, M.; Finkbeiner, F.; Fowler, J. W.; Hilton, G.; Irwin, K.; Kelley, R. L.; Moseley, S. J.; Porter, F. S.; Reintsema, C.; Sadleir, J.; Smith, S. J.; Swetz, D.; Ullom, J.

2014-01-01

NASA/GSFC and NIST-Boulder are collaborating on a program to advance superconducting transition-edge sensor (TES) microcalorimeter technology toward Technology Readiness Level (TRL) 6. The technology development for a TES imaging X-ray microcalorimeter spectrometer (TES microcalorimeter arrays and time-division multiplexed SQUID readout) is now at TRL 4, as evaluated by both NASA and the European Space Agency (ESA) during mission formulation for the International X-ray Observatory (IXO). We will present the status of the development program. The primary goal of the current project is to advance the core X-ray Microcalorimeter Spectrometer (XMS) detector-system technologies to a demonstration of TRL 5 in 2014. Additional objectives are to develop and demonstrate two important related technologies to at least TRL 4: position-sensitive TES devices and code-division multiplexing (CDM). These technologies have the potential to expand significantly the range of possible instrument optimizations; together they allow an expanded focal plane and higher per-pixel count rates without greatly increasing mission resources. The project also includes development of a design concept and critical technologies needed for the thermal, electrical, and mechanical integration of the detector and readout components into the focal-plane assembly. A verified design concept for the packaging of the focal-plane components will be needed for the detector system eventually to advance to TRL 6. Thus, the current project is a targeted development and demonstration program designed to make significant progress in advancing the XMS detector system toward TRL 6, establishing its readiness for a range of possible mission implementations.

Multiplexed Detection of Cytokines Based on Dual Bar-Code Strategy and Single-Molecule Counting.

PubMed

Li, Wei; Jiang, Wei; Dai, Shuang; Wang, Lei

2016-02-02

Cytokines play important roles in the immune system and have been regarded as biomarkers. While single cytokine is not specific and accurate enough to meet the strict diagnosis in practice, in this work, we constructed a multiplexed detection method for cytokines based on dual bar-code strategy and single-molecule counting. Taking interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) as model analytes, first, the magnetic nanobead was functionalized with the second antibody and primary bar-code strands, forming a magnetic nanoprobe. Then, through the specific reaction of the second antibody and the antigen that fixed by the primary antibody, sandwich-type immunocomplex was formed on the substrate. Next, the primary bar-code strands as amplification units triggered multibranched hybridization chain reaction (mHCR), producing nicked double-stranded polymers with multiple branched arms, which were served as secondary bar-code strands. Finally, the secondary bar-code strands hybridized with the multimolecule labeled fluorescence probes, generating enhanced fluorescence signals. The numbers of fluorescence dots were counted one by one for quantification with epi-fluorescence microscope. By integrating the primary and secondary bar-code-based amplification strategy and the multimolecule labeled fluorescence probes, this method displayed an excellent sensitivity with the detection limits were both 5 fM. Unlike the typical bar-code assay that the bar-code strands should be released and identified on a microarray, this method is more direct. Moreover, because of the selective immune reaction and the dual bar-code mechanism, the resulting method could detect the two targets simultaneously. Multiple analysis in human serum was also performed, suggesting that our strategy was reliable and had a great potential application in early clinical diagnosis.

Toward Large Field-of-View High-Resolution X-ray Imaging Spectrometers: Microwave Multiplexed Readout of 28 TES Microcalorimeters

NASA Astrophysics Data System (ADS)

Yoon, W.; Adams, J. S.; Bandler, S. R.; Becker, D.; Bennett, D. A.; Chervenak, J. A.; Datesman, A. M.; Eckart, M. E.; Finkbeiner, F. M.; Fowler, J. W.; Gard, J. D.; Hilton, G. C.; Kelley, R. L.; Kilbourne, C. A.; Mates, J. A. B.; Miniussi, A. R.; Moseley, S. H.; Noroozian, O.; Porter, F. S.; Reintsema, C. D.; Sadleir, J. E.; Sakai, K.; Smith, S. J.; Stevenson, T. R.; Swetz, D. S.; Ullom, J. N.; Vale, L. R.; Wakeham, N. A.; Wassell, E. J.; Wollack, E. J.

2018-04-01

We performed small-scale demonstrations 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 55 mK stage of our laboratory ADR. Using a ROACH2 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.

The energy spectrum of ultra-high-energy cosmic rays measured by the Telescope Array FADC fluorescence detectors in monocular mode

NASA Astrophysics Data System (ADS)

Abu-Zayyad, T.; Aida, R.; Allen, M.; Anderson, R.; Azuma, R.; Barcikowski, E.; Belz, J. W.; Bergman, D. R.; Blake, S. A.; Cady, R.; Cheon, B. G.; Chiba, J.; Chikawa, M.; Cho, E. J.; Cho, W. R.; Fujii, H.; Fujii, T.; Fukuda, T.; Fukushima, M.; Hanlon, W.; Hayashi, K.; Hayashi, Y.; Hayashida, N.; Hibino, K.; Hiyama, K.; Honda, K.; Iguchi, T.; Ikeda, D.; Ikuta, K.; Inoue, N.; Ishii, T.; Ishimori, R.; Ito, H.; Ivanov, D.; Iwamoto, S.; Jui, C. C. H.; Kadota, K.; Kakimoto, F.; Kalashev, O.; Kanbe, T.; Kasahara, K.; Kawai, H.; Kawakami, S.; Kawana, S.; Kido, E.; Kim, H. B.; Kim, H. K.; Kim, J. H.; Kim, J. H.; Kitamoto, K.; Kitamura, S.; Kitamura, Y.; Kobayashi, K.; Kobayashi, Y.; Kondo, Y.; Kuramoto, K.; Kuzmin, V.; Kwon, Y. J.; Lan, J.; Lim, S. I.; Lundquist, J. P.; Machida, S.; Martens, K.; Matsuda, T.; Matsuura, T.; Matsuyama, T.; Matthews, J. N.; Myers, I.; Minamino, M.; Miyata, K.; Murano, Y.; Nagataki, S.; Nakamura, T.; Nam, S. W.; Nonaka, T.; Ogio, S.; Ogura, J.; Ohnishi, M.; Ohoka, H.; Oki, K.; Oku, D.; Okuda, T.; Ono, M.; Oshima, A.; Ozawa, S.; Park, I. H.; Pshirkov, M. S.; Rodriguez, D. C.; Roh, S. Y.; Rubtsov, G.; Ryu, D.; Sagawa, H.; Sakurai, N.; Sampson, A. L.; Scott, L. M.; Shah, P. D.; Shibata, F.; Shibata, T.; Shimodaira, H.; Shin, B. K.; Shin, J. I.; Shirahama, T.; Smith, J. D.; Sokolsky, P.; Sonley, T. J.; Springer, R. W.; Stokes, B. T.; Stratton, S. R.; Stroman, T. A.; Suzuki, S.; Takahashi, Y.; Takeda, M.; Taketa, A.; Takita, M.; Tameda, Y.; Tanaka, H.; Tanaka, K.; Tanaka, M.; Thomas, S. B.; Thomson, G. B.; Tinyakov, P.; Tkachev, I.; Tokuno, H.; Tomida, T.; Troitsky, S.; Tsunesada, Y.; Tsutsumi, K.; Tsuyuguchi, Y.; Uchihori, Y.; Udo, S.; Ukai, H.; Vasiloff, G.; Wada, Y.; Wong, T.; Yamakawa, Y.; Yamane, R.; Yamaoka, H.; Yamazaki, K.; Yang, J.; Yoneda, Y.; Yoshida, S.; Yoshii, H.; Zollinger, R.; Zundel, Z.

2013-08-01

We present a measurement of the energy spectrum of ultra-high-energy cosmic rays performed by the Telescope Array experiment using monocular observations from its two new FADC-based fluorescence detectors. After a short description of the experiment, we describe the data analysis and event reconstruction procedures. Since the aperture of the experiment must be calculated by Monte Carlo simulation, we describe this calculation and the comparisons of simulated and real data used to verify the validity of the aperture calculation. Finally, we present the energy spectrum calculated from the merged monocular data sets of the two FADC-based detectors, and also the combination of this merged spectrum with an independent, previously published monocular spectrum measurement performed by Telescope Array's third fluorescence detector [T. Abu-Zayyad et al., The energy spectrum of Telescope Array's middle drum detector and the direct comparison to the high resolution fly's eye experiment, Astroparticle Physics 39 (2012) 109-119, http://dx.doi.org/10.1016/j.astropartphys.2012.05.012, Available from: ]. This combined spectrum corroborates the recently published Telescope Array surface detector spectrum [T. Abu-Zayyad, et al., The cosmic-ray energy spectrum observed with the surface detector of the Telescope Array experiment, ApJ 768 (2013) L1, http://dx.doi.org/10.1088/2041-8205/768/1/L1, Available from: ] with independent systematic uncertainties.

Modulation-frequency encoded multi-color fluorescent DNA analysis in an optofluidic chip.

PubMed

Dongre, Chaitanya; van Weerd, Jasper; Besselink, Geert A J; Vazquez, Rebeca Martinez; Osellame, Roberto; Cerullo, Giulio; van Weeghel, Rob; van den Vlekkert, Hans H; Hoekstra, Hugo J W M; Pollnau, Markus

2011-02-21

We introduce a principle of parallel optical processing to an optofluidic lab-on-a-chip. During electrophoretic separation, the ultra-low limit of detection achieved with our set-up allows us to record fluorescence from covalently end-labeled DNA molecules. Different sets of exclusively color-labeled DNA fragments-otherwise rendered indistinguishable by spatio-temporal coincidence-are traced back to their origin by modulation-frequency-encoded multi-wavelength laser excitation, fluorescence detection with a single ultrasensitive, albeit color-blind photomultiplier, and Fourier analysis decoding. As a proof of principle, fragments obtained by multiplex ligation-dependent probe amplification from independent human genomic segments, associated with genetic predispositions to breast cancer and anemia, are simultaneously analyzed.

A frequency and sensitivity tunable microresonator array for high-speed quantum processor readout

DOE Office of Scientific and Technical Information (OSTI.GOV)

Whittaker, J. D., E-mail: jwhittaker@dwavesys.com; Swenson, L. J.; Volkmann, M. H.

Superconducting microresonators have been successfully utilized as detection elements for a wide variety of applications. With multiplexing factors exceeding 1000 detectors per transmission line, they are the most scalable low-temperature detector technology demonstrated to date. For high-throughput applications, fewer detectors can be coupled to a single wire but utilize a larger per-detector bandwidth. For all existing designs, fluctuations in fabrication tolerances result in a non-uniform shift in resonance frequency and sensitivity, which ultimately limits the efficiency of bandwidth utilization. Here, we present the design, implementation, and initial characterization of a superconducting microresonator readout integrating two tunable inductances per detector. Wemore » demonstrate that these tuning elements provide independent control of both the detector frequency and sensitivity, allowing us to maximize the transmission line bandwidth utilization. Finally, we discuss the integration of these detectors in a multilayer fabrication stack for high-speed readout of the D-Wave quantum processor, highlighting the use of control and routing circuitry composed of single-flux-quantum loops to minimize the number of control wires at the lowest temperature stage.« less

Noise, fluctuation, and HADAMARD-transform spectrometry

NASA Astrophysics Data System (ADS)

Nitzsche, Guenter; Riesenberg, Rainer

2003-05-01

The HADAMARD principle is known in optics as a multiplex technique. It describes the mode with the most advantageous increase of the signal-to-noise ratio (SNR) in terms of scanning (Fellget advantage). The maximum increase of SNR, we call it gain, is (n+1)/(2On), where n is the number of multiplexing. It is valid in the case of pure detector noise. The multiplex encoding Hadamard pattern in case of n = 7 is 1110100, whereby 1 stands for a switched on channel performed by a field selector. The signals of all (switched on) channels are detected by a single detector. n measurement steps with a cyclic change of the pattern is necessary to perform the Hadamard transformation and to get the result of each individual channel. In case of n = 7 the theoretical gain is 1.51. For all possible multiplex pattern (1100000, 1110000 and so on) the gain is theoretically investigated. A multiplexing advantage (gain > 1) is reached only by the Hadamard pattern, the inverse Hadamard pattern and for (0111111)-pattern (gain=1.08). Most of the multiplex pattern are disadvantageous. The reason for maximum gain of the HADAMARD transformation is analysed theoretically. Signal fluctuations during the measurement caused by fluctuations of the illumination or by the object under test, reduce the multiplex gain, too. So the limits for realizing a gain are estimated theoretically. Essential is the transformation procedure and its influence on the error propagation. The results could be verified by experiments with array spectrometeres. Requirements are derived by numerical simulation concerning the stability of the signals to be multiplexed. It is simulated the needed stability of the signals with increasing order of multiplexing. So the increase of the multiplex gain is limited by signal fluctuations. A realized 96 channel spectral reader is presented as a modern application of an optical multiplexing arrangement. ! M. Harvid, N. J. A. Sloane, Hadamard Transform Optics, Academic Press, 1979 ! R.A. De Verse, R.M. Hammaker, W. G. Fately, J.A.Graham, J.D.Tate, "Spectrometry and imaging using a digital micromirror array" American Laboratory, Vol. 30, 21, pp. 112-120, 1998 ! R. Riesenberg, A. Wuttig, B. Harnisch, "Optical MEMS Technology for Multiplexing in High-End Micro-Scpectrometers", Proc. SPIE 4928, 6-14, 2002 ! A. Wuttig, R. Riesenberg, "Hyperspectral imager with a facile MEMS", Proc. SPIE 4881A, 2002, to be published ! R. Riesenberg, G. Nitzsche, W. Voigt, 'HADAMARD Encoding and other optical Multiplexing', VDI-Berichte 1694, pp. 345-350, 2002 ! A. Wuttig, R. Riesenberg, G. Nitzsche, "Subpixel Analysis of Double Array Grating Spectrometer", Proc. SPIE 4480, pp. 334-344, 2002 ! A. Wuttig, R. Riesenberg, G. Nitzsche, "Integral Field and Multi Object Spectrometry with MEMS", Proc. SPIE 4480, pp. 367-376, 2002 ! R. Riesenberg, G. Nitzsche, A. Wuttig, B. Harnisch, "Micro Spectrometer and MEMS for Space" in "Smaller Satellites: Bigger Business?", edited by M. Rycroft, N. Crosby, Kluwer Academic Publisher, pp. 403-406, 2002 ! R. Riesenberg, A. Wuttig, "Optical sensors with MEMS, slit masks and micromechanical devices", Proc. SPIE 4561, pp. 315-322, 2001 ! R. Riesenberg, "MicroMechanical Slit Positioning System as a transmissive spatial Light Modulator", Proc. SPIE 4457, pp.197-203, 2001 ! R. Riesenberg, J. Lonschinski, "HADAMARD-Minispectrometer made by a Micro Device", Proc. "3rd Round Table on Micro/NanoTechnologies for Space", ESTEC, Noordwijk, The Netherlands, pp. 291 - 297, 2000 ! R. Riesenberg, U. Dillner, "HADAMARD Imaging Spectrometers", Proc. SPIE 3753, pp. 203-213, 1999 ! R. Riesenberg, Th. Seifert, "Design of spatial Light Modulator Microdevices - Micro Slit Arrays", Proc. SPIE 3680, Part One, pp. 406-414, 1999 ! R. Riesenberg, W. Voigt, J. Schoneich, "Compact Spectrometers made by Micro System Technology", Sensor 97, Proc. Vol. 2, pp. 145-150,1997

High-speed wavelength-division multiplexing quantum key distribution system.

PubMed

Yoshino, Ken-ichiro; Fujiwara, Mikio; Tanaka, Akihiro; Takahashi, Seigo; Nambu, Yoshihiro; Tomita, Akihisa; Miki, Shigehito; Yamashita, Taro; Wang, Zhen; Sasaki, Masahide; Tajima, Akio

2012-01-15

A high-speed quantum key distribution system was developed with the wavelength-division multiplexing (WDM) technique and dedicated key distillation hardware engines. Two interferometers for encoding and decoding are shared over eight wavelengths to reduce the system's size, cost, and control complexity. The key distillation engines can process a huge amount of data from the WDM channels by using a 1 Mbit block in real time. We demonstrated a three-channel WDM system that simultaneously uses avalanche photodiodes and superconducting single-photon detectors. We achieved 12 h continuous key generation with a secure key rate of 208 kilobits per second through a 45 km field fiber with 14.5 dB loss.

Continuous-variable quantum key distribution with 1 Mbps secure key rate.

PubMed

Huang, Duan; Lin, Dakai; Wang, Chao; Liu, Weiqi; Fang, Shuanghong; Peng, Jinye; Huang, Peng; Zeng, Guihua

2015-06-29

We report the first continuous-variable quantum key distribution (CVQKD) experiment to enable the creation of 1 Mbps secure key rate over 25 km standard telecom fiber in a coarse wavelength division multiplexers (CWDM) environment. The result is achieved with two major technological advances: the use of a 1 GHz shot-noise-limited homodyne detector and the implementation of a 50 MHz clock system. The excess noise due to noise photons from local oscillator and classical data channels in CWDM is controlled effectively. We note that the experimental verification of high-bit-rate CVQKD in the multiplexing environment is a significant step closer toward large-scale deployment in fiber networks.

Analog bus driver and multiplexer

NASA Technical Reports Server (NTRS)

Pain, Bedabrata (Inventor); Hancock, Bruce (Inventor); Cunningham, Thomas J. (Inventor)

2012-01-01

For a source-follower signal chain, the ohmic drop in the selection switch causes unacceptable voltage offset, non-linearity, and reduced small signal gain. For an op amp signal chain, the required bias current and the output noise rises rapidly with increasing the array format due to a rapid increase in the effective capacitance caused by the Miller effect boosting up the contribution of the bus capacitance. A new switched source-follower signal chain circuit overcomes limitations of existing op-amp based or source follower based circuits used in column multiplexers and data readout. This will improve performance of CMOS imagers, and focal plane read-out integrated circuits for detectors of infrared or ultraviolet light.

Gallium arsenide quantum well-based far infrared array radiometric imager

NASA Technical Reports Server (NTRS)

Forrest, Kathrine A.; Jhabvala, Murzy D.

1991-01-01

We have built an array-based camera (FIRARI) for thermal imaging (lambda = 8 to 12 microns). FIRARI uses a square format 128 by 128 element array of aluminum gallium arsenide quantum well detectors that are indium bump bonded to a high capacity silicon multiplexer. The quantum well detectors offer good responsivity along with high response and noise uniformity, resulting in excellent thermal images without compensation for variation in pixel response. A noise equivalent temperature difference of 0.02 K at a scene temperature of 290 K was achieved with the array operating at 60 K. FIRARI demonstrated that AlGaAS quantum well detector technology can provide large format arrays with performance superior to mercury cadmium telluride at far less cost.

Development of a canopy Solar-induced chlorophyll fluorescence measurement instrument

NASA Astrophysics Data System (ADS)

Sun, G.; Wang, X.; Niu, Zh; Chen, F.

2014-02-01

A portable solar-induced chlorophyll fluorescence detecting instrument based on Fraunhofer line principle was designed and tested. The instrument has a valid survey area of 1.3 × 1.3 meter when the height was fixed to 1.3 meter. The instrument uses sunlight as its light source. The instrument is quipped with two sets of special photoelectrical detectors with the centre wavelength at 760 nm and 771 nm respectively and bandwidth less than 1nm. Both sets of detectors are composed of an upper detector which are used for detecting incidence sunlight and a bottom detector which are used for detecting reflex light from the canopy of crop. This instrument includes photoelectric detector module, signal process module, A/D convert module, the data storage and upload module and human-machine interface module. The microprocessor calculates solar-induced fluorescence value based on the A/D values get from detectors. And the value can be displayed on the instrument's LCD, stored in the flash memory of instrument and can also be uploaded to PC through the PC's serial interface. The prototype was tested in the crop field and the results demonstrate that the instrument can measure the solar-induced chlorophyll value exactly with the correlation coefficients was 0.9 compared to the values got from Analytical Spectral Devices FieldSpec Pro spectrometer. This instrument can diagnose the plant growth status by the acquired spectral response.

Techniques Suitable for a Portable Wear Metal Analyzer.

DTIC Science & Technology

1981-09-01

measured by a detector. Commonly used detectors are semiconductor detectors or proportional counters. b. Energy-Dispersive XRPS . In the energy-dispersive...because the sample must be charred before the analysis. C. X-Ray Fluorescence Spectroscopy. Normally the counting time for XRPS is 100 seconds

Recent Results of the Telescope Array Experiment

NASA Astrophysics Data System (ADS)

Ivanov, Dmitri

2015-04-01

The Telescope Array (TA) is the largest cosmic ray experiment in the northern hemisphere and covers 10 PeV to 100 EeV range. TA is a hybrid detector that uses air fluorescence detectors combined with a ground array. TA consists of 507 plastic scintillation counters on a 1.2km square grid, overlooked by 3 fluorescence detector stations, and measures cosmic rays above 1 EeV. TA has collected 6.5 years of data. Results from the TA low energy extension (TALE), which sees cosmic rays down to 10 PeV, will also be shown. This contribution will consist of three parts. First, we will present the cosmic ray energy spectrum measured over 4 decades in energy. Next, we will discuss the latest results of the measurements of cosmic ray mass composition by the TA fluorescence detectors. Finally, we will show the latest results of the TA anisotropy measurements at the highest energies, where we have seen a concentration of events, called the ``hotspot,'' centered in the Ursa Major. For the Telescope Array Collaboration. Done...processed 1261 records...10:46:59 Beginning APS data extraction...10:47:48

A portable fluorescence detector for fast ultra trace detection of explosive vapors

NASA Astrophysics Data System (ADS)

Xin, Yunhong; He, Gang; Wang, Qi; Fang, Yu

2011-10-01

This paper developed a portable detector based on a specific material-based fluorescent sensing film for an ultra trace detection of explosives, such as 2,4,6-trinitrotoluene (TNT) or its derivate 2,4-dinitrotoluene (DNT), in ambient air or on objects tainted by explosives. The fluorescent sensing films are based on single-layer chemistry and the signal amplification effect of conjugated polymers, which exhibited higher sensitivity and shorter response time to TNT or DNT at their vapor pressures. Due to application of the light emitting diode and the solid state photomultiplier and the cross-correlation-based circuit design technology, the device has the advantages of low-power, low-cost, small size, and an improved signal to noise ratio. The results of the experiments showed that the detector can real-time detect and identify of explosive vapors at extremely low levels; it is suitable for the identification of suspect luggage, forensic analyses, or battlefields clearing.

A portable fluorescence detector for fast ultra trace detection of explosive vapors.

PubMed

Xin, Yunhong; He, Gang; Wang, Qi; Fang, Yu

2011-10-01

This paper developed a portable detector based on a specific material-based fluorescent sensing film for an ultra trace detection of explosives, such as 2,4,6-trinitrotoluene (TNT) or its derivate 2,4-dinitrotoluene (DNT), in ambient air or on objects tainted by explosives. The fluorescent sensing films are based on single-layer chemistry and the signal amplification effect of conjugated polymers, which exhibited higher sensitivity and shorter response time to TNT or DNT at their vapor pressures. Due to application of the light emitting diode and the solid state photomultiplier and the cross-correlation-based circuit design technology, the device has the advantages of low-power, low-cost, small size, and an improved signal to noise ratio. The results of the experiments showed that the detector can real-time detect and identify of explosive vapors at extremely low levels; it is suitable for the identification of suspect luggage, forensic analyses, or battlefields clearing.

Performance of a multistep fluorescence-gated proportional counter for hard X-ray astronomy

NASA Technical Reports Server (NTRS)

Dietz, K. L.; Ramsey, B. D.; Weisskopf, M. C.

1992-01-01

Results from the first flight of our proportional counter in an imaging telescope led us to rebuild the detector. We have used a Penning gas mixture (xenon + 1 percent isobutylene) and introduced a preamplification region to improve the energy resolution. We have rebuilt the pressure vessel making novel use of molybdenum as the housing material in order to reduce the residual instrument background, particularly in the fluorescence-gated mode for which the detector design has been optimized. We have also increased the sensitive gas depth from 9 to 14 cm to further increase the sensitivity to both fluorescent pairs and conventional singles. Our calibrations have shown that the overall energy resolution of the detector has been enhanced by a factor of 2, and we predict that the sensitivity at float will increase by a factor of 3 in the 50-70 keV energy band.

Precolumn derivatization followed by liquid chromatographic separation and determination of tramiprosate in rat plasma by fluorescence detector: application to pharmacokinetics.

PubMed

Rao, R Nageswara; Maurya, Pawan K; Shinde, Dhananjay D; Khalid, Sara

2011-05-15

Alzheimer disease (AD) is characterized pathologically by extracellular amyloid deposits composed of amyloid β (Aβ) protein. A simple and rapid method using HPLC with fluorescence detector was developed and validated for determination of tramiprosate in rat plasma. Pre-column derivatization of the deproteinized rat plasma was carried out using o-phthaldialdehyde (OPA) as a fluorescent reagent in presence of 3-mercaptopropionic acid. The liquid chromatographic separation was achieved on a Kromasil C18 column using methanol:acetonitrile: 20 mM phosphate buffer pH 7.5 (8.0:17.5:74.5 v/v/v) as a mobile phase in an isocratic elution mode. The eluents were monitored by a fluorescence detector set at 330 and 450 nm of excitation and emission wavelength respectively. Vigabatrin was used as an internal standard. The method was linear within the range 30.0-1000.0 ng/mL. Design of experiments (DOE) was used to evaluate the robustness of the method. The developed method was applied to study the pharmacokinetics of tramiprosate in rats. Copyright © 2011. Published by Elsevier B.V.

Trends in hard X-ray fluorescence mapping: environmental applications in the age of fast detectors.

PubMed

Lombi, E; de Jonge, M D; Donner, E; Ryan, C G; Paterson, D

2011-06-01

Environmental samples are extremely diverse but share a tendency for heterogeneity and complexity. This heterogeneity poses methodological challenges when investigating biogeochemical processes. In recent years, the development of analytical tools capable of probing element distribution and speciation at the microscale have allowed this challenge to be addressed. Of these available tools, laterally resolved synchrotron techniques such as X-ray fluorescence mapping are key methods for the in situ investigation of micronutrients and inorganic contaminants in environmental samples. This article demonstrates how recent advances in X-ray fluorescence detector technology are bringing new possibilities to environmental research. Fast detectors are helping to circumvent major issues such as X-ray beam damage of hydrated samples, as dwell times during scanning are reduced. They are also helping to reduce temporal beamtime requirements, making particularly time-consuming techniques such as micro X-ray fluorescence (μXRF) tomography increasingly feasible. This article focuses on μXRF mapping of nutrients and metalloids in environmental samples, and suggests that the current divide between mapping and speciation techniques will be increasingly blurred by the development of combined approaches.

Radioiodine detector based on laser induced fluorescence

DOEpatents

McDonald, Jimmie R.; Baronavski, Andrew P.

1980-01-01

The invention involves the measurement of the concentration of the radioisotope .sup.129 I.sub.2 in the presence of a gas. The invention uses a laser to excite a sample of the .sup.129 I.sub.2 in a sample gas chamber and a reference sample of a known concentration of .sup.129 I.sub.2 in a reference gas chamber. The .sup.129 I.sub.2 in the sample and reference gas chamber each gives off fluorescence emissions which are received by photomultipliers which provide signals to a detector. The detector uses a ratioing technique to determine the concentration of .sup.129 I.sub.2 in the sample gas chamber.

Optical performance of prototype horn-coupled TES bolometer arrays for SAFARI

NASA Astrophysics Data System (ADS)

Audley, Michael D.; de Lange, Gert; Gao, Jian-Rong; Khosropanah, Pourya; Hijmering, Richard; Ridder, Marcel L.

2016-07-01

The SAFARI Detector Test Facility is an ultra-low background optical testbed for characterizing ultra-sensitive prototype horn-coupled TES bolmeters for SAFARI, the grating spectrometer on board the proposed SPICA satellite. The testbed contains internal cold and hot black-body illuminators and a light-pipe for illumination with an external source. We have added reimaging optics to facilitate array optical measurements. The system is now being used for optical testing of prototype detector arrays read out with frequency-domain multiplexing. We present our latest optical measurements of prototype arrays and discuss these in terms of the instrument performance.

Antenna-Coupled Superconducting Tunnel Junctions with Single-Electron Transistor Readout for Detection of Sub-mm Radiation

NASA Technical Reports Server (NTRS)

Stevenson, T. R.; Hsieh, W.-T.; Li, M. J.; Stahle, C. M.; Wollack, E. J.; Schoelkopf, R. J.; Teufel, J.; Krebs, Carolyn (Technical Monitor)

2002-01-01

Antenna-coupled superconducting tunnel junction detectors have the potential for photon-counting sensitivity at sub-mm wavelengths. The device consists of an antenna structure to couple radiation into a small superconducting volume and cause quasiparticle excitations, and a single-electron transistor to measure currents through tunnel junction contacts to the absorber volume. We will describe optimization of device parameters, and recent results on fabrication techniques for producing devices with high yield for detector arrays. We will also present modeling of expected saturation power levels, antenna coupling, and rf multiplexing schemes.

The application of multiplex fluorimetric sensor for the analysis of flavonoids content in the medicinal herbs family Asteraceae, Lamiaceae, Rosaceae.

PubMed

Sytar, Oksana; Bruckova, Klaudia; Hunkova, Elena; Zivcak, Marek; Konate, Kiessoun; Brestic, Marian

2015-01-16

The aim of our research work was to quantify total flavonoid contents in the leaves of 13 plant species family Asteraceae, 8 representatives of family Lamiaceae and 9 plant species belonging to family Rosaceae, using the multiplex fluorimetric sensor. Fluorescence was measured using optical fluorescence apparatus Multiplex(R) 3 (Force-A, France) for non-destructive flavonoids estimation. The content of total flavonoids was estimated by FLAV index (expressed in relative units), that is deduced from flavonoids UV absorbing properties. Among observed plant species, the highest amount of total flavonoids has been found in leaves of Helianthus multiflorus (1.65 RU) and Echinops ritro (1.27 RU), Rudbeckia fulgida (1.13 RU) belonging to the family Asteraceae. Lowest flavonoid content has been observed in the leaves of marigold (Calendula officinalis) (0.14 RU) also belonging to family Asteraceae. The highest content of flavonoids among experimental plants of family Rosaceae has been estimated in the leaves of Rosa canina (1.18 RU) and among plant species of family Lamiaceae in the leaves of Coleus blumei (0.90 RU). This research work was done as pre-screening of flavonoids content in the leaves of plant species belonging to family Asteraceae, Lamiaceae and Rosaceae. Results indicated that statistically significant differences (P > 0.05) in flavonoids content were observed not only between families, but also among individual plant species within one family.

Development and Validation of a Fluorescent Multiplexed Immunoassay for Measurement of Transgenic Proteins in Cotton (Gossypium hirsutum).

PubMed

Yeaman, Grant R; Paul, Sudakshina; Nahirna, Iryna; Wang, Yongcheng; Deffenbaugh, Andrew E; Liu, Zi Lucy; Glenn, Kevin C

2016-06-22

In order to provide farmers with better and more customized alternatives to improve yields, combining multiple genetically modified (GM) traits into a single product (called stacked trait crops) is becoming prevalent. Trait protein expression levels are used to characterize new GM products and establish exposure limits, two important components of safety assessment. Developing a multiplexed immunoassay capable of measuring all trait proteins in the same sample allows for higher sample throughput and savings in both time and expense. Fluorescent (bead-based) multiplexed immunoassays (FMI) have gained wide acceptance in mammalian research and in clinical applications. In order to facilitate the measurement of stacked GM traits, we have developed and validated an FMI assay that can measure five different proteins (β-glucuronidase, neomycin phosphotransferase II, Cry1Ac, Cry2Ab2, and CP4 5-enolpyruvyl-shikimate-3-phosphate synthase) present in cotton leaf from a stacked trait product. Expression levels of the five proteins determined by FMI in cotton leaf tissues have been evaluated relative to expression levels determined by enzyme-linked immunosorbent assays (ELISAs) of the individual proteins and shown to be comparable. The FMI met characterization requirements similar to those used for ELISA. Therefore, it is reasonable to conclude that FMI results are equivalent to those determined by conventional individual ELISAs to measure GM protein expression levels in stacked trait products but with significantly higher throughput, reduced time, and more efficient use of resources.

Populational analysis of suspended microtissues for high-throughput, multiplexed 3D tissue engineering

PubMed Central

Chen, Alice A.; Underhill, Gregory H.; Bhatia, Sangeeta N.

2014-01-01

Three-dimensional (3D) tissue models have significantly improved our understanding of structure/function relationships and promise to lead to new advances in regenerative medicine. However, despite the expanding diversity of 3D tissue fabrication methods, approaches for functional assessment have been relatively limited. Here, we describe the fabrication of microtissue (μ-tissue) suspensions and their quantitative evaluation with techniques capable of analyzing large sample numbers and performing multiplexed parallel analysis. We applied this platform to 3D μ-tissues representing multiple stages of liver development and disease including: embryonic stem cells, bipotential hepatic progenitors, mature hepatocytes, and hepatoma cells photoencapsulated in polyethylene glycol hydrogels. Multiparametric μ-tissue cytometry enabled quantitation of fluorescent reporter expression within populations of intact μ-tissues (n≥102-103) and sorting-based enrichment of subsets for subsequent studies. Further, 3D μ-tissues could be implanted in vivo, respond to systemic stimuli, retrieved and quantitatively assessed. In order to facilitate multiplexed ‘pooled’ experimentation, fluorescent labeling strategies were developed and utilized to investigate the impact of μ-tissue composition and exposure to soluble factors. In particular, examination of drug/gene interactions on collections of 3D hepatoma μ-tissues indicated synergistic influence of doxorubicin and knockdown of the anti-apoptotic gene BCL-XL. Collectively, these studies highlight the broad utility of μ-tissue suspensions as an enabling approach for high n, populational analysis of 3D tissue biology in vitro and in vivo. PMID:20820630

Non-stochastic sampling error in quantal analyses for Campylobacter species on poultry products

USDA-ARS?s Scientific Manuscript database

Using primers and fluorescent probes specific for the most common foodborne Campylobacter species (C. jejuni = Cj and C. coli = Cc), we developed a multiplex, most probable number (MPN) assay using quantitative PCR (qPCR) as the determinant for binomial detection: number of p positives out of n = 6 ...

Evaluation of Fluorescence Microsphere Immunoassay for the Detection of Antibodies to Rift Valley Fever Nucleocapsid Protein and Glycoproteins

USDA-ARS?s Scientific Manuscript database

Rift Valley Fever virus (RVFV) is a zoonotic virus that infects ruminants including cattle, sheep, goats, camels and buffalo. Multiplexing diagnostic assays that can simultaneously detect antibodies against multiple RVFV antigens offer a high throughput test for disease surveillance and vaccine eva...

The Mexican participation at the Pierre Auger Observatory: Recent results

NASA Astrophysics Data System (ADS)

Román, S.; Alcaráz, F.; Cantoral, E.; Castro, J.; Cordero, A.; Fernández, A.; López, R.; Pacheco, C.; Rubín, M.; Salazar, H.; Valdés, J.; Vargas, M.; Villaseñor, L.; Zepeda, A.

1998-02-01

In this work we present the participations of the Mexican group at development of the Pierre Auger Observatory. We have been working in both parts of the hybrid proposed for the Auger detector, the fluorescence and the surface detectors. In the part of fluorescence, we have analyzed the resolution of the Hi-Res optical design of the fluorescence detector observatory. We have found a heterogeneus image resolution. We propose to use a lensless Schmidt camera (with spherical image surface) to duplicate the field of view to 30×30 degrees and simultaneously guarantee a resolution of one degree over of the whole field of view. By the Surface Detector, a water Čerenkov detector (WCD) prototype of reduced dimensions (cylinder 1.54 diameter filled with purified water up to 1.20 m high) is used to obtain preliminary experimental results that validate the concept of remote calibration and monitoring of WCDs. We use muons that stop and decay inside the WCD and, in a complementary way, muons that croos the WCD. We used a moun telescope trigger in order to study the charge distribution of vertical muons, their pulse amplitude decay and the Cerenkov light attenuation length of those secondary cosmic muons we include the bacteria population content for the four months of operation to validate the monitoring method.

High-Performance Multiplex SNP Analysis of Three Hemochromatosis-Related Mutations With Capillary Array Electrophoresis Microplates

PubMed Central

Medintz, Igor; Wong, Wendy W.; Berti, Lorenzo; Shiow, Lawrence; Tom, Jennifer; Scherer, James; Sensabaugh, George; Mathies, Richard A.

2001-01-01

An assay is described for high-throughput single nucleotide polymorphism (SNP) genotyping on a microfabricated capillary array electrophoresis (CAE) microchip. The assay targets the three common variants at the HFE locus associated with the genetic disease hereditary hemochromatosis (HHC). The assay employs allele-specific PCR (ASPCR) for the C282Y (845g->a), H63D (187c->g), and S65C (193a->t) variants using fluorescently-labeled energy-transfer (ET) allele-specific primers. Using a 96-channel radial CAE microplate, the labeled ASPCR products generated from 96 samples in a reference Caucasian population are simultaneously separated with single-base-pair resolution and genotyped in under 10 min. Detection is accomplished with a laser-excited rotary four-color fluorescence scanner. The allele-specific amplicons are differentiated on the basis of both their size and the color of the label emission. This study is the first demonstration of the combined use of ASPCR with ET primers and microfabricated radial CAE microplates to perform multiplex SNP analyses in a clinically relevant population. PMID:11230165

Multichannel waveguides for the simultaneous detection of disease biomarkers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mukundan, Harshini; Price, Dominique Z; Grace, Wynne K

2009-01-01

The sensor team at the Los Alamos National Laboratory has developed a waveguide-based optical biosensor that has previously been used for the detection of biomarkers associated with diseases such as tuberculosis, breast cancer, anthrax and influenza in complex biological samples (e.g., serum and urine). However, no single biomarker can accurately predict disease. To address this issue, we developed a multiplex assay for the detection of components of the Bacillus anthracis lethal toxin on single mode planar optical waveguides with tunable quantum dots as the fluorescence reporter. This limited ability to multiplex is still insufficient for accurate detection of disease ormore » for monitoring prognosis. In this manuscript, we demonstrate for the first time, the design, fabrication and successful evaluation of a multichannel planar optical waveguide for the simultaneous detection of at least three unknown samples in quadruplicate. We demonstrate the simultaneous, rapid (30 min), quantitative (with internal standard) and sensitive (limit of detection of 1 pM) detection of protective antigen and lethal factor of Bacillus anthracis in complex biological samples (serum) using specific monoclonal antibodies labeled with quantum dots as the fluorescence reporter.« less

Cell Painting, a high-content image-based assay for morphological profiling using multiplexed fluorescent dyes

PubMed Central

Bray, Mark-Anthony; Singh, Shantanu; Han, Han; Davis, Chadwick T.; Borgeson, Blake; Hartland, Cathy; Kost-Alimova, Maria; Gustafsdottir, Sigrun M.; Gibson, Christopher C.; Carpenter, Anne E.

2016-01-01

In morphological profiling, quantitative data are extracted from microscopy images of cells to identify biologically relevant similarities and differences among samples based on these profiles. This protocol describes the design and execution of experiments using Cell Painting, a morphological profiling assay multiplexing six fluorescent dyes imaged in five channels, to reveal eight broadly relevant cellular components or organelles. Cells are plated in multi-well plates, perturbed with the treatments to be tested, stained, fixed, and imaged on a high-throughput microscope. Then, automated image analysis software identifies individual cells and measures ~1,500 morphological features (various measures of size, shape, texture, intensity, etc.) to produce a rich profile suitable for detecting subtle phenotypes. Profiles of cell populations treated with different experimental perturbations can be compared to suit many goals, such as identifying the phenotypic impact of chemical or genetic perturbations, grouping compounds and/or genes into functional pathways, and identifying signatures of disease. Cell culture and image acquisition takes two weeks; feature extraction and data analysis take an additional 1-2 weeks. PMID:27560178

The DCU: the detector control unit for SPICA-SAFARI

NASA Astrophysics Data System (ADS)

Clénet, Antoine; Ravera, Laurent; Bertrand, Bernard; den Hartog, Roland H.; Jackson, Brian D.; van Leeuven, Bert-Joost; van Loon, Dennis; Parot, Yann; Pointecouteau, Etienne; Sournac, Anthony

2014-08-01

IRAP is developing the warm electronic, so called Detector Control Unit" (DCU), in charge of the readout of the SPICA-SAFARI's TES type detectors. The architecture of the electronics used to readout the 3 500 sensors of the 3 focal plane arrays is based on the frequency domain multiplexing technique (FDM). In each of the 24 detection channels the data of up to 160 pixels are multiplexed in frequency domain between 1 and 3:3 MHz. The DCU provides the AC signals to voltage-bias the detectors; it demodulates the detectors data which are readout in the cold by a SQUID; and it computes a feedback signal for the SQUID to linearize the detection chain in order to optimize its dynamic range. The feedback is computed with a specific technique, so called baseband feedback (BBFB) which ensures that the loop is stable even with long propagation and processing delays (i.e. several µs) and with fast signals (i.e. frequency carriers at 3:3 MHz). This digital signal processing is complex and has to be done at the same time for the 3 500 pixels. It thus requires an optimisation of the power consumption. We took the advantage of the relatively reduced science signal bandwidth (i.e. 20 - 40 Hz) to decouple the signal sampling frequency (10 MHz) and the data processing rate. Thanks to this method we managed to reduce the total number of operations per second and thus the power consumption of the digital processing circuit by a factor of 10. Moreover we used time multiplexing techniques to share the resources of the circuit (e.g. a single BBFB module processes 32 pixels). The current version of the firmware is under validation in a Xilinx Virtex 5 FPGA, the final version will be developed in a space qualified digital ASIC. Beyond the firmware architecture the optimization of the instrument concerns the characterization routines and the definition of the optimal parameters. Indeed the operation of the detection and readout chains requires to properly define more than 17 500 parameters (about 5 parameters per pixel). Thus it is mandatory to work out an automatic procedure to set up these optimal values. We defined a fast algorithm which characterizes the phase correction to be applied by the BBFB firmware and the pixel resonance frequencies. We also defined a technique to define the AC-carrier initial phases in such a way that the amplitude of their sum is minimized (for a better use of the DAC dynamic range).

Effect of Shot Noise on Simultaneous Sensing in Frequency Division Multiplexed Diffuse Optical Tomographic Imaging Process.

PubMed

Jang, Hansol; Lim, Gukbin; Hong, Keum-Shik; Cho, Jaedu; Gulsen, Gultekin; Kim, Chang-Seok

2017-11-28

Diffuse optical tomography (DOT) has been studied for use in the detection of breast cancer, cerebral oxygenation, and cognitive brain signals. As optical imaging studies have increased significantly, acquiring imaging data in real time has become increasingly important. We have developed frequency-division multiplexing (FDM) DOT systems to analyze their performance with respect to acquisition time and imaging quality, in comparison with the conventional time-division multiplexing (TDM) DOT. A large tomographic area of a cylindrical phantom 60 mm in diameter could be successfully reconstructed using both TDM DOT and FDM DOT systems. In our experiment with 6 source-detector (S-D) pairs, the TDM DOT and FDM DOT systems required 6.18 and 1 s, respectively, to obtain a single tomographic data set. While the absorption coefficient of the reconstruction image was underestimated in the case of the FDM DOT, we experimentally confirmed that the abnormal region can be clearly distinguished from the background phantom using both methods.

Development of a Bioaerosol single particle detector (BIO IN) for the Fast Ice Nucleus CHamber FINCH

NASA Astrophysics Data System (ADS)

Bundke, U.; Reimann, B.; Nillius, B.; Jaenicke, R.; Bingemer, H.

2010-02-01

In this work we present the setup and first tests of our new BIO IN detector. This detector was constructed to classify atmospheric ice nuclei (IN) for their biological content. It is designed to be coupled to the Fast Ice Nucleus CHamber FINCH. If one particle acts as an ice nucleus, it will be at least partly covered with ice at the end of the development section of the FINCH chamber. The device combines an auto-fluorescence detector and a circular depolarization detector for simultaneous detection of biological material and discrimination between water droplets, ice crystals and non activated large aerosol particles. The excitation of biological material with UV light and analysis of auto-fluorescence is a common principle used for flow cytometry, fluorescence microscopy, spectroscopy and imaging. The detection of auto-fluorescence of airborne single particles demands some more experimental effort. However, expensive commercial sensors are available for special purposes, e.g. size distribution measurements. But these sensors will not fit the specifications needed for the FINCH IN counter (e.g. high sample flow of up 10 LPM). The newly developed -low cost- BIO IN sensor uses a single high-power UV LED for the electronic excitation instead of much more expensive UV lasers. Other key advantages of the new sensor are the low weight, compact size, and the little effect on the aerosol sample, which allows it to be coupled with other instruments for further analysis. The instrument will be flown on one of the first missions of the new German research aircraft "HALO" (High Altitude and LOng range).

Large Format, Background Limited Arrays of Kinetic Inductance Detectors for Sub-mm Astronomy

NASA Astrophysics Data System (ADS)

Baselmans, Jochem

2018-01-01

We present the development of large format imaging arrays for sub-mm astronomy based upon microwave Kinetic Inductance detectors and their read-out. In particular we focus on the arrays developed for the A-MKID instrument for the APEX telescope. AMKID contains 2 focal plane arrays, covering a field of view of 15?x15?. One array is optimized for the 350 GHz telluric window, the other for the 850 GHz window. Both arrays are constructed from four 61 x 61 mm detector chips, each of which contains up to 3400 detectors and up to 880 detectors per readout line. The detectors are lens antenna coupled MKIDs made from NbTiN and Aluminium that reach photon noise limited sensitivity in combination with a high optical coupling. The lens-antenna radiation coupling enables the use of 4K optics and Lyot stop due to the intrinsic directivity of the detector beam, allowing a simple cryogenic architecture. We discuss the pixel design and verification, detector packaging and the array performance. We will also discuss the readout system, which is a combination of a digital and analog back-end that can read-out up to 4000 pixels simultaneously using frequency division multiplexing.

A Rapid Turnaround Cryogenic Detector Characterization System

NASA Technical Reports Server (NTRS)

Benford, Dominic j.; Dipirro, Michael J.; Forgione, Joshua B.; Jackson, Clifton E.; Jackson, Michael L.; Kogut, Al; Moseley, S. Harvey; Shirron, Peter J.

2004-01-01

Upcoming major NASA missions such as the Einstein Inflation Probe and the Single Aperture Far-Infrared Observatory require arrays of detectors with thousands of elements, operating at temperatures near l00 mK and sensitive to wavelengths from approx. 100 microns to approx. 3 mm. Such detectors represent a substantial enabling technology for these missions, and must be demonstrated soon in order for them to proceed. In order to make rapid progress on detector development, the cryogenic testing cycle must be made convenient and quick. We have developed a cryogenic detector characterization system capable of testing superconducting detector arrays in formats up to 8 x 32, read out by SQUID multiplexers. The system relies on the cooling of a two-stage adiabatic demagnetization refrigerator immersed in a liquid helium bath. This approach permits a detector to be cooled from 300K to 50 mK in about 4 hours, so that a test cycle begun in the morning will be over by the end of the day. Tine system is modular, with two identical immersible units, so that while one unit is cooling, the second can be reconfigured for the next battery of tests. We describe the design, construction, and performance of this cryogenic detector testing facility.

Practical photon number detection with electric field-modulated silicon avalanche photodiodes.

PubMed

Thomas, O; Yuan, Z L; Shields, A J

2012-01-24

Low-noise single-photon detection is a prerequisite for quantum information processing using photonic qubits. In particular, detectors that are able to accurately resolve the number of photons in an incident light pulse will find application in functions such as quantum teleportation and linear optics quantum computing. More generally, such a detector will allow the advantages of quantum light detection to be extended to stronger optical signals, permitting optical measurements limited only by fluctuations in the photon number of the source. Here we demonstrate a practical high-speed device, which allows the signals arising from multiple photon-induced avalanches to be precisely discriminated. We use a type of silicon avalanche photodiode in which the lateral electric field profile is strongly modulated in order to realize a spatially multiplexed detector. Clearly discerned multiphoton signals are obtained by applying sub-nanosecond voltage gates in order to restrict the detector current.

Lab on a chip for multiplexed immunoassays to detect bladder cancer using multifunctional dielectrophoretic manipulations.

PubMed

Chuang, Cheng-Hsin; Wu, Ting-Feng; Chen, Cheng-Ho; Chang, Kai-Chieh; Ju, Jing-Wei; Huang, Yao-Wei; Van Nhan, Vo

2015-07-21

A multiplexed immunosensor has been developed for the detection of specific biomarkers Galectin-1 (Gal-1) and Lactate Dehydrogenase B (LDH-B) present in different grades of bladder cancer cell lysates. In order to immobilize nanoprobes with different antibodies on a single chip we employed three-step programmable dielectrophoretic manipulations for focusing, guiding and trapping to enhance the fluorescent response and reduce the interference between the two antibody arrays. The chip consisted of a patterned indium tin oxide (ITO) electrode for sensing and a middle fish bone shaped gold electrode for focusing and guiding. Using ITO electrodes for the sensing area can effectively eliminate the background noise of fluorescence response as compared to metal electrodes. It was also observed that the three step manipulation increased fluorescence response after immunosensing by about 4.6 times as compared to utilizing DEP for just trapping the nanoprobes. Two different-grade bladder cancer cell lysates (grade I: RT4 and grade III: T24) were individually analyzed for detecting the protein expression levels of Gal-1 and LDH-B. The fluorescence intensity observed for Gal-1 is higher than that of LDH-B in the T24 cell lysate; however the response observed in RT4 is higher for LDH-B as compared to Gal-1. Thus we can effectively identify the different grades of bladder cancer cells. In addition, the platform for DEP manipulation developed in this study can enable real time detection of multiple analytes on a single chip and provide more practical benefits for clinical diagnosis.

Distributed Antenna-Coupled TES for FIR Detectors Arrays

NASA Technical Reports Server (NTRS)

Day, Peter K.; Leduc, Henry G.; Dowell, C. Darren; Lee, Richard A.; Zmuidzinas, Jonas

2007-01-01

We describe a new architecture for a superconducting detector for the submillimeter and far-infrared. This detector uses a distributed hot-electron transition edge sensor (TES) to collect the power from a focal-plane-filling slot antenna array. The sensors lay directly across the slots of the antenna and match the antenna impedance of about 30 ohms. Each pixel contains many sensors that are wired in parallel as a single distributed TES, which results in a low impedance that readily matches to a multiplexed SQUID readout These detectors are inherently polarization sensitive, with very low cross-polarization response, but can also be configured to sum both polarizations. The dual-polarization design can have a bandwidth of 50The use of electron-phonon decoupling eliminates the need for micro-machining, making the focal plane much easier to fabricate than with absorber-coupled, mechanically isolated pixels. We discuss applications of these detectors and a hybridization scheme compatible with arrays of tens of thousands of pixels.

Conceptual design of a hybrid Ge:Ga detector array

NASA Technical Reports Server (NTRS)

Parry, C. M.

1984-01-01

For potential applications in space infrared astronomy missions such as the Space Infrared Telescope Facility and the Large Deployable Reflector, integrated arrays of long-wavelength detectors are desired. The results of a feasibility study which developed a design for applying integrated array techniques to a long-wavelength (gallium-doped germanium) material to achieve spectral coverage between 30 and 200 microns are presented. An approach which builds up a two-dimensional array by stacking linear detector modules is presented. The spectral response of the Ge:Ga detectors is extended to 200 microns by application of uniaxial stress to the stack of modules. The detectors are assembled with 1 mm spacing between the elements. Multiplexed readout of each module is accomplished with integration sampling of a metal-oxide-semiconductor (MOS) switch chip. Aspects of the overall design, including the anticipated level of particle effects on the array in the space environment, a transparent electrode design for 200 microns response, estimates of optical crosstalk, and mechanical stress design calculations are included.

Multiplexed wavelet transform technique for detection of microcalcification in digitized mammograms.

PubMed

Mini, M G; Devassia, V P; Thomas, Tessamma

2004-12-01

Wavelet transform (WT) is a potential tool for the detection of microcalcifications, an early sign of breast cancer. This article describes the implementation and evaluates the performance of two novel WT-based schemes for the automatic detection of clustered microcalcifications in digitized mammograms. Employing a one-dimensional WT technique that utilizes the pseudo-periodicity property of image sequences, the proposed algorithms achieve high detection efficiency and low processing memory requirements. The detection is achieved from the parent-child relationship between the zero-crossings [Marr-Hildreth (M-H) detector] /local extrema (Canny detector) of the WT coefficients at different levels of decomposition. The detected pixels are weighted before the inverse transform is computed, and they are segmented by simple global gray level thresholding. Both detectors produce 95% detection sensitivity, even though there are more false positives for the M-H detector. The M-H detector preserves the shape information and provides better detection sensitivity for mammograms containing widely distributed calcifications.

Multiplexed Analysis of Serum Breast and Ovarian Cancer Markers by Means of Suspension Bead-quantum Dot Microarrays

NASA Astrophysics Data System (ADS)

Brazhnik, Kristina; Sokolova, Zinaida; Baryshnikova, Maria; Bilan, Regina; Nabiev, Igor; Sukhanova, Alyona

Multiplexed analysis of cancer markers is crucial for early tumor diagnosis and screening. We have designed lab-on-a-bead microarray for quantitative detection of three breast cancer markers in human serum. Quantum dots were used as bead-bound fluorescent tags for identifying each marker by means of flow cytometry. Antigen-specific beads reliably detected CA 15-3, CEA, and CA 125 in serum samples, providing clear discrimination between the samples with respect to the antigen levels. The novel microarray is advantageous over the routine single-analyte ones due to the simultaneous detection of various markers. Therefore the developed microarray is a promising tool for serum tumor marker profiling.

Highly Stable Lyophilized Homogeneous Bead-Based Immunoassays for On-Site Detection of Bio Warfare Agents from Complex Matrices.

PubMed

Mechaly, Adva; Marx, Sharon; Levy, Orly; Yitzhaki, Shmuel; Fisher, Morly

2016-06-21

This study shows the development of dry, highly stable immunoassays for the detection of bio warfare agents in complex matrices. Thermal stability was achieved by the lyophilization of the complete, homogeneous, bead-based immunoassay in a special stabilizing buffer, resulting in a ready-to-use, simple assay, which exhibited long shelf and high-temperature endurance (up to 1 week at 100 °C). The developed methodology was successfully implemented for the preservation of time-resolved fluorescence, Alexa-fluorophores, and horse radish peroxidase-based bead assays, enabling multiplexed detection. The multiplexed assay was successfully implemented for the detection of Bacillus anthracis, botulinum B, and tularemia in complex matrices.

Improvement for identification of heterophile antibody interference and AFP hook effect in immunoassays with multiplex suspension bead array system.

PubMed

Wang, Yajie; Yu, Jinsheng; Ren, Yuan; Liu, Li; Li, Haowen; Guo, Anchen; Shi, Congning; Fang, Fang; Juehne, Twyla; Yao, Jianer; Yang, Enhuan; Zhou, Xuelei; Kang, Xixiong

2013-11-15

A variety of immunoassays including multiplex suspension bead array have been developed for tumor marker detections; however, these assays could be compromised in their sensitivity and specificity by well-known heterophile antibody interference and hook effect. Using Luminex® multiplex suspension bead arrays, we modified protocols with two newly-developed solutions that can identify heterophile antibody interference and AFP hook effect. Effectiveness of the two solutions was assessed in serum samples from patients. Concentrations of 9 tumor markers in heterophile antibody positive samples assayed with Solution A, containing murine monoclonal antibodies and mouse serum, were significantly reduced when compared with those false high signals assayed without Solution A (all p<0.01). With incorporation of Solution H (fluorescent beads linked with AFP antigen), a new strategy for identification of AFP hook effect was established, and with this strategy AFP hook effect was identified effectively in serum samples with very high levels of AFP. Two proprietary solutions improve the identification of heterophile antibody interference and AFP hook effect. With these solutions, multiplex suspension bead arrays provide more reliable testing results in tumor marker detection where complex clinical serum samples are used. © 2013.

Efficiency of semi-automated fluorescent multiplex PCRs with 11 microsatellite markers for genetic studies of deer populations.

PubMed

Bonnet, A; Thévenon, S; Maudet, F; Maillard, J C

2002-10-01

Thirty bovine and eight ovine microsatellite primer pairs were tested on four tropical deer species: Eld's and Swamp deer (highly threatened) and Rusa and Vietnamese Sika deer (economically important). Thirty markers gave an amplified product in all four species (78.9%). The number of polymorphic microsatellite markers varied among the species from 14 in Eld's deer (47%) to 20 in Swamp deer (67%). Among them, 11 microsatellite loci were multiplexed in three polymerase chain reactions (PCRs) and labelled with three different fluorochromes that can be loaded in one gel-lane. To test the efficiency of the multiplex, primary genetic studies (mean number of alleles, expected heterozygosities and Fis values) were carried out on four deer populations. Parentage exclusion probability and probability of identity were computed and discussed on a Swamp deer population. These multiplexes PCRs were also tested on several other deer species and subspecies. The aim of this study is to establish a tool useful for genetic studies of population structure and diversity in four tropical deer species which with few modifications can be applied to other species of the genus Cervus.

A multiplex branched DNA assay for parallel quantitative gene expression profiling.

PubMed

Flagella, Michael; Bui, Son; Zheng, Zhi; Nguyen, Cung Tuong; Zhang, Aiguo; Pastor, Larry; Ma, Yunqing; Yang, Wen; Crawford, Kimberly L; McMaster, Gary K; Witney, Frank; Luo, Yuling

2006-05-01

We describe a novel method to quantitatively measure messenger RNA (mRNA) expression of multiple genes directly from crude cell lysates and tissue homogenates without the need for RNA purification or target amplification. The multiplex branched DNA (bDNA) assay adapts the bDNA technology to the Luminex fluorescent bead-based platform through the use of cooperative hybridization, which ensures an exceptionally high degree of assay specificity. Using in vitro transcribed RNA as reference standards, we demonstrated that the assay is highly specific, with cross-reactivity less than 0.2%. We also determined that the assay detection sensitivity is 25,000 RNA transcripts with intra- and interplate coefficients of variance of less than 10% and less than 15%, respectively. Using three 10-gene panels designed to measure proinflammatory and apoptosis responses, we demonstrated sensitive and specific multiplex gene expression profiling directly from cell lysates. The gene expression change data demonstrate a high correlation coefficient (R(2)=0.94) compared with measurements obtained using the single-plex bDNA assay. Thus, the multiplex bDNA assay provides a powerful means to quantify the gene expression profile of a defined set of target genes in large sample populations.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sun, Yue; Gleber, Sophie-Charlotte; Jacobsen, Chris

Trace metals play critical roles in a variety of systems, ranging from cells to photovoltaics. X-Ray Fluorescence (XRF) microscopy using X-ray excitation provides one of the highest sensitivities available for imaging the distribution of trace metals at sub-100 nm resolution. With the growing availability and increasing performance of synchrotron light source based instruments and X-ray nanofocusing optics, and with improvements in energy-dispersive XRF detectors, what are the factors that limit trace element detectability? To address this question, we describe an analytical model for the total signal incident on XRF detectors with various geometries, including the spectral response of energy dispersivemore » detectors. This model agrees well with experimentally recorded X-ray fluorescence spectra, and involves much shorter calculation times than with Monte Carlo simulations. With such a model, one can estimate the signal when a trace element is illuminated with an X-ray beam, and when just the surrounding non-fluorescent material is illuminated. From this signal difference, a contrast parameter can be calculated and this can in turn be used to calculate the signal-to-noise ratio (S/N) for detecting a certain elemental concentration. We apply this model to the detection of trace amounts of zinc in biological materials, and to the detection of small quantities of arsenic in semiconductors. We conclude that increased detector collection solid angle is (nearly) always advantageous even when considering the scattered signal. However, given the choice between a smaller detector at 90° to the beam versus a larger detector at 180° (in a backscatter-like geometry), the 90° detector is better for trace element detection in thick samples, while the larger detector in 180° geometry is better suited to trace element detection in thin samples.« less

Nano metal-organic framework (NMOF)-based strategies for multiplexed microRNA detection in solution and living cancer cells

NASA Astrophysics Data System (ADS)

Wu, Yafeng; Han, Jianyu; Xue, Peng; Xu, Rong; Kang, Yuejun

2015-01-01

MiRNAs are an emerging type of biomarker for diagnostics and prognostics. A reliable sensing strategy that can monitor miRNA expression in living cancer cells would be critical in view of its extensive advantages for fundamental research related to miRNA-associated bioprocesses and biomedical applications. Conventional miRNA sensing methods include northern blot, microarrays and real-time quantitative PCR. However, none of them is able to monitor miRNA levels expressed in living cancer cells in a real-time fashion. Some fluorescennt biosensors developed recently from carbon nanomaterials, such as single-walled carbon nanotubes (SWNTs), graphene oxide (GO), and carbon nanoparticles, have been successfully used for assaying miRNA in vitro; however the preparation processes are often expensive, complicated and time-consuming, which have motivated the research on other substitute and novel materials. Herein we present a novel sensing strategy based on peptide nucleic acid (PNA) probes labeled with fluorophores and conjugated with an NMOF vehicle to monitor multiplexed miRNAs in living cancer cells. The NMOF works as a fluorescence quencher of the labelled PNA that is firmly bound with the metal center. In the presence of a target miRNA, PNA is hybridized and released from the NMOF leading to the recovery of fluorescence. This miRNA sensor not only enables the quantitative and highly specific detection of multiplexed miRNAs in living cancer cells, but it also allows the precise and in situ monitoring of the spatiotemporal changes of miRNA expression.MiRNAs are an emerging type of biomarker for diagnostics and prognostics. A reliable sensing strategy that can monitor miRNA expression in living cancer cells would be critical in view of its extensive advantages for fundamental research related to miRNA-associated bioprocesses and biomedical applications. Conventional miRNA sensing methods include northern blot, microarrays and real-time quantitative PCR. However, none of them is able to monitor miRNA levels expressed in living cancer cells in a real-time fashion. Some fluorescennt biosensors developed recently from carbon nanomaterials, such as single-walled carbon nanotubes (SWNTs), graphene oxide (GO), and carbon nanoparticles, have been successfully used for assaying miRNA in vitro; however the preparation processes are often expensive, complicated and time-consuming, which have motivated the research on other substitute and novel materials. Herein we present a novel sensing strategy based on peptide nucleic acid (PNA) probes labeled with fluorophores and conjugated with an NMOF vehicle to monitor multiplexed miRNAs in living cancer cells. The NMOF works as a fluorescence quencher of the labelled PNA that is firmly bound with the metal center. In the presence of a target miRNA, PNA is hybridized and released from the NMOF leading to the recovery of fluorescence. This miRNA sensor not only enables the quantitative and highly specific detection of multiplexed miRNAs in living cancer cells, but it also allows the precise and in situ monitoring of the spatiotemporal changes of miRNA expression. Electronic supplementary information (ESI) available: Extra figures and tables. See DOI: 10.1039/c4nr05447d

Optimized Detector Angular Configuration Increases the Sensitivity of X-ray Fluorescence Computed Tomography (XFCT).

PubMed

Ahmad, Moiz; Bazalova-Carter, Magdalena; Fahrig, Rebecca; Xing, Lei

2015-05-01

In this work, we demonstrated that an optimized detector angular configuration based on the anisotropic energy distribution of background scattered X-rays improves X-ray fluorescence computed tomography (XFCT) detection sensitivity. We built an XFCT imaging system composed of a bench-top fluoroscopy X-ray source, a CdTe X-ray detector, and a phantom motion stage. We imaged a 6.4-cm-diameter phantom containing different concentrations of gold solution and investigated the effect of detector angular configuration on XFCT image quality. Based on our previous theoretical study, three detector angles were considered. The X-ray fluorescence detector was first placed at 145 (°) (approximating back-scatter) to minimize scatter X-rays. XFCT image quality was compared to images acquired with the detector at 60 (°) (forward-scatter) and 90 (°) (side-scatter). The datasets for the three different detector positions were also combined to approximate an isotropically arranged detector. The sensitivity was optimized with detector in the 145 (°) back-scatter configuration counting the 78-keV gold Kβ1 X-rays. The improvement arose from the reduced energy of scattered X-ray at the 145 (°) position and the large energy separation from gold K β1 X-rays. The lowest detected concentration in this configuration was 2.5 mgAu/mL (or 0.25% Au with SNR = 4.3). This concentration could not be detected with the 60 (°) , 90 (°) , or isotropic configurations (SNRs = 1.3, 0, 2.3, respectively). XFCT imaging dose of 14 mGy was in the range of typical clinical X-ray CT imaging doses. To our knowledge, the sensitivity achieved in this experiment is the highest in any XFCT experiment using an ordinary bench-top X-ray source in a phantom larger than a mouse ( > 3 cm).

Automated fluorescent miscroscopic image analysis of PTBP1 expression in glioma

PubMed Central

Becker, Aline; Elder, Brad; Puduvalli, Vinay; Winter, Jessica; Gurcan, Metin

2017-01-01

Multiplexed immunofluorescent testing has not entered into diagnostic neuropathology due to the presence of several technical barriers, amongst which includes autofluorescence. This study presents the implementation of a methodology capable of overcoming the visual challenges of fluorescent microscopy for diagnostic neuropathology by using automated digital image analysis, with long term goal of providing unbiased quantitative analyses of multiplexed biomarkers for solid tissue neuropathology. In this study, we validated PTBP1, a putative biomarker for glioma, and tested the extent to which immunofluorescent microscopy combined with automated and unbiased image analysis would permit the utility of PTBP1 as a biomarker to distinguish diagnostically challenging surgical biopsies. As a paradigm, we utilized second resections from patients diagnosed either with reactive brain changes (pseudoprogression) and recurrent glioblastoma (true progression). Our image analysis workflow was capable of removing background autofluorescence and permitted quantification of DAPI-PTBP1 positive cells. PTBP1-positive nuclei, and the mean intensity value of PTBP1 signal in cells. Traditional pathological interpretation was unable to distinguish between groups due to unacceptably high discordance rates amongst expert neuropathologists. Our data demonstrated that recurrent glioblastoma showed more DAPI-PTBP1 positive cells and a higher mean intensity value of PTBP1 signal compared to resections from second surgeries that showed only reactive gliosis. Our work demonstrates the potential of utilizing automated image analysis to overcome the challenges of implementing fluorescent microscopy in diagnostic neuropathology. PMID:28282372

A superconducting focal plane array for ultraviolet, optical, and near-infrared astrophysics.

PubMed

Mazin, Benjamin A; Bumble, Bruce; Meeker, Seth R; O'Brien, Kieran; McHugh, Sean; Langman, Eric

2012-01-16

Microwave Kinetic Inductance Detectors, or MKIDs, have proven to be a powerful cryogenic detector technology due to their sensitivity and the ease with which they can be multiplexed into large arrays. A MKID is an energy sensor based on a photon-variable superconducting inductance in a lithographed microresonator, and is capable of functioning as a photon detector across the electromagnetic spectrum as well as a particle detector. Here we describe the first successful effort to create a photon-counting, energy-resolving ultraviolet, optical, and near infrared MKID focal plane array. These new Optical Lumped Element (OLE) MKID arrays have significant advantages over semiconductor detectors like charge coupled devices (CCDs). They can count individual photons with essentially no false counts and determine the energy and arrival time of every photon with good quantum efficiency. Their physical pixel size and maximum count rate is well matched with large telescopes. These capabilities enable powerful new astrophysical instruments usable from the ground and space. MKIDs could eventually supplant semiconductor detectors for most astronomical instrumentation, and will be useful for other disciplines such as quantum optics and biological imaging.

Kinetic inductance detectors for millimeter and submillimeter astronomy

NASA Astrophysics Data System (ADS)

Boudou, Nicolas; Benoit, Alain; Bourrion, Olivier; Calvo, Martino; Désert, François-Xavier; Macias-Perez, Juan; Monfardini, Alessandro; Roesch, Markus

2012-01-01

We present recent developments in Kinetic Inductance Detectors (KID) for large arrays of detectors. The main application is ground-based millimeter wave astronomy. We focus in particular, as a case study, on our own experiment: NIKA (Néel IRAM KID Arrays). NIKA is today the best in-the-field experiment using KID-based instruments, and consists of a dual-band imaging system designed for the IRAM 30 meter telescope at Pico Veleta. We describe in this article, after a general context introduction, the KID working principle and the readout electronics, crucial to take advantage of the intrinsic KID multiplexability. We conclude with a small subset of the astronomical sources observed simultaneously at 2 mm and 1.4 mm by NIKA during the last run, held in October 2010.

Recent developments in Förster resonance energy transfer (FRET) diagnostics using quantum dots.

PubMed

Geißler, Daniel; Hildebrandt, Niko

2016-07-01

The exceptional photophysical properties and the nanometric dimensions of colloidal semiconductor quantum dots (QD) have strongly attracted the bioanalytical community over the last approximately 20 y. In particular, the integration of QDs in the analysis of biological components and interactions, and the related diagnostics using Förster resonance energy transfer (FRET), have allowed researchers to significantly improve and diversify fluorescence-based biosensing. In this TRENDS article, we review some recent developments in QD-FRET biosensing that have implemented this technology in electronic consumer products, multiplexed analysis, and detection without light excitation for diagnostic applications. In selected examples of smartphone-based imaging, single- and multistep FRET, steady-state and time-resolved spectroscopy, and bio/chemiluminescence detection of QDs used as both FRET donors and acceptors, we highlight the advantages of QD-based FRET biosensing for multiplexed and sensitive diagnostics. Graphical Abstract Quantum dots (QDs) can be applied as donors and/or acceptors for Förster resonance energy transfer- (FRET-) based biosensing for multiplexed and sensitive diagnostics in various assay formats.

Single quantum dot analysis enables multiplexed point mutation detection by gap ligase chain reaction.

PubMed

Song, Yunke; Zhang, Yi; Wang, Tza-Huei

2013-04-08

Gene point mutations present important biomarkers for genetic diseases. However, existing point mutation detection methods suffer from low sensitivity, specificity, and a tedious assay processes. In this report, an assay technology is proposed which combines the outstanding specificity of gap ligase chain reaction (Gap-LCR), the high sensitivity of single-molecule coincidence detection, and the superior optical properties of quantum dots (QDs) for multiplexed detection of point mutations in genomic DNA. Mutant-specific ligation products are generated by Gap-LCR and subsequently captured by QDs to form DNA-QD nanocomplexes that are detected by single-molecule spectroscopy (SMS) through multi-color fluorescence burst coincidence analysis, allowing for multiplexed mutation detection in a separation-free format. The proposed assay is capable of detecting zeptomoles of KRAS codon 12 mutation variants with near 100% specificity. Its high sensitivity allows direct detection of KRAS mutation in crude genomic DNA without PCR pre-amplification. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Large format imaging arrays for the Atacama Cosmology Telescope

NASA Technical Reports Server (NTRS)

Chervenak, J. A.; Wollack, E. J.; Marraige, T.; Staggs, S.; Niemack, M.; Doriese, B.

2006-01-01

We describe progress in the fabrication, characterization, and production of detector arrays for the Atacama Cosmology Telescope (ACT). The completed ACT instrument is specified to image simultaneously at 145, 225, and 265 GHz using three 32x32 filled arrays of superconducting transition edge sensors (TES) read out with time-division-multiplexed SQUID amplifiers. We present details of the pixel design and testing including the optimization of the electrical parameters for multiplexed readout. Using geometric noise suppression and careful tuning of operation temperature and device bias resistance, the excess noise in the TES devices is balanced with detector speed for interfacing with the ACT optics. The design also accounts for practical tolerances such as transition temperature gradients and scatter that occur in the production of multiple wafers to populate fully the kilopixel cameras. We have developed an implanted absorber layer compatible with our silicon-on-insulator process that allows for tunable optical resistance with requisite on-wafer uniformity and wafer-to-wafer reproducibility. Arrays of 32 elements have been tested in the laboratory environment including electrical, optical, and multiplexed performance. Given this pixel design, optical tests and modeling are used to predict the performance of the filled array under anticipated viewing conditions. Integration of the filled array of pixels with a tuned backshort and dielectric plate in front of the array maximize absorption and the focal plane and suppress reflections. A mechanical design for the build of the full structure is completed and we report on progress toward the construction of a prototype array for first light on the ACT.

Integrated performance of a frequency domain multiplexing readout in the SPT-3G receiver

NASA Astrophysics Data System (ADS)

Bender, A. N.; Ade, P. A. R.; Anderson, A. J.; Avva, J.; Ahmed, Z.; Arnold, K.; Austermann, J. E.; Basu Thakur, R.; Benson, B. A.; Bleem, L. E.; Byrum, K.; Carlstrom, J. E.; Carter, F. W.; Chang, C. L.; Cho, H. M.; Cliche, J. F.; Crawford, T. M.; Cukierman, A.; Czaplewski, D. A.; Ding, J.; Divan, R.; de Haan, T.; Dobbs, M. A.; Dutcher, D.; Everett, W.; Gilbert, A.; Groh, J. C.; Guyser, R.; Halverson, N. W.; Harke-Hosemann, A.; Harrington, N. L.; Hattori, K.; Henning, J. W.; Hilton, G. C.; Holzapfel, W. L.; Huang, N.; Irwin, K. D.; Jeong, O.; Khaire, T.; Korman, M.; Kubik, D.; Kuo, C. L.; Lee, A. T.; Leitch, E. M.; Lendinez, S.; Meyer, S. S.; Miller, C. S.; Montgomery, J.; Nadolski, A.; Natoli, T.; Nguyen, H.; Novosad, V.; Padin, S.; Pan, Z.; Pearson, J.; Posada, C. M.; Rahlin, A.; Reichardt, C. L.; Ruhl, J. E.; Saliwanchik, B. R.; Sayre, J. T.; Shariff, J. A.; Shirley, Ian; Shirokoff, E.; Smecher, G.; Sobrin, J.; Stan, L.; Stark, A. A.; Story, K.; Suzuki, A.; Tang, Q. Y.; Thompson, K. L.; Tucker, C.; Vanderlinde, K.; Vieira, J. D.; Wang, G.; Whitehorn, N.; Yefremenko, V.; Yoon, K. W.

2016-07-01

The third generation receiver for the South Pole Telescope, SPT-3G, will make extremely deep, arcminuteresolution maps of the temperature and polarization of the cosmic microwave background. The SPT-3G maps will enable studies of the B-mode polarization signature, constraining primordial gravitational waves as well as the effect of massive neutrinos on structure formation in the late universe. The SPT-3G receiver will achieve exceptional sensitivity through a focal plane of 16,000 transition-edge sensor bolometers, an order of magnitude more than the current SPTpol receiver. SPT-3G uses a frequency domain multiplexing (fMux) scheme to read out the focal plane, combining the signals from 64 bolometers onto a single pair of wires. The fMux readout facilitates the large number of detectors in the SPT-3G focal plane by limiting the thermal load due to readout wiring on the 250 millikelvin cryogenic stage. A second advantage of the fMux system is that the operation of each bolometer can be optimized. In addition to these benefits, the fMux readout introduces new challenges into the design and operation of the receiver. The bolometers are operated at a range of frequencies up to 5 MHz, requiring control of stray reactances over a large bandwidth. Additionally, crosstalk between multiplexed detectors will inject large false signals into the data if not adequately mitigated. SPT-3G is scheduled to deploy to the South Pole Telescope in late 2016. Here, we present the pre-deployment performance of the fMux readout system with the SPT-3G focal plane.

Visualisation of blood and lymphatic vessels with increasing exposure time of the detector

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kalchenko, V V; Kuznetsov, Yu L; Meglinski, I V

2013-07-31

We describe the laser speckle contrast method for simultaneous noninvasive imaging of blood and lymphatic vessels of living organisms, based on increasing detector exposure time. In contrast to standard methods of fluorescent angiography, this technique of vascular bed imaging and lymphatic and blood vessel demarcation does not employ toxic fluorescent markers. The method is particularly promising with respect to the physiology of the cardiovascular system under in vivo conditions. (laser applications in biology and medicine)

Construction of a low-cost detector to identify dissolved metals in aqueous media by fluorescence spectroscopy: design and perspectives.

NASA Astrophysics Data System (ADS)

González, M.; Montaño, M.; Hoyo, C.

2017-01-01

We have constructed a low cost fluorescence detector model to determine the presence of some heavy metals in an aqueous medium. In particular, we focus on metals which cause public health problems in our country. We did the first tests with standard samples of Hg (II). The innovative features of this instrument are its small dimensions (9 dm3) and the low cost of materials used in its construction.

Pyrene As a New Detector for Determining the Composition of Silver Nanoparticle Dispersions in Aqueous Solutions

NASA Astrophysics Data System (ADS)

Romanovskaya, G. I.; Kazakova, S. Yu.; Koroleva, M. V.; Zuev, B. K.

2018-03-01

It is proposed that the fluorescence of monomeric molecules of pyrene in solid matrices or in concentrated micellar solutions be used as a detector for determining the compositional homogeneity of silver nanoparticle (NP) dispersions in aqueous solutions synthesized in different ways. It is found that the morphology of silver NPs affects the change in the fluorescence intensity of monomeric molecules of pyrene in a certain (violet or blue) region of the pyrene optical spectrum. The observed phenomenon is attributed to the resonance of electronic transitions in the monomeric molecules of pyrene in regions with plasmon oscillations in silver nanoparticles. A new way of obtaining fluorescent silver NPs is found.

Fluorescence calibration method for single-particle aerosol fluorescence instruments

NASA Astrophysics Data System (ADS)

Shipley Robinson, Ellis; Gao, Ru-Shan; Schwarz, Joshua P.; Fahey, David W.; Perring, Anne E.

2017-05-01

Real-time, single-particle fluorescence instruments used to detect atmospheric bioaerosol particles are increasingly common, yet no standard fluorescence calibration method exists for this technique. This gap limits the utility of these instruments as quantitative tools and complicates comparisons between different measurement campaigns. To address this need, we have developed a method to produce size-selected particles with a known mass of fluorophore, which we use to calibrate the fluorescence detection of a Wideband Integrated Bioaerosol Sensor (WIBS-4A). We use mixed tryptophan-ammonium sulfate particles to calibrate one detector (FL1; excitation = 280 nm, emission = 310-400 nm) and pure quinine particles to calibrate the other (FL2; excitation = 280 nm, emission = 420-650 nm). The relationship between fluorescence and mass for the mixed tryptophan-ammonium sulfate particles is linear, while that for the pure quinine particles is nonlinear, likely indicating that not all of the quinine mass contributes to the observed fluorescence. Nonetheless, both materials produce a repeatable response between observed fluorescence and particle mass. This procedure allows users to set the detector gains to achieve a known absolute response, calculate the limits of detection for a given instrument, improve the repeatability of the instrumental setup, and facilitate intercomparisons between different instruments. We recommend calibration of single-particle fluorescence instruments using these methods.

Designing a new type of neutron detector for neutron and gamma-ray discrimination via GEANT4.

PubMed

Shan, Qing; Chu, Shengnan; Ling, Yongsheng; Cai, Pingkun; Jia, Wenbao

2016-04-01

Design of a new type of neutron detector, consisting of a fast neutron converter, plastic scintillator, and Cherenkov detector, to discriminate 14-MeV fast neutrons and gamma rays in a pulsed n-γ mixed field and monitor their neutron fluxes is reported in this study. Both neutrons and gamma rays can produce fluorescence in the scintillator when they are incident on the detector. However, only the secondary charged particles of the gamma rays can produce Cherenkov light in the Cherenkov detector. The neutron and gamma-ray fluxes can be calculated by measuring the fluorescence and Cherenkov light. The GEANT4 Monte Carlo simulation toolkit is used to simulate the whole process occurring in the detector, whose optimum parameters are known. Analysis of the simulation results leads to a calculation method of neutron flux. This method is verified by calculating the neutron fluxes using pulsed n-γ mixed fields with different n/γ ratios, and the results show that the relative errors of all calculations are <5%. Copyright © 2016 Elsevier Ltd. All rights reserved.

Elemental mapping in a contemporary miniature by full-field X-ray fluorescence imaging with gaseous detector vs. scanning X-ray fluorescence imaging with polycapillary optics

NASA Astrophysics Data System (ADS)

Silva, A. L. M.; Cirino, S.; Carvalho, M. L.; Manso, M.; Pessanha, S.; Azevedo, C. D. R.; Carramate, L. F. N. D.; Santos, J. P.; Guerra, M.; Veloso, J. F. C. A.

2017-03-01

Energy dispersive X-ray imaging can be used in several research fields and industrial applications. Elemental mapping through energy dispersive X-ray imaging technique has become a promising method to obtain positional distribution of specific elements in a non-destructive way. To obtain the elemental distribution of a sample it is necessary to use instruments capable of providing a precise positioning together with a good energy resolution. Polycapillary beams together with silicon drift chamber detectors are used in several commercial systems and are considered state-of-the-art spectrometers, however they are usually very costly. A new concept of large energy dispersive X-ray imaging systems based on gaseous radiation detectors emerged in the last years enabling a promising 2D elemental detection at a very reduced price. The main goal of this work is to analyze a contemporary Indian miniature with both X-ray fluorescence imaging systems, the one based on a gaseous detector 2D-THCOBRA and the state-of-the-art spectrometer M4 Tornado, from Bruker. The performance of both systems is compared and evaluated in the context of the sample's analysis.

78 FR 28864 - Prospective Grant of Start-Up Exclusive Evaluation Option License Agreement: In Vitro Diagnostics...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-05-16

... Efficacy in Cancer and Other Angiogenesis-Mediated Diseases AGENCY: National Institutes of Health, HHS... Fluorescent Cells'', filed October 1, 2007 (HHS Ref. No. E-281-2007/0-US-01); U.S. Patent Application No. 12/ 060,752, entitled ``Multiplex Assay Method for Mixed Cell Populations'', filed April 1, 2008, (HHS Ref...

The hybrid energy spectrum of Telescope Array's Middle Drum Detector and surface array

NASA Astrophysics Data System (ADS)

Abbasi, R. U.; Abe, M.; Abu-Zayyad, T.; Allen, M. G.; Anderson, R.; Azuma, R.; Barcikowski, E.; Belz, J. W.; Bergman, D. R.; Blake, S. A.; Cady, R.; Chae, M. J.; Cheon, B. G.; Chiba, J.; Chikawa, M.; Cho, W. R.; Fujii, T.; Fukushima, M.; Goto, T.; Hanlon, W.; Hayashi, Y.; Hayashida, N.; Hibino, K.; Honda, K.; Ikeda, D.; Inoue, N.; Ishii, T.; Ishimori, R.; Ito, H.; Ivanov, D.; Jui, C. C. H.; Kadota, K.; Kakimoto, F.; Kalashev, O.; Kasahara, K.; Kawai, H.; Kawakami, S.; Kawana, S.; Kawata, K.; Kido, E.; Kim, H. B.; Kim, J. H.; Kim, J. H.; Kitamura, S.; Kitamura, Y.; Kuzmin, V.; Kwon, Y. J.; Lan, J.; Lim, S. I.; Lundquist, J. P.; Machida, K.; Martens, K.; Matsuda, T.; Matsuyama, T.; Matthews, J. N.; Minamino, M.; Mukai, K.; Myers, I.; Nagasawa, K.; Nagataki, S.; Nakamura, T.; Nonaka, T.; Nozato, A.; Ogio, S.; Ogura, J.; Ohnishi, M.; Ohoka, H.; Oki, K.; Okuda, T.; Ono, M.; Oshima, A.; Ozawa, S.; Park, I. H.; Pshirkov, M. S.; Rodriguez, D. C.; Rubtsov, G.; Ryu, D.; Sagawa, H.; Sakurai, N.; Sampson, A. L.; Scott, L. M.; Shah, P. D.; Shibata, F.; Shibata, T.; Shimodaira, H.; Shin, B. K.; Shin, H. S.; Smith, J. D.; Sokolsky, P.; Springer, R. W.; Stokes, B. T.; Stratton, S. R.; Stroman, T. A.; Suzawa, T.; Takamura, M.; Takeda, M.; Takeishi, R.; Taketa, A.; Takita, M.; Tameda, Y.; Tanaka, H.; Tanaka, K.; Tanaka, M.; Thomas, S. B.; Thomson, G. B.; Tinyakov, P.; Tkachev, I.; Tokuno, H.; Tomida, T.; Troitsky, S.; Tsunesada, Y.; Tsutsumi, K.; Uchihori, Y.; Udo, S.; Urban, F.; Vasiloff, G.; Wong, T.; Yamane, R.; Yamaoka, H.; Yamazaki, K.; Yang, J.; Yashiro, K.; Yoneda, Y.; Yoshida, S.; Yoshii, H.; Zollinger, R.; Zundel, Z.

2015-08-01

The Telescope Array experiment studies ultra high energy cosmic rays using a hybrid detector. Fluorescence telescopes measure the longitudinal development of the extensive air shower generated when a primary cosmic ray particle interacts with the atmosphere. Meanwhile, scintillator detectors measure the lateral distribution of secondary shower particles that hit the ground. The Middle Drum (MD) fluorescence telescope station consists of 14 telescopes from the High Resolution Fly's Eye (HiRes) experiment, providing a direct link back to the HiRes measurements. Using the scintillator detector data in conjunction with the telescope data improves the geometrical reconstruction of the showers significantly, and hence, provides a more accurate reconstruction of the energy of the primary particle. The Middle Drum hybrid spectrum is presented and compared to that measured by the Middle Drum station in monocular mode. Further, the hybrid data establishes a link between the Middle Drum data and the surface array. A comparison between the Middle Drum hybrid energy spectrum and scintillator Surface Detector (SD) spectrum is also shown.

Image scanning fluorescence emission difference microscopy based on a detector array.

PubMed

Li, Y; Liu, S; Liu, D; Sun, S; Kuang, C; Ding, Z; Liu, X

2017-06-01

We propose a novel imaging method that enables the enhancement of three-dimensional resolution of confocal microscopy significantly and achieve experimentally a new fluorescence emission difference method for the first time, based on the parallel detection with a detector array. Following the principles of photon reassignment in image scanning microscopy, images captured by the detector array were arranged. And by selecting appropriate reassign patterns, the imaging result with enhanced resolution can be achieved with the method of fluorescence emission difference. Two specific methods are proposed in this paper, showing that the difference between an image scanning microscopy image and a confocal image will achieve an improvement of transverse resolution by approximately 43% compared with that in confocal microscopy, and the axial resolution can also be enhanced by at least 22% experimentally and 35% theoretically. Moreover, the methods presented in this paper can improve the lateral resolution by around 10% than fluorescence emission difference and 15% than Airyscan. The mechanism of our methods is verified by numerical simulations and experimental results, and it has significant potential in biomedical applications. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

New target and detection methods: active detectors

NASA Astrophysics Data System (ADS)

Mittig, W.; Savajols, H.; Demonchy, C. E.; Giot, L.; Roussel-Chomaz, P.; Wang, H.; Ter-Akopian, G.; Fomichev, A.; Golovkov, M. S.; Stepansov, S.; Wolski, R.; Alamanos, N.; Drouart, A.; Gillibert, A.; Lapoux, V.; Pollacco, E.

2003-07-01

The study of nuclei far from stability interacting with simple target nuclei, such as protons, deuterons, 3He and 4He implies the use of inverse kinematics. The very special kinematics, together with the low intensities of the beams calls for special techniques. In july 2002 we tested a new detector, in which the detector gas is the target. This allows in principle a 4π solid angle of the detection, and a big effective target thickness without loss of resolution. The detector developped, called Maya, used isobuthane C4H10 as gas in present tests, and other gases are possible. The multiplexed electronics of more than 1000channels allows the reconstruction of the events occuring between the incoming particle and the detector gas atoms in 3D. Here we were interested in the elastic scattering of 8He on protons for the study of the isobaric analogue states (IAS) of 9He. The beam, in this case, is stopped in the detector. The resonance energy is determined by the place of interaction and the energy of the recoiling proton. The design of the detector is shown, together with some preliminary results are discussed.

Efficient heralding of O-band passively spatial-multiplexed photons for noise-tolerant quantum key distribution.

PubMed

Liu, Mao Tong; Lim, Han Chuen

2014-09-22

When implementing O-band quantum key distribution on optical fiber transmission lines carrying C-band data traffic, noise photons that arise from spontaneous Raman scattering or insufficient filtering of the classical data channels could cause the quantum bit-error rate to exceed the security threshold. In this case, a photon heralding scheme may be used to reject the uncorrelated noise photons in order to restore the quantum bit-error rate to a low level. However, the secure key rate would suffer unless one uses a heralded photon source with sufficiently high heralding rate and heralding efficiency. In this work we demonstrate a heralded photon source that has a heralding efficiency that is as high as 74.5%. One disadvantage of a typical heralded photon source is that the long deadtime of the heralding detector results in a significant drop in the heralding rate. To counter this problem, we propose a passively spatial-multiplexed configuration at the heralding arm. Using two heralding detectors in this configuration, we obtain an increase in the heralding rate by 37% and a corresponding increase in the heralded photon detection rate by 16%. We transmit the O-band photons over 10 km of noisy optical fiber to observe the relation between quantum bit-error rate and noise-degraded second-order correlation function of the transmitted photons. The effects of afterpulsing when we shorten the deadtime of the heralding detectors are also observed and discussed.

10Gbps monolithic silicon FTTH transceiver without laser diode for a new PON configuration.

PubMed

Zhang, Jing; Liow, Tsung-Yang; Lo, Guo-Qiang; Kwong, Dim-Lee

2010-03-01

A new passive optical network (PON) configuration and a novel silicon photonic transceiver architecture for optical network unit (ONU) are proposed, eliminating the need for an internal laser source in ONU. The Si transceiver is fully monolithic, includes integrated wavelength division multiplexing (WDM) filters, modulators (MOD) and photo-detectors (PD), and demonstrates low-cost high volume manufacturability.

Engineering cell-fluorescent ion track hybrid detectors.

PubMed

Niklas, Martin; Greilich, Steffen; Melzig, Claudius; Akselrod, Mark S; Debus, Jürgen; Jäkel, Oliver; Abdollahi, Amir

2013-06-11

The lack of sensitive biocompatible particle track detectors has so far limited parallel detection of physical energy deposition and biological response. Fluorescent nuclear track detectors (FNTDs) based on Al₂O₃:C,Mg single crystals combined with confocal laser scanning microscopy (CLSM) provide 3D information on ion tracks with a resolution limited by light diffraction. Here we report the development of next generation cell-fluorescent ion track hybrid detectors (Cell-Fit-HD). The biocompatibility of FNTDs was tested using six different cell lines, i.e. human non-small cell lung carcinoma (A549), glioblastoma (U87), androgen independent prostate cancer (PC3), epidermoid cancer (A431) and murine (VmDk) glioma SMA-560. To evaluate cell adherence, viability and conformal coverage of the crystals different seeding densities and alternative coating with extracellular matrix (fibronectin) was tested. Carbon irradiation was performed in Bragg peak (initial 270.55 MeV u⁻¹). A series of cell compartment specific fluorescence stains including nuclear (HOECHST), membrane (Glut-1), cytoplasm (Calcein AM, CM-DiI) were tested on Cell-Fit-HDs and a single CLSM was employed to co-detect the physical (crystal) as well as the biological (cell layer) information. The FNTD provides a biocompatible surface. Among the cells tested, A549 cells formed the most uniform, viable, tightly packed epithelial like monolayer. The ion track information was not compromised in Cell-Fit-HD as compared to the FNTD alone. Neither cell coating and culturing, nor additional staining procedures affected the properties of the FNTD surface to detect ion tracks. Standard immunofluorescence and live staining procedures could be employed to co-register cell biology and ion track information. The Cell-Fit-Hybrid Detector system is a promising platform for a multitude of studies linking biological response to energy deposition at high level of optical microscopy resolution.

A portable and autonomous multichannel fluorescence detector for on-line and in situ explosive detection in aqueous phase.

PubMed

Xin, Yunhong; Wang, Qi; Liu, Taihong; Wang, Lingling; Li, Jia; Fang, Yu

2012-11-21

A multichannel fluorescence detector used to detect nitroaromatic explosives in aqueous phase has been developed, which is composed of a five-channel sample-sensor unit, a measurement and control unit, a microcontroller, and a communication unit. The characteristics of the detector as developed are mainly embedded in the sensor unit, and each sensor consists of a fluorescent sensing film, a light emitting diode (LED), a multi-pixel photon counter (MPPC), and an optical module with special bandpass optical filters. Due to the high sensitivity of the sensing film, the small size and low cost of LED and MPPC, the developed detector not only has a better detecting performance and small size, but also has a very low cost - it is an alternative to the device made with an expensive high power lamp and photomultiplier tube. The wavelengths of the five sensors covered extend from the upper UV through the visible spectrum, 370-640 nm, and thereby it possesses the potential to detect a variety of explosives and other hazardous materials in aqueous phase. An additional function of the detector is its ability to function via a wireless network, by which the data recorded by the detector can be sent to the host computer, and at the same time the instructions can be sent to the detector from the host computer. By means of the powerful computing ability of the host computer, and utilizing the classical principal component analysis (PCA) algorithm, effective classification of the analytes is achieved. Furthermore, the detector has been tested and evaluated using NB, PA, TNT and DNT as the analytes, and toluene, benzene, methanol and ethanol as interferent compounds (concentration various from 10 and 60 μM). It has been shown that the detector can detect the four nitroaromatics with high sensitivity and selectivity.

Application of GEM-based detectors in full-field XRF imaging

NASA Astrophysics Data System (ADS)

Dąbrowski, W.; Fiutowski, T.; Frączek, P.; Koperny, S.; Lankosz, M.; Mendys, A.; Mindur, B.; Świentek, K.; Wiącek, P.; Wróbel, P. M.

2016-12-01

X-ray fluorescence spectroscopy (XRF) is a commonly used technique for non-destructive elemental analysis of cultural heritage objects. It can be applied to investigations of provenance of historical objects as well as to studies of art techniques. While the XRF analysis can be easily performed locally using standard available equipment there is a growing interest in imaging of spatial distribution of specific elements. Spatial imaging of elemental distrbutions is usually realised by scanning an object with a narrow focused X-ray excitation beam and measuring characteristic fluorescence radiation using a high energy resolution detector, usually a silicon drift detector. Such a technique, called macro-XRF imaging, is suitable for investigation of flat surfaces but it is time consuming because the spatial resolution is basically determined by the spot size of the beam. Another approach is the full-field XRF, which is based on simultaneous irradiation and imaging of large area of an object. The image of the investigated area is projected by a pinhole camera on a position-sensitive and energy dispersive detector. The infinite depth of field of the pinhole camera allows one, in principle, investigation of non-flat surfaces. One of possible detectors to be employed in full-field XRF imaging is a GEM based detector with 2-dimensional readout. In the paper we report on development of an imaging system equipped with a standard 3-stage GEM detector of 10 × 10 cm2 equipped with readout electronics based on dedicated full-custom ASICs and DAQ system. With a demonstrator system we have obtained 2-D spatial resolution of the order of 100 μm and energy resolution at a level of 20% FWHM for 5.9 keV . Limitations of such a detector due to copper fluorescence radiation excited in the copper-clad drift electrode and GEM foils is discussed and performance of the detector using chromium-clad electrodes is reported.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wilson, Matthew D., E-mail: Matt.Wilson@stfc.ac.uk; Seller, Paul; Veale, Matthew C.

A novel, “single-shot” fluorescence imaging technique has been demonstrated on the B16 beamline at the Diamond Light Source synchrotron using the HEXITEC energy dispersive imaging detector. A custom made furnace with 200µm thick metal alloy samples was positioned in a white X-ray beam with a hole made in the furnace walls to allow the transmitted beam to be imaged with a conventional X-ray imaging camera consisting of a 500 µm thick single crystal LYSO scintillator, mirror and lens coupled to an AVT Manta G125B CCD sensor. The samples were positioned 45° to the incident beam to enable simultaneous transmission andmore » fluorescence imaging. The HEXITEC detector was positioned at 90° to the sample with a 50 µm pinhole 13 cm from the sample and the detector positioned 2.3m from pinhole. The geometric magnification provided a field of view of 1.1×1.1mm{sup 2} with one of the 80×80 pixels imaging an area equivalent to 13µm{sup 2}. Al-Cu alloys doped with Zr, Ag and Mo were imaged in transmission and fluorescence mode. The fluorescence images showed that the dopant metals could be simultaneously imaged with sufficient counts on all 80x80 pixels within 60 s, with the X-ray flux limiting the fluorescence imaging rate. This technique demonstrated that it is possible to simultaneously image and identify multiple elements on a spatial resolution scale ~10µm or higher without the time consuming need to scan monochromatic energies or raster scan a focused beam of X-rays. Moving to high flux beamlines and using an array of detectors could improve the imaging speed of the technique with element specific imaging estimated to be on a 1 s timescale.« less

Effect of Clouds on Apertures of Space-based Air Fluorescence Detectors

NASA Technical Reports Server (NTRS)

Sokolsky, P.; Krizmanic, J.

2003-01-01

Space-based ultra-high-energy cosmic ray detectors observe fluorescence light from extensive air showers produced by these particles in the troposphere. Clouds can scatter and absorb this light and produce systematic errors in energy determination and spectrum normalization. We study the possibility of using IR remote sensing data from MODIS and GOES satellites to delimit clear areas of the atmosphere. The efficiency for detecting ultra-high-energy cosmic rays whose showers do not intersect clouds is determined for real, night-time cloud scenes. We use the MODIS SST cloud mask product to define clear pixels for cloud scenes along the equator and use the OWL Monte Carlo to generate showers in the cloud scenes. We find the efficiency for cloud-free showers with closest approach of three pixels to a cloudy pixel is 6.5% exclusive of other factors. We conclude that defining a totally cloud-free aperture reduces the sensitivity of space-based fluorescence detectors to unacceptably small levels.

EUSO-TA prototype telescope

NASA Astrophysics Data System (ADS)

Bisconti, Francesca; JEM-EUSO Collaboration

2016-07-01

EUSO-TA is one of the prototypes developed for the JEM-EUSO project, a space-based large field-of-view telescope to observe the fluorescence light emitted by cosmic ray air showers in the atmosphere. EUSO-TA is a ground-based prototype located at the Telescope Array (TA) site in Utah, USA, where an Electron Light Source and a Central Laser Facility are installed. The purpose of the EUSO-TA project is to calibrate the prototype with the TA fluorescence detector in presence of well-known light sources and cosmic ray air showers. In 2015, the detector started the first measurements and tests using the mentioned light sources have been performed successfully. A first cosmic ray candidate has been observed, as well as stars of different magnitude and color index. Since Silicon Photo-Multipliers (SiPMs) are very promising for fluorescence telescopes of next generation, they are under consideration for the realization of a new prototype of EUSO Photo Detector Module (PDM). The response of this sensor type is under investigation through simulations and laboratory experimentation.

Full-field fan-beam x-ray fluorescence computed tomography system design with linear-array detectors and pinhole collimation: a rapid Monte Carlo study

NASA Astrophysics Data System (ADS)

Zhang, Siyuan; Li, Liang; Li, Ruizhe; Chen, Zhiqiang

2017-11-01

We present the design concept and initial simulations for a polychromatic full-field fan-beam x-ray fluorescence computed tomography (XFCT) device with pinhole collimators and linear-array photon counting detectors. The phantom is irradiated by a fan-beam polychromatic x-ray source filtered by copper. Fluorescent photons are stimulated and then collected by two linear-array photon counting detectors with pinhole collimators. The Compton scatter correction and the attenuation correction are applied in the data processing, and the maximum-likelihood expectation maximization algorithm is applied for the image reconstruction of XFCT. The physical modeling of the XFCT imaging system was described, and a set of rapid Monte Carlo simulations was carried out to examine the feasibility and sensitivity of the XFCT system. Different concentrations of gadolinium (Gd) and gold (Au) solutions were used as contrast agents in simulations. Results show that 0.04% of Gd and 0.065% of Au can be well reconstructed with the full scan time set at 6 min. Compared with using the XFCT system with a pencil-beam source or a single-pixel detector, using a full-field fan-beam XFCT device with linear-array detectors results in significant scanning time reduction and may satisfy requirements of rapid imaging, such as in vivo imaging experiments.

First upper limits on the radar cross section of cosmic-ray induced extensive air showers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Abbasi, R. U.; Abe, M.; Abou Bakr Othman, M.

TARA (Telescope Array Radar) is a cosmic ray radar detection experiment colocated with Telescope Array, the conventional surface scintillation detector (SD) and fluorescence telescope detector (FD) near Delta, Utah, U.S.A. Furthermore, the TARA detector combines a 40 kW, 54.1 MHz VHF transmitter and high-gain transmitting antenna which broadcasts the radar carrier over the SD array and within the FD field of view, towards a 250 MS/s DAQ receiver. TARA has been collecting data since 2013 with the primary goal of observing the radar signatures of extensive air showers (EAS). Simulations indicate that echoes are expected to be short in durationmore » (~10 µs) and exhibit rapidly changing frequency, with rates on the order 1 MHz/µs. The EAS radar cross-section (RCS) is currently unknown although it is the subject of over 70 years of speculation. One novel signal search technique is described in which the expected radar echo of a particular air shower is used as a matched filter template and compared to waveforms obtained by triggering the radar DAQ using the Telescope Array fluorescence detector. No evidence for the scattering of radio frequency radiation by EAS is obtained to date. Finally, we report the first quantitative RCS upper limits using EAS that triggered the Telescope Array Fluorescence Detector.« less

First upper limits on the radar cross section of cosmic-ray induced extensive air showers

DOE PAGES

Abbasi, R. U.; Abe, M.; Abou Bakr Othman, M.; ...

2016-11-19

TARA (Telescope Array Radar) is a cosmic ray radar detection experiment colocated with Telescope Array, the conventional surface scintillation detector (SD) and fluorescence telescope detector (FD) near Delta, Utah, U.S.A. Furthermore, the TARA detector combines a 40 kW, 54.1 MHz VHF transmitter and high-gain transmitting antenna which broadcasts the radar carrier over the SD array and within the FD field of view, towards a 250 MS/s DAQ receiver. TARA has been collecting data since 2013 with the primary goal of observing the radar signatures of extensive air showers (EAS). Simulations indicate that echoes are expected to be short in durationmore » (~10 µs) and exhibit rapidly changing frequency, with rates on the order 1 MHz/µs. The EAS radar cross-section (RCS) is currently unknown although it is the subject of over 70 years of speculation. One novel signal search technique is described in which the expected radar echo of a particular air shower is used as a matched filter template and compared to waveforms obtained by triggering the radar DAQ using the Telescope Array fluorescence detector. No evidence for the scattering of radio frequency radiation by EAS is obtained to date. Finally, we report the first quantitative RCS upper limits using EAS that triggered the Telescope Array Fluorescence Detector.« less

Submicrometre geometrically encoded fluorescent barcodes self-assembled from DNA

NASA Astrophysics Data System (ADS)

Lin, Chenxiang; Jungmann, Ralf; Leifer, Andrew M.; Li, Chao; Levner, Daniel; Church, George M.; Shih, William M.; Yin, Peng

2012-10-01

The identification and differentiation of a large number of distinct molecular species with high temporal and spatial resolution is a major challenge in biomedical science. Fluorescence microscopy is a powerful tool, but its multiplexing ability is limited by the number of spectrally distinguishable fluorophores. Here, we used (deoxy)ribonucleic acid (DNA)-origami technology to construct submicrometre nanorods that act as fluorescent barcodes. We demonstrate that spatial control over the positioning of fluorophores on the surface of a stiff DNA nanorod can produce 216 distinct barcodes that can be decoded unambiguously using epifluorescence or total internal reflection fluorescence microscopy. Barcodes with higher spatial information density were demonstrated via the construction of super-resolution barcodes with features spaced by ˜40 nm. One species of the barcodes was used to tag yeast surface receptors, which suggests their potential applications as in situ imaging probes for diverse biomolecular and cellular entities in their native environments.

Realizing up-conversion fluorescence tuning in lanthanide-doped nanocrystals by femtosecond pulse shaping method

PubMed Central

Zhang, Shian; Yao, Yunhua; Shuwu, Xu; Liu, Pei; Ding, Jingxin; Jia, Tianqing; Qiu, Jianrong; Sun, Zhenrong

2015-01-01

The ability to tune color output of nanomaterials is very important for their applications in laser, optoelectronic device, color display and multiplexed biolabeling. Here we first propose a femtosecond pulse shaping technique to realize the up-conversion fluorescence tuning in lanthanide-doped nanocrystals dispersed in the glass. The multiple subpulse formation by a square phase modulation can create different excitation pathways for various up-conversion fluorescence generations. By properly controlling these excitation pathways, the multicolor up-conversion fluorescence can be finely tuned. This color tuning by the femtosecond pulse shaping technique is realized in single material by single-color laser field, which is highly desirable for further applications of the lanthanide-doped nanocrystals. This femtosecond pulse shaping technique opens an opportunity to tune the color output in the lanthanide-doped nanocrystals, which may bring a new revolution in the control of luminescence properties of nanomaterials. PMID:26290391

Sub-micrometer Geometrically Encoded Fluorescent Barcodes Self-Assembled from DNA

PubMed Central

Lin, Chenxiang; Jungmann, Ralf; Leifer, Andrew M.; Li, Chao; Levner, Daniel; Church, George M.; Shih, William M.; Yin, Peng

2012-01-01

The identification and differentiation of a large number of distinct molecular species with high temporal and spatial resolution is a major challenge in biomedical science. Fluorescence microscopy is a powerful tool, but its multiplexing ability is limited by the number of spectrally distinguishable fluorophores. Here we use DNA-origami technology to construct sub-micrometer nanorods that act as fluorescent barcodes. We demonstrate that spatial control over the positioning of fluorophores on the surface of a stiff DNA nanorod can produce 216 distinct barcodes that can be unambiguously decoded using epifluorescence or total internal reflection fluorescence (TIRF) microscopy. Barcodes with higher spatial information density were demonstrated via the construction of super-resolution barcodes with features spaced by ~40 nm. One species of the barcodes was used to tag yeast surface receptors, suggesting their potential applications as in situ imaging probes for diverse biomolecular and cellular entities in their native environments. PMID:23000997

Multiphoton versus confocal high resolution z-sectioning of enhanced green fluorescent microtubules: increased multiphoton photobleaching within the focal plane can be compensated using a Pockels cell and dual widefield detectors.

PubMed

Drummond, D R; Carter, N; Cross, R A

2002-05-01

Multiphoton excitation was originally projected to improve live cell fluorescence imaging by minimizing photobleaching effects outside the focal plane, yet reports suggest that photobleaching within the focal plane is actually worse than with one photon excitation. We confirm that when imaging enhanced green fluorescent protein, photobleaching is indeed more acute within the multiphoton excitation volume, so that whilst fluorescence increases as predicted with the square of the excitation power, photobleaching rates increase with a higher order relationship. Crucially however, multiphoton excitation also affords unique opportunities for substantial improvements to fluorescence detection. By using a Pockels cell to minimize exposure of the specimen together with multiple nondescanned detectors we show quantitatively that for any particular bleach rate multiphoton excitation produces significantly more signal than one photon excitation confocal microscopy in high resolution Z-axis sectioning of thin samples. Both modifications are readily implemented on a commercial multiphoton microscope system.

Small fluorescence-activating and absorption-shifting tag for tunable protein imaging in vivo

PubMed Central

Plamont, Marie-Aude; Billon-Denis, Emmanuelle; Maurin, Sylvie; Gauron, Carole; Pimenta, Frederico M.; Specht, Christian G.; Shi, Jian; Quérard, Jérôme; Pan, Buyan; Rossignol, Julien; Moncoq, Karine; Morellet, Nelly; Volovitch, Michel; Lescop, Ewen; Chen, Yong; Triller, Antoine; Vriz, Sophie; Le Saux, Thomas; Jullien, Ludovic; Gautier, Arnaud

2016-01-01

This paper presents Yellow Fluorescence-Activating and absorption-Shifting Tag (Y-FAST), a small monomeric protein tag, half as large as the green fluorescent protein, enabling fluorescent labeling of proteins in a reversible and specific manner through the reversible binding and activation of a cell-permeant and nontoxic fluorogenic ligand (a so-called fluorogen). A unique fluorogen activation mechanism based on two spectroscopic changes, increase of fluorescence quantum yield and absorption red shift, provides high labeling selectivity. Y-FAST was engineered from the 14-kDa photoactive yellow protein by directed evolution using yeast display and fluorescence-activated cell sorting. Y-FAST is as bright as common fluorescent proteins, exhibits good photostability, and allows the efficient labeling of proteins in various organelles and hosts. Upon fluorogen binding, fluorescence appears instantaneously, allowing monitoring of rapid processes in near real time. Y-FAST distinguishes itself from other tagging systems because the fluorogen binding is highly dynamic and fully reversible, which enables rapid labeling and unlabeling of proteins by addition and withdrawal of the fluorogen, opening new exciting prospects for the development of multiplexing imaging protocols based on sequential labeling. PMID:26711992

Small fluorescence-activating and absorption-shifting tag for tunable protein imaging in vivo.

PubMed

Plamont, Marie-Aude; Billon-Denis, Emmanuelle; Maurin, Sylvie; Gauron, Carole; Pimenta, Frederico M; Specht, Christian G; Shi, Jian; Quérard, Jérôme; Pan, Buyan; Rossignol, Julien; Moncoq, Karine; Morellet, Nelly; Volovitch, Michel; Lescop, Ewen; Chen, Yong; Triller, Antoine; Vriz, Sophie; Le Saux, Thomas; Jullien, Ludovic; Gautier, Arnaud

2016-01-19

This paper presents Yellow Fluorescence-Activating and absorption-Shifting Tag (Y-FAST), a small monomeric protein tag, half as large as the green fluorescent protein, enabling fluorescent labeling of proteins in a reversible and specific manner through the reversible binding and activation of a cell-permeant and nontoxic fluorogenic ligand (a so-called fluorogen). A unique fluorogen activation mechanism based on two spectroscopic changes, increase of fluorescence quantum yield and absorption red shift, provides high labeling selectivity. Y-FAST was engineered from the 14-kDa photoactive yellow protein by directed evolution using yeast display and fluorescence-activated cell sorting. Y-FAST is as bright as common fluorescent proteins, exhibits good photostability, and allows the efficient labeling of proteins in various organelles and hosts. Upon fluorogen binding, fluorescence appears instantaneously, allowing monitoring of rapid processes in near real time. Y-FAST distinguishes itself from other tagging systems because the fluorogen binding is highly dynamic and fully reversible, which enables rapid labeling and unlabeling of proteins by addition and withdrawal of the fluorogen, opening new exciting prospects for the development of multiplexing imaging protocols based on sequential labeling.

High-Resolution Detector For X-Ray Diffraction

NASA Technical Reports Server (NTRS)

Carter, Daniel C.; Withrow, William K.; Pusey, Marc L.; Yost, Vaughn H.

1988-01-01

Proposed x-ray-sensitive imaging detector offers superior spatial resolution, counting-rate capacity, and dynamic range. Instrument based on laser-stimulated luminescence and reusable x-ray-sensitive film. Detector scans x-ray film line by line. Extracts latent image in film and simultaneously erases film for reuse. Used primarily for protein crystallography. Principle adapted to imaging detectors for electron microscopy and fluorescence spectroscopy and general use in astronomy, engineering, and medicine.

Recent results of synchrotron radiation induced total reflection X-ray fluorescence analysis at HASYLAB, beamline L

NASA Astrophysics Data System (ADS)

Streli, C.; Pepponi, G.; Wobrauschek, P.; Jokubonis, C.; Falkenberg, G.; Záray, G.; Broekaert, J.; Fittschen, U.; Peschel, B.

2006-11-01

At the Hamburger Synchrotronstrahlungslabor (HASYLAB), Beamline L, a vacuum chamber for synchrotron radiation-induced total reflection X-ray fluorescence analysis, is now available which can easily be installed using the adjustment components for microanalysis present at this beamline. The detector is now in the final version of a Vortex silicon drift detector with 50-mm 2 active area from Radiant Detector Technologies. With the Ni/C multilayer monochromator set to 17 keV extrapolated detection limits of 8 fg were obtained using the 50-mm 2 silicon drift detector with 1000 s live time on a sample containing 100 pg of Ni. Various applications are presented, especially of samples which are available in very small amounts: As synchrotron radiation-induced total reflection X-ray fluorescence analysis is much more sensitive than tube-excited total reflection X-ray fluorescence analysis, the sampling time of aerosol samples can be diminished, resulting in a more precise time resolution of atmospheric events. Aerosols, directly sampled on Si reflectors in an impactor were investigated. A further application was the determination of contamination elements in a slurry of high-purity Al 2O 3. No digestion is required; the sample is pipetted and dried before analysis. A comparison with laboratory total reflection X-ray fluorescence analysis showed the higher sensitivity of synchrotron radiation-induced total reflection X-ray fluorescence analysis, more contamination elements could be detected. Using the Si-111 crystal monochromator also available at beamline L, XANES measurements to determine the chemical state were performed. This is only possible with lower sensitivity as the flux transmitted by the crystal monochromator is about a factor of 100 lower than that transmitted by the multilayer monochromator. Preliminary results of X-ray absorption near-edge structure measurements for As in xylem sap from cucumber plants fed with As(III) and As(V) are reported. Detection limits of 170 ng/l of As in xylem sap were achieved.

Fluorescence-suppressed time-resolved Raman spectroscopy of pharmaceuticals using complementary metal-oxide semiconductor (CMOS) single-photon avalanche diode (SPAD) detector.

PubMed

Rojalin, Tatu; Kurki, Lauri; Laaksonen, Timo; Viitala, Tapani; Kostamovaara, Juha; Gordon, Keith C; Galvis, Leonardo; Wachsmann-Hogiu, Sebastian; Strachan, Clare J; Yliperttula, Marjo

2016-01-01

In this work, we utilize a short-wavelength, 532-nm picosecond pulsed laser coupled with a time-gated complementary metal-oxide semiconductor (CMOS) single-photon avalanche diode (SPAD) detector to acquire Raman spectra of several drugs of interest. With this approach, we are able to reveal previously unseen Raman features and suppress the fluorescence background of these drugs. Compared to traditional Raman setups, the present time-resolved technique has two major improvements. First, it is possible to overcome the strong fluorescence background that usually interferes with the much weaker Raman spectra. Second, using the high photon energy excitation light source, we are able to generate a stronger Raman signal compared to traditional instruments. In addition, observations in the time domain can be performed, thus enabling new capabilities in the field of Raman and fluorescence spectroscopy. With this system, we demonstrate for the first time the possibility of recording fluorescence-suppressed Raman spectra of solid, amorphous and crystalline, and non-photoluminescent and photoluminescent drugs such as caffeine, ranitidine hydrochloride, and indomethacin (amorphous and crystalline forms). The raw data acquired by utilizing only the picosecond pulsed laser and a CMOS SPAD detector could be used for identifying the compounds directly without any data processing. Moreover, to validate the accuracy of this time-resolved technique, we present density functional theory (DFT) calculations for a widely used gastric acid inhibitor, ranitidine hydrochloride. The obtained time-resolved Raman peaks were identified based on the calculations and existing literature. Raman spectra using non-time-resolved setups with continuous-wave 785- and 532-nm excitation lasers were used as reference data. Overall, this demonstration of time-resolved Raman and fluorescence measurements with a CMOS SPAD detector shows promise in diverse areas, including fundamental chemical research, the pharmaceutical setting, process analytical technology (PAT), and the life sciences.

The Development of a 30-125 Micron Array for Airborne Astronomy

NASA Technical Reports Server (NTRS)

Mason, C. G.; Dotson, J. L.; Erickson, E. F.; Farhoomand, J.; Haas, M. R.; Koerber, C. T.; Prasad, A.; Sisson, D.; Witteborn, F. C.; DeVincenzi, Donald (Technical Monitor)

2002-01-01

The development of a 30-125 micron Ge:Sb photoconductor array for AIRES (Airborne Infra-Red Echelle Spectrometer) is described. The prototype array is a 2x24 module which can be close-stacked to provide larger two-dimensional formats. Light is focused onto each detector using a collecting cone with a 2 mm pitch. The array is read out by two Raytheon SBRC-190 cryogenic multiplexers that also provide a CTIA (capacitive transimpedance amplifier) unit cell for each detector. We discuss our results from a test series conducted to measure the array performance and to evaluate its suitability for airborne astronomy.

The Mid-Infrared Instrument for the James Webb Space Telescope, VIII: The MIRI Focal Plane System

NASA Astrophysics Data System (ADS)

Ressler, M. E.; Sukhatme, K. G.; Franklin, B. R.; Mahoney, J. C.; Thelen, M. P.; Bouchet, P.; Colbert, J. W.; Cracraft, Misty; Dicken, D.; Gastaud, R.; Goodson, G. B.; Eccleston, Paul; Moreau, V.; Rieke, G. H.; Schneider, Analyn

2015-07-01

We describe the layout and unique features of the focal plane system for MIRI. We begin with the detector array and its readout integrated circuit (combining the amplifier unit cells and the multiplexer), the electronics, and the steps by which the data collection is controlled and the output signals are digitized and delivered to the JWST spacecraft electronics system. We then discuss the operation of this MIRI data system, including detector readout patterns, operation of subarrays, and data formats. Finally, we summarize the performance of the system, including remaining anomalies that need to be corrected in the data pipeline.

Apparatus for eliminating background interference in fluorescence measurements

DOEpatents

Martin, J.C.; Jett, J.H.

1984-01-06

The disclosure is directed to an apparatus for eliminating background interference during fluorescence measurements in a multiple laser flow cytometer. A biological particle stained with fluorescent dyes is excited by a laser. A fluorescence detector detects the fluorescence. The particle scatters light and a gate signal is generated and delayed until the biological particle reaches the next laser. The delayed signal turns on this next laser which excites a different stained component of the same biological particle.

Apparatus for eliminating background interference in fluorescence measurements

DOEpatents

Martin, John C.; Jett, James H.

1986-01-01

The disclosure is directed to an apparatus for eliminating background interference during fluorescence measurements in a multiple laser flow cytometer. A biological particle stained with fluorescent dyes is excited by a laser. A fluorescence detector detects the fluorescence. The particle scatters light and a gate signal is generated and delayed until the biological particle reaches the next laser. The delayed signal turns on this next laser, which excites a different stained component of the same biological particle.

Apparatus for eliminating background interference in fluorescence measurements

DOEpatents

Martin, J.C.; Jett, J.H.

1986-03-04

The disclosure is directed to an apparatus for eliminating background interference during fluorescence measurements in a multiple laser flow cytometer. A biological particle stained with fluorescent dyes is excited by a laser. A fluorescence detector detects the fluorescence. The particle scatters light and a gate signal is generated and delayed until the biological particle reaches the next laser. The delayed signal turns on this next laser, which excites a different stained component of the same biological particle. 8 figs.

Six orders of magnitude dynamic range in capillary electrophoresis with ultrasensitive laser-induced fluorescence detection

PubMed Central

Whitmore, Colin D.; Essaka, David; Dovichi, Norman J.

2009-01-01

An ultrasensitive laser-induced fluorescence detector was used with capillary electrophoresis for the study of 5-carboxy-tetramethylrhodamine. The raw signal from the detector provided roughly three orders of magnitude dynamic range. The signal saturated at high analyte concentrations due to the dead time associated with the single-photon counting avalanche photodiode employed in the detector. The signal can be corrected for the detector dead time, providing an additional order of magnitude dynamic range. To further increase dynamic range, two fiber-optic beam-splitters were cascaded to generate a primary signal and two attenuated signals, each monitored by a single-photon counting avalanche photodiode. The combined signals from the three photodiodes are reasonably linear from the concentration detection limit of 3 pM to 10 μM, the maximum concentration investigated, a range of 3,000,000. Mass detection limits were 150 yoctomoles injected onto the capillary. PMID:19836546

Detectors for single-molecule fluorescence imaging and spectroscopy

PubMed Central

MICHALET, X.; SIEGMUND, O.H.W.; VALLERGA, J.V.; JELINSKY, P.; MILLAUD, J.E.; WEISS, S.

2010-01-01

Single-molecule observation, characterization and manipulation techniques have recently come to the forefront of several research domains spanning chemistry, biology and physics. Due to the exquisite sensitivity, specificity, and unmasking of ensemble averaging, single-molecule fluorescence imaging and spectroscopy have become, in a short period of time, important tools in cell biology, biochemistry and biophysics. These methods led to new ways of thinking about biological processes such as viral infection, receptor diffusion and oligomerization, cellular signaling, protein-protein or protein-nucleic acid interactions, and molecular machines. Such achievements require a combination of several factors to be met, among which detector sensitivity and bandwidth are crucial. We examine here the needed performance of photodetectors used in these types of experiments, the current state of the art for different categories of detectors, and actual and future developments of single-photon counting detectors for single-molecule imaging and spectroscopy. PMID:20157633

Small Angle X-Ray Scattering Detector

DOEpatents

Hessler, Jan P.

2004-06-15

A detector for time-resolved small-angle x-ray scattering includes a nearly constant diameter, evacuated linear tube having an end plate detector with a first fluorescent screen and concentric rings of first fiber optic bundles for low angle scattering detection and an annular detector having a second fluorescent screen and second fiber optic bundles concentrically disposed about the tube for higher angle scattering detection. With the scattering source, i.e., the specimen under investigation, located outside of the evacuated tube on the tube's longitudinal axis, scattered x-rays are detected by the fiber optic bundles, to each of which is coupled a respective photodetector, to provide a measurement resolution, i.e., dq/q, where q is the momentum transferred from an incident x-ray to an x-ray scattering specimen, of 2% over two (2) orders of magnitude in reciprocal space, i.e., q.sub.max /q.sub.min.congruent.100.

Highly multiplexed single-cell analysis of formalin-fixed, paraffin-embedded cancer tissue

PubMed Central

Gerdes, Michael J.; Sevinsky, Christopher J.; Sood, Anup; Adak, Sudeshna; Bello, Musodiq O.; Bordwell, Alexander; Can, Ali; Corwin, Alex; Dinn, Sean; Filkins, Robert J.; Hollman, Denise; Kamath, Vidya; Kaanumalle, Sireesha; Kenny, Kevin; Larsen, Melinda; Lazare, Michael; Lowes, Christina; McCulloch, Colin C.; McDonough, Elizabeth; Pang, Zhengyu; Rittscher, Jens; Santamaria-Pang, Alberto; Sarachan, Brion D.; Seel, Maximilian L.; Seppo, Antti; Shaikh, Kashan; Sui, Yunxia; Zhang, Jingyu; Ginty, Fiona

2013-01-01

Limitations on the number of unique protein and DNA molecules that can be characterized microscopically in a single tissue specimen impede advances in understanding the biological basis of health and disease. Here we present a multiplexed fluorescence microscopy method (MxIF) for quantitative, single-cell, and subcellular characterization of multiple analytes in formalin-fixed paraffin-embedded tissue. Chemical inactivation of fluorescent dyes after each image acquisition round allows reuse of common dyes in iterative staining and imaging cycles. The mild inactivation chemistry is compatible with total and phosphoprotein detection, as well as DNA FISH. Accurate computational registration of sequential images is achieved by aligning nuclear counterstain-derived fiducial points. Individual cells, plasma membrane, cytoplasm, nucleus, tumor, and stromal regions are segmented to achieve cellular and subcellular quantification of multiplexed targets. In a comparison of pathologist scoring of diaminobenzidine staining of serial sections and automated MxIF scoring of a single section, human epidermal growth factor receptor 2, estrogen receptor, p53, and androgen receptor staining by diaminobenzidine and MxIF methods yielded similar results. Single-cell staining patterns of 61 protein antigens by MxIF in 747 colorectal cancer subjects reveals extensive tumor heterogeneity, and cluster analysis of divergent signaling through ERK1/2, S6 kinase 1, and 4E binding protein 1 provides insights into the spatial organization of mechanistic target of rapamycin and MAPK signal transduction. Our results suggest MxIF should be broadly applicable to problems in the fields of basic biological research, drug discovery and development, and clinical diagnostics. PMID:23818604

Highly multiplexed single-cell analysis of formalin-fixed, paraffin-embedded cancer tissue.

PubMed

Gerdes, Michael J; Sevinsky, Christopher J; Sood, Anup; Adak, Sudeshna; Bello, Musodiq O; Bordwell, Alexander; Can, Ali; Corwin, Alex; Dinn, Sean; Filkins, Robert J; Hollman, Denise; Kamath, Vidya; Kaanumalle, Sireesha; Kenny, Kevin; Larsen, Melinda; Lazare, Michael; Li, Qing; Lowes, Christina; McCulloch, Colin C; McDonough, Elizabeth; Montalto, Michael C; Pang, Zhengyu; Rittscher, Jens; Santamaria-Pang, Alberto; Sarachan, Brion D; Seel, Maximilian L; Seppo, Antti; Shaikh, Kashan; Sui, Yunxia; Zhang, Jingyu; Ginty, Fiona

2013-07-16

Limitations on the number of unique protein and DNA molecules that can be characterized microscopically in a single tissue specimen impede advances in understanding the biological basis of health and disease. Here we present a multiplexed fluorescence microscopy method (MxIF) for quantitative, single-cell, and subcellular characterization of multiple analytes in formalin-fixed paraffin-embedded tissue. Chemical inactivation of fluorescent dyes after each image acquisition round allows reuse of common dyes in iterative staining and imaging cycles. The mild inactivation chemistry is compatible with total and phosphoprotein detection, as well as DNA FISH. Accurate computational registration of sequential images is achieved by aligning nuclear counterstain-derived fiducial points. Individual cells, plasma membrane, cytoplasm, nucleus, tumor, and stromal regions are segmented to achieve cellular and subcellular quantification of multiplexed targets. In a comparison of pathologist scoring of diaminobenzidine staining of serial sections and automated MxIF scoring of a single section, human epidermal growth factor receptor 2, estrogen receptor, p53, and androgen receptor staining by diaminobenzidine and MxIF methods yielded similar results. Single-cell staining patterns of 61 protein antigens by MxIF in 747 colorectal cancer subjects reveals extensive tumor heterogeneity, and cluster analysis of divergent signaling through ERK1/2, S6 kinase 1, and 4E binding protein 1 provides insights into the spatial organization of mechanistic target of rapamycin and MAPK signal transduction. Our results suggest MxIF should be broadly applicable to problems in the fields of basic biological research, drug discovery and development, and clinical diagnostics.

Experimental verification of the kinetic theory of FRET using optical microspectroscopy and obligate oligomers.

PubMed

Patowary, Suparna; Pisterzi, Luca F; Biener, Gabriel; Holz, Jessica D; Oliver, Julie A; Wells, James W; Raicu, Valerică

2015-04-07

Förster resonance energy transfer (FRET) is a nonradiative process for the transfer of energy from an optically excited donor molecule (D) to an acceptor molecule (A) in the ground state. The underlying theory predicting the dependence of the FRET efficiency on the sixth power of the distance between D and A has stood the test of time. In contrast, a comprehensive kinetic-based theory developed recently for FRET efficiencies among multiple donors and acceptors in multimeric arrays has waited for further testing. That theory has been tested in the work described in this article using linked fluorescent proteins located in the cytoplasm and at the plasma membrane of living cells. The cytoplasmic constructs were fused combinations of Cerulean as donor (D), Venus as acceptor (A), and a photo-insensitive molecule (Amber) as a nonfluorescent (N) place holder: namely, NDAN, NDNA, and ADNN duplexes, and the fully fluorescent quadruplex ADAA. The membrane-bound constructs were fused combinations of GFP2 as donor (D) and eYFP as acceptor (A): namely, two fluorescent duplexes (i.e., DA and AD) and a fluorescent triplex (ADA). According to the theory, the FRET efficiency of a multiplex such as ADAA or ADA can be predicted from that of analogs containing a single acceptor (e.g., NDAN, NDNA, and ADNN, or DA and AD, respectively). Relatively small but statistically significant differences were observed between the measured and predicted FRET efficiencies of the two multiplexes. While elucidation of the cause of this mismatch could be a worthy endeavor, the discrepancy does not appear to question the theoretical underpinnings of a large family of FRET-based methods for determining the stoichiometry and quaternary structure of complexes of macromolecules in living cells. Copyright © 2015 Biophysical Society. Published by Elsevier Inc. All rights reserved.

The First Multichroic Polarimeter Array on the Atacama Cosmology Telescope: Characterization and Performance

NASA Technical Reports Server (NTRS)

Ho, S. P.; Pappas, C. G.; Austermann, J.; Beall, J. A.; Becker, D.; Choi, S. K.; Datta, R.; Duff, S. M.; Gallardo, P. A.; Grace, E.;

The First Multichroic Polarimeter Array on the Atacama Cosmology Telescope: Characterization and Performance

NASA Astrophysics Data System (ADS)

Ho, S. P.; Pappas, C. G.; Austermann, J.; Beall, J. A.; Becker, D.; Choi, S. K.; Datta, R.; Duff, S. M.; Gallardo, P. A.; Grace, E.; Hasselfield, M.; Henderson, S. W.; Hilton, G. C.; Hubmayr, J.; Koopman, B. J.; Lanen, J. V.; Li, D.; McMahon, J.; Nati, F.; Niemack, M. D.; Niraula, P.; Salatino, M.; Schillaci, A.; Schmitt, B. L.; Simon, S. M.; Staggs, S. T.; Stevens, J. R.; Ward, J. T.; Wollack, E. J.; Vavagiakis, E. M.

2016-08-01

The Atacama Cosmology Telescope Polarimeter (ACTPol) is a polarization sensitive receiver for the 6-m Atacama Cosmology Telescope (ACT) and measures the small angular scale polarization anisotropies in the cosmic microwave background (CMB). The full focal plane is composed of three detector arrays, containing over 3000 transition edge sensors (TES detectors) in total. The first two detector arrays, observing at 146 GHz, were deployed in 2013 and 2014, respectively. The third and final array is composed of multichroic pixels sensitive to both 90 and 146 GHz and saw first light in February 2015. Fabricated at NIST, this dichroic array consists of 255 pixels, with a total of 1020 polarization sensitive bolometers and is coupled to the telescope with a monolithic array of broad-band silicon feedhorns. The detectors are read out using time-division SQUID multiplexing and cooled by a dilution refrigerator at 110 mK. We present an overview of the assembly and characterization of this multichroic array in the lab, and the initial detector performance in Chile. The detector array has a TES detector electrical yield of 85 %, a total array sensitivity of less than 10 \\upmu K√{ {s}}, and detector time constants and saturation powers suitable for ACT CMB observations.

Detectors for Tomorrow's Instruments

NASA Technical Reports Server (NTRS)

Moseley, Harvey

2009-01-01

Cryogenically cooled superconducting detectors have become essential tools for a wide range of measurement applications, ranging from quantum limited heterodyne detection in the millimeter range to direct searches for dark matter with superconducting phonon detectors operating at 20 mK. Superconducting detectors have several fundamental and practical advantages which have resulted in their rapid adoption by experimenters. Their excellent performance arises in part from reductions in noise resulting from their low operating temperatures, but unique superconducting properties provide a wide range of mechanisms for detection. For example, the steep dependence of resistance with temperature on the superconductor/normal transition provides a sensitive thermometer for calorimetric and bolometric applications. Parametric changes in the properties of superconducting resonators provides a mechanism for high sensitivity detection of submillimeter photons. From a practical point of view, the use of superconducting detectors has grown rapidly because many of these devices couple well to SQUID amplifiers, which are easily integrated with the detectors. These SQUID-based amplifiers and multiplexers have matured with the detectors; they are convenient to use, and have excellent noise performance. The first generation of fully integrated large scale superconducting detection systems are now being deployed. I will discuss the prospects for a new generation of instruments designed to take full advantage of the revolution in detector technology.

Time-Resolved Measurements in Optoelectronic Microbioanalysis

NASA Technical Reports Server (NTRS)

Bearman, Gregory; Kossakovski, Dmitri

2003-01-01

A report presents discussion of time-resolved measurements in optoelectronic microbioanalysis. Proposed microbioanalytical laboratory-on-a-chip devices for detection of microbes and toxic chemicals would include optoelectronic sensors and associated electronic circuits that would look for fluorescence or phosphorescence signatures of multiple hazardous biomolecules in order to detect which ones were present in a given situation. The emphasis in the instant report is on gating an active-pixel sensor in the time domain, instead of filtering light in the wavelength domain, to prevent the sensor from responding to a laser pulse used to excite fluorescence or phosphorescence while enabling the sensor to respond to the decaying fluorescence or phosphorescence signal that follows the laser pulse. The active-pixel sensor would be turned on after the laser pulse and would be used to either integrate the fluorescence or phosphorescence signal over several lifetimes and many excitation pulses or else take time-resolved measurements of the fluorescence or phosphorescence. The report also discusses issues of multiplexing and of using time-resolved measurements of fluorophores with known different fluorescence lifetimes to distinguish among them.

The NSLS 100 element solid state array detector

PubMed Central

Furenlid, L.R.; Kraner, H.W.; Rogers, L.C.; Cramer, S.P.; Stephani, D.; Beuttenmuller, R.H.; Beren, J.

2015-01-01

X-ray absorption studies of dilute samples require fluorescence detection techniques. Since signal-to-noise ratios are governed by the ratio of fluorescent to scattered photons counted by a detector, solid state detectors which can discriminate between fluorescence and scattered photons have become the instruments of choice for trace element measurements. Commercially available 13 element Ge array detectors permitting total count rates < 500000 counts per second are now in routine use. Since X-ray absorption beamlines at high brightness synchrotron sources can already illuminate most dilute samples with enough flux to saturate the current generation of solid state detectors, the development of next-generation instruments with significantly higher total count rates is essential. We present the design and current status of the 100 element Si array detector being developed in a collaboration between the NSLS and the Instrumentation Division at Brookhaven National Laboratory. The detecting array consists of a 10×10 matrix of 4 mm×4 mm elements laid out on a single piece of ultrahigh purity silicon mounted at the front end of a liquid nitrogen dewar assembly. A matrix of charge sensitive integrating preamplifiers feed signals to an array of shaping amplifiers, single channel analyzers, and scalers. An electronic switch, delay amplifier, linear gate, digital scope, peak sensing A/D converter, and histogramining memory module provide for complete diagnostics and channel calibration. The entire instrument is controlled by a LabView 2 application on a MacII ci; the software also provides full control over beamline hardware and performs the data collection. PMID:26722135