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At least 19 records

Data for "In vitro and in vivo NIR Fluorescence Lifetime Imaging with a time-gated SPAD camera"

Near-infrared (NIR) fluorescence lifetime imaging (FLI) provides a unique contrast mechanism to monitor biological parameters and molecular events in vivo. Single-photon avalanche diode (SPAD) cameras have been recently demonstrated in FLI microscopy (FLIM) applications, but their suitability for in vivo macroscopic FLI (MFLI) in deep tissues remains to be demonstrated. Herein, we report in vivo NIR MFLI measurement with SwissSPAD2, a large time-gated SPAD camera. We first benchmark its performance in well-controlled in vitro experiments, ranging from monitoring environmental effects on fluorescence lifetime, to quantifying Förster resonant energy transfer (FRET) between dyes. Next, we use it for in vivo studies of target-drug engagement in live and intact tumor xenografts using FRET. Information obtained with SwissSPAD2 was successfully compared to that obtained with a gated intensified charge-coupled device (ICCD) camera, using two different approaches. Our results demonstrate that SPAD cameras offer a powerful technology for in vivo preclinical applications in the NIR window.

2DG↗

Coupling a recurrent neural network to SPAD TCSPC systems for real-time fluorescence lifetime imaging

Fluorescence lifetime imaging (FLI) has been receiving increased attention in recent years as a powerful diagnostic technique in biological and medical research. However, existing FLI systems often suffer from a tradeoff between processing speed, accuracy, and robustness. Inspired by the concept of Edge Artificial Intelligence (Edge AI), we propose a robust approach that enables fast FLI with no degradation of accuracy. This approach couples a recurrent neural network (RNN), which is trained to estimate the fluorescence lifetime directly from raw timestamps without building histograms, to SPAD TCSPC systems, thereby drastically reducing transfer data volumes and hardware resource utilization, and enabling real-time FLI acquisition. We train two variants of the RNN on a synthetic dataset and compare the results to those obtained using center-of-mass method (CMM) and least squares fitting (LS fitting). Results demonstrate that two RNN variants, gated recurrent unit (GRU) and long short-term memory (LSTM), are comparable to CMM and LS fitting in terms of accuracy, while outperforming them in the presence of background noise by a large margin. To explore the ultimate limits of the approach, we derive the Cramer-Rao lower bound of the measurement, showing that RNN yields lifetime estimations with near-optimal precision. To demonstrate real-time operation, we build a FLI microscope based on an existing SPAD TCSPC system comprising a 32 x 32 SPAD sensor named Piccolo. Four quantized GRU cores, capable of processing up to 4 million photons per second, are deployed on the Xilinx Kintex-7 FPGA that controls the Piccolo. Powered by the GRU, the FLI setup can retrieve real-time fluorescence lifetime images at up to 10 frames per second. The proposed FLI system is promising and ideally suited for biomedical applications, including biological imaging, biomedical diagnostics, and fluorescence-assisted surgery, etc.

47 OTHER INSTRUMENTATION↗

In vitro and in vivo NIR fluorescence lifetime imaging with a time-gated SPAD camera

Near-infrared (NIR) fluorescence lifetime imaging (FLI) provides a unique contrast mechanism to monitor biological parameters and molecular events in vivo . Single-photon avalanche diode (SPAD) cameras have been recently demonstrated in FLI microscopy (FLIM) applications, but their suitability for in vivo macroscopic FLI (MFLI) in deep tissues remains to be demonstrated. Herein, we report in vivo NIR MFLI measurement with SwissSPAD2, a large time-gated SPAD camera. We first benchmark its performance in well-controlled in vitro experiments, ranging from monitoring environmental effects on fluorescence lifetime, to quantifying Förster resonant energy transfer (FRET) between dyes. Next, we use it for in vivo studies of target-drug engagement in live and intact tumor xenografts using FRET. Information obtained with SwissSPAD2 was successfully compared to that obtained with a gated intensified charge-coupled device (ICCD) camera, using two different approaches. Our results demonstrate that SPAD cameras offer a powerful technology for in vivo preclinical applications in the NIR window.

47 OTHER INSTRUMENTATION↗

Tracking capabilities of SPADs for laser ranging

The spatial sensitivity of Single-Photon Avalanche Diodes (SPADs) can be exploited in laser ranging measurements to finely tune the laser spot in the center of the detector sensitive area. We report the performance of a SPAD with l00 micron diameter. It features a time resolution better than 80 ps rms when operated 4V above V(b) at minus 30 C, and a spatial sensitivity better than 20 microns to radial displacements of the laser spot. New SPAD structures with auxiliary delay detectors are proposed. These improved devices could allow a two dimensional sensitivity, that could be employed for the design of pointing servos.

Zappa, F.↗

NIR fluorescence lifetime macroscopic imaging with a time-gated SPAD camera

The performance of SwissSPAD2 (SS2), a large scale, widefield time-gated CMOS SPAD imager developed for fluorescence lifetime imaging, has recently been described in the context of visible range and fluorescence lifetime imaging microscopy (FLIM) of dyes with lifetimes in the 2.5 – 4 ns range. Here, we explore its capabilities in the NIR regime relevant for small animal imaging, where its sensitivity is lower and typical NIR fluorescent dye lifetimes are much shorter (1 ns or less). We carry out this study in a simple macroscopic imaging setup based on a compact NIR picosecond pulsed laser, an engineered diffuser-based illumination optics, and NIR optimized imaging lens suitable for well-plate or small animal imaging. Because laser repetition rates can vary between models, but the synchronization signal frequency accepted by SS2 is fixed to 20 MHz, we first checked that a simple frequency-division scheme enables data recording for different laser repetition rates. Next, we acquired data using different time gate widths, including gates with duration longer than the laser period, and analyzed the resulting data using both standard nonlinear least-square fit (NLSF) and phasor analysis. We show that the fixed synchronization rate and large gate widths characterizing SS2 (10 ns and over) are not an obstacle to accurately extracting lifetime in the 1 ns range and to distinguishing between close lifetimes. As a result, SS2 and similar very large gated SPAD imagers appear as a versatile alternative to other widefield time-resolved detectors for NIR fluorescence lifetime imaging, including preclinical molecular applications.

FLIM↗

How to squeeze high quantum efficiency and high time resolution out of a SPAD

We address the issue whether Single-Photon Avalanche Diodes (SPADs) can be suitably designed to achieve a trade-off between quantum efficiency and time resolution performance. We briefly recall the physical mechanisms setting the time resolution of avalanche photodiodes operated in single-photon counting, and we give some criteria for the design of SPADs with a quantum efficiency better than l0 percent at 1064 nm together with a time resolution below 50 ps rms.

Lacaita, A.↗

NIR fluorescence lifetime macroscopic imaging with a novel time-gated SPAD camera

SwissSPAD3 is the latest of a family of widefield time-gated SPAD imagers developed for fluorescence lifetime imaging (FLI) applications. Its distinctive features are (i) the ability to define shorter gates than its predecessors (width W < 1 ns), (ii) support for laser repetition rates up to at least 80 MHz and (iii) a dual-gate architecture providing an effective duty cycle of 100%. We present widefield macroscopic FLI measurements of short lifetime NIR dyes, analyzed using the phasor approach. Furthermore, the results are compared with those previously obtained with SwissSPAD2 and to theoretical predictions.

47 OTHER INSTRUMENTATION↗

The Use of a Chlorophyll Meter (SPAD-502) for Field Determinations of Red Mangrove (Rhizophora Mangle L.) Leaf Chlorophyll Amount

The red mangrove Rhizophora mangle L., is a halophytic woody spermatophyte common to the land-sea interface of tropical and subtropical intertidal zones. It has been reported that 60 to 75% of the coastline of the earth's tropical regions are lined with mangroves. Mangroves help prevent shoreline erosion, provide breeding, nesting and feeding areas for many marine animals and birds. Mangroves are important contributors of primary production in the coastal environment, and this is largely proportional to the standing crop of leaf chlorophylls. Higher intensities of ultraviolet radiation, resulting from stratospheric ozone depletion, can lead to a reduction of chlorophyll in terrestrial plants. Since the most common method for determining chlorophyll concentration is by extraction and this is labor intensive and time consuming, few studies on photosynthetic pigments of mangroves have been reported. Chlorophyll meter readings have been related to leaf chlorophyll content in apples and maples. It has also been correlated to nitrogen status in corn and cotton. Peterson et al., (1993) used a chlorophyll meter to detect nitrogen deficiency in crops and in determining the need for additional nitrogen fertilizer. Efforts to correlate chlorophyll meter measurements to chlorophyll content of mangroves have not been reported. This paper describes the use of a hand-held chlorophyll meter (Minolta SPAD-502) to determine the amount of red mangrove foliar chlorophyll present in the field.

Connelly, Xana M.↗

Two wavelength satellite laser ranging using SPAD

When ranging to satellites with lasers, there are several principal contributions to the error budget: from the laser ranging system on the ground, from the satellite retroarray geometry, and from the atmosphere. Using a single wavelength, we have routinely achieved a ranging precision of 8 millimeters when ranging to the ERS-1 and Starlette satellites. The systematic error of the atmosphere, assuming the existing dispersion models, is expected to be of the order of 1 cm. Multiple wavelengths ranging might contribute to the refinement of the existing models. Taking into account the energy balance, the existing picosecond lasers and the existing receiver and detection technology, several pairs or multiple wavelengths may be considered. To be able to improve the atmospheric models to the subcentimeter accuracy level, the differential time interval (DTI) has to be determined within a few picoseconds depending on the selected wavelength pair. There exist several projects based on picosecond lasers as transmitters and on two types of detection techniques: one is based on photodetectors, like photomultipliers or photodiodes connected to the time interval meters. Another technique is based on the use of a streak camera as an echo signal detector, temporal analyzer, and time interval vernier. The temporal analysis at a single wavelength using the streak camera showed the complexity of the problem.

Prochazka, Ivan↗

Photon-to-Digital Converters for Neutron Imaging Instruments (Final Report)

Oak Ridge National Laboratory (ORNL) collaborated with the Université de Sherbrooke (UdeS) to design a modular and scalable tile of innovative digital photodetectors called Photon-to-Digital Converters (PDC) and the required subsystems to demonstrate the feasibility of large area single-photon detectors to support high-resolution, depth-of-interaction, fast neutron radiography based on scintillation detectors. The goal is to demonstrate the advantages of using PDC over conventional Silicon Photomultiplier (SiPM) for neutron imaging systems. The basic building blocks of analog SiPM and PDC are Single-Photon Avalanche Photodiodes (SPAD). The main difference between conventional SiPMs and PDCs, is in the fact that analog SiPMs sum the charge produced by individual SPAD passively, while in PDCs each SPAD is read out individually by an active electronic circuit. Hence, PDCs provide a direct photon-to-bit conversion where a logic “1” means that there was a detection in a given SPAD. Conversely, an analog SiPM requires a sophisticated preamplifier (current amplifiers or transimpedance amplifier) followed by a shaping amplifier and an analog-to-digital converter. Moreover, since the charge from each SPAD varies slightly, the passive sum of the analog SiPM will have signal fluctuations for the same amount of photon detected. This is one of many issues that is completely eliminated by individual one-to-one SPAD read out in PDCs. The large area required for scintillator readouts for neutron imaging is such that the output capacitance of a large area of SiPM arrays is very high. This creates a burden on the signal-to-noise optimization with respect to the power budget. For PDC, the power consumption is dictated by rate of the incident photon flux, or in photon-starved environments, the dark noise rate of the SPAD array. In other words, the power consumption in the absence of events is extremely low. Hence, the power consumption of a PDC read out system is much lower than its analog counterpart. Further, the large SiPM capacitance may introduce signal distortions on the scintillator fast rise or decay time with undesired effects on timing and pulse shape discrimination. The problem is absent in PDCs, as their readout is independent on device capacitance. UdeS is the world leader in 3D vertical integration of frontside illuminated SPAD arrays over CMOS readouts. These capabilities allow future production of “3D PDC” with maximum photosensitive fill factor and electronic functionalities tailored to the application. UdeS' vast knowledge of SPADs and CMOS design made the team an ideal collaborator in the development of large area high-resolution fast neutron radiography.

42 ENGINEERING↗

Radiation Tests of Single Photon Avalanche Diode for Space Applications

Single photon avalanche diodes (SPADs) have been recently studied as photodetectors for applications in space missions. In this presentation we report the results of radiation hardness test on large area SPAD (actual results refer to SPADs having 500 micron diameter). Dark counts rate as low as few kHz at −10 degC has been obtained for the 500 micron devices, before irradiation. We performed bulk damage and total dose radiation tests with protons and gamma-rays in order to evaluate their radiation hardness properties and their suitability for application in a Low Earth Orbit (LEO) space mission. With this aim SPAD devices have been irradiated using up to 20 krad total dose with gamma-rays and 5 krad with protons. The test performed show that large area SPADs are very sensitive to proton doses as low as 2×10(exp 8) (1 MeV eq) n/cm2 with a significant increase in dark counts rate (DCR) as well as in the manifestation of the "random telegraph signal" effect. Annealing studies at room temperature (RT) and at 80 degC have been carried out, showing a high decrease of DCR after 24-48 h at RT. Lower protons doses in the range 1-10×10(exp 7) (1 MeV eq) n/cm(exp 2) result in a lower increase of DCR suggesting that the large-area SPADs tested in this study are well suitable for application in low-inclination LEO, particularly useful for gamma-ray astrophysics.

radiation hardness↗

Scintillation event imaging with a single photon avalanche diode camera

Position and time measurements of scintillation events encode information about the radiation source. Single photon avalanche diode (SPAD) arrays offer multiple-megapixel spatial resolution and tens of picoseconds temporal resolution for detecting single photons. Current lensless designs for measuring scintillation events use sensors that are lower in spatial resolution. Camera-based designs use sensors that are lower in temporal resolution or readout rate and cannot image individual interactions. Here we propose to image scintillation events in a thick, monolithic scintillator using a high-resolution SPAD camera. We demonstrate that a commercial SPAD camera is able to gather sufficient signal to image individual scintillation events and observe 3D shifts in their spatial distribution. Simulations show that a SPAD camera can localize individual scintillation events in 3D. We report direct imaging of gamma-ray interactions in a scintillator with a SPAD camera. The proposed design may allow to measure complex signatures of individual particles interacting in the scintillator.

47 OTHER INSTRUMENTATION↗

InGaAsP Avalanche Photodetectors for Non-Gated 1.06 micrometer Photon-Counting Receivers

The efficient detection of single photons at 1.06 micron is of considerable interest for lidar/ladar systems designed for remote sensing an d ranging as well as for free-space optical transmission in photon-st arved applications. However, silicon-based single photon avalanche diodes (SPADs) used at shorter wavelengths have very low single photon d etection efficiency (approximately 1 - 2%) at 1.06 micron, and InP/In GaAs SPADs designed for telecommunications wavelengths near 1.5 micro n exhibit high dark count rates that generally inhibit non-gated (free-running) operation. To bridge this "single photon detection gap" for wavelengths just beyond 1 micron, we have developed high performance , large area (80 - 200 micron diameter) InP-based InGaAsP quaternary absorber SPADs optimized for operation at 1.06 micron and based on a highly reliable planar geometry avalanche photodiode structure. We wil l show that dark count rates are sufficiently low to allow for non-ga ted operation while achieving detection efficiencies far surpassing t hose found for Si SPADs. At a detection efficiency of 10%, 80 micron diameter devices exhibit dark count rates below 1000 Hz and count rate s of at least 3 MHz when operated at -40 C. Significantly higher dete ction efficiencies (30 - 50%) are achievable with acceptable tradeoff s in dark count rate. In this paper, we will also discuss performance modeling for these devices and compare their behavior with longer wav elength InP-based InGaAs ternary absorber SPADs fabricated on a relat ed device design platform.

Itzler, Mark A.↗

User's operating procedures. Volume 1: Scout project information programs

A review of the user's operating procedures for the Scout Project Automatic Data System, called SPADS is given. SPADS is the result of the past seven years of software development on a Prime minicomputer located at the Scout Project Office. SPADS was developed as a single entry, multiple cross reference data management and information retrieval system for the automation of Project office tasks, including engineering, financial, managerial, and clerical support. The instructions to operate the Scout Project Information programs in data retrieval and file maintenance via the user friendly menu drivers is presented.

Harris, C. G.↗

User's operating procedures. Volume 2: Scout project financial analysis program

A review is presented of the user's operating procedures for the Scout Project Automatic Data system, called SPADS. SPADS is the result of the past seven years of software development on a Prime mini-computer located at the Scout Project Office, NASA Langley Research Center, Hampton, Virginia. SPADS was developed as a single entry, multiple cross-reference data management and information retrieval system for the automation of Project office tasks, including engineering, financial, managerial, and clerical support. This volume, two (2) of three (3), provides the instructions to operate the Scout Project Financial Analysis program in data retrieval and file maintenance via the user friendly menu drivers.

Harris, C. G.↗