A Single-Photon Avalanche Camera for Fluorescence Lifetime Imaging Microscopy and Correlation Spectroscopy

Vitali, Marco; Bronzi, Danilo; Krmpot, Aleksandar J.; Nikolić, Stanko N.; Schmitt, Franz Josef; Junghans, Cornelia; Tisa, Simone; Friedrich, Thomas; Vukojević, Vladana; Terenius, Lars; Zappa, Franco; Rigler, Rudolf

doi:10.1109/jstqe.2014.2333238

Cited by 47 publications

(45 citation statements)

References 41 publications

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“…Vitali et al also used a multibeam approach with a 32×32 array (square pixels of 100 µm with a 20 µm circular SPAD and 8-bit counters) and measured FLIM of eGFP in living HEK293-T cells [87]. The SPAD sensor was implemented in a standard CMOS process and included time-gating with minimum gate width of 1.5 ns and delay steps below 100 ps.…”

Section: Multibeam Flimmentioning

confidence: 99%

Single-photon avalanche diode imagers in biophotonics: review and outlook

et al. 2019

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Single-photon avalanche diode (SPAD) arrays are solid-state detectors that offer imaging capabilities at the level of individual photons, with unparalleled photon counting and time-resolved performance. This fascinating technology has progressed at a very fast pace in the past 15 years, since its inception in standard CMOS technology in 2003. A host of architectures have been investigated, ranging from simpler implementations, based solely on off-chip data processing, to progressively “smarter” sensors including on-chip, or even pixel level, time-stamping and processing capabilities. As the technology has matured, a range of biophotonics applications have been explored, including (endoscopic) FLIM, (multibeam multiphoton) FLIM-FRET, SPIM-FCS, super-resolution microscopy, time-resolved Raman spectroscopy, NIROT and PET. We will review some representative sensors and their corresponding applications, including the most relevant challenges faced by chip designers and end-users. Finally, we will provide an outlook on the future of this fascinating technology.

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Section: Multibeam Flimmentioning

confidence: 99%

Single-photon avalanche diode imagers in biophotonics: review and outlook

et al. 2019

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“…1 High detection efficiency, insensitivity to magnetic fields, low cost, compactness, and high gain characteristics have led to a growing attention for these detectors in high-energy physics, 2,3 fluorescence lifetime measurement, 4 and fluorescence lifetime imaging microscopy. 5,6 A SiPM detector is usually made up of a large number of parallel-connected single photon avalanche diodes (SPADs) or Geiger-mode avalanche photodiodes (GM-APDs) each one with its integrated quenching resistor. Each SPAD and its quenching resistor are called a microcell.…”

Section: Introductionmentioning

confidence: 99%

A comprehensive circuit model for evaluating the response of silicon photomultipliers in continuous wave light regime

Siminfar

Shoaei

2019

Circuit Theory & Apps

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Summary In this paper, a comprehensive dynamic model for silicon photomultiplier (SiPM) detectors is proposed to investigate their response when exposed to the light of any wavelength and relatively high intensity. Also, a procedure is proposed to perform reliable circuit‐level simulations to predict the response of these detectors under various photon rates. The approach of dynamic model is based on the state of microcells in terms of overvoltage, elapsed recharging time, and their effect on the triggering probability with which the detector current and voltage can be accurately simulated. Using the proposed model, the SiPM behaviour is examined under various continuous illumination power in simulation. The effectiveness of the proposed model and the simulation method are verified by experimental results extracted from two SiPM detectors.

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“…In such an arrangement, it becomes necessary to focus the fluorescence from each spot onto a single SPAD. This was first achieved with a small, fully integrated 2 Â 2 CMOS SPAD array in (12), then extended in (13) to the detection in 8 Â 8 spots of bright 100-nm-diameter fluorescent beads in solutions and subsequently to multifocal FCS measurements of quantum dot diffusion in solution (14)(15)(16) for the measurement of fluorescence decay kinetics. In the latter case, a diffractive-optical-element-based optical setup allowed the generation of 32 Â 32 spots with a pitch of 100 mm and a diameter of 12.5 mm at the image plane.…”

Section: Introductionmentioning

confidence: 99%

Widefield High Frame Rate Single-Photon SPAD Imagers for SPIM-FCS

Buchholz

Krieger

Bruschini

et al. 2018

Biophysical Journal

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Photon-counting sensors based on standard complementary metal-oxide-semiconductor single-photon avalanche diodes (SPADs) represent an emerging class of imagers that enable the counting and/or timing of single photons at zero readout noise (better than high-speed electron-multiplying charge-coupling devices) and over large arrays. They have seen substantial progress over the last 15 years, increasing their spatial resolution, timing accuracy, and sensitivity while reducing spurious signals such as afterpulsing and dark counts. They are increasingly being applied for time-resolved applications with the added advantage of enabling real-time options such as autocorrelation. We report in this study on the use of such a state-of-the-art 512 × 128 SPAD array, capable of a time resolution of 10-10 s for full frames while retaining acceptable photosensitivity thanks to the use of dedicated microlenses, in a selective plane illumination-fluorescence correlation spectroscopy setup. The latter allows us to perform thousands of fluorescence-correlation spectroscopy measurements simultaneously in a two-dimensional slice of the sample. This high-speed SPAD imager enables the measurement of molecular motion of small fluorescent particles such as single chemical dye molecules. Inhomogeneities in the molecular detection efficiency were compensated for by means of a global fit of the auto- and cross-correlation curves, which also made a calibration-free measurement of various samples possible. The afterpulsing effect could also be mitigated, making the measurement of the diffusion of Alexa-488 possible, and the overall result quality was further improved by spatial binning. The particle concentrations in the focus tend to be overestimated by a factor of 1.7 compared to a confocal setup; a calibration is thus required if absolute concentrations need to be measured. The first high-speed selective plane illumination-fluorescence correlation spectroscopy in vivo measurements to our knowledge were also recorded: although two-component fit models could not be employed because of noise, the diffusion of eGFP oligomers in HeLa cells could be measured. Sensitivity and noise will be further improved in the next generation of SPAD-based widefield sensors, which are currently under testing.

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A Single-Photon Avalanche Camera for Fluorescence Lifetime Imaging Microscopy and Correlation Spectroscopy

Cited by 47 publications

References 41 publications

Single-photon avalanche diode imagers in biophotonics: review and outlook

Single-photon avalanche diode imagers in biophotonics: review and outlook

A comprehensive circuit model for evaluating the response of silicon photomultipliers in continuous wave light regime

Widefield High Frame Rate Single-Photon SPAD Imagers for SPIM-FCS

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