The performance of 2D array detectors for light sheet based fluorescence correlation spectroscopy

Singh, Annette; Krieger, Jan; Buchholz, Jan; Charbon, Edoardo; Langowski, Jörg; Wohland, Thorsten

doi:10.1364/oe.21.008652

Cited by 67 publications

(74 citation statements)

References 67 publications

(52 reference statements)

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“…This SPAD array was successfully used in wide-field fluorescence-lifetime imaging (FLIM) [12] experiments in the blue/green wavelength range [109]. A 32×32 SPAD array [107] was used in combination with a custom selective plane illumination microscope (SPIM) [111] for performing FCS measurements [112][113]. However, no true single-molecule experiments have been reported so far with these advanced SPAD array detectors, likely due in part to their limited sensitivity.…”

Section: Available Spad Technologiesmentioning

confidence: 99%

Silicon Photon-Counting Avalanche Diodes for Single-Molecule Fluorescence Spectroscopy

Michalet

Ingargiola

Colyer

et al. 2014

IEEE J. Select. Topics Quantum Electron.

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Solution-based single-molecule fluorescence spectroscopy is a powerful experimental tool with applications in cell biology, biochemistry and biophysics. The basic feature of this technique is to excite and collect light from a very small volume and work in a low concentration regime resulting in rare burst-like events corresponding to the transit of a single molecule. Detecting photon bursts is a challenging task: the small number of emitted photons in each burst calls for high detector sensitivity. Bursts are very brief, requiring detectors with fast response time and capable of sustaining high count rates. Finally, many bursts need to be accumulated to achieve proper statistical accuracy, resulting in long measurement time unless parallelization strategies are implemented to speed up data acquisition. In this paper we will show that silicon single-photon avalanche diodes (SPADs) best meet the needs of single-molecule detection. We will review the key SPAD parameters and highlight the issues to be addressed in their design, fabrication and operation. After surveying the state-of-the-art SPAD technologies, we will describe our recent progress towards increasing the throughput of single-molecule fluorescence spectroscopy in solution using parallel arrays of SPADs. The potential of this approach is illustrated with single-molecule Förster resonance energy transfer measurements.

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Section: Available Spad Technologiesmentioning

confidence: 99%

Silicon Photon-Counting Avalanche Diodes for Single-Molecule Fluorescence Spectroscopy

Michalet

Ingargiola

Colyer

et al. 2014

IEEE J. Select. Topics Quantum Electron.

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“…With a large number of pixels, each of them capable of detecting single photons and measuring their arrival time, it is possible to explore phenomena which would be otherwise difficult to study. Among the different approaches that can take advantage of this capability, time-correlated single-photon counting (TCSPC) techniques, which is used for fluorescence lifetime imaging microscopy (FLIM), figure prominently and have been demonstrated using SPAD-based imaging systems [7,8].…”

Section: Introductionmentioning

confidence: 99%

Architecture and applications of a high resolution gated SPAD image sensor

Burri¹,

Maruyama²,

Michalet³

et al. 2014

Opt. Express

Self Cite

104

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Abstract:We present the architecture and three applications of the largest resolution image sensor based on single-photon avalanche diodes (SPADs) published to date. The sensor, fabricated in a high-voltage CMOS process, has a resolution of 512 x 128 pixels and a pitch of 24 μm. The fill-factor of 5% can be increased to 30% with the use of microlenses. For precise control of the exposure and for time-resolved imaging, we use fast global gating signals to define exposure windows as small as 4 ns. The uniformity of the gate edges location is~140 ps (FWHM) over the whole array, while in-pixel digital counting enables frame rates as high as 156 kfps. Currently, our camera is used as a highly sensitive sensor with high temporal resolution, for applications ranging from fluorescence lifetime measurements to fluorescence correlation spectroscopy and generation of true random numbers.

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“…As for SPT applications, the orthogonal illumination used in LSs produces ideal imaging conditions, in terms of signal contrast and SNR, required for measuring the mobility and reaction parameters of molecules inside living cells by FCS measurements in 3D samples [191,192]. Interaction between proteins can also be studied by extending the FCS methodology to fluorescence cross-correlation spectroscopy using two colors acquired simultaneously [193].…”

Section: 2c Wavefront Codingmentioning

confidence: 99%

Light-sheet microscopy: a tutorial

et al. 2018

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This paper is intended to give a comprehensive review of light-sheet (LS) microscopy from an optics perspective. As such, emphasis is placed on the advantages that LS microscope configurations present, given the degree of freedom gained by uncoupling the excitation and detection arms. The new imaging properties are first highlighted in terms of optical parameters and how these have enabled several biomedical applications. Then, the basics are presented for understanding how a LS microscope works. This is followed by a presentation of a tutorial for LS microscope designs, each working at different resolutions and for different applications. Then, based on a numerical Fourier analysis and given the multiple possibilities for generating the LS in the microscope (using Gaussian, Bessel, and Airy beams in the linear and nonlinear regimes), a systematic comparison of their optical performance is presented. Finally, based on advances in optics and photonics, the novel optical implementations possible in a LS microscope are highlighted.

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The performance of 2D array detectors for light sheet based fluorescence correlation spectroscopy

Cited by 67 publications

References 67 publications

Silicon Photon-Counting Avalanche Diodes for Single-Molecule Fluorescence Spectroscopy

Silicon Photon-Counting Avalanche Diodes for Single-Molecule Fluorescence Spectroscopy

Architecture and applications of a high resolution gated SPAD image sensor

Light-sheet microscopy: a tutorial

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