Optical crosstalk in SPAD arrays for high-throughput single-molecule fluorescence spectroscopy

Antonino Ingargiola; Maya Segal; Angelo Gulinatti; Ivan Rech; Ivan Labanca; Piera Maccagnani; Massimo Ghioni; Shimon Weiss; Xavier Michalet

doi:10.1016/j.nima.2017.11.070

Optical crosstalk in SPAD arrays for high-throughput single-molecule fluorescence spectroscopy

Antonino Ingargiola, Maya Segal, Angelo Gulinatti, Ivan Rech, Ivan Labanca, Piera Maccagnani, Massimo Ghioni, Shimon Weiss, Xavier Michalet

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

Abstract

Single-molecule fluorescence spectroscopy (SMFS), based on the detection of individual molecules freely diffusing through the excitation spot of a confocal microscope, has allowed unprecedented insights into biological processes at the molecular level, but suffers from limited throughput. We have recently introduced a multispot version of SMFS, which allows achieving high-throughput SMFS by virtue of parallelization, and relies on custom silicon single-photon avalanche diode (SPAD) detector arrays. Here, we examine the premise of this parallelization approach, which is that data acquired from different spots is uncorrelated. In particular, we measure the optical crosstalk characteristics of the two 48-pixel SPAD arrays used in our recent SMFS studies, and demonstrate that it is negligible (crosstalk probability ≤1.110⁻³) and undetectable in cross-correlation analysis of actual single-molecule fluorescence data.

Original language	English
Pages (from-to)	255-258
Number of pages	4
Journal	Nuclear Instruments and Methods in Physics Research, Section A: Accelerators, Spectrometers, Detectors and Associated Equipment
Volume	912
DOIs	https://doi.org/10.1016/j.nima.2017.11.070
State	Published - 21 Dec 2018
Externally published	Yes

Bibliographical note

Funding Information:
This work was funded by NIH Grant R01 GM095904 . S. Weiss discloses intellectual property used in the research reported here. The work at UCLA was conducted in Dr. Weiss’s Laboratory. M. Ghioni discloses equity in Micro Photon Devices S.r.l. (MPD). No resources or personnel from MPD were involved in this work. We are grateful to our colleagues at UCLA and Polimi for their contributions to this project over the years.

Funding Information:
This work was funded by NIH Grant R01 GM095904. S. Weiss discloses intellectual property used in the research reported here. The work at UCLA was conducted in Dr. Weiss's Laboratory. M. Ghioni discloses equity in Micro Photon Devices S.r.l. (MPD). No resources or personnel from MPD were involved in this work. We are grateful to our colleagues at UCLA and Polimi for their contributions to this project over the years.

Publisher Copyright:
© 2017

Keywords

Fluorescence
Optical crosstalk
SPAD array
Single-molecule
Spectroscopy

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10.1016/j.nima.2017.11.070

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Ingargiola, A., Segal, M., Gulinatti, A., Rech, I., Labanca, I., Maccagnani, P., Ghioni, M., Weiss, S., & Michalet, X. (2018). Optical crosstalk in SPAD arrays for high-throughput single-molecule fluorescence spectroscopy. Nuclear Instruments and Methods in Physics Research, Section A: Accelerators, Spectrometers, Detectors and Associated Equipment, 912, 255-258. https://doi.org/10.1016/j.nima.2017.11.070

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abstract = "Single-molecule fluorescence spectroscopy (SMFS), based on the detection of individual molecules freely diffusing through the excitation spot of a confocal microscope, has allowed unprecedented insights into biological processes at the molecular level, but suffers from limited throughput. We have recently introduced a multispot version of SMFS, which allows achieving high-throughput SMFS by virtue of parallelization, and relies on custom silicon single-photon avalanche diode (SPAD) detector arrays. Here, we examine the premise of this parallelization approach, which is that data acquired from different spots is uncorrelated. In particular, we measure the optical crosstalk characteristics of the two 48-pixel SPAD arrays used in our recent SMFS studies, and demonstrate that it is negligible (crosstalk probability ≤1.110−3) and undetectable in cross-correlation analysis of actual single-molecule fluorescence data.",

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Ingargiola, A, Segal, M, Gulinatti, A, Rech, I, Labanca, I, Maccagnani, P, Ghioni, M, Weiss, S & Michalet, X 2018, 'Optical crosstalk in SPAD arrays for high-throughput single-molecule fluorescence spectroscopy', Nuclear Instruments and Methods in Physics Research, Section A: Accelerators, Spectrometers, Detectors and Associated Equipment, vol. 912, pp. 255-258. https://doi.org/10.1016/j.nima.2017.11.070

Optical crosstalk in SPAD arrays for high-throughput single-molecule fluorescence spectroscopy. / Ingargiola, Antonino; Segal, Maya; Gulinatti, Angelo et al.
In: Nuclear Instruments and Methods in Physics Research, Section A: Accelerators, Spectrometers, Detectors and Associated Equipment, Vol. 912, 21.12.2018, p. 255-258.

Research output: Contribution to journal › Article › peer-review

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AU - Gulinatti, Angelo

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AU - Labanca, Ivan

AU - Maccagnani, Piera

AU - Ghioni, Massimo

AU - Weiss, Shimon

AU - Michalet, Xavier

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Optical crosstalk in SPAD arrays for high-throughput single-molecule fluorescence spectroscopy

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Keywords

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