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

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

Keywords

Fluorescence
Optical crosstalk
SPAD array
Single-molecule
Spectroscopy

All Science Journal Classification (ASJC) codes

Nuclear and High Energy Physics
Instrumentation

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10.1016/j.nima.2017.11.070

Cite this

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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title = "Optical crosstalk in SPAD arrays for high-throughput single-molecule fluorescence spectroscopy",

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.",

keywords = "Fluorescence, Optical crosstalk, SPAD array, Single-molecule, Spectroscopy",

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

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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

TY - JOUR

T1 - Optical crosstalk in SPAD arrays for high-throughput single-molecule fluorescence spectroscopy

AU - Ingargiola, Antonino

AU - Segal, Maya

AU - Gulinatti, Angelo

AU - Rech, Ivan

AU - Labanca, Ivan

AU - Maccagnani, Piera

AU - Ghioni, Massimo

AU - Weiss, Shimon

AU - Michalet, Xavier

PY - 2018/12/21

Y1 - 2018/12/21

N2 - 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.

AB - 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.

KW - Fluorescence

KW - Optical crosstalk

KW - SPAD array

KW - Single-molecule

KW - Spectroscopy

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U2 - 10.1016/j.nima.2017.11.070

DO - 10.1016/j.nima.2017.11.070

M3 - مقالة

C2 - 31223178

SN - 0168-9002

VL - 912

SP - 255

EP - 258

JO - Nuclear Instruments and Methods in Physics Research, Section A: Accelerators, Spectrometers, Detectors and Associated Equipment

JF - Nuclear Instruments and Methods in Physics Research, Section A: Accelerators, Spectrometers, Detectors and Associated Equipment

ER -

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

Abstract

Keywords

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