16-Ch time-resolved single-molecule spectroscopy using line excitation

Antonino Ingargiola; Pietro Peronio; Eitan Lerner; Angelo Gulinatti; Ivan Rech; Massimo Ghioni; Shimon Weiss; Xavier Michalet

doi:10.1117/12.2256367

16-Ch time-resolved single-molecule spectroscopy using line excitation

Antonino Ingargiola, Pietro Peronio, Eitan Lerner, Angelo Gulinatti, Ivan Rech, Massimo Ghioni, Shimon Weiss, Xavier Michalet

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution › peer-review

2 Scopus citations

Abstract

Single-molecule spectroscopy on freely-diffusing molecules allows detecting conformational changes of biomolecules without perturbation from surface immobilization. Resolving fluorescence lifetimes increases the sensitivity in detecting conformational changes and overcomes artifacts common in intensity-based measurements. Common to all freely-diffusing techniques, however, are the long acquisition times. We report a time-resolved multispot system employing a 16-channel SPAD array and TCSPC electronics, which overcomes the throughput issue. Excitation is obtained by shaping a 532 nm pulsed laser into a line, matching the linear SPAD array geometry. We show that the line-excitation is a robust and cost-effective approach to implement multispot systems based on linear detector arrays.

Original language	English
Title of host publication	Single Molecule Spectroscopy and Superresolution Imaging X
Editors	Zygmunt Karol Gryczynski, Jorg Enderlein, Ingo Gregor, Felix Koberling, Rainer Erdmann, Zygmunt Karol Gryczynski
Publisher	SPIE
ISBN (Electronic)	9781510605831
DOIs	https://doi.org/10.1117/12.2256367
State	Published - 28 Jan 2017
Externally published	Yes
Event	Single Molecule Spectroscopy and Superresolution Imaging X - San Francisco, United States Duration: 28 Jan 2017 → 29 Jan 2017

Publication series

Name	Progress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume	10071
ISSN (Print)	1605-7422

Conference

Conference	Single Molecule Spectroscopy and Superresolution Imaging X
Country/Territory	United States
City	San Francisco
Period	28/01/17 → 29/01/17

Bibliographical note

Publisher Copyright:
© 2017 SPIE.

Funding

This work was supported by NIH grants R01 GM095904 & R01 GM069709 and NSF grant MCB 1244175.

Funders	Funder number
National Science Foundation	MCB 1244175
National Institutes of Health	R01 GM069709, R01 GM095904

Keywords

Fluorescence
High-throughput
SPAD array
Single-molecule
TCSPC

Access to Document

10.1117/12.2256367

Cite this

Ingargiola, A., Peronio, P., Lerner, E., Gulinatti, A., Rech, I., Ghioni, M., Weiss, S., & Michalet, X. (2017). 16-Ch time-resolved single-molecule spectroscopy using line excitation. In Z. K. Gryczynski, J. Enderlein, I. Gregor, F. Koberling, R. Erdmann, & Z. K. Gryczynski (Eds.), Single Molecule Spectroscopy and Superresolution Imaging X Article 100710Q (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 10071). SPIE. https://doi.org/10.1117/12.2256367

Ingargiola, Antonino ; Peronio, Pietro ; Lerner, Eitan et al. / 16-Ch time-resolved single-molecule spectroscopy using line excitation. Single Molecule Spectroscopy and Superresolution Imaging X. editor / Zygmunt Karol Gryczynski ; Jorg Enderlein ; Ingo Gregor ; Felix Koberling ; Rainer Erdmann ; Zygmunt Karol Gryczynski. SPIE, 2017. (Progress in Biomedical Optics and Imaging - Proceedings of SPIE).

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abstract = "Single-molecule spectroscopy on freely-diffusing molecules allows detecting conformational changes of biomolecules without perturbation from surface immobilization. Resolving fluorescence lifetimes increases the sensitivity in detecting conformational changes and overcomes artifacts common in intensity-based measurements. Common to all freely-diffusing techniques, however, are the long acquisition times. We report a time-resolved multispot system employing a 16-channel SPAD array and TCSPC electronics, which overcomes the throughput issue. Excitation is obtained by shaping a 532 nm pulsed laser into a line, matching the linear SPAD array geometry. We show that the line-excitation is a robust and cost-effective approach to implement multispot systems based on linear detector arrays.",

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author = "Antonino Ingargiola and Pietro Peronio and Eitan Lerner and Angelo Gulinatti and Ivan Rech and Massimo Ghioni and Shimon Weiss and Xavier Michalet",

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doi = "10.1117/12.2256367",

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Ingargiola, A, Peronio, P, Lerner, E, Gulinatti, A, Rech, I, Ghioni, M, Weiss, S & Michalet, X 2017, 16-Ch time-resolved single-molecule spectroscopy using line excitation. in ZK Gryczynski, J Enderlein, I Gregor, F Koberling, R Erdmann & ZK Gryczynski (eds), Single Molecule Spectroscopy and Superresolution Imaging X., 100710Q, Progress in Biomedical Optics and Imaging - Proceedings of SPIE, vol. 10071, SPIE, Single Molecule Spectroscopy and Superresolution Imaging X, San Francisco, United States, 28/01/17. https://doi.org/10.1117/12.2256367

16-Ch time-resolved single-molecule spectroscopy using line excitation. / Ingargiola, Antonino; Peronio, Pietro; Lerner, Eitan et al.
Single Molecule Spectroscopy and Superresolution Imaging X. ed. / Zygmunt Karol Gryczynski; Jorg Enderlein; Ingo Gregor; Felix Koberling; Rainer Erdmann; Zygmunt Karol Gryczynski. SPIE, 2017. 100710Q (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 10071).

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution › peer-review

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AU - Ingargiola, Antonino

AU - Peronio, Pietro

AU - Lerner, Eitan

AU - Gulinatti, Angelo

AU - Rech, Ivan

AU - Ghioni, Massimo

AU - Weiss, Shimon

AU - Michalet, Xavier

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AB - Single-molecule spectroscopy on freely-diffusing molecules allows detecting conformational changes of biomolecules without perturbation from surface immobilization. Resolving fluorescence lifetimes increases the sensitivity in detecting conformational changes and overcomes artifacts common in intensity-based measurements. Common to all freely-diffusing techniques, however, are the long acquisition times. We report a time-resolved multispot system employing a 16-channel SPAD array and TCSPC electronics, which overcomes the throughput issue. Excitation is obtained by shaping a 532 nm pulsed laser into a line, matching the linear SPAD array geometry. We show that the line-excitation is a robust and cost-effective approach to implement multispot systems based on linear detector arrays.

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Ingargiola A, Peronio P, Lerner E, Gulinatti A, Rech I, Ghioni M et al. 16-Ch time-resolved single-molecule spectroscopy using line excitation. In Gryczynski ZK, Enderlein J, Gregor I, Koberling F, Erdmann R, Gryczynski ZK, editors, Single Molecule Spectroscopy and Superresolution Imaging X. SPIE. 2017. 100710Q. (Progress in Biomedical Optics and Imaging - Proceedings of SPIE). doi: 10.1117/12.2256367

16-Ch time-resolved single-molecule spectroscopy using line excitation

Abstract

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Bibliographical note

Funding

Keywords

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