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 |
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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 | |
State | Published - 21 Dec 2018 |
Externally published | Yes |
Bibliographical note
Publisher Copyright:© 2017
Funding
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. 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.
Funders | Funder number |
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National Institutes of Health | R01 GM095904 |
University of California, Los Angeles |
Keywords
- Fluorescence
- Optical crosstalk
- SPAD array
- Single-molecule
- Spectroscopy