Development of a full field-of-view spectroscopic single-molecule fluorescence instrument with improved methods for calibration

We have developed a high-throughput, full field-of-view (FOV) spectroscopic single-molecule fluorescence instrument, overcoming distortion issues with gratings and prisms. Our calibration techniques ensure spectral uniformity across the whole FOV, and our two-step Gaussian fit method enables better...

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Main Authors: Deqi Yi, Shangyuan Deng, Chuang Tan, Jie Ma
Format: Article
Language:English
Published: IOP Publishing 2025-01-01
Series:Applied Physics Express
Subjects:
Online Access:https://doi.org/10.35848/1882-0786/ada10b
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author Deqi Yi
Shangyuan Deng
Chuang Tan
Jie Ma
author_facet Deqi Yi
Shangyuan Deng
Chuang Tan
Jie Ma
author_sort Deqi Yi
collection DOAJ
description We have developed a high-throughput, full field-of-view (FOV) spectroscopic single-molecule fluorescence instrument, overcoming distortion issues with gratings and prisms. Our calibration techniques ensure spectral uniformity across the whole FOV, and our two-step Gaussian fit method enables better than 2 nm precision in spectral peak determination for accurate molecule identification, making it a valuable tool for multi-color single-molecule detection and analysis.
format Article
id doaj-art-da81ef1794694ca5ad3770a7f6171591
institution Kabale University
issn 1882-0786
language English
publishDate 2025-01-01
publisher IOP Publishing
record_format Article
series Applied Physics Express
spelling doaj-art-da81ef1794694ca5ad3770a7f61715912025-01-14T16:11:13ZengIOP PublishingApplied Physics Express1882-07862025-01-0118101200310.35848/1882-0786/ada10bDevelopment of a full field-of-view spectroscopic single-molecule fluorescence instrument with improved methods for calibrationDeqi Yi0Shangyuan Deng1Chuang Tan2Jie Ma3https://orcid.org/0000-0002-7570-9554School of Physics, Sun Yat-sen University , Guangzhou 510275, People’s Republic of China; State Key Laboratory of Optoelectronic Materials and Technologies, Sun Yat-sen University , Guangzhou 510275, People’s Republic of ChinaSchool of Physics, Sun Yat-sen University , Guangzhou 510275, People’s Republic of China; State Key Laboratory of Optoelectronic Materials and Technologies, Sun Yat-sen University , Guangzhou 510275, People’s Republic of ChinaSchool of Physics, Sun Yat-sen University , Guangzhou 510275, People’s Republic of China; State Key Laboratory of Optoelectronic Materials and Technologies, Sun Yat-sen University , Guangzhou 510275, People’s Republic of ChinaSchool of Physics, Sun Yat-sen University , Guangzhou 510275, People’s Republic of China; State Key Laboratory of Optoelectronic Materials and Technologies, Sun Yat-sen University , Guangzhou 510275, People’s Republic of ChinaWe have developed a high-throughput, full field-of-view (FOV) spectroscopic single-molecule fluorescence instrument, overcoming distortion issues with gratings and prisms. Our calibration techniques ensure spectral uniformity across the whole FOV, and our two-step Gaussian fit method enables better than 2 nm precision in spectral peak determination for accurate molecule identification, making it a valuable tool for multi-color single-molecule detection and analysis.https://doi.org/10.35848/1882-0786/ada10bsingle-molecule fluorescence spectroscopyspectrometer calibrationdistortion correctionsingle-molecule spectral analysis
spellingShingle Deqi Yi
Shangyuan Deng
Chuang Tan
Jie Ma
Development of a full field-of-view spectroscopic single-molecule fluorescence instrument with improved methods for calibration
Applied Physics Express
single-molecule fluorescence spectroscopy
spectrometer calibration
distortion correction
single-molecule spectral analysis
title Development of a full field-of-view spectroscopic single-molecule fluorescence instrument with improved methods for calibration
title_full Development of a full field-of-view spectroscopic single-molecule fluorescence instrument with improved methods for calibration
title_fullStr Development of a full field-of-view spectroscopic single-molecule fluorescence instrument with improved methods for calibration
title_full_unstemmed Development of a full field-of-view spectroscopic single-molecule fluorescence instrument with improved methods for calibration
title_short Development of a full field-of-view spectroscopic single-molecule fluorescence instrument with improved methods for calibration
title_sort development of a full field of view spectroscopic single molecule fluorescence instrument with improved methods for calibration
topic single-molecule fluorescence spectroscopy
spectrometer calibration
distortion correction
single-molecule spectral analysis
url https://doi.org/10.35848/1882-0786/ada10b
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AT shangyuandeng developmentofafullfieldofviewspectroscopicsinglemoleculefluorescenceinstrumentwithimprovedmethodsforcalibration
AT chuangtan developmentofafullfieldofviewspectroscopicsinglemoleculefluorescenceinstrumentwithimprovedmethodsforcalibration
AT jiema developmentofafullfieldofviewspectroscopicsinglemoleculefluorescenceinstrumentwithimprovedmethodsforcalibration