Bessel Beam Illumination Reduces Random and Systematic Errors in Quantitative Functional Studies Using Light-Sheet Microscopy
Light-sheet microscopy (LSM), in combination with intrinsically transparent zebrafish larvae, is a method of choice to observe brain function with high frame rates at cellular resolution. Inherently to LSM, however, residual opaque objects cause stripe artifacts, which obscure features of interest a...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-09-01
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| Series: | Frontiers in Cellular Neuroscience |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fncel.2018.00315/full |
| _version_ | 1811319035033288704 |
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| author | M. Caroline Müllenbroich M. Caroline Müllenbroich Lapo Turrini Ludovico Silvestri Ludovico Silvestri Tommaso Alterini Ali Gheisari Natascia Tiso Francesco Vanzi Francesco Vanzi Leonardo Sacconi Leonardo Sacconi Francesco S. Pavone Francesco S. Pavone Francesco S. Pavone |
| author_facet | M. Caroline Müllenbroich M. Caroline Müllenbroich Lapo Turrini Ludovico Silvestri Ludovico Silvestri Tommaso Alterini Ali Gheisari Natascia Tiso Francesco Vanzi Francesco Vanzi Leonardo Sacconi Leonardo Sacconi Francesco S. Pavone Francesco S. Pavone Francesco S. Pavone |
| author_sort | M. Caroline Müllenbroich |
| collection | DOAJ |
| description | Light-sheet microscopy (LSM), in combination with intrinsically transparent zebrafish larvae, is a method of choice to observe brain function with high frame rates at cellular resolution. Inherently to LSM, however, residual opaque objects cause stripe artifacts, which obscure features of interest and, during functional imaging, modulate fluorescence variations related to neuronal activity. Here, we report how Bessel beams reduce streaking artifacts and produce high-fidelity quantitative data demonstrating a fivefold increase in sensitivity to calcium transients and a 20-fold increase in accuracy in the detection of activity correlations in functional imaging. Furthermore, using principal component analysis, we show that measurements obtained with Bessel beams are clean enough to reveal in one-shot experiments correlations that can not be averaged over trials after stimuli as is the case when studying spontaneous activity. Our results not only demonstrate the contamination of data by systematic and random errors through conventional Gaussian illumination and but,furthermore, quantify the increase in fidelity of such data when using Bessel beams. |
| first_indexed | 2024-04-13T12:35:22Z |
| format | Article |
| id | doaj.art-62f1a6b5464749bea8828570e1d57b2d |
| institution | Directory Open Access Journal |
| issn | 1662-5102 |
| language | English |
| last_indexed | 2024-04-13T12:35:22Z |
| publishDate | 2018-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular Neuroscience |
| spelling | doaj.art-62f1a6b5464749bea8828570e1d57b2d2022-12-22T02:46:40ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022018-09-011210.3389/fncel.2018.00315388732Bessel Beam Illumination Reduces Random and Systematic Errors in Quantitative Functional Studies Using Light-Sheet MicroscopyM. Caroline Müllenbroich0M. Caroline Müllenbroich1Lapo Turrini2Ludovico Silvestri3Ludovico Silvestri4Tommaso Alterini5Ali Gheisari6Natascia Tiso7Francesco Vanzi8Francesco Vanzi9Leonardo Sacconi10Leonardo Sacconi11Francesco S. Pavone12Francesco S. Pavone13Francesco S. Pavone14National Institute of Optics, National Research Council, Sesto Fiorentino, ItalyEuropean Laboratory for Non-linear Spectroscopy, LENS, Sesto Fiorentino, ItalyEuropean Laboratory for Non-linear Spectroscopy, LENS, Sesto Fiorentino, ItalyNational Institute of Optics, National Research Council, Sesto Fiorentino, ItalyEuropean Laboratory for Non-linear Spectroscopy, LENS, Sesto Fiorentino, ItalyEuropean Laboratory for Non-linear Spectroscopy, LENS, Sesto Fiorentino, ItalyEuropean Laboratory for Non-linear Spectroscopy, LENS, Sesto Fiorentino, ItalyDepartment of Biology, University of Padova, Padua, ItalyEuropean Laboratory for Non-linear Spectroscopy, LENS, Sesto Fiorentino, ItalyDepartment of Biology, University of Padova, Padua, ItalyNational Institute of Optics, National Research Council, Sesto Fiorentino, ItalyEuropean Laboratory for Non-linear Spectroscopy, LENS, Sesto Fiorentino, ItalyNational Institute of Optics, National Research Council, Sesto Fiorentino, ItalyEuropean Laboratory for Non-linear Spectroscopy, LENS, Sesto Fiorentino, ItalyDepartment of Physics and Astronomy, University of Florence, Sesto Fiorentino, ItalyLight-sheet microscopy (LSM), in combination with intrinsically transparent zebrafish larvae, is a method of choice to observe brain function with high frame rates at cellular resolution. Inherently to LSM, however, residual opaque objects cause stripe artifacts, which obscure features of interest and, during functional imaging, modulate fluorescence variations related to neuronal activity. Here, we report how Bessel beams reduce streaking artifacts and produce high-fidelity quantitative data demonstrating a fivefold increase in sensitivity to calcium transients and a 20-fold increase in accuracy in the detection of activity correlations in functional imaging. Furthermore, using principal component analysis, we show that measurements obtained with Bessel beams are clean enough to reveal in one-shot experiments correlations that can not be averaged over trials after stimuli as is the case when studying spontaneous activity. Our results not only demonstrate the contamination of data by systematic and random errors through conventional Gaussian illumination and but,furthermore, quantify the increase in fidelity of such data when using Bessel beams.https://www.frontiersin.org/article/10.3389/fncel.2018.00315/fullspontaneous activityzebrafishprinciple component analysislight-sheet microscopyfunctional imagingBessel beams |
| spellingShingle | M. Caroline Müllenbroich M. Caroline Müllenbroich Lapo Turrini Ludovico Silvestri Ludovico Silvestri Tommaso Alterini Ali Gheisari Natascia Tiso Francesco Vanzi Francesco Vanzi Leonardo Sacconi Leonardo Sacconi Francesco S. Pavone Francesco S. Pavone Francesco S. Pavone Bessel Beam Illumination Reduces Random and Systematic Errors in Quantitative Functional Studies Using Light-Sheet Microscopy Frontiers in Cellular Neuroscience spontaneous activity zebrafish principle component analysis light-sheet microscopy functional imaging Bessel beams |
| title | Bessel Beam Illumination Reduces Random and Systematic Errors in Quantitative Functional Studies Using Light-Sheet Microscopy |
| title_full | Bessel Beam Illumination Reduces Random and Systematic Errors in Quantitative Functional Studies Using Light-Sheet Microscopy |
| title_fullStr | Bessel Beam Illumination Reduces Random and Systematic Errors in Quantitative Functional Studies Using Light-Sheet Microscopy |
| title_full_unstemmed | Bessel Beam Illumination Reduces Random and Systematic Errors in Quantitative Functional Studies Using Light-Sheet Microscopy |
| title_short | Bessel Beam Illumination Reduces Random and Systematic Errors in Quantitative Functional Studies Using Light-Sheet Microscopy |
| title_sort | bessel beam illumination reduces random and systematic errors in quantitative functional studies using light sheet microscopy |
| topic | spontaneous activity zebrafish principle component analysis light-sheet microscopy functional imaging Bessel beams |
| url | https://www.frontiersin.org/article/10.3389/fncel.2018.00315/full |
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