Bessel beam illumination reduces random and systematic errors in quantitative functional studies using light-sheet microscopy

Muellenbroich, M. C. , Turrini, L., Silvestri, L., Alterini, T., Gheisari, A., Vanzi, F., Sacconi, L. and Pavone, F. S. (2018) Bessel beam illumination reduces random and systematic errors in quantitative functional studies using light-sheet microscopy. Frontiers in Cellular Neuroscience, 12, 315. (doi: 10.3389/fncel.2018.00315) (PMID:30294262) (PMCID:PMC6158350)

Abstract

Light-sheet microscopy (LSM), in combination with intrinsically transparent zebrafish larvae, is a choice method 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.

• Item Type: Articles
• Additional Information: This project received funding from the European Union’s H2020 research and innovation programme under grant agreements No. 720270 (Human Brain Project) and 654148 (Laserlab-Europe), and from the EU programme H2020 EXCELLENT SCIENCEEuropean Research Council (ERC) under grant agreement ID n.692943 (BrainBIT). The project has also been supported by the Italian Ministry for Education, University, and Research in the framework of the Flagship Project NanoMAX and of Eurobioimaging Italian Nodes (ESFRI research infrastructure), and by Ente Cassa di Risparmio di Firenze (private foundation).
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Muellenbroich, Dr Caroline
• Authors: Muellenbroich, M. C., Turrini, L., Silvestri, L., Alterini, T., Gheisari, A., Vanzi, F., Sacconi, L., and Pavone, F. S.
• College/School: College of Science and Engineering > School of Physics and Astronomy
• Journal Name: Frontiers in Cellular Neuroscience
• Publisher: Frontiers Research Foundation
• ISSN: 1662-5102
• ISSN (Online): 1662-5102
• Copyright Holders: Copyright © 2018 Müllenbroich, Turrini, Silvestri, Alterini, Gheisari, Vanzi, Sacconi and Pavone
• First Published: First published in Frontiers in Cellular Neuroscience 12: 315
• Publisher Policy: Reproduced under a Creative Commons License