ZFIN ID: ZDB-PUB-181009-1
Bessel Beam Illumination Reduces Random and Systematic Errors in Quantitative Functional Studies Using Light-Sheet Microscopy
Müllenbroich, M.C., Turrini, L., Silvestri, L., Alterini, T., Gheisari, A., Vanzi, F., Sacconi, L., Pavone, F.S.
Date: 2018
Source: Frontiers in Cellular Neuroscience   12: 315 (Journal)
Registered Authors: Tiso, Natascia, Turrini, Lapo, Vanzi, Francesco
Keywords: Bessel beams, flickering artifacts, functional imaging, light-sheet microscopy, principle component analysis, spontaneous activity, striping, zebrafish
MeSH Terms: none
PubMed: 30294262 Full text @ Front. Cell. Neurosci.
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ABSTRACT
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.
ADDITIONAL INFORMATION
ERRATA and NOTES
This article is corrected by ZDB-PUB-190312-6.