High-Accuracy Detection of Neuronal Ensemble Activity in Two-Photon Functional Microscopy Using Smart Line Scanning
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High-Accuracy Detection of Neuronal Ensemble Activity in Two-Photon Functional Microscopy Using Smart Line Scanning. / Brondi, Marco; Moroni, Monica; Vecchia, Dania; Molano-Mazón, Manuel; Panzeri, Stefano; Fellin, Tommaso.
In: CELL REP, Vol. 30, No. 8, 25.02.2020, p. 2567-2580.e6.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - High-Accuracy Detection of Neuronal Ensemble Activity in Two-Photon Functional Microscopy Using Smart Line Scanning
AU - Brondi, Marco
AU - Moroni, Monica
AU - Vecchia, Dania
AU - Molano-Mazón, Manuel
AU - Panzeri, Stefano
AU - Fellin, Tommaso
N1 - Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.
PY - 2020/2/25
Y1 - 2020/2/25
N2 - Two-photon functional imaging using genetically encoded calcium indicators (GECIs) is one prominent tool to map neural activity. Under optimized experimental conditions, GECIs detect single action potentials in individual cells with high accuracy. However, using current approaches, these optimized conditions are never met when imaging large ensembles of neurons. Here, we developed a method that substantially increases the signal-to-noise ratio (SNR) of population imaging of GECIs by using galvanometric mirrors and fast smart line scan (SLS) trajectories. We validated our approach in anesthetized and awake mice on deep and dense GCaMP6 staining in the mouse barrel cortex during spontaneous and sensory-evoked activity. Compared to raster population imaging, SLS led to increased SNR, higher probability of detecting calcium events, and more precise identification of functional neuronal ensembles. SLS provides a cheap and easily implementable tool for high-accuracy population imaging of neural GCaMP6 signals by using galvanometric-based two-photon microscopes.
AB - Two-photon functional imaging using genetically encoded calcium indicators (GECIs) is one prominent tool to map neural activity. Under optimized experimental conditions, GECIs detect single action potentials in individual cells with high accuracy. However, using current approaches, these optimized conditions are never met when imaging large ensembles of neurons. Here, we developed a method that substantially increases the signal-to-noise ratio (SNR) of population imaging of GECIs by using galvanometric mirrors and fast smart line scan (SLS) trajectories. We validated our approach in anesthetized and awake mice on deep and dense GCaMP6 staining in the mouse barrel cortex during spontaneous and sensory-evoked activity. Compared to raster population imaging, SLS led to increased SNR, higher probability of detecting calcium events, and more precise identification of functional neuronal ensembles. SLS provides a cheap and easily implementable tool for high-accuracy population imaging of neural GCaMP6 signals by using galvanometric-based two-photon microscopes.
KW - Action Potentials/physiology
KW - Animals
KW - Artifacts
KW - Calcium/metabolism
KW - Image Processing, Computer-Assisted
KW - Mice
KW - Microscopy, Fluorescence, Multiphoton
KW - Motion
KW - Neurons/physiology
KW - Neuropil Threads/physiology
KW - Wakefulness
U2 - 10.1016/j.celrep.2020.01.105
DO - 10.1016/j.celrep.2020.01.105
M3 - SCORING: Journal article
C2 - 32101736
VL - 30
SP - 2567-2580.e6
JO - CELL REP
JF - CELL REP
SN - 2211-1247
IS - 8
ER -
