Method to quantify accuracy of position feedback signals of a three-dimensional two-photon laser-scanning microscope

Kummer, Michael; Kirmse, Knut; Witte, Otto W.; Haueisen, Jens; Holthoff, Knut

doi:10.1364/boe.6.003678

Cited by 7 publications

(5 citation statements)

References 23 publications

(37 reference statements)

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“…Imaging was performed using a Movable Objective Microscope (Sutter Instrument) equipped with two galvanometric scan mirrors (6210 H, MicroMax 673 XX Dual Axis Servo Driver, Cambridge Technology) and a piezo focusing unit (P-725.4CD PIFOC, E-665.CR ampliﬁer, Physik Instrumente) controlled by a custom-made software written in LabVIEW 2010 (National Instruments; Kummer et al, 2015 ) and MPScope ( Nguyen et al, 2006 ). Fluorescence excitation at 920 nm was provided by a tunable Ti:Sapphire laser (Chameleon Ultra II, Coherent) using a 20×/1.0 NA water immersion objective (XLUMPLFLN 20XW, Olympus).…”

Section: Methodsmentioning

confidence: 99%

Network instability dynamics drive a transient bursting period in the developing hippocampus in vivo

Graf

Rahmati

et al. 2022

eLife

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Spontaneous correlated activity is a universal hallmark of immature neural circuits. However, the cellular dynamics and intrinsic mechanisms underlying network burstiness in the intact developing brain are largely unknown. Here, we use two-photon Ca2+ imaging to comprehensively map the developmental trajectories of spontaneous network activity in the hippocampal area CA1 of mice in vivo. We unexpectedly find that network burstiness peaks after the developmental emergence of effective synaptic inhibition in the second postnatal week. We demonstrate that the enhanced network burstiness reflects an increased functional coupling of individual neurons to local population activity. However, pairwise neuronal correlations are low, and network bursts (NBs) recruit CA1 pyramidal cells in a virtually random manner. Using a dynamic systems modeling approach, we reconcile these experimental findings and identify network bi-stability as a potential regime underlying network burstiness at this age. Our analyses reveal an important role of synaptic input characteristics and network instability dynamics for NB generation. Collectively, our data suggest a mechanism, whereby developing CA1 performs extensive input-discrimination learning prior to the onset of environmental exploration.

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Section: Methodsmentioning

confidence: 99%

Network instability dynamics drive a transient bursting period in the developing hippocampus in vivo

Graf

Rahmati

et al. 2022

eLife

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“…Imaging was performed using a Movable Objective Microscope (Sutter Instrument) equipped with two galvanometric scan mirrors (6210H, MicroMax 673XX Dual Axis Servo Driver, Cambridge Technology) and a piezo focusing unit (P-725.4CD PIFOC, E-665.CR amplifier, Physik Instrumente) controlled by a custom-made software written in LabVIEW 2010 (National Instruments) (Kummer et al, 2015) and MPScope (Nguyen et al, 2006). Fluorescence excitation at 920 nm was provided by a tunable Ti:Sapphire laser (Chameleon Ultra II, Coherent) using a 20×/1.0 NA water immersion objective (XLUMPLFLN 20XW, Olympus).…”

Section: Methodsmentioning

confidence: 99%

Network instability dynamics drive a transient bursting period in the developing hippocampus in vivo

Graf

Rahmati

et al. 2021

Preprint

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Spontaneous correlated activity is a universal hallmark of immature neural circuits. However, the cellular dynamics and intrinsic mechanisms underlying neuronal synchrony in the intact developing brain are largely unknown. Here, we use two-photon Ca2+ imaging to comprehensively map the developmental trajectories of spontaneous network activity in hippocampal area CA1 in vivo. We unexpectedly find that synchronized activity peaks after the developmental emergence of effective synaptic inhibition in the second postnatal week. We demonstrate that the enhanced network synchrony reflects an increased functional coupling of individual neurons to local population activity. However, pairwise neuronal correlations are low, and network bursts recruit CA1 pyramidal cells in a virtually random manner. Using a dynamic systems modeling approach, we reconcile these experimental findings and identify network bi-stability as a potential regime underlying network burstiness at this age. Our analyses reveal an important role of synaptic input characteristics and network instability dynamics for the emergence of neuronal synchrony. Collectively, our data suggest a mechanism, whereby developing CA1 performs extensive input-discrimination learning prior to the onset of environmental exploration.

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“…The actual movement of the galvanometric mirrors may lag behind the signal that controls them (command signal; Kummer et al, 2015). In many galvanometric mirror types, including the ones used in this study, the actual mirror movement can be monitored through the signal of the feedback output (feedback signal).…”

Section: Stability Of Trajectory Computation and Precision Of Slsmentioning

confidence: 99%

High-Accuracy Detection of Neuronal Ensemble Activity in Two-Photon Functional Microscopy Using Smart Line Scanning

et al. 2020

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Graphical AbstractHighlights d Smart line scanning (SLS) restricts imaging to the most informative image pixels d SLS substantially increases the signal-to-noise ratio of population GCaMP6 imaging d SLS leads to higher probability of detecting calcium events in population imaging d SLS achieves more precise identification of functional neuronal ensembles SUMMARY 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 galvanometricbased two-photon microscopes.

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Method to quantify accuracy of position feedback signals of a three-dimensional two-photon laser-scanning microscope

Cited by 7 publications

References 23 publications

Network instability dynamics drive a transient bursting period in the developing hippocampus in vivo

Network instability dynamics drive a transient bursting period in the developing hippocampus in vivo

Network instability dynamics drive a transient bursting period in the developing hippocampus in vivo

High-Accuracy Detection of Neuronal Ensemble Activity in Two-Photon Functional Microscopy Using Smart Line Scanning

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