High‐Speed Multineuron Calcium Imaging Using Nipkow‐Type Confocal Microscopy

Oba, Shigeyuki; Yukinawa, Naoto; Ujita, Sakiko; Mizunuma, Mika; Matsuki, Norio; Ishii, Shin; Ikegaya, Yuji

doi:10.1002/0471142301.ns0214s57

Cited by 15 publications

(21 citation statements)

References 8 publications

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“…, various combinations of activated neurons) [24], [34], [35]; however, LFP recordings alone cannot determine whether the dopamine-increased occurrence of SWs reflected an increase in the repetitions of the same SW patterns or an increase in the SW pattern variability. To address this question, we used fMCI, an optical technique that records spikes through action potential-evoked calcium elevations in the cell bodies of the focused neurons [36], because unit activities were not evident in our LFP recordings. A drawback of the fMCI technique is its low time resolution due to slow calcium signal kinetics, so that we could not resolve individual spikes during a SW/ripple event.…”

Section: Resultsmentioning

confidence: 99%

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Dopamine Receptor Activation Reorganizes Neuronal Ensembles during Hippocampal Sharp Waves In Vitro

et al. 2014

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Hippocampal sharp wave (SW)/ripple complexes are thought to contribute to memory consolidation. Previous studies suggest that behavioral rewards facilitate SW occurrence in vivo. However, little is known about the precise mechanism underlying this enhancement. Here, we examined the effect of dopaminergic neuromodulation on spontaneously occurring SWs in acute hippocampal slices. Local field potentials were recorded from the CA1 region. A brief (1 min) treatment with dopamine led to a persistent increase in the event frequency and the magnitude of SWs. This effect lasted at least for our recording period of 45 min and did not occur in the presence of a dopamine D1/D5 receptor antagonist. Functional multineuron calcium imaging revealed that dopamine-induced SW augmentation was associated with an enriched repertoire of the firing patterns in SW events, whereas the overall tendency of individual neurons to participate in SWs and the mean number of cells participating in a single SW were maintained. Therefore, dopaminergic activation is likely to reorganize cell assemblies during SWs.

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Section: Resultsmentioning

confidence: 99%

“…fMCI was conducted using acute slices loaded locally with OGB1-AM [36]. Fluorophores were excited at 488 nm and visualized using a 507-nm long-pass emission filter.…”

Section: Methodsmentioning

confidence: 99%

Dopamine Receptor Activation Reorganizes Neuronal Ensembles during Hippocampal Sharp Waves In Vitro

et al. 2014

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“…2A). fMCI detects action potentials through action potential-evoked transient calcium elevations in the cell bodies of individual neurons13; simultaneous recording of cell-attached recordings and fMCI from the same cells revealed that spikes were tightly associated with individual somatic ΔF/F transients (Fig. 2B).…”

Section: Resultsmentioning

confidence: 99%

“…Here, we combined local field potential (LFP) recording with a functional multineuronal calcium imaging (fMCI) technique13 and monitored SW-R-relevant spiking of subicular neuron activity with cellular resolution in acute slices of the mouse hippocampal formation. We unexpectedly found that a fraction of subicular activation occurred before hippocampal SW-R events.…”

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confidence: 99%

Subicular activation preceding hippocampal ripples in vitro

Norimoto

Matsumoto

Miyawaki

et al. 2013

Sci Rep

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Sharp wave-ripple complexes (SW-Rs), a transient form of high-frequency field oscillations observed in the hippocampus, are thought to mediate memory consolidation. They are initiated mainly in hippocampal CA3 area and propagate to the entorhinal cortex through the subiculum; however, little is known about how SW-Rs are initiated and propagate. Here, we used functional multineuronal calcium imaging to monitor SW-R-relevant neuronal activity from the subiculum at single-cell resolution. An unexpected finding was that a subset of subicular neurons was activated immediately before hippocampal SW-Rs. The SW-R-preceding activity was not abolished by surgical lesion of the CA1-to-subiculum projection, and thus, it probably arose from entorhinal inputs. Therefore, SW-Rs are likely to be triggered by entorhinal-to-CA3/CA1 inputs. Moreover, the subiculum is not merely a passive intermediate region that SW-Rs pass through, but rather, it seems to contribute to an active modification of neural information related to SW-Rs.

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“…Cells in organotypic slices were bulk labeled with OGB-AM1 via glass electrode as described previously (22). Briefly, a 2 mM stock solution of OGB-AM1 in 10% Pluronic F-127/DMSO was diluted in ACSF to a final concentration of 200 mM and loaded into a patch pipette.…”

Section: Fluorescent Probe Labelingmentioning

confidence: 99%

Multifocal Fluorescence Microscope for Fast Optical Recordings of Neuronal Action Potentials

Shtrahman

Aharoni

Hardy

et al. 2015

Biophysical Journal

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In recent years, optical sensors for tracking neural activity have been developed and offer great utility. However, developing microscopy techniques that have several kHz bandwidth necessary to reliably capture optically reported action potentials (APs) at multiple locations in parallel remains a significant challenge. To our knowledge, we describe a novel microscope optimized to measure spatially distributed optical signals with submillisecond and near diffraction-limit resolution. Our design uses a spatial light modulator to generate patterned illumination to simultaneously excite multiple user-defined targets. A galvanometer driven mirror in the emission path streaks the fluorescence emanating from each excitation point during the camera exposure, using unused camera pixels to capture time varying fluorescence at rates that are ∼1000 times faster than the camera's native frame rate. We demonstrate that this approach is capable of recording Ca(2+) transients resulting from APs in neurons labeled with the Ca(2+) sensor Oregon Green Bapta-1 (OGB-1), and can localize the timing of these events with millisecond resolution. Furthermore, optically reported APs can be detected with the voltage sensitive dye DiO-DPA in multiple locations within a neuron with a signal/noise ratio up to ∼40, resolving delays in arrival time along dendrites. Thus, the microscope provides a powerful tool for photometric measurements of dynamics requiring submillisecond sampling at multiple locations.

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High‐Speed Multineuron Calcium Imaging Using Nipkow‐Type Confocal Microscopy

Cited by 15 publications

References 8 publications

Dopamine Receptor Activation Reorganizes Neuronal Ensembles during Hippocampal Sharp Waves In Vitro

Dopamine Receptor Activation Reorganizes Neuronal Ensembles during Hippocampal Sharp Waves In Vitro

Subicular activation preceding hippocampal ripples in vitro

Multifocal Fluorescence Microscope for Fast Optical Recordings of Neuronal Action Potentials

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