Random-access scanning microscopy for 3D imaging in awake behaving animals

Nadella, K. M. Naga Srinivas; Roš, Hana; Baragli, Chiara; Griffiths, Victoria A.; Konstantinou, George; Koimtzis, Theo; Evans, Geoffrey J.; Kirkby, Paul A.; Silver, R. Angus

doi:10.1038/nmeth.4033

Cited by 126 publications

(129 citation statements)

References 22 publications

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“…1b,c) by comparing, at equal laser power, the SNR of calcium signals generated by single APs and trains of few APs in the same individual GCaMP6-expressing neurons under the two experimental configurations (scanning and scanless). Random-access two-photon imaging is currently considered the gold standard for high-speed two-photon imaging in the mouse brain in vivo 910485152. Although random-access systems could theoretically scan ~167009 or ~5430010 different positions in one second, thus reaching the performances achieved in the present study (i.e., 47 cells at 1 kHz), previous studies have not achieved these performances at extremely high acquisition rates (e.g., 1 kHz)9104851.…”

Section: Discussionmentioning

confidence: 53%

Simultaneous high-speed imaging and optogenetic inhibition in the intact mouse brain

Bovetti

Moretti

Zucca

et al. 2017

Sci Rep

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Genetically encoded calcium indicators and optogenetic actuators can report and manipulate the activity of specific neuronal populations. However, applying imaging and optogenetics simultaneously has been difficult to establish in the mammalian brain, even though combining the techniques would provide a powerful approach to reveal the functional organization of neural circuits. Here, we developed a technique based on patterned two-photon illumination to allow fast scanless imaging of GCaMP6 signals in the intact mouse brain at the same time as single-photon optogenetic inhibition with Archaerhodopsin. Using combined imaging and electrophysiological recording, we demonstrate that single and short bursts of action potentials in pyramidal neurons can be detected in the scanless modality at acquisition frequencies up to 1 kHz. Moreover, we demonstrate that our system strongly reduces the artifacts in the fluorescence detection that are induced by single-photon optogenetic illumination. Finally, we validated our technique investigating the role of parvalbumin-positive (PV) interneurons in the control of spontaneous cortical dynamics. Monitoring the activity of cellular populations on a precise spatiotemporal scale while manipulating neuronal activity with optogenetics provides a powerful tool to causally elucidate the cellular mechanisms underlying circuit function in the intact mammalian brain.

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Section: Discussionmentioning

confidence: 53%

Simultaneous high-speed imaging and optogenetic inhibition in the intact mouse brain

Bovetti

Moretti

Zucca

et al. 2017

Sci Rep

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“…"The limitation to speed is the speed of sound across crystal, basically, " Silver says. The technique still isn't ideal for quickly imaging every neuron in a volume, he says, but it can move from one region to the next in about 25 microseconds 7 . That makes it useful for viewing all of a sparse population, such as inhibitory interneurons in a volume of brain, he suggests.…”

Section: Advancing Microscopymentioning

confidence: 99%

The real-time technicolour living brain

Dance¹

2016

Nature

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“…The major advancement in measuring Ca 2+ signals was the invention and the application of two-photon microscopy in the nervous system (Denk et al, 1990; Yuste and Denk, 1995). Over many years, particularly with the help of the continuous development of Ca 2+ indicators, two-photon Ca 2+ imaging has become widely used for detecting neural activities on multiple scales ranging from networks to single synapses in both anesthetized and behaving animals (Stosiek et al, 2003; Chen et al, 2011, 2013; Nadella et al, 2016; Szalay et al, 2016). Another commonly used approach for in vivo brain Ca 2+ imaging is based on the use of charged coupled detector/complementary metal-oxide-semiconductor-based cameras, which are particularly useful for recording large-field Ca 2+ dynamics in the superficial cortical layers (Berger et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Locomotion-Related Population Cortical Ca2+ Transients in Freely Behaving Mice

Zhang

Yao

et al. 2017

Front. Neural Circuits

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Locomotion involves complex neural activity throughout different cortical and subcortical networks. The primary motor cortex (M1) receives a variety of projections from different brain regions and is responsible for executing movements. The primary visual cortex (V1) receives external visual stimuli and plays an important role in guiding locomotion. Understanding how exactly the M1 and the V1 are involved in locomotion requires recording the neural activities in these areas in freely moving animals. Here, we used an optical fiber-based method for the real-time monitoring of neuronal population activities in freely moving mice. We combined the bulk loading of a synthetic Ca2+ indicator and the optical fiber-based Ca2+ recordings of neuronal activities. An optical fiber 200 μm in diameter can detect the coherent activity of a subpopulation of neurons. In layer 5 of the M1 and V1, we showed that population Ca2+ transients reliably occurred preceding the impending locomotion. Interestingly, the M1 Ca2+ transients started ~100 ms earlier than that in V1. Furthermore, the population Ca2+ transients were robustly correlated with head movements. Thus, our work provides a simple but efficient approach for monitoring the cortical Ca2+ activity of a local cluster of neurons during locomotion in freely moving animals.

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Random-access scanning microscopy for 3D imaging in awake behaving animals

Cited by 126 publications

References 22 publications

Simultaneous high-speed imaging and optogenetic inhibition in the intact mouse brain

Simultaneous high-speed imaging and optogenetic inhibition in the intact mouse brain

The real-time technicolour living brain

Locomotion-Related Population Cortical Ca2+ Transients in Freely Behaving Mice

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