Extended field-of-view and increased-signal 3D holographic illumination with time-division multiplexing

Yang, Samuel J.; Allen, William E.; Kauvar, Isaac; Andalman, Aaron S.; Young, Noah P.; Kim, Christina K.; Marshel, James H.; Wetzstein, Gordon; Deisseroth, Karl

doi:10.1364/oe.23.032573

Cited by 49 publications

(32 citation statements)

References 22 publications

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“…Instead of exciting the sample with a whole 2D plane, a computer-generated hologram is built with an SLM leading to multiple focal excitation spots. A two-photon laser pulse is split into multiple beamlets 33–35 that then target many neurons (>100) simultaneously in 3D (Fig. 2c).…”

Section: Transparent Samplesmentioning

confidence: 99%

“…By placing a cubic phase mask (that modifies the spatial phase of the light) in the detection path, targeted neurons at different depths (which can span hundreds of micrometers) 33,34 are focused on the camera, and their activity is recorded simultaneously 34 . By switching hologram patterns, different groups of neurons can be sequentially imaged 35 . EOF holography enables sparse excitation that reduces background, alleviating the effects of scattering and reducing photodamage.…”

Section: Transparent Samplesmentioning

confidence: 99%

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In vivo imaging of neural activity

2017

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Since the introduction of calcium imaging to monitor neuronal activity with single-cell resolution, optical imaging methods have revolutionized neuroscience by enabling systematic recordings of neuronal circuits in living animals. The plethora of methods for functional neural imaging can be daunting to the nonexpert to navigate. Here we review advanced microscopy techniques for in vivo functional imaging and offer guidelines for which technologies are best suited for particular applications.

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Section: Transparent Samplesmentioning

confidence: 99%

Section: Transparent Samplesmentioning

confidence: 99%

In vivo imaging of neural activity

2017

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“…However, while such 3D AOD-based systems provide high sampling speeds at isolated points within the sample 18–20 , they typically require a-priori knowledge of neuronal positions and are typically prone to errors from sample movement. Other approaches have utilized spatial 21 and temporal 22,23 multiplexing strategies to simultaneously image from several laterally or axially separated planes, in some cases in combination with holography for excitation, and signal demixing during post-processing of the recorded signals 24,25 . However, these approaches are currently restricted to a few planes at a time and would be eventually limited by signal statistics and crosstalk, i.e.…”

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confidence: 99%

Fast volumetric calcium imaging across multiple cortical layers using sculpted light

et al. 2016

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While whole-organism calcium imaging in small and semi-transparent animals has been demonstrated, capturing the functional dynamics of large-scale neuronal circuits in awake, behaving mammals at high speed and resolution has remained one of the main frontiers in systems neuroscience. Here we present a novel method based on light sculpting that enables unbiased single and dual-plane high-speed (up to 160 Hz) calcium imaging, as well as in vivo volumetric calcium imaging of a mouse cortical column (0.5 × 0.5 × 0.5 mm) at single-cell resolution and fast volume rates (3 – 6 Hz). This is achieved by tailoring the point-spread function of our microscope to the structures of interest, and by maximizing the signal-to-noise ratio by using a home-built fiber laser amplifier and synchronizing its pulses to the imaging voxel speed. This has enabled in-vivo recording of calcium dynamics of several thousand neurons across cortical layers and in the hippocampus of awake behaving mice.

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“…Two-photon imaging through patterned illumination has been previously applied to perform calcium imaging in vitro , in small organisms212233343536 or in the mouse brain in vivo 3738. Here, our aim was to develop an efficient yet simple microscope design to achieve high-speed imaging of neuronal activity in the intact mouse brain while allowing simultaneous optogenetic manipulation of specific neuronal networks.…”

Section: Discussionmentioning

confidence: 99%

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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Extended field-of-view and increased-signal 3D holographic illumination with time-division multiplexing

Cited by 49 publications

References 22 publications

In vivo imaging of neural activity

In vivo imaging of neural activity

Fast volumetric calcium imaging across multiple cortical layers using sculpted light

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

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