Scanless two-photon excitation of channelrhodopsin-2

Papagiakoumou, Eirini; Anselmi, Francesca; Bègue, Aurélien; Sars, Vincent de; Glückstad, Jesper; Isacoff, Ehud Y.; Emiliani, Valentina

doi:10.1038/nmeth.1505

Cited by 387 publications

(397 citation statements)

References 38 publications

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“…Integrated with patterned illumination, the system could further be applied for functional excitation [17,28,29,31], 3D fabrication [30], and structured illumination purposes [39][40][41][42]. In this paper, we have demonstrated that the AEC would vary with different patterns at the same single spatial frequency and different orientations, and could reach nearly 29 % difference, e. g., from À11 % to + 18 % with 1.09 mm À1 spatial frequency patterns at the 908 and 08 orientations.…”

Section: Discussionmentioning

confidence: 90%

Temporal focusing‐based widefield multiphoton microscopy with spatially modulated illumination for biotissue imaging

Chang

Lin

et al. 2017

Journal of Biophotonics

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A developed temporal focusing-based multiphoton excitation microscope (TFMPEM) has a digital micromirror device (DMD) which is adopted not only as a blazed grating for light spatial dispersion but also for patterned illumination simultaneously. Herein, the TFMPEM has been extended to implement spatially modulated illumination at structured frequency and orientation to increase the beam coverage at the back-focal aperture of the objective lens. The axial excitation confinement (AEC) of TFMPEM can be condensed from 3.0 mm to 1.5 mm for a 50 % improvement. By using the TFMPEM with HiLo technique as two structured illuminations at the same spatial frequency but different orientation, reconstructed biotissue images according to the condensed AEC structured illumination are shown obviously superior in contrast and better scattering suppression. Picture: TPEF images of the eosin-stained mouse cerebellar cortex by conventional TFMPEM (left), and the TFMPEM with HiLo technique as 1.09 mm À1 spatially modulated illumination at 908 (center) and 08 (right) orientations.

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Section: Discussionmentioning

confidence: 90%

Temporal focusing‐based widefield multiphoton microscopy with spatially modulated illumination for biotissue imaging

Chang

Lin

et al. 2017

Journal of Biophotonics

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“…Because shift-averaging removes speckle deterministically 28 , and the display's projection rate is 2,000 frames per second, effective patch de-speckling requires only a few milliseconds (2-12 ms in these experiments), allowing for very high effective temporal resolution, which would practically probably be limited by other factors (for example, video processing, hologram computation and optogenetic probe response times). Holographic speckle has emerged as an important issue in the implementation of diffractive photostimulation systems 31 , and it is important to note several alternative designs for circumventing this issue including the use of Generalized Phase Contrast 26 , hybrid holographicmechanical projection systems (that is, rotating diffusers 32 or mechanically vibrated smearing of point patterns 33 ). Other possibilities for speckle reduction are the use of temporally and/ or spatially incoherent light sources.…”

Section: Resultsmentioning

confidence: 99%

“…Diffractive wavefront shaping techniques including computergenerated holography (CGH) and generalized phase contrast are an emerging tool for dynamic patterned photo-excitation [20][21][22][23] that were shown to allow structured two-(2D) and threedimensional (3D) excitation of dendritic arbors 20,24 and neurons 25 using neurotransmitter photolysis as well as twophoton optogenetic stimulation of several neurons in brain slices 26 . The principle advantage of CGH systems 22,23 is that they naturally combine the high intensity, efficiency and resolution that are characteristic of sequential laser deflection methods (such as acousto-optical deflectors) with the capacity for scan-less simultaneous parallel illumination of multiple locations of microdisplay array projectors, but without their respective limitations.…”

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confidence: 99%

Holographic optogenetic stimulation of patterned neuronal activity for vision restoration

et al. 2013

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When natural photoreception is disrupted, as in outer-retinal degenerative diseases, artificial stimulation of surviving nerve cells offers a potential strategy for bypassing compromised neural circuits. Recently, light-sensitive proteins that photosensitize quiescent neurons have generated unprecedented opportunities for optogenetic neuronal control, inspiring early development of optical retinal prostheses. Selectively exciting large neural populations are essential for eliciting meaningful perceptions in the brain. Here we provide the first demonstration of holographic photo-stimulation strategies for bionic vision restoration. In blind retinas, we demonstrate reliable holographically patterned optogenetic stimulation of retinal ganglion cells with millisecond temporal precision and cellular resolution. Holographic excitation strategies could enable flexible control over distributed neuronal circuits, potentially paving the way towards high-acuity vision restoration devices and additional medical and scientific neuro-photonics applications.

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“…Indeed, with adaptive optics we can now image to >400 μm deep into the brain with resolution similar to that normally seen at the superficial depth in brain slices. As a result, adaptive optics would play an important role in the application of many physiological techniques in vivo, such as two-photon uncaging (19), two-photon imaging of dendrites, spines, and axons (20), multiphoton ablation of neuronal structures (21), optogenetic stimulation (22,23), and superresolution imaging (24).…”

Section: Resultsmentioning

confidence: 99%

Characterization and adaptive optical correction of aberrations during in vivo imaging in the mouse cortex

Satō

Betzig

2011

Proc. Natl. Acad. Sci. U.S.A.

183

158

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The signal and resolution during in vivo imaging of the mouse brain is limited by sample-induced optical aberrations. We find that, although the optical aberrations can vary across the sample and increase in magnitude with depth, they remain stable for hours. As a result, two-photon adaptive optics can recover diffraction-limited performance to depths of 450 μm and improve imaging quality over fields of view of hundreds of microns. Adaptive optical correction yielded fivefold signal enhancement for small neuronal structures and a threefold increase in axial resolution. The corrections allowed us to detect smaller neuronal structures at greater contrast and also improve the signal-to-noise ratio during functional Ca 2 imaging in single neurons.T he ability to visualize biological systems in vivo has been a major attraction of optical microscopy, because studying biological systems as they evolve in their natural, physiological state provides relevant information that in vitro preparations often do not allow (1). However, for conventional optical microscopes to achieve their optimal, diffraction-limited resolution, the specimen needs to have identical optical properties to those of the immersion media for which the microscope objective is designed. For example, one of the most widely applied microscopy techniques for in vivo imaging, two-photon fluorescence microscopy, often uses water-dipping objectives. Because biological samples are comprised of structures (i.e., proteins, nuclear acids, and lipids) with refractive indices different from that of water, they induce optical aberrations to the incoming excitation wave and result in an enlarged focal spot within the sample and a concomitant deterioration of signal and resolution (2, 3). As a result, the resolution and contrast of optical microscopes is compromised in vivo, especially deep in tissue.Many questions related to how the brain processes information on both the neuronal circuit level and the cell biological level can be addressed by observing the morphology and activity of neurons inside a living and, preferably, awake and behaving mouse (1). In a typical experiment, an area of the skull is surgically removed and replaced with a cover glass to provide optical access to the underlying structure of interest (4). For imaging during behavior, the cover glass is often attached to an optically transparent plug embedded in the skull to improve mechanical stability and to prevent the skull from growing back and blocking the optical access (5, 6) (Fig. 1A). Before the excitation light of a two-photon microscope reaches the desired focal plane inside the brain, it has to traverse first the cranial window and then the brain tissue, both with optical properties different from water. Thus, they both impart optical aberrations on the excitation light, which leads to a distorted focus, even at the surface of the brain.These sample-induced aberrations can be corrected with adaptive optics (AO) to recover diffraction-limited resolution. In AO, a wavefront-shaping device m...

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Scanless two-photon excitation of channelrhodopsin-2

Cited by 387 publications

References 38 publications

Temporal focusing‐based widefield multiphoton microscopy with spatially modulated illumination for biotissue imaging

Temporal focusing‐based widefield multiphoton microscopy with spatially modulated illumination for biotissue imaging

Holographic optogenetic stimulation of patterned neuronal activity for vision restoration

Characterization and adaptive optical correction of aberrations during in vivo imaging in the mouse cortex

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