SLM Microscopy: Scanless Two-Photon Imaging and Photostimulation with Spatial Light Modulators.
Volodymyr Nikolenko,Brendon O. Watson,Roberto Araya,Alan Woodruff,Darcy S. Peterka,Rafael Yuste +5 more
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TLDR
A “scanless” microscope that uses a diffractive spatial light modulator (SLM) to shape an incoming two-photon laser beam into any arbitrary light pattern, which allows the simultaneous imaging or photostimulation of different regions of a sample with three-dimensional precision.Abstract:
Laser microscopy has generally poor temporal resolution, caused by the serial scanning of each pixel. This is a significant problem for imaging or optically manipulating neural circuits, since neuronal activity is fast. To help surmount this limitation, we have developed a “scanless” microscope that does not contain mechanically moving parts. This microscope uses a diffractive spatial light modulator (SLM) to shape an incoming two-photon laser beam into any arbitrary light pattern. This allows the simultaneous imaging or photostimulation of different regions of a sample with three-dimensional precision. To demonstrate the usefulness of this microscope, we perform two-photon uncaging of glutamate to activate dendritic spines and cortical neurons in brain slices. We also use it to carry out fast (60 Hz) two-photon calcium imaging of action potentials in neuronal populations. Thus, SLM microscopy appears to be a powerful tool for imaging and optically manipulating neurons and neuronal circuits. Moreover, the use of SLMs expands the flexibility of laser microscopy, as it can substitute traditional simple fixed lenses with any calculated lens function.read more
Citations
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Simultaneous Denoising, Deconvolution, and Demixing of Calcium Imaging Data
Eftychios A. Pnevmatikakis,Daniel Soudry,Yuanjun Gao,Timothy A. Machado,Josh Merel,David Pfau,Thomas Reardon,Thomas Reardon,Yu Mu,Clay Lacefield,Weijian Yang,Misha B. Ahrens,Randy M. Bruno,Thomas M. Jessell,Thomas M. Jessell,Darcy S. Peterka,Rafael Yuste,Liam Paninski +17 more
TL;DR: This work presents a modular approach for analyzing calcium imaging recordings of large neuronal ensembles that relies on a constrained nonnegative matrix factorization that expresses the spatiotemporal fluorescence activity as the product of a spatial matrix that encodes the spatial footprint of each neurons in the optical field and a temporal matrix that characterizes the calcium concentration of each neuron over time.
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Simultaneous all-optical manipulation and recording of neural circuit activity with cellular resolution in vivo
TL;DR: This approach extends the optogenetic toolkit beyond the specificity obtained with genetic or viral approaches, enabling high-throughput, flexible and long-term optical interrogation of functionally defined neural circuits with single-cell and single-spike resolution in the mouse brain in vivo.
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high-speed in vivo calcium imaging reveals neuronal network activity with near-millisecond precision
TL;DR: In vivo imaging in mouse neocortex is reported with greatly improved temporal resolution using random-access scanning with acousto-optic deflectors, uncovering spatiotemporal trial-to-trial variability of sensory responses in barrel cortex and visual cortex.
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What spatial light modulators can do for optical microscopy
TL;DR: In this article, a review of high-resolution miniature spatial light modulators (SLMs) in optical microscopy has been presented, which can be used to control and shape the sample illumination, or they can act as spatial Fourier filters in the imaging path.
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In vivo imaging of neural activity
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TL;DR: Advanced microscopy techniques for in vivo functional imaging are reviewed and guidelines for which technologies are best suited for particular applications are offered.
References
More filters
Journal ArticleDOI
Two-Photon Laser Scanning Fluorescence Microscopy
TL;DR: The fluorescence emission increased quadratically with the excitation intensity so that fluorescence and photo-bleaching were confined to the vicinity of the focal plane as expected for cooperative two-photon excitation.
Journal ArticleDOI
Millisecond-timescale, genetically targeted optical control of neural activity.
TL;DR: In this paper, the authors adapted the naturally occurring algal protein Channelrhodopsin-2, a rapidly gated light-sensitive cation channel, by using lentiviral gene delivery in combination with high-speed optical switching to photostimulate mammalian neurons.
Journal ArticleDOI
A new microscopic principle.
TL;DR: An improvement of the resolution by one decimal wotild require a correction of the objective to four decimals, a practically hopeless task.
Journal ArticleDOI
Neuronal ensemble control of prosthetic devices by a human with tetraplegia
Leigh R. Hochberg,Leigh R. Hochberg,Mijail D. Serruya,Gerhard Friehs,Gerhard Friehs,Jon A. Mukand,Jon A. Mukand,Maryam Saleh,Abraham H. Caplan,Almut Branner,David Chen,Richard D. Penn,John P. Donoghue +12 more
TL;DR: Initial results for a tetraplegic human using a pilot NMP suggest that NMPs based upon intracortical neuronal ensemble spiking activity could provide a valuable new neurotechnology to restore independence for humans with paralysis.
Journal ArticleDOI
Channelrhodopsin-2, a directly light-gated cation-selective membrane channel.
Georg Nagel,Tanjef Szellas,Wolfram Huhn,Suneel Kateriya,Nona Adeishvili,Peter Berthold,Doris Ollig,Peter Hegemann,Ernst Bamberg +8 more
TL;DR: It is demonstrated by functional expression, both in oocytes of Xenopus laevis and mammalian cells, that ChR2 is a directly light-switched cation-selective ion channel, and may be used to depolarize small or large cells, simply by illumination.