DeepCGH: 3D computer-generated holography using deep learning.

The goal of computer-generated holography (CGH) is to synthesize custom illumination patterns by modulating a coherent light beam. CGH algorithms typically rely on iterative optimization with a built-in trade-off between computation speed and hologram accuracy that limits performance in advanced applications such as optogenetic photostimulation. We introduce a non-iterative algorithm, DeepCGH, that relies on a convolutional neural network with unsupervised learning to compute accurate holograms with fixed computational complexity. Simulations show that our method generates holograms orders of magnitude faster and with up to 41% greater accuracy than alternate CGH techniques. Experiments in a holographic multiphoton microscope show that DeepCGH substantially enhances two-photon absorption and improves performance in photostimulation tasks without requiring additional laser power.

Social threat alters the behavioral structure of social motivation and reshapes functional brain connectivity.

Traumatic social experiences redefine socially motivated behaviors to enhance safety and survival. Although many brain regions have been implicated in signaling a social threat, the mechanisms by which global neural networks regulate such motivated behaviors remain unclear. To address this issue, we first combined traditional and modern behavioral tracking techniques in mice to assess both approach and avoidance, as well as sub-second behavioral changes, during a social threat learning task. We were able to identify previously undescribed body and tail movements during social threat learning and recognition that demonstrate unique alterations into the behavioral structure of social motivation. We then utilized inter-regional correlation analysis of brain activity after a mouse recognizes a social threat to explore functional communication amongst brain regions implicated in social motivation. Broad brain activity changes were observed within the nucleus accumbens, the paraventricular thalamus, the ventromedial hypothalamus, and the nucleus of reuniens. Inter-regional correlation analysis revealed a reshaping of the functional connectivity across the brain when mice recognize a social threat. Altogether, these findings suggest that reshaping of functional brain connectivity may be necessary to alter the behavioral structure of social motivation when a social threat is encountered.

Three-dimensional multi-site random access photostimulation (3D-MAP).

Optical control of neural ensemble activity is crucial for understanding brain function and disease, yet no technology can achieve optogenetic control of very large numbers of neurons at an extremely fast rate over a large volume. State-of-the-art multiphoton holographic optogenetics requires high-power illumination that only addresses relatively small populations of neurons in parallel. Conversely, one-photon holographic techniques can stimulate more neurons with two to three orders lower power, but with limited resolution or addressable volume. Perhaps most problematically, two-photon holographic optogenetic systems are extremely expensive and sophisticated which has precluded their broader adoption in the neuroscience community. To address this technical gap, we introduce a new one-photon light sculpting technique, three-dimensional multi-site random access photostimulation (3D-MAP), that overcomes these limitations by modulating light dynamically, both in the spatial and in the angular domain at multi-kHz rates. We use 3D-MAP to interrogate neural circuits in 3D and demonstrate simultaneous photostimulation and imaging of dozens of user-selected neurons in the intact mouse brain in vivo with high spatio-temporal resolution. 3D-MAP can be broadly adopted for high-throughput all-optical interrogation of brain circuits owing to its powerful combination of scale, speed, simplicity, and cost.

Advances in computer-generated holography for targeted neuronal modulation.

Genetically encoded calcium indicators and optogenetics have revolutionized neuroscience by enabling the detection and modulation of neural activity with single-cell precision using light. To fully leverage the immense potential of these techniques, advanced optical instruments that can place a light on custom ensembles of neurons with a high level of spatial and temporal precision are required. Modern light sculpting techniques that have the capacity to shape a beam of light are preferred because they can precisely target multiple neurons simultaneously and modulate the activity of large ensembles of individual neurons at rates that match natural neuronal dynamics. The most versatile approach, computer-generated holography (CGH), relies on a computer-controlled light modulator placed in the path of a coherent laser beam to synthesize custom three-dimensional (3D) illumination patterns and illuminate neural ensembles on demand. Here, we review recent progress in the development and implementation of fast and spatiotemporally precise CGH techniques that sculpt light in 3D to optically interrogate neural circuit functions.

Spectral fiber photometry derives hemoglobin concentration changes for accurate measurement of fluorescent sensor activity.

Fiber photometry is an emerging technique for recording fluorescent sensor activity in the brain. However, significant hemoglobin absorption artifacts in fiber photometry data may be misinterpreted as sensor activity changes. Because hemoglobin exists widely in the brain, and its concentration varies temporally, such artifacts could impede the accuracy of photometry recordings. Here we present use of spectral photometry and computational methods to quantify photon absorption effects by using activity-independent fluorescence signals, which can be used to derive oxy- and deoxy-hemoglobin concentration changes. Although these changes are often temporally delayed compared with the fast-responding fluorescence spikes, we found that erroneous interpretation may occur when examining pharmacology-induced sustained changes and that sometimes hemoglobin absorption could flip the GCaMP signal polarity. We provide hemoglobin-based correction methods to restore fluorescence signals and compare our results with other commonly used approaches. We also demonstrated the utility of spectral fiber photometry for delineating regional differences in hemodynamic response functions.

Optimizing holographic data storage using a fractional Fourier transform.

We demonstrate a method to optimize the reconstruction of a hologram when the storage device has a limited dynamic range and a minimum grain size. The optimal solution at the recording plane occurs when the object wave has propagated an intermediate distance between the near and far fields. This distance corresponds to an optimal order and magnification of the fractional Fourier transform of the object.

A micromirror array with annular partitioning for high-speed random-access axial focusing.

Dynamic axial focusing functionality has recently experienced widespread incorporation in microscopy, augmented/virtual reality (AR/VR), adaptive optics and material processing. However, the limitations of existing varifocal tools continue to beset the performance capabilities and operating overhead of the optical systems that mobilize such functionality. The varifocal tools that are the least burdensome to operate (e.g. liquid crystal, elastomeric or optofluidic lenses) suffer from low (≈100 Hz) refresh rates. Conversely, the fastest devices sacrifice either critical capabilities such as their dwelling capacity (e.g. acoustic gradient lenses or monolithic micromechanical mirrors) or low operating overhead (e.g. deformable mirrors). Here, we present a general-purpose random-access axial focusing device that bridges these previously conflicting features of high speed, dwelling capacity and lightweight drive by employing low-rigidity micromirrors that exploit the robustness of defocusing phase profiles. Geometrically, the device consists of an 8.2 mm diameter array of piston-motion and 48-µm-pitch micromirror pixels that provide 2π phase shifting for wavelengths shorter than 1100 nm with 10-90% settling in 64.8 µs (i.e., 15.44 kHz refresh rate). The pixels are electrically partitioned into 32 rings for a driving scheme that enables phase-wrapped operation with circular symmetry and requires <30 V per channel. Optical experiments demonstrated the array's wide focusing range with a measured ability to target 29 distinct resolvable depth planes. Overall, the features of the proposed array offer the potential for compact, straightforward methods of tackling bottlenecked applications, including high-throughput single-cell targeting in neurobiology and the delivery of dense 3D visual information in AR/VR.

All-Silicon Broadband Ultraviolet Metasurfaces.

Featuring high photon energy and short wavelength, ultraviolet (UV) light enables numerous applications such as high-resolution imaging, photolithography, and sensing. In order to manipulate UV light, bulky optics are usually required, and hence do not meet the fast-growing requirements of integration in compact systems. Recently, metasurfaces have shown unprecedented control of light, enabling substantial miniaturization of photonic devices from terahertz to visible regions. However, material challenges have hampered the realization of such functionalities at shorter wavelengths. Herein, it is experimentally demonstrated that all-silicon (Si) metasurfaces with thicknesses of only one-tenth of the working wavelength can be designed and fabricated to manipulate broadband UV light with efficiencies comparable to plasmonic metasurface performance in infrared (IR). Also, for the first time, photolithography enabled by metasurface-generated UV holograms is shown. Such performance enhancement is attributed to increased scattering cross sections of Si antennas in the UV range, which is adequately modeled via a circuit. The new platform introduced here will deepen the understanding of light-matter interactions and introduce even more material options to broadband metaphotonic applications, including those in integrated photonics and holographic lithography technologies.

Precise multimodal optical control of neural ensemble activity.

Understanding brain function requires technologies that can control the activity of large populations of neurons with high fidelity in space and time. We developed a multiphoton holographic approach to activate or suppress the activity of ensembles of cortical neurons with cellular resolution and sub-millisecond precision. Since existing opsins were inadequate, we engineered new soma-targeted (ST) optogenetic tools, ST-ChroME and IRES-ST-eGtACR1, optimized for multiphoton activation and suppression. Employing a three-dimensional all-optical read-write interface, we demonstrate the ability to simultaneously photostimulate up to 50 neurons distributed in three dimensions in a 550 × 550 × 100-µm3 volume of brain tissue. This approach allows the synthesis and editing of complex neural activity patterns needed to gain insight into the principles of neural codes.

UC Berkeley Sculpted Light in the Brain 2017 debates future technologies to communicate with the brain.

This article summarizes presentations at Sculpted Light in the Brain 2017.

Three-dimensional scanless holographic optogenetics with temporal focusing (3D-SHOT).

Optical methods capable of manipulating neural activity with cellular resolution and millisecond precision in three dimensions will accelerate the pace of neuroscience research. Existing approaches for targeting individual neurons, however, fall short of these requirements. Here we present a new multiphoton photo-excitation method, termed three-dimensional scanless holographic optogenetics with temporal focusing (3D-SHOT), which allows precise, simultaneous photo-activation of arbitrary sets of neurons anywhere within the addressable volume of a microscope. This technique uses point-cloud holography to place multiple copies of a temporally focused disc matching the dimensions of a neuron's cell body. Experiments in cultured cells, brain slices, and in living mice demonstrate single-neuron spatial resolution even when optically targeting randomly distributed groups of neurons in 3D. This approach opens new avenues for mapping and manipulating neural circuits, allowing a real-time, cellular resolution interface to the brain.

Flow-scanning optical tomography.

We present a 3D tomography technique for in vivo observation of microscopic samples. The method combines flow in a microfluidic channel, illumination through a slit aperture, and a Fourier lens for simultaneous acquisition of multiple perspective angles in the phase-space domain. The technique is non-invasive and naturally robust to parasitic sample motion. 3D absorption is retrieved using standard back-projection algorithms, here a limited-domain inverse radon transform. Simultaneously, 3D differential phase contrast images are obtained by computational refocusing and comparison of complementary illumination angles. We implement the technique on a modified glass slide which can be mounted directly on existing optical microscopes. We demonstrate both amplitude and phase tomography on live, freely swimming C. elegans nematodes.

Three-dimensional deconvolution microfluidic microscopy using a tilted channel.

We have developed a microfluidic device that enables the computation of three-dimensional (3-D) images of flowing samples. Using a microfluidic channel that is tilted along the optical axis, we record several progressively defocused images of the flowing sample as it passes across the focal plane. The resulting focal stack is then deconvolved to generate 3-D images. Experimental results on flowing yeast cells reveal both volume and surface profile information. The microfluidic channel eliminates the need for a precise translation stage to control defocusing and enables high sample throughput in an insulated, nontoxic, liquid environment. The experimental device can be implemented in all existing microscopes as a modified slide stage and is ideally suited for 3-D profiling in flow cytometers.