Fast neural population dynamics in primate V1 captured by a genetically-encoded voltage indicator.

This review addresses the challenges of obtaining high-quality quantitative data in the optical imaging of membrane voltage and calcium dynamics. The paper provides a comprehensive overview and systematization of recent studies that analyze factors limiting signal fidelity and propose strategies to enhance data quality. The primary sources of signal degradation in biological optical imaging, with an emphasis on membrane voltage and calcium imaging, are systematically explored across four major indicator classes: voltage-sensitive dyes (VSDs), genetically encoded voltage indicators (GEVIs), calcium-sensitive dyes (CSDs), and genetically encoded calcium indicators (GECIs). Common mechanisms that compromise data quality are classified into three main categories: fundamental photon shot noise, device-related errors, and sample-related measurement errors. For each class of limitation, its physical or biological origin and characteristic manifestations are described, which are followed by an analysis of available mitigation strategies, including hardware optimization, choice of sensors, sample preparation and experimental design, post-processing and computational correction methods.

We have developed a non-linear optical microscope for recording 3D neuronal activity in a cortical column, from awake head fixed behaving mice, with unrivalled temporal resolution.This new technique, called Custom-Address Serial Holography (CASH), uses acousto-optic deflectors (AODs) synchronized with a regenerative amplifier operating at 40 kHz.In this configuration, we have shown that the AODs function as ultrafast phase and amplitude modulators, enabling each laser pulse to be spatially shaped at 40kHz.Spatial modulation of laser pulses enables digital holography of extended volumes in the microscope's focal plane for fluorescence excitation.We will demonstrate the possibility of recording the activity of neurons labeled with Genetically Encoded Calcium Indicators (GECIs) such as GCaMP6f with holographic volumes covering their cell bodies.Using holographic volumes covering only cell membranes, we will demonstrate the possibility of using Genetically Encoded Voltage Indicators (GEVIs) to record the activity of 3D neuronal networks.By updating aberration and diffusion correction synchronously with the scanning, we have also performed trans-cranial imaging over fields of view that go beyond the scattering memory effect.Finally, we will discuss the implementation of temporal focusing to improve the optical sectioning of the 3D-CASH holographic microscope.

Voltage imaging with Genetically Encoded Voltage Indicators (GEVIs) is a promising approach for optically recording membrane potential dynamics with single-cell or subcellular resolution and cell type specificity. We recently reported that a new GEVI named Jellyfish-derived Electricity-reporting Designer Indicator for 2 Photon microscopy (JEDI-2P) could delineate individual action potentials in awake behaving mice for extended durations (> 30 min). However, there remains a need for a broader palette of GEVIs optimized for different illumination modalities and to relay different types of voltage signals.

Measuring the transduction of electrical signals within neurons is a key capability in neuroscience. Fluorescent voltage sensitive dyes (VSDs) were early tools that complemented classical electrophysiology by enabling the optical recording of membrane potential changes from many cells simultaneously. Recent advances in the VSD field have led to bright and highly sensitive sensors that can be targeted to the desired cell populations in live brain tissue. Despite this progress, recently, protein-based genetically encoded voltage indicators (GEVIs) have become the goto tools for targeted voltage imaging in complex environments. In this Perspective, we summarize progress in developing targetable VSDs, discuss areas where these synthetic sensors are or could become relevant, and outline hurdles that need to be overcome to promote the routine use of targetable VSDs in neuroscience research.

Improving the flexibility of genetically encoded voltage indicators via intermolecular FRET

.The pioneering work of Amiram Grinvald established voltage-sensitive dye imaging (VSDI) in the mammalian cortex in the 1980s and inspired decades of cortical voltage imaging and the associated technological developments. The recent conception and development of genetically encoded voltage indicators (GEVIs) overcome many of the limitations of classical VSDI, and open experimental approaches that provide accruing support for orchestrated neuronal circuit dynamics of spatially distributed neuronal circuit underlying behaviors. We will review recent achievements using GEVIs to optically monitor the cortical activity in mammalian brains in vivo and provide a perspective for potential future directions.

Neuronal activity occurs simultaneously and in a highly coordinated fashion in many different areas across the brain. Real-time visualization of large-scale neural dynamics in whole mammalian brains is hindered with the existing neuroimaging methods that are limited in their ability to image large tissue volumes at high speeds. Genetically encoded calcium indicators (GECIs) that modulate their fluorescence intensity as a function of intracellular calcium concentrations are powerful tools for the observation of large neuronal networks. Optoacoustic imaging has been shown capable of real-time three-dimensional imaging of multiple cerebral hemodynamic parameters in rodents. However, optoacoustic imaging of calcium activity deep in mammalian brain is hampered by strong blood absorption in the visible light spectrum as well as lack of activity labels excitable in the near-infrared window. We developed and validated an isolated whole mouse brain preparation labelled with genetically encoded calcium indicator GCaMP6f, which can closely resemble in vivo conditions and exhibit functional activity for several hours to several days. An optoacoustic imaging system coupled to a superfusion system was further devised and used for rapid volumetric monitoring of calcium dynamics in the brain evoked using an epilepsy-inducing drug. The new technique captures calcium fluxes as true 3D information across the entire brain with temporal resolution of 10ms and spatial resolution of 150µm, thus enabling large-scale neural recording at penetration depths and spatio-temporal resolution scales not covered with the existing neuroimaging techniques. The system could be readily adapted to work with future generations of far-red- and near-infrared GECIs.

A temporal femtosecond pulse shaping device, based on all-diffractive method, is designed for arbitrary waveform generation. The key components of the device are micro-gratings arranged in line. By changing the period and phase pattern of each grating, the diffraction angle, phase and amplitude of the first order diffraction light can be modulated. Experimental results are consistent well with simulation results, which indicate that arbitrary temporal waveforms can be gained by using micro-gratings array. Additionally, the configuration of the device allows for multiple outputs and can operate over a large wavelength range from ultraviolet to infrared pulse.

We report advances in time-domain continuous-variable photonic quantum computing, demonstrating a programmable quantum processor capable of processing non-Gaussian states and a programmable quantum light source that generates various non-Gaussian states with arbitrary temporal waveforms, both crucial for universal quantum computation.

.Toward the goal of understanding cutaneous sensory integration during manual behavior, we used voltage-sensitive dye (VSD) imaging to study the organization and dynamics of anesthetized monkey primary somatosensory cortex (SI) in response to single and multidigit tactile stimulation. We find that in both macaque and squirrel monkey SI, VSD reveals clear focal digit topography consistent with previous electrophysiological and intrinsic signal imaging studies. VSD also reveals interactions in SI in response to multidigit stimulation. With a tactile funneling paradigm in areas 3b and 1 in squirrel monkeys, VSD reveals two-digit induction of subthreshhold influences, consistent with lateral intracortical inhibition. In response to tactile apparent motion stimuli, VSD reveals preferential response to motion stimuli over static tactile stimuli in both areas 1 and 3b. Comparison of the response at different digit locations to “toward digit” stimuli suggests the presence of direction-selective response in area 1; however, further study is needed. These exciting results indicate that VSD constitutes a powerful tool for studying somatosensory cortical processing in nonhuman primates and should be further developed for future somatosensory studies in awake behaving monkeys.

‘Real-motion’ cells in the primary visual cortex of macaque monkeys

Optical recordings of membrane potential using genetically targeted voltage-sensitive fluorescent proteins

Chapter 7 New views of primate retinal function

The primate hippocampus, crucial for both episodic memory and spatial navigation, remains an enigma regarding whether these functions share the same neural substrates. We investigated how identical hippocampal neurons in macaque monkeys dynamically shifted their representations between tasks. In a recognition memory task, a notable fraction of hippocampal neurons showed that rate modulation strongly correlated with recognition performance. During free navigation in an open arena, spatial view, rather than position, predominantly influenced the spatial selectivity of hippocampal neurons. Neurons selective for recognition memory displayed minimal spatial tuning, while spatially tuned neurons exhibited limited memory-related activity. These neural correlates of recognition memory and space were more pronounced in the anterior and posterior portions of the hippocampus, respectively. These opposing gradients extended further into the anterior and posterior neocortices. Overall, our findings suggest the presence of orthogonal long-axis gradients between recognition memory and spatial navigation in the hippocampal-neocortical networks of macaque monkeys.

Author SummaryBrain–machine interfaces (BMIs) have the potential to revolutionize the care of neurologically impaired patients. Numerous studies have now shown the feasibility of direct “brain control” of a neuroprosthetic device, yet it remains unclear whether the neural representation for prosthetic control can become consolidated and remain stable over time. This question is especially intriguing given the evidence demonstrating that the neural representation for natural movements can be unstable: BMIs provide a window into the plasticity of cortical circuits in awake-behaving subjects. Here, we show that long-term neuroprosthetic control leads to the formation of a remarkably stable cortical map. Interestingly, this map has the putative attributes of a memory trace, namely, it is stable across time, readily recalled, and resistant to the storage of a second map. The demonstration of such a cortical map for prosthetic control indicates that neuroprosthetic devices could eventually be controlled through the effortless recall of motor memory in a manner that mimics natural skill acquisition and motor control.