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Register a new userNon-genetic acoustic stimulation of single neurons by a tapered fiber optoacoustic emitter
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As an emerging technology, transcranial focused ultrasound has been demonstrated to successfully evoke motor responses in mice, rabbits, and sensory/motor responses in humans. Yet, the spatial resolution of ultrasound does not allow for high-precision stimulation. Here, we developed a tapered fiber optoacoustic emitter (TFOE) for optoacoustic stimulation of neurons with an unprecedented spatial resolution of 20 microns, enabling selective activation of single neurons or subcellular structures, such as axons and dendrites. A single acoustic pulse of 1 microsecond converted by the TFOE from a single laser pulse of 3 nanoseconds is shown as the shortest acoustic stimuli so far for successful neuron activation. The highly localized ultrasound generated by the TFOE made it possible to integrate the optoacoustic stimulation and highly stable patch clamp recording on single neurons. Direct measurements of electrical response of single neurons to acoustic stimulation, which is difficult for conventional ultrasound stimulation, have been demonstrated for the first time. By coupling TFOE with ex vivo brain slice electrophysiology, we unveil cell-type-specific response of excitatory and inhibitory neurons to acoustic stimulation. These results demonstrate that TFOE is a non-genetic single-cell and sub-cellular modulation technology, which could shed new insights into the mechanism of neurostimulation.
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Optoacoustic image formation is conventionally based upon ultrasound time-of-flight readings from multiple detection positions. Herein, we exploit acoustic scattering to physically encode the position of optical absorbers in the acquired signals, thus reduce the amount of data required to reconstruct an image from a single waveform. This concept is experimentally tested by including a random distribution of scatterers between the sample and an ultrasound detector array. Ultrasound transmission through a randomized scattering medium was calibrated by raster scanning a light-absorbing microparticle across a Cartesian grid. Image reconstruction from a single time-resolved signal was then enabled with a regularized model-based iterative algorithm relying on the calibration signals. The signal compression efficiency is facilitated by the relatively short acquisition time window needed to capture the entire scattered wavefield. The demonstrated feasibility to form an image using a single recorded optoacoustic waveform paves a way to the development of faster and affordable optoacoustic imaging systems.
A diamond nano-crystal hosting a single nitrogen vacancy (NV) center is optically selected with a confocal scanning microscope and positioned deterministically onto the subwavelength-diameter waist of a tapered optical fiber (TOF) with the help of an atomic force microscope. Based on this nano-manipulation technique we experimentally demonstrate the evanescent coupling of single fluorescence photons emitted by a single NV-center to the guided mode of the TOF. By comparing photon count rates of the fiber-guided and the free-space modes and with the help of numerical FDTD simulations we determine a lower and upper bound for the coupling efficiency of (9.5+/-0.6)% and (10.4+/-0.7)%, respectively. Our results are a promising starting point for future integration of single photon sources into photonic quantum networks and applications in quantum information science.
Focused ultrasound has attracted great attention in minimally invasive therapy, gene delivery, brain stimulation, etc. Frequency below 1 MHz has been identified preferable for high-efficacy drug delivery, gene transfection and neurostimulation due to minimized tissue heating and cell fragmentation. However, the poor spatial resolution of several millimeters and the large device diameter of ~25 mm of current sub-MHz ultrasound technology severely hinders its further applications for effective, precise, safe and wearable biomedical studies and clinical use. To address this issue, we report the development of a novel fiber-based optoacoustic emitter (FOE). The FOE, a new miniaturized ultrasound source, is composed of an optical diffusion coating layer and an expansion coating layer at an optical fiber distal end with a diameter of approximately 500 microns. Taking advantage of the fiber size and diffusive nanoparticles introduced, the ultrasound generated by the FOEs showed a spatial confinement of sub-millimeter. The optoacoustic conversion efficiency was maximized through choosing absorbing nanomaterials and thermal expansion matrix. Controllable frequencies in the range of 0.083 MHz to 5.500 MHz were achieved through using the diffusion layer as a damping material or modifying the nano-composition in the expansion layer. This sub-MHz frequency controllability allows FOEs to be used as a localized ultrasound source for precise cell modulation. We demonstrated optoacoustic cell membrane sonoporation with a localization of sub-millimeter and negligible heat deposition, implicating its broad biomedical applications, including region-specific drug delivery, gene transfection as well as localized neuron stimulation.
Deep supervised neural networks trained to classify objects have emerged as popular models of computation in the primate ventral stream. These models represent information with a high-dimensional distributed population code, implying that inferotemporal (IT) responses are also too complex to interpret at the single-neuron level. We challenge this view by modelling neural responses to faces in the macaque IT with a deep unsupervised generative model, beta-VAE. Unlike deep classifiers, beta-VAE disentangles sensory data into interpretable latent factors, such as gender or hair length. We found a remarkable correspondence between the generative factors discovered by the model and those coded by single IT neurons. Moreover, we were able to reconstruct face images using the signals from just a handful of cells. This suggests that the ventral visual stream may be optimising the disentangling objective, producing a neural code that is low-dimensional and semantically interpretable at the single-unit level.
The cerebrospinal fluid (CSF) constitutes an interface through which chemical cues can reach and modulate the activity of neurons located at the epithelial boundary within the entire nervous system. Here, we investigate the role and functional connectivity of a class of GABAergic sensory neurons contacting the CSF in the vertebrate spinal cord and referred to as CSF-cNs. The remote activation of CSF-cNs was shown to trigger delayed slow locomotion in the zebrafish larva, suggesting that these cells modulate components of locomotor central pattern generators (CPGs). Combining anatomy, electrophysiology, and optogenetics in vivo, we show that CSF-cNs form active GABAergic synapses onto V0-v glutamatergic interneurons, an essential component of locomotor CPGs. We confirmed that activating CSF-cNs at rest induced delayed slow locomotion in the fictive preparation. In contrast, the activation of CSF-cNs promptly inhibited ongoing slow locomotion. Moreover, selective activation of rostral CSF-cNs during ongoing activity disrupted rostrocaudal propagation of descending excitation along the spinal cord, indicating that CSF-cNs primarily act at the premotor level. Altogether, our results demonstrate how a spinal GABAergic sensory neuron can tune the excitability of locomotor CPGs in a state-dependent manner by projecting onto essential components of the excitatory premotor pool.
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