Acoustic Gradient Index Research Articles

Dual-resonant scanning multiphoton microscope with ultrasound lens and resonant mirror for rapid volumetric imaging

A dual-resonant scanning multiphoton (DRSM) microscope incorporating a tunable acoustic gradient index of refraction lens with a resonant mirror is developed for high-speed volumetric imaging. In the proposed microscope, the pulse train signal of a femtosecond laser is used to trigger an embedded field programmable gate array to sample the multiphoton excited fluorescence signal at the rate of one pixel per laser pulse. It is shown that a frame rate of around 8000 Hz can be obtained in the x–z plane for an image region with a size of 256 × 80 pixels. Moreover, a volumetric imaging rate of over 30 Hz can be obtained for a large image volume of 343 × 343 × 120 μm3 with an image size of 256 × 256 × 80 voxels. Moreover, a volumetric imaging rate of over 30 Hz can be obtained for a large image volume of 256 × 256 × 80 voxels, which represents 343 × 343 × 120 μm3 in field-of-view. The rapid volumetric imaging rate eliminates the aliasing effect for observed temporal frequencies lower than 15 Hz. The practical feasibility of the DRSM microscope is demonstrated by observing the mushroom bodies of a drosophila brain and performing 3D dynamic observations of moving 10-μm fluorescent beads.

Three-dimensional-generator U-net for dual-resonant scanning multiphoton microscopy image inpainting and denoising.

A dual-resonant scanning multiphoton (DRSM) microscope incorporating a tunable acoustic gradient index of refraction lens and a resonant mirror is developed for rapid volumetric bioimaging. It is shown that the microscope achieves a volumetric imaging rate up to 31.25 volumes per second (vps) for a scanning volume of up to 200 × 200 × 100 µm3 with 256 × 256 × 128 voxels. However, the volumetric images have a severe negative signal-to-noise ratio (SNR) as a result of a large number of missing voxels for a large scanning volume and the presence of Lissajous patterning residuals. Thus, a modified three-dimensional (3D)-generator U-Net model trained using simulated microbead images is proposed and used to inpaint and denoise the images. The performance of the 3D U-Net model for bioimaging applications is enhanced by training the model with high-SNR in-vitro drosophila brain images captured using a conventional point scanning multiphoton microscope. The trained model shows the ability to produce clear in-vitro drosophila brain images at a rate of 31.25 vps with a SNR improvement of approximately 20 dB over the original images obtained by the DRSM microscope. The training convergence time of the modified U-Net model is just half that of a general 3D U-Net model. The model thus has significant potential for 3D in-vivo bioimaging transfer learning. Through the assistance of transfer learning, the model can be extended to the restoration of in-vivo drosophila brain images with a high image quality and a rapid training time.

Axial line-scanning stimulated emission depletion fluorescence correlation spectroscopy.

Investigating the dynamics and interactions of biomolecules within or attached to membranes of living cells is crucial for understanding biology at the molecular level. In this pursuit, classical, diffraction-limited optical fluorescence microscopy is widely used, but it faces limitations due to (1) the heterogeneity of biomembranes on the nanoscale and (2) the intrinsic motion of membranes with respect to the focus. Here we introduce a new confocal microscopy-based fluctuation spectroscopy technique aimed at alleviating these two problems, called axial line-scanning stimulated emission depletion fluorescence correlation spectroscopy (axial ls-STED-FCS). Axial line scanning by means of a tunable acoustic gradient index of refraction lens provides a time resolution of a few microseconds, which is more than two orders of magnitude greater than that of conventional, lateral line-scanning fluorescence correlation spectroscopy (typically around 1 ms). Using STED excitation, the observation area on the membrane can be reduced 10-100 fold, resulting in sub-diffraction spatial resolution and the ability to study samples with densely labeled membranes. Due to these attractive properties, we expect that the axial ls-STED-FCS will find wide application, especially in the biomolecular sciences.

Axial Line-Scanning STED-FCS

We present axial line-scanning stimulated emission depletion fluorescence correlation spectroscopy (axial ls-STED-FCS), a super-resolution method to investigate the dynamics and interactions of biomolecules associated with the plasma membrane of live cells. Intrinsic membrane fluctuations are detrimental for FCS, and line scanning of the confocal spot through the membrane is an effective tool to cope with this problem. In contrast to earlier lateral (within the focal area) line scanning approaches, we employ axial (along the optical axis) line scanning at ultra-high speed by using a tunable acoustic gradient index of refraction (TAG) lens. This device allows us to sweep the observation volume through the membrane within a few microseconds and thus about two orders of magnitude faster than with lateral line scanning using galvanometric scanners. A key advantage of axial lsFCS is that its observation area on the membrane is circular whereas the one of lateral lsFCS is elliptical and larger due to the lower axial resolution of the confocal microscope. As a result, the correlation decay is significantly faster with axial lsFCS, allowing shorter overall measurement times to reach a comparable precision, and its simpler model function makes fitting more robust. Moreover, it is easier to find suitable spots in live imaging of cultured cells. With STED excitation, the observation area can be markedly reduced, as was shown with giant unilamellar vesicles (GUVs) labeled with Atto 647N. Axial ls-STED-FCS enables measurements on smaller spatial scales and with greater biomolecule densities and, therefore, is a valuable extension of standard axial lsFCS, which we have recently introduced for precise measurements of ligand-membrane receptor binding affinities in the (sub)nanomolar range.

Dual GRIN lens two-photon endoscopy for high-speed volumetric and deep brain imaging.

Studying neural connections and activities in vivo is fundamental to understanding brain functions. Given the cm-size brain and three-dimensional neural circuit dynamics, deep-tissue, high-speed volumetric imaging is highly desirable for brain study. With sub-micrometer spatial resolution, intrinsic optical sectioning, and deep-tissue penetration capability, two-photon microscopy (2PM) has found a niche in neuroscience. However, the current 2PM typically relies on a slow axial scan for volumetric imaging, and the maximal penetration depth is only about 1 mm. Here, we demonstrate that by integrating a gradient-index (GRIN) lens and a tunable acoustic GRIN (TAG) lens into 2PM, both penetration depth and volume-imaging rate can be significantly improved. Specifically, an ∼ 1-cm long GRIN lens allows imaging relay from any target region of a mouse brain, while a TAG lens provides a sub-second volume rate via a 100 kHz ∼ 1 MHz axial scan. This technique enables the study of calcium dynamics in cm-deep brain regions with sub-cellular and sub-second spatiotemporal resolution, paving the way for interrogating deep-brain functional connectome.

Optimization of ultrafast axial scanning parameters for efficient pulsed laser micro-machining

Abstract In order to achieve high-resolution micro-machined features, tight focusing is traditionally adopted in laser micro-machining systems, at the cost of a narrow axial machining range. As the laser energy is confined in a single, fixed focal spot, machining throughput is significantly constrained by the writing speed of either translation stages or galvanometer scanners. We study an ultrafast axial scanning technique that asynchronously distributes laser pulses along the axial direction. Oscillation of the focal spot induced by the tunable acoustic gradient index lens is visualized in borosilicate glass with femtosecond laser pulses. By tuning frequency and amplitude of the acoustic field, machining parameters can be optimized, as demonstrated by the case of line machining on silicon. Multi-pass groove machining with axial scanning on silicon as well as femtosecond laser cutting of battery separators exhibit an extended effective machining range and an improved machining efficiency for both focused and defocused surfaces.

Quasi-simultaneous multi-focus imaging using a lock-in pixel image sensor and TAG lens.

In this paper, a quasi-simultaneous multi-focus imaging technique named simulfocus imaging is reported. This technique was developed for measuring an entire object distributed in the depth direction beyond the depth of field (DOF) with high resolution in a single shot. Simulfocus imaging can acquire multiple focal planes in one shot by synchronizing a tunable acoustic gradient index (TAG) lens and a lock-in pixel image sensor. The TAG lens is a tunable-focus lens whose focal position can be changed at a high speed of several tens to several hundreds of kilohertz. The lock-in pixel image sensor is a special image sensor that can execute multiple exposures at an arbitrary timing during a single shooting. The sensor includes a number of photoelectron storage units in each pixel, and the units where the photoelectrons generated by each exposure are stored can be freely selected. Since an image can be acquired for a single storage unit, and the lock-in pixel image sensor has a number of storage units, the lock-in pixel image sensor can acquire multiple images in one shot. By assigning a specific exposure timing to each unit and synchronizing the exposure timing with the focus fluctuation of the TAG lens, it is possible to simultaneously acquire images in different focal planes. To evaluate the system, we conducted experiments to show the effectiveness of simulfocus imaging in microscope and telescope configurations. From the experimental results, it was confirmed that simulfocus was effective in both configurations.

Achromatic acoustic gradient-index phononic crystal lens for broadband focusing

The aim of this study is to realize an achromatic acoustic gradient-index (GRIN) phononic crystal (PC) lens system with a spatially invariant focal length over a broad operating frequency range. To this end, we propose an approach of introducing thin achromatic coating layers that can be easily assembled into the front and rear regions of the acoustic GRIN PC lens. A systematic design method based on topology optimization (TO) is developed to inversely design the achromatic coating components. The topology-optimized achromatic coating components are fabricated using 3D printing and coupled with the acoustic GRIN PC lens for acoustic characterization. Both numerical simulation and experimental characterization demonstrate the achromatic focusing capabilities of the GRIN PC lens with the designed achromatic coating layers in a wide range of frequencies (2.5 kHz–5.5 kHz). The proposed concept of applying achromatic coating layers along with the TO-based design method is expected to provide remarkable versatility to design GRIN PC lens-based applications such as energy harvesting, acoustic imaging, and acoustic wireless power transfer in broadband operation.

Broadband high-quality airy beams via lossy acoustic gradient-index metasurfaces

In this study, we show that fine manipulation of sound waves can be achieved through lossy acoustic gradient-index metasurfaces by utilizing the energy losses. This work, provides a practical route for achieving independent and arbitrary modulation of the amplitude and phase of sound. We demonstrate this mechanism by producing broadband high-quality airy beams. Furthermore, we realize the multifocal focusing based on this lossy acoustic gradient-index metasurfaces. Our findings increase the freedom and flexibility of the control of sound transmission.

All-Optical Volumetric Physiology for Connectomics in Dense Neuronal Structures.

SummaryAll-optical physiology (AOP) manipulates and reports neuronal activities with light, allowing for interrogation of neuronal functional connections with high spatiotemporal resolution. However, contemporary high-speed AOP platforms are limited to single-depth or discrete multi-plane recordings that are not suitable for studying functional connections among densely packed small neurons, such as neurons in Drosophila brains. Here, we constructed a 3D AOP platform by incorporating single-photon point stimulation and two-photon high-speed volumetric recordings with a tunable acoustic gradient-index (TAG) lens. We demonstrated the platform effectiveness by studying the anterior visual pathway (AVP) of Drosophila. We achieved functional observation of spatiotemporal coding and the strengths of calcium-sensitive connections between anterior optic tubercle (AOTU) sub-compartments and >70 tightly assembled 2-μm bulb (BU) microglomeruli in 3D coordinates with a single trial. Our work aids the establishment of in vivo 3D functional connectomes in neuron-dense brain areas.

Wavefront shaping in underwater with soft gradient-index metasurfaces at ultrasonic frequencies

Metasurfaces are planar metamaterials with a subwavelength thickness that allows wavefront shaping by introducing in-plane variations, namely, gradients, in the spatial wave response of these flat structures. Most of the recently reported acoustic metasurfaces were designed to control air-borne waves. Here, we report a new class of acoustic gradient-index (GRIN) metasurfaces engineered from soft graded-porous silicone rubber with a high and tunable acoustic index for broadband ultrasonic 3-D wavefront shaping in aqueous media. The functionalities of these soft flat lenses are illustrated through various experiments, which demonstrate beam steering and beam focusing, as well as vortex beam generation in free water. These new GRIN metasurfaces may have important applications in various domains using designed ultrasonic fields (biomedical imaging, industrial non-destructive testing, contactless particle manipulation), since their fabrication is very straightforward with common polymer science engineering. Y. Jin, R. Kumar, O. Poncelet, O. Mondain-Monval, and T. Brunet, Nat. Commun. 10, 143 (2019).

Millisecond two-photon optical ribbon imaging for small-animal functional connectome study.

We developed a high-speed two-photon optical ribbon imaging system, which combines galvo-mirrors for an arbitrary curve scan on a lateral plane and a tunable acoustic gradient-index lens for a 100kHz-1MHz axial scan. The system provides micrometer/millisecond spatiotemporal resolutions, which enable continuous readout of functional dynamics from small and densely packed neurons in a living adult Drosophila brain. Compared to sparse sampling techniques, the ribbon imaging modality avoids motion artifacts. Combined with a Drosophila anatomical connectome database, which is the most complete among all model animals, this technique paves the way toward establishing whole-brain functional connectome.

Fabrication and experimental demonstration of a hybrid resonant acoustic gradient index metasurface at 40 kHz

Over the past few years, acoustic gradient index metasurfaces (GIMs) have been actively studied for the numerous wave control capabilities that they facilitate. Previous research, however, has primarily focused on GIMs that operate in the audible frequency range, due to the difficulties in fabricating such intricate structures at the millimeter and submillimeter scales, for ultrasonic applications. In this work, we design, fabricate, and experimentally demonstrate the working of a hybrid resonant acoustic gradient index metasurface for airborne ultrasound at 40 kHz. The fabrication of such a GIM is made possible by projection microstereolithography, an emerging additive manufacturing technique. Numerical simulations were conducted to verify the metasurface design, and experiments were performed to corroborate these simulations. The stronger dissipation associated with airborne ultrasound is highlighted in this paper. The experimental demonstration of such a metasurface for airborne ultrasound could further its prospects as a candidate for miniaturized acoustic devices.

Retrieving continuously varying effective properties of non-resonant acoustic metamaterials

We exploit non-resonant inclusions under the metamaterial scheme to enable full analysis of the effective properties with different shapes and lattice structures using the S-parameter retrieval method. Various constraints on the extraction of effective properties are investigated to quantify their dependence on the number of unit cells, the frequency, and the lattice structure. Moreover, distinct effective parameters are confirmed with respect to the size parameter and packing factor of the proposed metamaterials. By this approach, continuously varying effective properties in a broad range of frequencies are achieved, resulting in the realization of acoustic gradient-index devices with appropriate inclusions with non-resonant aspects.

Fast Volumetric Feedback under Microscope by Temporally Coded Exposure Camera.

We developed a temporally coded exposure (TeCE) camera that can cope with high-speed focus variations of a tunable acoustic gradient index (TAG) lens. The TeCE camera can execute a very short exposure multiple times at an arbitrary timing during one shot. Furthermore, by accumulating the photoelectrons generated by each exposure, it is possible to maintain the brightness even with a short exposure time. By synchronously driving the TeCE camera and the TAG lens, different focal planes of an observation target can be acquired at high speed. As a result, high-speed three-dimensional measurement becomes possible, and this can be used for feedback of three-dimensional information. In the work described in this paper, we conducted a focus tracking experiment to evaluate the feedback performance of the TeCE camera. From the experimental results, we confirmed the feedback capability of the TeCE camera.

Flat acoustics with soft gradient-index metasurfaces

Recently, metasurfaces have been proven to be effective and compact devices for the design of arbitrary wavefronts. Metasurfaces are planar metamaterials with a subwavelength thickness that allows wavefront shaping by introducing in-plane variations, namely, gradients, in the spatial wave response of these flat structures. Here we report a new class of acoustic gradient-index (GRIN) metasurfaces engineered from soft graded-porous silicone rubber with a high acoustic index for broadband ultrasonic three-dimensional wavefront shaping in water. The functionalities of these soft flat lenses are illustrated through various experiments, which demonstrate beam steering and beam focusing, as well as vortex beam generation in free space. These new GRIN metasurfaces may have important applications in various domains using designed ultrasonic fields (biomedical imaging, industrial non-destructive testing, contactless particle manipulation), since their fabrication is very straightforward with common polymer science engineering.

Dual-mode phase and fluorescence imaging with a confocal laser scanning microscope

We present dual-mode phase and fluorescence imaging in a confocal laser scanning microscopy (CLSM) system. For phase imaging, the depth of field of the CLSM system is extended by fast axial scanning with a tunable acoustic gradient index of refraction lens. Under transillumination, intensity images of the sample are recorded at a few different defocusing distances. The phase image is reconstructed from these intensity images by using the transport-of-intensity equation. The 3D fluorescence image is obtained by confocal scanning. The dual-mode images with pixel-to-pixel correspondence yield complementary quantitative structural and functional information. Combination of the two imaging modalities enables standalone determination of the refractive index of live cells.

Metasurface-based angle-selective multichannel acoustic refractor

We theoretically study the angle-selective refractions of an impedance-matched acoustic gradient-index metasurface, which is integrated with a rigid bar array of a deep subwavelength period. An interesting refraction order appears under the all-angle incidence despite the existence of a critical angle, and notably, the odevity of the phase-discretization level apparently selects the transmitted diffraction orders. We utilize the strategy of multilayered media design to realize a three-channel acoustic refractor, which shows good promise for constructing multifunctional diffractive acoustic elements for acoustic communication.

Acoustic metacages for sound shielding with steady air flow

Conventional sound shielding structures typically prevent fluid transport between the exterior and interior. A design of a two-dimensional acoustic metacage with subwavelength thickness which can shield acoustic waves from all directions while allowing steady fluid flow is presented in this paper. The structure is designed based on acoustic gradient-index metasurfaces composed of open channels and shunted Helmholtz resonators. In-plane sound at an arbitrary angle of incidence is reflected due to the strong parallel momentum on the metacage surface, which leads to low sound transmission through the metacage. The performance of the proposed metacage is verified by numerical simulations and measurements on a three-dimensional printed prototype. The acoustic metacage has potential applications in sound insulation where steady fluid flow is necessary or advantageous.

Acoustic gradient index lens design using gradient based optimization

We demonstrate a gradient based optimization of the absolute acoustic pressure at a focal point by incrementally repositioning a set of cylindrical obstacles so that they eventually act as an effective gradient index (GRIN) lens. The gradient-based optimization algorithm maximizes the absolute pressure at the focal point by analytically evaluating its derivative with respect to the cylinder positions and then perturbatively optimizing the position of each cylinder in the GRIN lens device while taking into account acoustic multiple scattering between the cylinders. The method is presented by examples of sets of hard cylinders and sets of elastic cylindrical shells of uniform size. Designs using available cylindrical shells enable easy and realistic fabrication of acoustic GRIN lenses. Computations are performed on MATLAB using parallel optimization algorithms and a multistart optimization solver, and supplying the gradients of absolute pressure at the focal point with respect to position vectors. Providing the analytic form of the gradients enhances modeling by allowing the use of exact gradients with optimization algorithms and parallel computing. This results in reducing number of function calls and time needed to converge, and improving solution accuracy for large scale optimization problems especially at high frequencies and with a large number of scatterers.