Simultaneous photoacoustic microscopy of microvascular anatomy, oxygen saturation, and blood flow.

Capitalizing on the optical absorption of hemoglobin, photoacoustic microscopy (PAM) is uniquely capable of anatomical and functional characterization of the intact microcirculation in vivo. However, PAM of the metabolic rate of oxygen (MRO2) at the microscopic level remains an unmet challenge, mainly due to the inability to simultaneously quantify microvascular diameter, oxygen saturation of hemoglobin (sO2), and blood flow at the same spatial scale. To fill this technical gap, we have developed a multi-parametric PAM platform. By analyzing both the sO2-encoded spectral dependence and the flow-induced temporal decorrelation of photoacoustic signals generated by the raster-scanned mouse ear vasculature, we demonstrated-for the first time-simultaneous wide-field PAM of all three parameters down to the capillary level in vivo.

High-Frequency Ultrasound Elastography to Assess the Nonlinear Elastic Properties of the Cornea and Ciliary Body.

Mechanical properties of the anterior anatomical structures of the eye, such as the cornea and ciliary body, play a key role in the ocular function and homeostasis. However, measuring the biomechanical properties of the anterior ocular structures, especially deeper structures, such as the ciliary body, remains a challenge due to the lack of high-resolution imaging tools. Herein, we implement a mechanical shaker-based high-frequency ultrasound elastography technique that can track the induced elastic wave propagation to assess the linear and nonlinear elastic properties of anterior ocular structures. The findings of this study advance our understanding of the role of anterior ocular structures in the pathogenesis of different ocular disorders, such as glaucoma.

Ultrasonic elastography to assess biomechanical properties of the optic nerve head and peripapillary sclera of the eye.

PURPOSE: To quantitatively investigate both optic nerve head (ONH) and peripapillary sclera (PPS) biomechanical properties of porcine eyes through an ultrasonic elastography imaging system in response to both increasing and decreasing intraocular pressure (IOP). METHODS: The Young's modulus of the ONH and PPS were assessed using our high resolution ultrasonic imaging system which utilized a mechanical shaker to induce shear waves and an off-axis aligned 40 MHz needle transducer to track micron-level displacement along the direction of wave propagation. In this study, imaging on a total of 8 ex vivo porcine eyes preloaded with IOPs from 6 mmHg to 30 mmHg was performed. To have a better understanding of the effect of varying IOP on biomechanics, both increasing and decreasing IOPs were investigated. RESULTS: The increase of the Young's modulus of ONH (92.4 ± 13.9 kPa at 6 mmHg to 224.7 ± 71.1 kPa at 30 mmHg) and PPS (176.8 ± 14.3 kPa at 6 mmHg to 573.5 ± 64.4 kPa at 30 mmHg) following IOP elevation could be observed in the reconstructed Young's modulus of the shear wave elasticity (SWE) imaging while the B-mode structural images remained almost unchanged. In addition, for the same IOP level, both ONH and PPS have a tendency to be stiffer with decreasing IOP as compared to increasing IOP. CONCLUSIONS: Our results demonstrate the feasibility of using our ultrasonic elastography system to investigate the stiffness mapping of posterior eye with high resolution in both increasing and decreasing IOPs, making this technique potentially useful for glaucoma.

Layer-specific ultrasound elastography using a multi-layered shear wave dispersion model for assessing the viscoelastic properties.

Mechanical properties of biological tissues are significant biomarkers for diagnosing various diseases. Assessing the viscoelastic properties of multi-layer tissues has remained challenging for a long time. Some shear wave models have been proposed to estimate thin-layer tissues' viscoelasticity recently. However, the potential applications of these models are highly restricted since few biological tissues are single-layered. Here we proposed a multi-layer model for layer-specific viscoelasticity estimation of biological tissues. Integrating the theoretical model and ultrasonic micro-elastography imaging system, the viscoelasticity of both layers was assessed. Dual-layer phantoms and ex vivo porcine eyes were used to verify the proposed model. Results obtained from the mechanical test and shear wave rheological model using bulk phantoms were provided as validation criteria. The representative phantom had two layers with elastic moduli of 1.6 ± 0.2 kPa and 18.3 ± 1.1 kPa, and viscosity moduli of 0.56 ± 0.16 Pa·s and 2.11 ± 0.28 Pa·s, respectively. The estimated moduli using the proposed model were 1.3 ± 0.2 kPa and 16.20 ± 1.8 kPa, and 0.80 ± 0.31 Pa·s and 1.87 ± 0.67 Pa·s, more consistent with the criteria (one-tailed t-test, p < 0.1). By contrast, other methods, including the group velocity method and single-layer Rayleigh-Lamb model, generate significant errors in their estimates. For the ex vivo porcine eye, the estimated viscoelasticity was 23.2 ± 8.3 kPa and 1.0 ± 0.4 Pa·s in the retina, and 158.0 ± 17.6 kPa and 1.2 ± 0.4 Pa·s in the sclera. This study demonstrated the potential of the proposed method to significantly improve accuracy and expand clinical applications of shear wave elastography.

Transcranial Focused Ultrasound for Noninvasive Neuromodulation of the Visual Cortex.

Currently, blindness cannot be cured and patients' living quality can be compromised severely. Ultrasonic (US) neuromodulation is a promising technology for the development of noninvasive cortical visual prosthesis. We investigated the feasibility of transcranial focused ultrasound (tFUS) for noninvasive stimulation of the visual cortex (VC) to develop improved visual prosthesis. tFUS was used to successfully evoke neural activities in the VC of both normal and retinal degenerate (RD) blind rats. Our results showed that blind rats showed more robust responses to ultrasound stimulation when compared with normal rats. ( , two-sample t-test). Three different types of ultrasound waveforms were used in the three experimental groups. Different types of cortical activities were observed when different US waveforms were used. In all rats, when stimulated with continuous ultrasound waves, only short-duration responses were observed at "US on and off" time points. In comparison, pulsed waves (PWs) evoked longer low-frequency responses. Testing different parameters of PWs showed that a pulse repetition frequency higher than 100 Hz is required to obtain the low-frequency responses. Based on the observed cortical activities, we inferred that acoustic radiation force (ARF) is the predominant physical mechanism of ultrasound neuromodulation.

Coded excitation using biphase-coded pulse with mismatched filters for high-frequency ultrasound imaging.

A scheme of using phase-coded excitation and mismatched filter compression for high-frequency ultrasound imaging is presented in this paper. Biphase-coded pulses were constructed to excite the transducer. Received signals were compressed with mismatched filters optimized by minimizing peak-sidelobe-level (PSL). Both simulation and experiments were carried out to demonstrate the advantage of this technique. The simulation results demonstrated a possible sidelobe reduction (<-90 dB) with a slightly decrease of the signal-to-noise ratio of less than 1 dB compared with the compression using matched filters alone. The experimental results showed about 14 dB SNR improvement as well as -40 dB sidelobe level when the Barker-13 code excitation with 3-cycle sinusoidal wave carrier was used.

High-Frequency Ultrasound Array Designed for Ultrasound-Guided Breast Biopsy.

This paper describes the development of a miniaturized high-frequency linear array that can be integrated within a core biopsy needle to improve tissue sampling accuracy during breast cancer biopsy procedures. The 64-element linear array has an element width of [Formula: see text], kerf width of [Formula: see text], element length of 1 mm, and element thickness of [Formula: see text]. The 2-2 array composite was fabricated using deep reactive ion etching of lead magnesium niobate-lead titanate (PMN-PT) single crystal material. The array composite fabrication process as well as a novel high-density electrical interconnect solution are presented and discussed. Array performance measurements show that the array had a center frequency and fractional bandwidth ([Formula: see text]) of 59.1 MHz and 29.4%, respectively. Insertion loss and adjacent element crosstalk at the center frequency were -41.0 and [Formula: see text], respectively. A B-mode image of a tungsten wire target phantom was captured using a synthetic aperture imaging system and the imaging test results demonstrate axial and lateral resolutions of 33.2 and [Formula: see text], respectively.

Contactless microparticle control via ultrahigh frequency needle type single beam acoustic tweezers.

This paper reports on contactless microparticle manipulation including single-particle controlled trapping, transportation, and patterning via single beam acoustic radiation forces. As the core component of single beam acoustic tweezers, a needle type ultrasonic transducer was designed and fabricated with center frequency higher than 300 MHz and -6 dB fractional bandwidth as large as 64%. The transducer was built for an f-number close to 1.0, and the desired focal depth was achieved by press-focusing technology. Its lateral resolution was measured to be better than 6.7 µm by scanning a 4 µm tungsten wire target. Tightly focused acoustic beam produced by the transducer was shown to be capable of manipulating individual microspheres as small as 3 µm. "USC" patterning with 15 µm microspheres was demonstrated without affecting nearby microspheres. These promising results may expand the applications in biomedical and biophysical research of single beam acoustic tweezers.

Ultrasound-aided Multi-parametric Photoacoustic Microscopy of the Mouse Brain.

High-resolution quantitative imaging of cerebral oxygen metabolism in mice is crucial for understanding brain functions and formulating new strategies to treat neurological disorders, but remains a challenge. Here, we report on our newly developed ultrasound-aided multi-parametric photoacoustic microscopy (PAM), which enables simultaneous quantification of the total concentration of hemoglobin (CHb), the oxygen saturation of hemoglobin (sO2), and cerebral blood flow (CBF) at the microscopic level and through the intact mouse skull. The three-dimensional skull and vascular anatomies delineated by the dual-contrast (i.e., ultrasonic and photoacoustic) system provide important guidance for dynamically focused contour scan and vessel orientation-dependent correction of CBF, respectively. Moreover, bi-directional raster scan allows determining the direction of blood flow in individual vessels. Capable of imaging all three hemodynamic parameters at the same spatiotemporal scale, our ultrasound-aided PAM fills a critical gap in preclinical neuroimaging and lays the foundation for high-resolution mapping of the cerebral metabolic rate of oxygen (CMRO2)-a quantitative index of cerebral oxygen metabolism. This technical innovation is expected to shed new light on the mechanism and treatment of a broad spectrum of neurological disorders, including Alzheimer's disease and ischemic stroke.

High speed intravascular photoacoustic imaging with fast optical parametric oscillator laser at 1.7 µm.

Intravascular photoacoustic imaging at 1.7 µm spectral band has shown promising capabilities for lipid-rich vulnerable atherosclerotic plaque detection. In this work, we report a high speed catheter-based integrated intravascular photoacoustic/intravascular ultrasound (IVPA/IVUS) imaging system with a 500 Hz optical parametric oscillator laser at 1725 nm. A lipid-mimicking phantom and atherosclerotic rabbit abdominal aorta were imaged at 1 frame per second, which is two orders of magnitude faster than previously reported in IVPA imaging with the same wavelength. Clear photoacoustic signals by the absorption of lipid rich deposition demonstrated the ability of the system for high speed vulnerable atherosclerotic plaques detection.

High-frequency ultrasound imaging for breast cancer biopsy guidance.

Image-guided core needle biopsy is the current gold standard for breast cancer diagnosis. Microcalcifications, an important radiographic finding on mammography suggestive of early breast cancer such as ductal carcinoma in situ, are usually biopsied under stereotactic guidance. This procedure, however, is uncomfortable for patients and requires the use of ionizing radiation. It would be preferable to biopsy microcalcifications under ultrasound guidance since it is a faster procedure, more comfortable for the patient, and requires no radiation. However, microcalcifications cannot reliably be detected with the current standard ultrasound imaging systems. This study is motivated by the clinical need for real-time high-resolution ultrasound imaging of microcalcifications, so that biopsies can be accurately performed under ultrasound guidance. We have investigated how high-frequency ultrasound imaging can enable visualization of microstructures in ex vivo breast tissue biopsy samples. We generated B-mode images of breast tissue and applied the Nakagami filtering technique to help refine image output so that microcalcifications could be better assessed during ultrasound-guided core biopsies. We describe the preliminary clinical results of high-frequency ultrasound imaging of ex vivo breast biopsy tissue with microcalcifications and without Nakagami filtering and the correlation of these images with the pathology examination by hematoxylin and eosin stain and whole slide digital scanning.

Single crystal PZN/PT-polymer composites for ultrasound transducer applications.

Single crystal relaxor ferroelectrics of PZN-8%PT were investigated for potential application in ultrasound transducers. The full set of electromechanical properties was determined using combined resonance and laser interferometry techniques. Ultra-high length extensional coupling (k(33)) of 0.94 was observed, a 25% increase over Navy Type VI PZT ceramics. The thickness extensional coupling (k(t)) of 0.48 was comparable to PZT compositions, and the compliance S(33)(E) was a factor of six greater. To maximize height extensional coupling (k'(33)), while minimizing length extensional coupling k(31) in array elements, it was necessary to align the elements along the 100 crystallographic direction in the x-y plane. Mode coupling plots and test samples for array elements determined that width-to-height ratios of less than 0.5 were desired, similar to the requirement for polycrystalline PZT ceramics. Modeling of 1-3 composites and experimental results demonstrated that thickness coupling greater than 0.80 could be achieved with a 40% to 70% volume fraction of PZN-PT. Although this is a substantial increase over PZT 1-3 composites, with a thickness coupling coefficient of 0.66, it represents a smaller fraction of the length extensional coupling k(33). This reduction may be a consequence of the increased compliance of PZN-PT, which results in significant clamping by the polymer matrix. Ultrasonic transducers fabricated using PZN-8%PT 1-3 composites achieved experimental bandwidths as high as 141%. The pulse-echo responses displayed good agreement with modeled results using the Redwood equivalent circuit.

Non-contact multi-particle annular patterning and manipulation with ultrasound microbeam.

Multiparticle-trapping offers diverse opportunities and applications in biotechnology. It can be applied to creating various functional materials or organizing reactive particles. In this paper, we demonstrate that it is possible to trap and manipulate multi-particles in an annular pattern with a 24 MHz focused ring-type single element ultrasound transducer. Acoustic ring trap can be useful in undertaking biotropism studies due to an equal-distance condition from the center. Also, this ring trap could serve as a force shield to protect analysis area from other cells. The experimental results showed the capability of the proposed method as a multi-cell manipulator in formatting specific patterns of small cells like sperms.

Confocal acoustic radiation force optical coherence elastography using a ring ultrasonic transducer.

We designed and developed a confocal acoustic radiation force optical coherence elastography system. A ring ultrasound transducer was used to achieve reflection mode excitation and generate an oscillating acoustic radiation force in order to generate displacements within the tissue, which were detected using the phase-resolved optical coherence elastography method. Both phantom and human tissue tests indicate that this system is able to sense the stiffness difference of samples and quantitatively map the elastic property of materials. Our confocal setup promises a great potential for point by point elastic imaging in vivo and differentiation of diseased tissues from normal tissue.

Systematic study of high-frequency ultrasonic transducer design for laser-scanning photoacoustic ophthalmoscopy.

Photoacoustic ophthalmoscopy (PAOM) is a high-resolution in vivo imaging modality that is capable of providing specific optical absorption information for the retina. A high-frequency ultrasonic transducer is one of the key components in PAOM, which is in contact with the eyelid through coupling gel during imaging. The ultrasonic transducer plays a crucial role in determining the image quality affected by parameters such as spatial resolution, signal-to-noise ratio, and field of view. In this paper, we present the results of a systematic study on a high-frequency ultrasonic transducer design for PAOM. The design includes piezoelectric material selection, frequency selection, and the fabrication process. Transducers of various designs were successfully applied for capturing images of biological samples in vivo. The performances of these designs are compared and evaluated.

A simple method for evaluating the trapping performance of acoustic tweezers.

The purpose of this paper is to present a rapid and simple method to evaluate the trapping performance of high frequency focused ultrasonic transducers for acoustic tweezer applications. The method takes into consideration the friction between the particle to be trapped and the surface that it resides on. As a result it should be more reliable and accurate than the methods proposed previously. The trapping force produced by a 70-MHz press-focused transducer was measured to evaluate the performance of this approach. This method demonstrates its potential in optimizing the excitation conditions for acoustic tweezer applications and the design of acoustic tweezers.

Resonant acoustic radiation force optical coherence elastography.

We report on a resonant acoustic radiation force optical coherence elastography (ARF-OCE) technique that uses mechanical resonant frequency to characterize and identify tissues of different types. The linear dependency of the resonant frequency on the square root of Young's modulus was validated on silicone phantoms. Both the frequency response spectrum and the 3D imaging results from the agar phantoms with hard inclusions confirmed the feasibility of deploying the resonant frequency as a mechanical contrast for tissue imaging. Furthermore, the results of resonant ARF-OCE imaging of a post-mortem human coronary artery with atherosclerosis demonstrate the potential of the resonant ARF-OCE as a non-invasive method for imaging and characterizing vulnerable plaques.

Enhanced piezoelectric performance of composite sol-gel thick films evaluated using piezoresponse force microscopy.

Conventional composite sol-gel method has been modified to enhance the piezoelectric performance of ceramic thick films. Lead zirconate titanate (PZT) and lead magnesium niobate-lead titanate (PMN-PT) thick films were fabricated using the modified sol-gel method for ultrasonic transducer applications. In this work, piezoresponse force microscopy was employed to evaluate the piezoelectric characteristics of PZT and PMN-PT composite sol-gel thick films. The images of the piezoelectric response and the strain-electric field hysteresis loop behavior were measured. The effective piezoelectric coefficient (d33,eff) of the films was determined from the measured loop data. It was found that the effective local piezoelectric coefficient of both PZT and PMN-PT composite films is comparable to that of their bulk ceramics. The promising results suggest that the modified composite sol-gel method is a promising way to prepare the high-quality, crack-free ceramic thick films.

Miniature optical coherence tomography-ultrasound probe for automatically coregistered three-dimensional intracoronary imaging with real-time display.

We have developed a novel miniature integrated optical coherence tomography (OCT)-intravascular ultrasound (IVUS) probe, with a 1.5-mm-long rigid part and 0.9-mm outer diameter, for real-time intracoronary imaging of atherosclerotic plaques and guiding of interventional procedures. By placing the OCT ball lens and IVUS transducer back-to-back at the same axial position, this probe can provide automatically coregistered, coaxial OCT-IVUS imaging. To demonstrate its real-time capability, three-dimensional OCT-IVUS imaging of a pig's coronary artery displaying in polar coordinates, as well as images of three major types of atherosclerotic plaques in human cadaver coronary segments, were obtained using this probe and our upgraded system. Histology validation is also presented.

Focused high frequency needle transducer for ultrasonic imaging and trapping.

A miniature focused needle transducer (<1 mm) was fabricated using the press-focusing technique. The measured pulse-echo waveform showed the transducer had center frequency of 57.5 MHz with 54% bandwidth and 14 dB insertion loss. To evaluate the performance of this type of transducer, invitro ultrasonic biomicroscopy imaging on the rabbit eye was obtained. Moreover, a single beam acoustic trapping experiment was performed using this transducer. Trapping of targeted particle size smaller than the ultrasonic wavelength was observed. Potential applications of these devices include minimally invasive measurements of retinal blood flow and single beam acoustic trapping of microparticles.