Junhui Shi

Single-impulse panoramic photoacoustic computed tomography of small-animal whole-body dynamics at high spatiotemporal resolution

Imaging of small animals has played an indispensable role in preclinical research by providing high dimensional physiological, pathological, and phenotypic insights with clinical relevance. Yet pure optical imaging suffers from either shallow penetration (up to ~1–2 mm) or a poor depth-to-resolution ratio (~1/3), and non-optical techniques for whole-body imaging of small animals lack either spatiotemporal resolution or functional contrast. Here, we demonstrate that standalone single-impulse photoacoustic computed tomography (SIP-PACT) mitigates these limitations by combining high spatiotemporal resolution (125-µm in-plane resolution, 50 µs / frame data acquisition and 50-Hz frame rate), deep penetration (48-mm cross-sectional width in vivo), anatomical, dynamical and functional contrasts, and full-view fidelity. By using SIP-PACT, we imaged in vivo whole-body dynamics of small animals in real time and obtained clear sub-organ anatomical and functional details. We tracked unlabeled circulating melanoma cells and imaged the vasculature and functional connectivity of whole rat brains. SIP-PACT holds great potential for both pre-clinical imaging and clinical translation.

Lead zirconate titanate thick film with enhanced electrical properties for high frequency transducer applications

Piezoelectric Pb(Zr0.52Ti0.48)O3 thick film with the thickness around 10 mum has been deposited on the (111) Pt/Ti/SiO2/Si substrate using a ceramic powder/sol-gel solution modified composite method. X-ray diffraction analysis and scanning electron microscope revealed that the film was in the well-crystallized perovskite phase and cracked free. At 1 KHz, The dielectric constant and the loss were 1925 and 0.015, respectively. The remnant polarization was 42.0 muC/cm2 at room temperature. A high frequency single element acoustic transducer fabricated with this film showed a bandwidth at -6 dB of 50% at 156 MHz.

Focusing light inside dynamic scattering media with millisecond digital optical phase conjugation

Wavefront shaping based on digital optical phase conjugation (DOPC) focuses light through or inside scattering media, but the low speed of DOPC prevents it from being applied to thick, living biological tissue. Although a fast DOPC approach was recently developed, the reported single-shot wavefront measurement method does not work when the goal is to focus light inside, instead of through, highly scattering media. Here, using a ferroelectric liquid crystal based spatial light modulator, we develop a simpler but faster DOPC system that focuses light not only through, but also inside scattering media. By controlling 2.6 × 105 optical degrees of freedom, our system focused light through 3 mm thick moving chicken tissue, with a system latency of 3.0 ms. Using ultrasound-guided DOPC, along with a binary wavefront measurement method, our system focused light inside a scattering medium comprising moving tissue with a latency of 6.0 ms, which is one to two orders of magnitude shorter than those of previous digital wavefront shaping systems. Since the demonstrated speed approaches tissue decorrelation rates, this work is an important step toward in vivo deep-tissue non-invasive optical imaging, manipulation, and therapy.

Handheld optical-resolution photoacoustic microscopy

Abstract. Optical-resolution photoacoustic microscopy (OR-PAM) offers label-free in vivo imaging with high spatial resolution by acoustically detecting optical absorption contrasts via the photoacoustic effect. We developed a compact handheld OR-PAM probe for fast photoacoustic imaging. Different from benchtop microscopes, the handheld probe provides flexibility in imaging various anatomical sites. Resembling a cup in size, the probe uses a two-axis water-immersible microelectromechanical system mirror to scan both the illuminating optical beam and resultant acoustic beam. The system performance was tested in vivo by imaging the capillary bed in a mouse ear and both the capillary bed and a mole on a human volunteer.

Lock-in camera based heterodyne holography for ultrasound-modulated optical tomography inside dynamic scattering media

Ultrasound-modulated optical tomography (UOT) images optical contrast deep inside scattering media. Heterodyne holography based UOT is a promising technique that uses a camera for parallel speckle detection. In previous works, the speed of data acquisition was limited by the low frame rates of conventional cameras. In addition, when the signal-to-background ratio was low, these cameras wasted most of their bits representing an informationless background, resulting in extremely low efficiencies in the use of bits. Here, using a lock-in camera, we increase the bit efficiency and reduce the data transfer load by digitizing only the signal after rejecting the background. Moreover, compared with the conventional four-frame based amplitude measurement method, our single-frame method is more immune to speckle decorrelation. Using lock-in camera based UOT with an integration time of 286 μs, we imaged an absorptive object buried inside a dynamic scattering medium exhibiting a speckle correlation time ([Formula: see text]) as short as 26 μs. Since our method can tolerate speckle decorrelation faster than that found in living biological tissue ([Formula: see text] ∼ 100-1000 μs), it is promising for in vivo deep tissue non-invasive imaging.

In vivo label-free photoacoustic flow cytography and on-the-spot laser killing of single circulating melanoma cells

Metastasis causes as many as 90% of cancer-related deaths, especially for the deadliest skin cancer, melanoma. Since hematogenous dissemination of circulating tumor cells is the major route of metastasis, detection and destruction of circulating tumor cells are vital for impeding metastasis and improving patient prognosis. Exploiting the exquisite intrinsic optical absorption contrast of circulating melanoma cells, we developed dual-wavelength photoacoustic flow cytography coupled with a nanosecond-pulsed melanoma-specific laser therapy mechanism. We have successfully achieved in vivo label-free imaging of rare single circulating melanoma cells in both arteries and veins of mice. Further, the photoacoustic signal from a circulating melanoma cell immediately hardware-triggers a lethal pinpoint laser irradiation to kill it on the spot in a thermally confined manner without causing collateral damage. A pseudo-therapy study including both in vivo and in vitro experiments demonstrated the performance and the potential clinical value of our method, which can facilitate early treatment of metastasis by clearing circulating tumor cells from vasculature.

Bessel-beam Grueneisen relaxation photoacoustic microscopy with extended depth of field

Abstract. The short focal depth of a Gaussian beam limits the volumetric imaging speed of optical resolution photoacoustic microscopy (OR-PAM). A Bessel beam, which is diffraction free, provides a long focal depth, but its side lobes deteriorate image quality when the Bessel beam is directly employed to excite photoacoustic (PA) signals in OR-PAM. We present a nonlinear approach based on the Grueneisen relaxation effect to suppress the side-lobe artifacts in PA imaging. This method extends the focal depth of OR-PAM and speeds up volumetric imaging. We experimentally demonstrated a 1-mm focal depth with a 7-μm lateral resolution and volumetrically imaged a carbon fiber and red blood cell samples.

High-resolution deep functional imaging of the whole mouse brain by photoacoustic computed tomography in vivo

Photoacoustic computed tomography (PACT) is a non-invasive imaging technique offering high contrast, high resolution, and deep penetration in biological tissues. We report a PACT system equipped with a high frequency linear transducer array for mapping the microvascular network of a whole mouse brain with the skull intact and studying its hemodynamic activities. The linear array was scanned in the coronal plane to collect data from different angles, and full-view images were synthesized from the limited-view images in which vessels were only partially revealed. We investigated spontaneous neural activities in the deep brain by monitoring the concentration of hemoglobin in the blood vessels and observed strong interhemispherical correlations between several chosen functional regions, both in the cortical layer and in the deep regions. We also studied neural activities during an epileptic seizure and observed the epileptic wave spreading around the injection site and the wave propagating in the opposite hemisphere.

Label-free automated three-dimensional imaging of whole organs by microtomy-assisted photoacoustic microscopy

Three-dimensional (3D) optical imaging of whole biological organs with microscopic resolution has remained a challenge. Most versions of such imaging techniques require special preparation of the tissue specimen. Here we demonstrate microtomy-assisted photoacoustic microscopy (mPAM) of mouse brains and other organs, which automatically acquires serial distortion-free and registration-free images with endogenous absorption contrasts. Without tissue staining or clearing, mPAM generates micrometer-resolution 3D images of paraffin- or agarose-embedded whole organs with high fidelity, achieved by label-free simultaneous sensing of DNA/RNA, hemoglobins, and lipids. mPAM provides histology-like imaging of cell nuclei, blood vessels, axons, and other anatomical structures, enabling the application of histopathological interpretation at the organelle level to analyze a whole organ. Its deep tissue imaging capability leads to less sectioning, resulting in negligible sectioning artifact. mPAM offers a new way to better understand complex biological organs.The state-of-the-art three-dimensional biomedical imaging often requires specific tissue preparation that may alter the physical properties of the specimen causing loss of information. Here Wong et al. develop a microtomy-assisted photoacoustic microscopy that allows imaging of biological samples without labelling agents and with reduced sectioning.

Piezoelectric nonlinear nanomechanical temperature and acceleration insensitive clocks

This work presents the development of high frequency mechanical oscillators based on non-linear laterally vibrating aluminum nitride (AlN) piezoelectric resonators. Our efforts are focused on harnessing non-linear dynamics in resonant mechanical devices to devise frequency sources operating around 1 GHz and capable of outperforming state-of-the-art oscillators in terms of phase noise and size. To this extent, we have identified the thermal and mechanical origin of non-linearities in micro and nanomechanical AlN resonators and developed a theory that describes the optimal operating point for non-linear oscillators. Based on these considerations, we have devised 1 GHz oscillators that exhibit phase noise of < -90 dBc/Hz at 1 kHz offset and < -160 dBc/Hz at 10 MHz offset. In order to attain thermally stable oscillators showing few ppm shifts from - 40 to + 85 °C, we have implemented an embedded ovenization technique that consumes only few mW of power. By means of simple microfabrication techniques, we have included a serpentine heater in the body of the resonator and exploited it to heat it and monitor its temperature without degrading its electromechanical performance. The ovenized devices have resulted in high frequency stability with just few ppm of shift over the temperature range of interest. Finally, few of these oscillators were tested according to military standards for acceleration sensitivity and exhibited a frequency sensitivity lower than 30 ppb/G. These ultra stable oscillators with low jitter and phase noise will ultimately benefit military as well as commercial communication systems.