Roger J. Zemp

Photoacoustic imaging of lacZ gene expression in vivo

In the postgenomic era, imaging techniques are playing an important role in visualizing gene expression in vivo. This work represents the first demonstration of photoacoustic tomography (PAT) for reporter gene imaging. Rats inoculated with 9L/lacZ gliosarcoma tumor cells are imaged with PAT before and after injection of X-gal, a colorimetric assay for the lacZ-encoded enzyme beta-galactosidase. Using far-red optical illumination, the genetically tagged tumors in rats are clearly visualized by PAT following the assay. The spatial resolution is quantified to be less than 400 microm, while 500-nM-level sensitivity is demonstrated. With the future development of new absorption-based reporter gene systems, it is anticipated that photoacoustic technology will provide a valuable tool for molecular imaging research.

Modeling of nonlinear ultrasound propagation in tissue from array transducers

A computationally efficient model capable of simulating finite-amplitude ultrasound beam propagation in water and in tissue from phased linear arrays and other transducers of arbitrary quasiplanar geometry is described. It is based on a second-order operator splitting approach [Tavakkoli et al., J. Acoust. Soc. Am. 104, 2061-2072 (1998)], with a fractional step-marching scheme, whereby the effects of diffraction, attenuation, and nonlinearity can be computed independently over incremental steps. This approach is an extension to that of Christopher and Parker [J. Acoust. Soc. Am. 90, 507-521; 90, 488-499 (1991)], wherein linear and nonlinear effects are propagated separately over incremental steps, and the computation of the diffractive substeps are based on an angular spectrum technique with a modified sampling scheme for accurate and efficient implementation of diffractive propagation from nonradially symmetric sources. Results of the model are compared with published data. Predicted field profiles for nonlinear propagation in tissue from realistic array transducers using the pulse inversion method are presented.

Photoacoustic imaging of the microvasculature with a high-frequency ultrasound array transducer

Visualization of microvascular networks could provide new information about function and disease. We demonstrate the capabilities of a 30-MHz ultrasound array system for photoacoustic microscopy of small (< or = 300 microm) vessels in a rat. 3D images obtained by translating the array in the elevation direction are compared with photographs of excised skin. The system is shown to have 100-microm lateral resolution, 25-microm axial resolution, and 3-mm imaging depth. To our knowledge this is the first report on photoacoustic microscopy of the microvasculature with a high-frequency array transducer. It is anticipated that the system can be used for studying and diagnosing a number of diseases including cancer, atherosclerosis, dermatological disorders, and peripheral microvascular complications in diabetes.

Quantitative photoacoustic tomography with multiple optical sources

Quantitative imaging of optical properties of biological tissues with high resolution has been a long-sought-after goal of many research groups. Photoacoustic imaging is a hybrid bio-optical imaging technique offering optical absorption contrast with ultrasonic spatial resolution. While photoacoustic methods offer significant promise for high-resolution optical imaging, quantification has thus far proved challenging. In this paper, a noniterative reconstruction technique for producing quantitative photoacoustic images of absorption perturbations is introduced for the case when the optical properties of the turbid background are known and when multiple optical illumination locations are used. Through theoretical developments and computational examples it is demonstrated that multiple-optical-source photoacoustic imaging can produce quantitative optical absorption reconstructions. The combination of optical and photoacoustic measurements is shown to yield improved reconstruction stability.

Tyrosinase as a dual reporter gene for both photoacoustic and magnetic resonance imaging

Reporter genes are useful scientific tools for analyzing promoter activity, transfection efficiency, and cell migration. The current study has validated the use of tyrosinase (involved in melanin production) as a dual reporter gene for magnetic resonance and photoacoustic imaging. MCF-7 cells expressing tyrosinase appear brown due to melanin. Magnetic resonance imaging of tyrosinase-expressing MCF-7 cells in 300 μL plastic tubes displayed a 34 to 40% reduction in T1 compared to normal MCF-7 cells when cells were incubated with 250 μM ferric citrate. Photoacoustic imaging of tyrosinase-expressing MCF-7 cells in 700 μm plastic tubes displayed a 20 to 57-fold increase in photoacoustic signal compared to normal MCF-7 cells. The photoacoustic signal from tyrosinase-expressing MCF-7 cells was significantly greater than blood at 650 nm, suggesting that tyrosinase-expressing cells can be differentiated from the vasculature with in vivo photoacoustic imaging. The imaging results suggest that tyrosinase is a useful reporter gene for both magnetic resonance and photoacoustic imaging.

Realtime photoacoustic microscopy in vivo with a 30-MHz ultrasound array transducer

We present a novel high-frequency photoacoustic microscopy system capable of imaging the microvasculature of living subjects in realtime to depths of a few mm. The system consists of a high-repetition-rate Q-switched pump laser, a tunable dye laser, a 30-MHz linear ultrasound array transducer, a multichannel high-frequency data acquisition system, and a shared-RAM multi-core-processor computer. Data acquisition, beamforming, scan conversion, and display are implemented in realtime at 50 frames per second. Clearly resolvable images of 6-microm-diameter carbon fibers are experimentally demonstrated at 80 microm separation distances. Realtime imaging performance is demonstrated on phantoms and in vivo with absorbing structures identified to depths of 2.5-3 mm. This work represents the first high-frequency realtime photoacoustic imaging system to our knowledge.

Linear system models for ultrasonic imaging: application to signal statistics

Linear equations for modeling echo signals from shift-variant systems forming ultrasonic B-mode, Doppler, and strain images are analyzed and extended. The approach is based on a solution to the homogeneous wave equation for random inhomogeneous media. When the system is shift-variant, the spatial sensitivity function-defined as a spatial weighting function that determines the scattering volume for a fixed point of time-has advantages over the point-spread function traditionally used to analyze ultrasound systems. Spatial sensitivity functions are necessary for determining statistical moments in the context of rigorous image quality assessment, and they are time-reversed copies of point-spread functions for shift variant systems. A criterion is proposed to assess the validity of a local shift-invariance assumption. The analysis reveals realistic situations in which in-phase signals are correlated to the corresponding quadrature signals, which has strong implications for assessing lesion detectability. Also revealed is an opportunity to enhance near- and far-field spatial resolution by matched filtering unfocused beams. The analysis connects several well-known approaches to modeling ultrasonic echo signals.

Observer efficiency in discrimination tasks Simulating Malignant and benign breast lesions imaged with ultrasound

We investigate and extend the ideal observer methodology developed by Smith and Wagner to detection and discrimination tasks related to breast sonography. We provide a numerical approach for evaluating the ideal observer acting on radio frequency (RF) frame data, which involves inversion of large nonstationary covariance matrices, and we describe a power-series approach to computing this inverse. Considering a truncated power series suggests that the RF data be Wiener-filtered before forming the final envelope image. We have compared human performance for Wiener-filtered and conventional B-mode envelope images using psychophysical studies for 5 tasks related to breast cancer classification. We find significant improvements in visual detection and discrimination efficiency in four of these five tasks. We also use the Smith-Wagner approach to distinguish between human and processing inefficiencies, and find that generally the principle limitation comes from the information lost in computing the final envelope image.

Combined photoacoustic and ultrasound biomicroscopy

We report on the development of an imaging system capable of combined ultrasound and photoacoustic imaging based on a fast-scanning single-element 25-MHz ultrasound transducer and a unique light-delivery system. The system is capable of 20 ultrasound frames per second and slower photoacoustic frame rates limited by laser pulse-repetition rates. Laser and ultrasound pulses are interlaced for co-registration of photoacoustic and ultrasound images. In vivo imaging of a human finger permits ultrasonic visualization of vessel structures and speckle changes indicative of blood flow, while overlaid photoacoustic images highlight some small vessels that are not clear from the ultrasound scan. Photoacoustic images provide optical absorption contrast co-registered in the structural and blood-flow context of ultrasound with high-spatial resolution and may prove important for clinical diagnostics and basic science of the microvasculature.

Estimating optical absorption, scattering, and Grueneisen distributions with multiple-illumination photoacoustic tomography

While photoacoustic methods offer significant promise for high-resolution optical contrast imaging, quantification has thus far proved challenging. In this paper, a noniterative reconstruction technique for producing quantitative photoacoustic images of both absorption and scattering perturbations is introduced for the case when the optical properties of the turbid background are known and multiple optical illumination locations are used. Through theoretical developments and computational examples, it is demonstrated that multiple-illumination photoacoustic tomography (MI-PAT) can alleviate ill-posedness due to absorption-scattering nonuniqueness and produce quantitative high-resolution reconstructions of optical absorption, scattering, and Gruneisen parameter distributions. While numerical challenges still exist, we show that the linearized MI-PAT framework that we propose has orders of magnitude improved condition number compared with CW diffuse optical tomography.