We present the development of a photoacoustic tomography (PAT) imaging system with the demonstrated capability of obtaining high-throughput scans at a sustained rate of under 1 minute per animal using integrated robotics to assist in 3D PAT collection. This is a considerable achievement as there is currently no existing commercial or research PAT whole-body imaging system capable of high-throughput applications (15-20 animals per hour). High-throughput experimentation is imperative in the development, characterization, and use of rodent models of human diseases as it increases the number of animals that can be evaluated within a single experiment and may reduce the time under anesthesia for each animal, thereby improving the stability, duration, and confidence of longitudinal studies The developed system features coordinated automation for robotic animal manipulation, anesthesia distribution, temperature regulation, water management, laser excitation, and photoacoustic detection. Furthermore, as shown in validation studies using phantoms and live murine models, the prototype imaging platform demonstrates high-throughput performance while retaining high sensitivity and high resolution.
SignificanceTo effectively study preclinical animal models, medical imaging technology must be developed with a high enough resolution and sensitivity to perform anatomical, functional, and molecular assessments. Photoacoustic (PA) tomography provides high resolution and specificity, and fluorescence (FL) molecular tomography provides high sensitivity; the combination of these imaging modes will enable a wide range of research applications to be studied in small animals.AimWe introduce and characterize a dual-modality PA and FL imaging platform using in vivo and phantom experiments.ApproachThe imaging platform’s detection limits were characterized through phantom studies that determined the PA spatial resolution, PA sensitivity, optical spatial resolution, and FL sensitivity.ResultsThe system characterization yielded a PA spatial resolution of 173 ± 17 μm in the transverse plane and 640 ± 120 μm in the longitudinal axis, a PA sensitivity detection limit not less than that of a sample with absorption coefficient μa = 0.258 cm − 1, an optical spatial resolution of 70 μm in the vertical axis and 112 μm in the horizontal axis, and a FL sensitivity detection limit not <0.9 μM concentration of IR-800. The scanned animals displayed in three-dimensional renders showed high-resolution anatomical detail of organs.ConclusionsThe combined PA and FL imaging system has been characterized and has demonstrated its ability to image mice in vivo, proving its suitability for biomedical imaging research applications.
KEYWORDS: Spatial resolution, Imaging systems, Ultrasonography, Photoacoustic imaging, In vivo imaging, Signal detection, Video, Functional imaging, Doppler effect, Data acquisition
We present the continued development a clinical ultrasound (US) imaging device with a photoacoustic (PA) mode. The combined USPA imaging platform is designed around a compact US component capable of B-mode, M-mode, color Doppler, and pulsed wave Doppler US imaging modes with a 128 element US probe. The PA mode can support a 256 element PA probe with real-time 2D imaging up to 20 Hz. The PA signals are amplified by a 40 dB pre-amplifier while US mode signals bypass the circuit, resulting in high quality PA images. We demonstrate the USPA platform’s capabilities using tissue-mimicking phantoms.
PhotoSound Technologies specializes in the development of electronics solutions for massive parallel data acquisition applicable to the fields of photoacoustics (PA), X-ray acoustics, including 3D dosimetry, and ultrasound. PhotoSound’s Legion ADC256 R1.1, released in 2018, is a 256-channel 12-bit ADC with a sampling rate of 40 MHz. The ADC256’s average data bandwidth is limited by its USB3 PC interface, which has a data rate up to 3 Gbps per board. Multiple ADC256 boards can operate fully in parallel. On software level configurations, multiple ADC256 boards are represented as a single ADC board with increased number of channels. The incoming ultrasound (US) upgrades and modifications of ADC256 will enable combination and alternation of US and PA modes using the same probe. PhotoSound MoleculUS is a medical-grade Telemed US system combined with a PA-optimized ADC. MoleculUS utilizes clinical US probes to produce US images which can be interleaved with PA imaging by enabling optical fiber illumination. The other ADC256 modification, advanced PAUS oriented for research, will have PCIe PC interface for raw PA and US data and arbitrary software control over beamformer profiles, limited by high-voltage power only. The data in ultrasound and photoacoustics modes is user accessible in raw format and can be delivered to CUDA GPU using MATLAB parallel computing (CUDA) toolbox or other tools. Multiple PAUS boards can work in parallel in both PA and US modes.
KEYWORDS: Skin, Luminescence, Photoacoustic spectroscopy, Tissues, 3D image processing, Animal model studies, Preclinical imaging, Spatial resolution, In vivo imaging, Acquisition tracking and pointing
We report on the development of a preclinical 3D imaging platform integrating photoacoustic tomography and fluorescence (PAFT). The proposed multimodal imaging concept addresses known deficiencies in sensitivity, anatomical registration, and spatial resolution of the individual imaging modalities. Multi-view photoacoustic and optical projections of the studied animal are utilized to reconstruct large (27 cm3) volumes showing vascular network and blood-rich tissues, as well as regions with induced optical/fluorescence contrast with 3D resolution exceeding 150 μm. An additional 532-nm low-energy pulsed laser excitation is implemented as a separate imaging channel for registration over skin topography and superficial vasculature. PAFT technology enables functional and molecular volumetric imaging using wide range of fluorescent and luminescent biomarkers, nanoparticles, and other photosensitive constructs mapped with high fidelity over robust anatomical structures of the studied animal model. We demonstrated the PAFT performance using phantoms and by in vivo imaging of preclinical murine models.
KEYWORDS: Data acquisition, Clocks, Capacitance, Interfaces, Electromagnetic coupling, Amplifiers, Connectors, Field effect transistors, Signal to noise ratio, Human-machine interfaces
Large number of simultaneously acquired spatially distinct pressure signals is required to improve quality of real-time photoacoustic and x-ray acoustic biomedical images [1]. In the past this approach was limited by availability of commercial multi-channel analog-to-digital converter (ADC) systems and ability to operate multiple ADC boards with synchronized clock and trigger source. The new Legion series single-board 256-channel ADC (ADC256) was designed by PhotoSound for massive parallel data acquisition utilized in photoacoustic, laser-induced ultrasound, and X-ray acoustic real-time imaging applications. ADC256 is a 12-bit ADC with a sampling rate up to 40 MHz and a USB3 computer interface. It can run at 200 Hz frame rate with 4096 points per trigger acquired by each channel. Higher trigger rates without data loss are possible with smaller number of points per trigger. ADC256 has an integrated amplifier with programmable gain up to 51 dB. Additionally, it can be equipped with a matching photoacoustic preamplifier. The system architecture is scalable to 1024 channels using four synchronized boards with a single trigger source. The clock and the trigger can be delivered from the master ADC256 board (daisy chain) or from the clock and trigger server (star topology). The data collected by each ADC board has trigger and board stamps allowing to (a) use multiple computers for data acquisition, and (b) detection of lost data events, even if the trigger rate exceeds its maximum allowed value.
We have developed a preclinical 3D imaging instrument integrating photoacoustic tomography and fluorescence (PAFT) addressing known deficiencies in sensitivity and spatial resolution of the individual imaging components. PAFT is designed for simultaneous acquisition of photoacoustic and fluorescence orthogonal projections at each rotational position of a biological object, enabling direct registration of the two imaging modalities. Orthogonal photoacoustic projections are utilized to reconstruct large (21 cm3 ) volumes showing vascularized anatomical structures and regions of induced optical contrast with spatial resolution exceeding 100 µm. The major advantage of orthogonal fluorescence projections is significant reduction of background noise associated with transmitted or backscattered photons. The fluorescence imaging component of PAFT is used to boost detection sensitivity by providing low-resolution spatial constraint for the fluorescent biomarkers. PAFT performance characteristics were assessed by imaging optical and fluorescent contrast agents in tissue mimicking phantoms and in vivo. The proposed PAFT technology will enable functional and molecular volumetric imaging using fluorescent biomarkers, nanoparticles, and other photosensitive constructs mapped with high fidelity over robust anatomical structures, such as skin, central and peripheral vasculature, and internal organs.
We introduce a preclinical imaging platform – a 3D photoacoustic/fluorescence tomography (PAFT) instrument augmented with an environmentally responsive dual-contrast biocompatible nanoprobe. The PAFT instrument was designed for simultaneous acquisition of photoacoustic and fluorescence orthogonal projections at each rotational position of a biological object, enabling direct co-registration of the two imaging modalities. The nanoprobe was based on liposomes loaded with J-aggregates of indocyanine green (PAtrace). Once PAtrace interacts with the environment, a transition from J-aggregate to monomeric ICG is induced. The subsequent recovery of monomeric ICG is characterized by dramatic changes in the optical absorption spectrum and reinstated fluorescence. In the activated state, PAtrace can be simultaneously detected by both imaging modes of the PAFT instrument using 780 nm excitation and fluorescence detection at 810 nm. The fluorescence imaging component is used to boost detection sensitivity by providing lowresolution map of activated nanoprobes, which are then more precisely mapped in 3D by the photoacoustic imaging component. Activated vs non-activated particles can be distinguished based on their different optical absorption peaks, removing the requirements for complex image registration between reference and detection scans. Preliminary phantom and in vivo animal imaging results showed successful activation and visualization of PAtrace with high sensitivity and resolution. The proposed PAFT-PAtrace imaging platform could be used in various functional and molecular imaging applications including multi-point in vivo assessment of early metastasis.
We describe the ongoing development and performance of a high-pulse-energy wavelength-cycling laser system for three-dimensional optoacoustic tomography of the breast. Joule-level energies are desired for achieving the required penetration depths while maintaining safe fluence levels. Wavelength cycling provides a pulse sequence which repeatedly alternates between two wavelengths (approximately 756 and 797 nm) that provide differential imaging. This improves co-registration of captured differential images and quantification of blood oxygen saturation. New design features have been developed for and incorporated into a clinical prototype laser system, to improve efficacy and ease of use in the clinic. We describe the benefits of these features for operation with a clinical pilot optoacoustic / ultrasound dual-modality three-dimensional imaging system.
We report on the optoacoustic (OA) imaging of the whole mouse body using a biocompatible contrast agent - highly
purified, pegylated gold nanorods (GNR) - which has strong optical absorption in the near-infrared region and low level
of toxicity. In vitro toxicity studies showed no significant change in survival rates of the cultured normal epithelium
IEC-6 cells when incubated for 24 hours with up to 1 nM of GNR. In vivo toxicity studies in nude mice showed no
pathological changes in liver 1 month after the IV injection of GNR with intra-body concentration around 0.25-0.50 nM.
In order to study the enhancement of the OA contrast and accumulation of GNR in different tissues, we performed 3D
OA imaging of live nude mice with IV-injected GNR. The enhancement of the OA contrast in comparison with the
images of the untreated mice was visible starting 1 hour after the GNR injection. The OA contrast of kidneys, liver, and
spleen peaked at about 2-3 days after the administration of the GNR, and then was gradually reducing.
Coregistered optoacoustic (OA) and ultrasound (US) images obtained using a dual modality optoacoustic/ultrasonic
breast imaging system are used together for enhanced diagnostic capabilities in comparison to each individual
technology. Therefore, an operator-independent delineation of diagnostically relevant objects (in our case breast tumors)
with subsequent automatic analysis of image features is required. We developed the following procedure: 1) Image
filtering is implemented on a US image to minimize speckle noise and simultaneously preserve the sharpness of the
boundaries of the extended objects; 2) Boundaries of the objects of interest are automatically delineated starting with an
initial guess made by an operator; 3) Both US and OA images are analyzed using the detected boundaries (contrast,
boundary sharpness, homogeneity of the objects and background, boundary morphology parameters are calculated).
Calculated image characteristics can be used for statistically independent evaluation of structural information (US data)
and vascularization (OA data) of the studied breast tissues. Operator-independent delineation of the objects of interest
(e.g. tumors and blood vessels) is essential in clinical OA spectroscopy (using multiple laser wavelengths to quantify
concentrations of particular tissue chromophores, such as oxy- and deoxy- hemoglobin, water, and lipids). Another
potential application of the suggested image analysis algorithm could be in OA imaging system design, when system
performance should be evaluated in terms of quality of the images reconstructed from the well-defined objects of
interest. The discussed principles of image analysis are illustrated by using real clinical US and OA data.
We introduce a novel experimental design for non-invasive scanning optoacoustic microscopy that utilizes a parabolic
surface for ultrasound focusing. We demonstrate that off-axis parabolic mirrors made of sufficiently high acoustic
impedance materials work as ideal reflectors in a wide range of apertures and provide lossless conversion of a spherical
acoustic wavefront into a plane wave. We further test the performance of a custom optoacoustic imaging setup which
was developed and built based on these principles. The achieved resolution limit of 0.3 mm, with NA of 0.5 and the
transducer bandwidth of 5 MHz, matches the resolution limit defined by diffraction. Although further improvements of
current experimental setup are required to achieve resolution similar to leading microscopy systems, this proof-of-concept
work demonstrates the viability of the proposed design for optoacoustic microscopy applications.
We used a three-dimensional optical tomogaphy system that was previously developed to create high contrast maps of
optical absorbance of mice tissues. In this study, animals were scanned before and after injection of gold nanorods
(GNRs) at different time periods. As-synthesized GNRs were purified from hexadecyltrimethylammonium bromide
(CTAB) and coated with polyethylene glycol (PEG) to obtain GNR-PEG complexes suitable for in vivo applications.
Intravenous administration of the purified GNR-PEG complexes to mice resulted in an enhanced contrast of normal
tissues and blood vessels as compared to ordinary nude mice. In parallel with optoacoustic imaging we investigated the
accumulation of GNRs in liver using invasive analytical techniques. Maximum levels of GNRs in liver macrophages
were observed after 48-72 hours post-injection, followed by slow clearance trend after 8 days. Optoacoustic imaging
revealed redistribution of GNR in mouse organ and tissues: in the initial hours, accumulation of GNRs is seen
predominantly in the periphery of the mouse, while a gradual increase of GNR levels in liver, spleen and kidneys is seen
in 1 and 24 hours.
We present our findings from a real-time laser optoacoustic imaging system (LOIS). The system utilizes a Q-switched Nd:YAG laser; a standard 128-channel ultrasonic linear array probe; custom electronics and custom software to collect, process, and display optoacoustic (OA) images at 10 Hz. We propose that this system be used during preoperative mapping of forearm vessels for hemodialysis treatment. To demonstrate the real-time imaging capabilities of the system, we show OA images of forearm vessels in a volunteer and compare our results to ultrasound images of the same region. Our OA images show blood vessels in high contrast. Manipulations with the probe enable us to locate and track arteries and veins of a forearm in real time. We also demonstrate the ability to combine a series of OA image slices into a volume for spatial representation of the vascular network. Finally, we use frame-by-frame analysis of the recorded OA video to measure dynamic changes of the crossection of the ulnar artery.
Optoacoustic (OA) tomography has demonstrated utility in identifying blood-rich malignancies in breast tissue. We describe the development and characterization of a laser OA imaging system for the prostate (LOIS-P). The system consists of a fiber-coupled Q-switched laser operating at 757 nm, a commercial 128-channel ultrasonic probe, a digital signal processor, and software that uses the filtered radial back-projection algorithm for image reconstruction. The system is used to reconstruct OA images of a blood-rich lesion induced in vivo in a canine prostate. OA images obtained in vivo are compared to images acquired using ultrasound, the current gold standard for guiding biopsy of the prostate. Although key structural features such as the urethra could be identified with both imaging techniques, a bloody lesion representing a highly vascularized tumor could only be clearly identified in OA images. The advantages and limitations of both forward and backward illumination modes are also evaluated by collecting OA images of phantoms simulating blood vessels within tissue. System resolution is estimated to be 0.2 mm in the radial direction of the acoustic array. The minimum detectable pressure signal is 1.83 Pa. Our results encourage further development toward a dual-modality OA/ultrasonic system for prostate imaging and image-guided biopsy.
A 3-D optoacoustic imaging system was used to visualize thermal lesions produced in vivo using high-intensity focused ultrasound (HIFU). A 7.5-MHz, surgical, focused transducer with a radius of curvature of 35 mm and an aperture diameter of 23 mm was used to generate HIFU. A pulsed laser, which could operate at 755 nm and 1064 nm, was used to illuminate excised tissue and mice using a bifurcated fiber bundle resulting in two wide beams of light. Tomographic images were obtained while the specimens were rotated within a sphere outlined by a concave arc-shaped array of 64 piezo-composite transducers. These images were then combined to reconstruct 3-D volume images (voxel resolution 0.5 mm), which were acquired before and after HIFU exposure. In vivo optoacoustic images acquired at 1064 nm provided visualization of HIFU lesions. The lesion was indicated by a negative optoacoustic contrast. The molecular nature of such contrast may possibly be associated with reduction of the optical absorption due to reduced concentration of blood, tissue dehydration, denaturation of proteins and porphyrins, and reduction of thermoacoustic efficiency in the thermally treated tissue. These preliminary results demonstrate the potential of optoacoustic imaging to assess and monitor the progress of HIFU therapy.
In this work we modified light illumination of the laser optoacoustic (OA) imaging system to improve the 3D
visualization of human forearm vasculature. The computer modeling demonstrated that the new illumination design that
features laser beams converging on the surface of the skin in the imaging plane of the probe provides superior OA
images in comparison to the images generated by the illumination with parallel laser beams. We also developed the
procedure for vein/artery differentiation based on OA imaging with 690 nm and 1080 nm laser wavelengths. The
procedure includes statistical analysis of the intensities of OA images of the neighboring blood vessels. Analysis of the
OA images generated by computer simulation of a human forearm illuminated at 690 nm and 1080 nm resulted in
successful differentiation of veins and arteries. In vivo scanning of a human forearm provided high contrast 3D OA
image of a forearm skin and a superficial blood vessel. The blood vessel image contrast was further enhanced after it was
automatically traced using the developed software. The software also allowed evaluation of the effective blood vessel
diameter at each step of the scan. We propose that the developed 3D OA imaging system can be used during
preoperative mapping of forearm vessels that is essential for hemodialysis treatment.
In this report we demonstrate improved three-dimensional optoacoustic tomography in test samples. High quality
tomographic data and images were obtained from phantom of mice being 2.5 cm in diameter. Capillaries filled with
cupric sulfate, ferrous sulfate and nickel sulfate solutions, and immersed in a scattering medium were used for these
tests. The brightness of reconstructed phantom images was found to match accurately the absorption profiles of test
solutions. Hence, optoacoustic imaging can be applied in preclinical research to perform in vivo absorptivity
measurements to deduce functional information on blood oxygen levels or concentration of contrast agents.
A 3-D optoacoustic imaging system was used to visualize thermal lesions produced in excised tissue
specimens and in vivo mice using high intensity focused ultrasound (HIFU). A 7.5 MHz surgical,
focused transducer with a radius of curvature of 35 mm and an aperture diameter of 23 mm was used
to generate HIFU. A pulsed laser, which could operate at 755 nm and 1064 nm, was used to
illuminate excised tissue and mice using a bifurcated fiber bundle resulting in two wide beams of
light. Tomographic images were obtained while the specimens were rotated within a sphere outlined
by a concave arc-shaped array of 64 piezo-composite transducers. These images were then combined
to reconstruct 3-D volume images (voxel resolution 0.5 mm), which were acquired before and after
HIFU exposure. Optoacoustic images acquired at 1064 nm provided visualization of HIFU lesions.
The lesion in excised tissue was indicated by an increase in the optoacoustic signal; the in vivo lesion
was indicated by a decrease in the optoacoustic signal. The location and the extent of the lesions
were confirmed upon dissection. The discrepancy between the ex vivo and the in vivo results might
be attributed to the different effective thermal deposition in the two cases. These preliminary results
demonstrate the potential of optoacoustic imaging to assess and monitor the progress of HIFU
therapy.
We present results from a clinical case study on imaging breast cancer using a real-time interleaved two laser
optoacoustic imaging system co-registered with ultrasound. The present version of Laser Optoacoustic Ultrasonic
Imaging System (LOUIS) utilizes a commercial linear ultrasonic transducer array, which has been modified to include
two parallel rectangular optical bundles, to operate in both ultrasonic (US) and optoacoustic (OA) modes. In OA mode,
the images from two optical wavelengths (755 nm and 1064 nm) that provide opposite contrasts for optical absorption of
oxygenated vs deoxygenated blood can be displayed simultaneously at a maximum rate of 20 Hz. The real-time aspect of
the system permits probe manipulations that can assist in the detection of the lesion. The results show the ability of
LOUIS to co-register regions of high absorption seen in OA images with US images collected at the same location with
the dual modality probe. The dual wavelength results demonstrate that LOUIS can potentially provide breast cancer
diagnostics based on different intensities of OA images of the lesion obtained at 755 nm and 1064 nm. We also present
new data processing based on deconvolution of the LOUIS impulse response that helps recover original optoacoustic
pressure profiles. Finally, we demonstrate the image analysis tool that provides automatic detection of the tumor
boundary and quantitative metrics of the optoacoustic image quality. Using a blood vessel phantom submerged in a
tissue-like milky background solution we show that the image contrast is minimally affected by the phantom distance
from the LOUIS probe until about 60-65 mm. We suggest using the image contrast for quantitative assessment of an OA
image of a breast lesion, as a part of the breast cancer diagnostics procedure.
Optoacoustic Tomography (OAT) is a hybrid imaging modality that combines the advantages of both optical
imaging and ultrasound imaging techniques. Most existing reconstruction algorithms for OAT assume pointlike
transducers, which may result in conspicous image blurring and distortions in certain applications. In this
work, a new imaging model that incorporates the transducer response is employed for image reconstruction.
Computer-simulation studies demonstrate that the new reconstruction method can effectively compensate for
image resolution degradation associated with the transducer response.
We have investigated the limitations of our laser ultrasonic plane wave δ-source. In theory, the device is capable of
producing an acoustic impulse with a bandwidth exceeding 30 MHz. However, a bandwidth of 12 MHz is measured with
a calibrated wideband hydrophone. A test setup was designed and built. It permits the investigation of experimental
parameters that alter the generated acoustic impulse: laser pulse duration, laser spatial profile, and absorber opacity.
Laser energy spatial profile is the main contributor to the narrowing of the frequency band. Our findings are presented,
along with further justifications for a device with very large effective area.
We develop a system for three-dimensional whole-body optoacoustic tomography of small animals for applications in preclinical research. The tomographic images are obtained while the objects of study (phantoms or mice) are rotated within a sphere outlined by a concave arc-shaped array of 64 piezocomposite transducers. Two pulsed lasers operating in the near-IR spectral range (755 and 1064 nm) with an average pulsed energy of about 100 mJ, a repetition rate of 10 Hz, and a pulse duration of 15 to 75 ns are used as optical illumination sources. During the scan, the mouse is illuminated orthogonally to the array with two wide beams of light from a bifurcated fiber bundle. The system is capable of generating images of individual organs and blood vessels through the entire body of a mouse with spatial resolution of ~0.5 mm.
We demonstrated the ability to detect surface antigens, associated with pathogens, utilizing laser optoacoustic
spectroscopy with antibody-coupled gold nanorods (GNR) as a contrast agent specifically targeted to the antigen of
interest. The sensitivity of the technique has been assessed by determining the minimum detectable concentration of a
surface antigen from biological samples. We compared the sensitivity and applicability of two different methods for
detecting optoacoustic responses, using either optical or piezoelectric sensors. Biological samples were adsorbed to the
inside walls of detachable, flat-bottomed plastic micro-wells, and then probed with appropriate antibodies conjugated
with gold nanorods. If the target antigens were present, the antibody-nanoparticle conjugates were bound, while any
nonadsorbed nanoparticles were washed out of the wells. Optoacoustic signals were generated from the bound nanorods
using a pulsed pump laser at wavelengths corresponding to one of the peak absorptions of the nanorods. Optoacoustic
responses were obtained from the samples using both detection modalities. The sensitivity, suitability, ease of use of
each method were assessed and compared. Initial results indicate that optical detection gives comparable sensitivity as
the piezoelectric method, and further enhancement of the detection sensitivity is possible for both methods. An
advantage of the piezoelectric detection method is that it may be implemented in a more compact assembly, compared to
the optical method, however, the optical method may be less sensitive to external electromagnetic and acoustic noise.
Further evaluation will be required to refine these measurements. The results with both methods indicate that the use of
antibody-targeted nanorod contrast agents, with laser-optoacoustic detection, is a promising technology for the
development of rapid in vitro diagnostic tests.
KEYWORDS: Transducers, Acoustics, Sensors, Calibration, Wavefronts, Ultrasonography, Signal attenuation, Pulsed laser operation, Image restoration, Signal to noise ratio
Comprehensive characterization of optoacoustic transducers is achieved through the analysis of their frequency
response using a procedure of measuring angular dependence of the transducer sensitivity to the ultrawide-band
delta pulse. The testing was performed under standard repeatable operating conditions. Back-illumination of a
blackened, acoustically matched, planar surface with a short laser pulse creates an acoustic impulse which was used
as an ultrawide-band ultrasonic source. The bandwidth of such a source extends well over 10 MHz (6dB point at 16
MHz for illumination with a 16 ns pulse) and the low frequency
roll-off is around 300 kHz. Analysis of the angular
dependence of the frequency response yields invaluable directivity information about the detector under study,
which in turn permits accurate forward and inverse problem models.
We developed a 3D whole-body optoacoustic tomography system for applications in preclinical research on mice. The
system is capable of generating images with resolution better than 0.6 mm. Two pulsed lasers, an Alexandrite laser
operating at 755 nm and a Nd:YAG laser operating at 532 nm and 1064nm were used for light delivery. The
tomographic images were obtained while the objects of study (phantoms or mice) were rotated within a sphere outlined
by a concave arc-shaped array of 64 piezo-composite transducers. During the scan, the mouse was illuminated
orthogonally to the array with two wide beams of light from a bifurcated fiber bundle. Illumination at 532 nm showed
superficial vasculature, but limited penetration depth at this wavelength prevented the detection of deeper structures.
Illumination at 755 and 1064 nm showed organs and blood vessels, respectively. Filtering of the optoacoustic signals
using high frequency enhancing wavelets further emphasized the smaller blood vessels.
We describe two laser optoacoustic imaging systems for breast cancer detection based on arrays of acoustic detectors
operated manually in a way similar to standard ultrasonic breast imaging. The systems have the advantages of standard
light illumination (regardless of the interrogated part of the breast), the ability to visualize any part of the breast, and
convenience in operation. The first system could work in both ultrasonic and optoacoustic mode, and was developed
based on a linear ultrasonic breast imaging probe with two parallel rectangular optical bundles. We used it in a pilot
clinical study to provide for the first time demonstration that the boundaries of the tumors visualized on the optoacoustic
and ultrasonic images matched. Such correlation of coregistered images proves that the objects on both images
represented indeed the same tumor. In the optoacoustic mode we were also able to visualize blood vessels located in the
neighborhood of the tumor. The second system was proposed as a circular array of acoustic transducers with an axisymmetric
laser beam in the center. It was capable of 3D optoacoustic imaging with minimized optoacoustic artifacts
caused by the distribution of the absorbed optical energy within the breast tissue. The distribution of optical energy
absorbed in the bulk tissue of the breast was removed from the image by implementing the principal component analysis
on the measured signals. The computer models for optoacoustic imaging using these two handheld probes were
developed. The models included three steps: (1) Monte Carlo simulations of the light distribution within the breast
tissue, (2) generation of optoacoustic signals by convolving
N-shaped pressure signals from spherical voxels with the
shape of individual transducers, and (3) back-projecting processed optoacoustic signals onto spherical surfaces for image
reconstruction. Using the developed models we demonstrated the importance of the included spatial impulse response of
the optoacoustic imaging system.
Ultrasound imaging is the current gold standard for guiding biopsy of prostate. Optoacoustic imaging yields higher
contrast in detection of malignant tissues. The two techniques provide complementary information. We are currently
developing a hybrid laser optoacoustic and ultrasound imaging system for prostate tumor detection (LOUIS-P). The
optoacoustic part consists of a fiber-coupled Q-switched laser operating at either 757 nm or 1064 nm attached to a
commercially-available 128-channel ultrasonic probe modified for optimal detection of optoacoustic signals, a digital
signal processor with 128 independent channels, and software that uses the radial (filtered) backprojection algorithm to
reconstruct tomographic images. We evaluated system-imaging performance using test objects submerged in milky
water, and poly(vinyl-chloride) plastisol tissue phantoms simulating malignant lesions. LOUIS-P demonstrates potential
as a clinical technique for minimally invasive imaging and diagnosis of prostate cancer.
Optoacoustic imaging, a novel noninvasive modality, combines the advantages of optical methods and the ultrasound
technique. The optoacoustic technique is based on tissue irradiation with nanosecond laser pulses and detection of ultrasound waves generated due to thermo-elastic expansion. Using a modified Monte Carlo technique and solution of wave equation for velocity potential, we modeled optoacoustic signals from cylindrical blood vessels with varying oxygenation and varying total hemoglobin concentration. A specially designed computer code was used for reconstruction of images of absorbed energy in the blood vessels and surrounding tissues. Then we performed a set of experiments with our optoacoustic system and phantoms that simulate blood vessels such as veins and arteries at depths of up to 2 cm. The optoacoustic signals from the phantoms were used for reconstruction of 2-D cross-section images and
correlated well with geometry and optical properties of the phantoms. The obtained data suggest that the developed optoacoustic imaging approach can be used for accurate mapping of blood oxygenation and hemoglobin concentration in blood vessels.
We present a rapid, robust method of signal processing useful for optoacoustic monitoring of total hemoglobin concentration ([THb]) and oxygen saturation level in small blood vessels. Our method includes the wavelet-based regularization of the difference operator which is a typical discrete approximation of the derivative. The optimal degree of regularization is defined by the signal-to-noise ratio (SNR). We applied the proposed method to Monte Carlo-modeled signals from a cylinder simulating the human radial artery (diameter 1.6 mm, depth from skin 2 mm, and [THb] varied in a wide range from 4 - 16 g/dL). We obtained N-shaped signals and found that the maximum of the first derivative between the front and rear walls systematically correlates with the actual value of [THb]. We estimated the accuracy of [THb] reconstruction from the maximum of the first derivative as 0.32 ± 0.18 g/dL (mean value ± SD) at an SNR typical for our in vivo experiments at the wavelength of 1064 nm. We also demonstrated that the difference between the maxima of the first derivative of the signals obtained at 700 nm and 1000 nm depends on oxygen saturation level.
Continuous monitoring of cerebral blood oxygenation is critically important for successful treatment of patients with severe traumatic brain injury. At present, the techniques for monitoring blood oxygenation are invasive. Recently we proposed noninvasive monitoring of cerebral blood oxygenation by using optoacoustic probing of blood circulating in the internal jugular vein (IJV). A major source of error in the optoacoustic measurement with a single-element optoacoustic probe is the spatial misalignment between the probe and the IJV. We built a LabView®-based scanning system that automatically moves our optoacoustic probe across the IJV while continuously taking measurements. Automatic signal processing determines the signal with the best probe-vessel alignment which then is used for further processing. The scanning system was tested in phantoms using solutions with different absorption coefficients and with blood with various levels of blood oxygenation. Amplitudes and profiles of the optoacoustic signals recorded from the phantoms closely followed the blood oxygenation changes in accordance with blood optical properties. These data indicate that the scanning system is capable of improving the accuracy of non-invasive monitoring of blood oxygenation by minimizing errors associated with lateral misalignment of the probe with respect to blood vessels.
The mechanical characteristics of the outer hair cells (OHCs) plasma membrane was studied by forming tethers with optical tweezers. The average force to and from a plasma membrane tether at the lateral wall of the OHC was about 3.5 times greater than that at the basal end of the cell, consistent with the presence of a more extensive cytoskeleton support beneath the PM at the site of the lateral wall. The apparent viscosity of the PM was measured by pulling tethers at different rates while continuously recording the tether force, and was estimated in the range of 13-33 pN*s/micrometers .
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