In vivo three-dimensional photoacoustic imaging based on a clinical matrix array ultrasound probe.

We present an integrated photoacoustic and ultrasonic three-dimensional (3-D) volumetric imaging system based on a two-dimensional (2-D) matrix array ultrasound probe. A wavelength-tunable dye laser pumped by a Q-switched Nd:YAG laser serves as the light source and a modified commercial ultrasound imaging system (iU22, Philips Healthcare) with a 2-D array transducer (X7-2, Philips Healthcare) detects both the pulse-echo ultrasound and photoacoustic signals. A multichannel data acquisition system acquires the RF channel data. The imaging system enables rendering of co-registered 3-D ultrasound and photoacoustic images without mechanical scanning. The resolution along the azimuth, elevation, and axial direction are measured to be 0.69, 0.90 and 0.84 mm for photoacoustic imaging. In vivo 3-D photoacoustic mapping of the sentinel lymph node was demonstrated in a rat model using methylene blue dye. These results highlight the clinical potential of 3-D PA imaging for identification of sentinel lymph nodes for cancer staging in humans.

Performance characterization of an integrated ultrasound, photoacoustic, and thermoacoustic imaging system.

We developed a novel trimodality system for human breast imaging by integrating photoacoustic (PA) and thermoacoustic (TA) imaging techniques into a modified commercial ultrasound scanner. Because light was delivered with an optical assembly placed within the microwave antenna, no mechanical switching between the microwave and laser sources was needed. Laser and microwave excitation pulses were interleaved to enable PA and TA data acquisition in parallel at a rate of 10 frames per second. A tube (7 mm inner diameter) filled with oxygenated bovine blood or 30 mM methylene blue dye was successfully detected in PA images in chicken breast tissue at depths of 6.6 and 8.4 cm, respectively, for the first time. The SNRs at these depths reached ∼24 and ∼15 dB, respectively, by averaging 200 signal acquisitions. Similarly, a tube (13 mm inner diameter) filled with saline solution (0.9%) at a depth of 4.4 cm in porcine fat tissue was successfully detected in TA images. The PA axial, lateral, and elevational resolutions were 640 µm, 720 µm, and 3.5 mm, respectively, suitable for breast cancer imaging. A PA noise-equivalent sensitivity to methylene blue solution of 260 nM was achieved in chicken tissue at a depth of 3.4 cm.

Performance benchmarks of an array-based hand-held photoacoustic probe adapted from a clinical ultrasound system for non-invasive sentinel lymph node imaging.

Clinical translation of photoacoustic (PA) imaging can be facilitated by integration with commercial ultrasound (US) scanners to enable dual-modality imaging. An array-based US scanner was modified for hand-held PA imaging. The performance was benchmarked in terms of signal-to-noise ratio (SNR), axial spatial resolution and sensitivity. PA images of a tube, filled with methylene blue (MB; approx. 30 mM) and placed at various depths in chicken tissue, were acquired. A 5 cm penetration depth was achieved with an 18.6 dB SNR using a laser fluence of 3 mJ cm(-2), only one-seventh of the safety limit (20 mJ cm(-2)). An axial resolution of approximately 400 µm was maintained at all imaging depths. The PA sensitivity to MB placed 2.3 cm deep in chicken tissue was less than 100 µM. Further, after intradermal injection of MB (approx. 30 mM), a rat sentinel lymph node was clearly identified in vivo, beneath a 3.8 cm thick layer of chicken breast. The accumulated concentration of MB in the node was estimated to be approximately 7 mM. The noise-equivalent sensitivities (approx. 2 cm depth) were 17 and 85 µM, ex vivo and in vivo, respectively. These results support the use of this PA system for non-invasive mapping and image-guided needle biopsy of sentinel nodes in breast cancer patients.

Handheld array-based photoacoustic probe for guiding needle biopsy of sentinel lymph nodes.

By modifying a clinical ultrasound array system, we develop a novel handheld photoacoustic probe for image-guided needle biopsy. The integration of optical fiber bundles for pulsed laser light delivery enables photoacoustic image-guided insertion of a needle into rat axillary lymph nodes with accumulated indocyanine green (ICG). Strong photoacoustic contrast of the needle is achieved. After subcutaneous injection of the dye in the left forepaw, sentinel lymph nodes are easily detected, in vivo and in real time, beneath 2-cm-thick chicken breast overlaying the axillary region. ICG uptake in axillary lymph nodes is confirmed with fluorescence imaging both in vivo and ex vivo. These results demonstrate the clinical potential of this handheld photoacoustic system for facile identification and needle biopsy of sentinel lymph nodes for cancer staging and metastasis detection in humans.

Sentinel lymph nodes in the rat: noninvasive photoacoustic and US imaging with a clinical US system.

PURPOSE: To evaluate in vivo sentinel lymph node (SLN) mapping by using photoacoustic and ultrasonographic (US) imaging with a modified clinical US imaging system. MATERIALS AND METHODS: Animal protocols were approved by the Animal Studies Committee. Methylene blue dye accumulation in axillary lymph nodes of seven healthy Sprague-Dawley rats was imaged by using a photoacoustic imaging system adapted from a clinical US imaging system. To investigate clinical translation, the imaging depth was extended up to 2.5 cm by adding chicken or turkey breast on top of the rat skin surface. Three-dimensional photoacoustic images were acquired by mechanically scanning the US transducer and light delivery fiber bundle along the elevational direction. RESULTS: Photoacoustic images of rat SLNs clearly help visualization of methylene blue accumulation, whereas coregistered photoacoustic/US images depict lymph node positions relative to surrounding anatomy. Twenty minutes following methylene blue injection, photoacoustic signals from SLN regions increased nearly 33-fold from baseline signals in preinjection images, and mean contrast between SLNs and background tissue was 76.0 +/- 23.7 (standard deviation). Methylene blue accumulation in SLNs was confirmed photoacoustically by using the optical absorption spectrum of the dye. Three-dimensional photoacoustic images demonstrate dynamic accumulation of methylene blue in SLNs after traveling through lymph vessels. CONCLUSION: In vivo photoacoustic and US mapping of SLNs was successfully demonstrated with a modified clinical US scanner. These results raise confidence that photoacoustic and US imaging can be used clinically for accurate, noninvasive imaging of SLNs for axillary lymph node staging in breast cancer patients.

Deeply penetrating in vivo photoacoustic imaging using a clinical ultrasound array system.

Using a hand-held photoacoustic probe integrated with a clinical ultrasound array system, we successfully imaged objects deeply positioned in biological tissues. The optical contrasts were enhanced by methylene blue with a concentration of ~30 mM. The penetration depth reached ~5.2 cm in chicken breast tissue by using 650-nm wavelength, which is ~4.7 times the 1/e optical penetration depth. This imaging depth was achieved using a laser fluence on the tissue surface of only 3 mJ/cm(2), which is 1/7 of the American National Standards Institute (ANSI) safety limit (20 mJ/cm(2)). The noise equivalent sensitivity at this depth was ~11 mM. Further, after intradermal injection of methylene blue in a rat, a sentinel lymph node was easily detected in vivo, beneath a 2-cm thick layer of chicken breast. Also, blood located 3.5 cm deep in the rat was clearly imaged with intrinsic contrast. We have photoacoustically guided insertion of a needle into a rat sentinel lymph node with accumulated methylene blue. These results highlight the clinical potential of photoacoustic image-guided identification and needle biopsy of sentinel lymph nodes for axillary staging in breast cancer patients.

Acoustic fingerprints of dye-labeled protein submicrosphere photoacoustic contrast agents.

Dye-labeled protein microspheres, submicron in size and capable of producing thermoelastically generated ultrasound in response to laser stimulation, are presented as contrast agents for photoacoustic imaging. Incident laser energy absorbed by fluorescein isothiocyanate (FITC)-labeled elastin submicrospheres results in thermoelastically generated sound production. Plotted A-line graphs reveal a distinctive morphology and a greater than two orders of magnitude increase in signal amplitude subsequent to converting FITC elastin into submicrospheres (despite a four orders of magnitude decrease in concentration). Evidence of nonlinearity and enhancement of ultrasound backscatter indicate a potential use in contrast-enhanced harmonic imaging. Photoacoustic and ultrasound imaging of FITC-elastin submicrospheres in a water-filled phantom vessel shows enhanced contrast at low concentration and clear delineation of the phantom vessel wall.

Quadratic soliton collisions.

The details of two soliton collision processes were investigated in detail in a 1 cm long periodically poled KTP crystal for the case when the solitons were excited by inputting only the fundamental beam. The effects on the collision outcomes of the distance of the collision into the sample, collision angle and phase mismatch were measured for different relative phases between the input beams. At small angles (around 0.4 degrees ) fusion, repulsion and energy transfer processes were observed, while at the collision angles approaching 3.2 degrees the two output soliton beams were essentially unaffected by the interaction. The phase mismatch was varied from 3.5 to -1.5pi for the 0.4 degrees collision angle case. The output solitons separation at pi input phase difference showed strongly asymmetric behavior with phase mismatch. In general, the measurements indicate a decrease in the interaction strength with increasing phase mismatch. All collision processes were performed in the vicinity of a non-critical phase matching.

Quadratic soliton self-reflection at a quadratically nonlinear interface.

The reflection of bulk quadratic solutions incident onto a quadratically nonlinear interface in periodically poled potassium titanyl phosphate was observed. The interface consisted of the boundary between two quasi-phase-matched regions displaced from each other by a half-period. At high intensities and small angles of incidence the soliton is reflected.

Weak beam control of multiple quadratic soliton generation.

Theoretical predictions and experimental observations show that a weak harmonic seed beam can control the process of generating multiple quadratic solitons in periodically poled KTiOPO4 by inputting only a fundamental beam. This all-optical switching does not depend on the relative phase of the input beams, an unusual property for such coherent solitons.

Tellurite glasses with peak absolute Raman gain coefficients up to 30 times that of fused silica.

An experimental system has been assembled to measure the absolute values of the Raman gain spectrum for millimeter-thick glass samples. Results are reported for two new oxide glasses with Raman gain coefficients as much as 30 times larger than that of fused silica and more than twice its spectral coverage.

Birth of solitons in quadratic spatial soliton collisions.

We demonstrate experimentally and theoretically the creation of a third soliton in two soliton collision processes in type I noncritically phase-matched KNbO3. The output pattern in the collision process is phase dependent, but the total energy and the relative ratio of the fundamental to the second harmonic in each soliton remain essentially unchanged to within experimental accuracy.

Observation of multiple soliton generation mediated by amplification of asymmetries.

We report the experimental observation of the formation of multiple optical quadratic solitons in a process mediated by the amplification of minute asymmetries in the diffraction properties of the input light. Experiments were conducted in phase-matched second-harmonic generation in a bulk crystal of periodically poled potassium titanyl phosphate pumped at 1064 nm. The different mechanisms that influence the process were investigated numerically, and the pulsed nature of the pump light was found to play a key role in the observed light distributions.

Quadratic spatial solitons in periodically poled KTiOPO4.

The evolution of fundamental beams into quadratic spatial solitons was investigated in a periodically poled bulk KTiOPO4 crystal near and at phase matching. Various soliton properties, such as trapping efficiency as a function of phase mismatch, poling-induced walk-off, and the effects of pulse width, beam width, and input beam quality, were measured. Effects attributed to the presence of cubic nonlinearities were also observed.

Properties of quadratic multi-soliton generation near phase-match in periodically poled potassium titanyl phosphate.

The properties of the multi-quadratic-soliton generation process have been investigated both theoretically and experimentally near and on phase-match in non-critically-phase-matched, periodically poled, potassium titanyl phosphate (PPKTP). It was found that multi-soliton generation occurs primarily due to asymmetry in the input beam and at phase-matching. The number of solitons generated depended on the input intensity in a non-trivial way.

Complex soliton-like pattern generation in Potassium Niobate due to noisy, high intensity, input beams.

We report the generation of complex soliton-like patterns in noncritically-phase-matched potassium niobate which occur in random spatial patterns from shot-to-shot. Up to five spots have been generated at input intensities of 10's GW/cm2, many times the single soliton threshold. The mechanism which leads to the symmetry breaking required for the complex patterns is interpreted to be random noise imprinted on the input light.

Two-dimensional type I quadratic spatial solitons in KNbO(3) near noncritical phase matching.

Experiments on the properties of quadratic, two-dimensional spatial soliton properties in KNbO(3) near and at noncritical phase matching (NCPM) are reported. The NCPM geometry leads to unique features such as a large angular bandwidth for soliton generation, weak dependence of soliton composition on intensity and incidence angle, and unique multisoliton-generation properties.

Effects of anisotropic diffraction on quadratic multisoliton excitation in noncritically p hase-matched crystals.

The generation of multiple quadratic spatial solitons aligned approximately along a crystal axis in a biaxial noncritically phase-matched crystal is shown theoretically and experimentally to be the consequence of anisotropic diffraction.