Ladislav Jankovic

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/cm2, which is 1/7 of the American National Standards Institute (ANSI) safety limit (20 mJ/cm2). 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.

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.

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.

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.

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.

Raman gain measurements in bulk glass samples

Increased interest in highly nonlinear glasses for use as Raman amplifiers has encouraged the development of an experimental apparatus to measure the material Raman gain coefficient on millimeter-thick bulk glass samples. Apparatus design considerations, details of the apparatus, and justification for the data analysis employed are provided. The apparatus is a powerful tool offering the ability to quickly screen glass samples over a wide range of compositions without the time and cost of fiberizing candidate materials into a guiding geometry to directly measure Raman gain.

Real-time photoacoustic data acquisition with Philips iU22 ultrasound scanner

A one of a kind photoacoustic system has been built around a Philips iU22 ultrasound scanner. The modified channel board architecture allows access to the raw per-channel photoacoustic data, while keeping all of the imaging capabilities of an actual commercial ultrasound scanner. A captured photoacoustic data frame is Fourier beamformed to generate a single laser shot photoacoustic image. In addition to the photoacoustic data, the system supplies the beamformed ultrasound data, providing a truly dual-modality imaging capability. A tunable OPO laser system (700-900nm), pumped by an Nd:YAG solid state laser, is used as an illumination source with 5ns long pulses. An FPGA-based electronic board synchronizes the iU22 start of frame with the laser firing, currently permitting photoacoustic imaging at a rate of 10 Hz (laser repetition rate limit). At that imaging frame rate the photoacoustic system, consisting of a PC modified with 32 Gbytes of acquisition memory and an FPGA array, is able to store several minutes of continuously captured data, enabling monitoring and off-line analysis of dynamic photoacoustic events and/or fast scanning for performing pseudo-3D imaging. The system can use all of the standard iU22 array transducers both for photoacoustic imaging, and in all of the ultrasound imaging modes.

Three-dimensional photoacoustic imaging with a clinical two-dimensional matrix ultrasound transducer

Photoacoustic tomography provides both structural and functional imaging in vivo based on optical absorption contrast. A novel imaging system that incorporates a two-dimensional matrix ultrasound probe for combined photoacoustic and ultrasonic three-dimensional (3D) volumetric imaging is presented. The system consists of a tunable dye laser pumped by a Nd:YAG laser, a commercial ultrasound imaging system (Philips iU22) with a two-dimensional matrix transducer (Philips X7-2, 2500 elements, 2-7 MHz), and a multichannel data acquisition system which allows us to acquire RF channel data. Compared with alternative 3D techniques, this system is attractive because it can generate co-registered 3D photoacoustic and ultrasound images without mechanical scanning. Moreover, the lateral resolution along the azimuth and elevational directions are measured to be 0.77 ± 0.06 mm and 0.96 ± 0.06 mm, respectively, based on reconstructed photoacoustic images of phantoms containing individual human hairs. Finally, in vivo 3D photoacoustic sentinel lymph node mapping using methylene blue dye in a rat model is demonstrated.

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.