Tumor angiogenesis, O2 saturation, glucose and amino acid metabolisms study using functional imaging

This research is primarily focused on the study of tumors in experimental animal models using functional imaging in the presence of various contrast agents. The study of malignant tumor angiogenesis, oxygen saturation, glucose and amino acid metabolisms will lead to better methods for cancer detection as well as diagnosing and managing cancer. Non invasive in vivo diagnostic imaging technique is an area of great clinical interest in present days. In this study, noninvasive in vivo photoacoustic tomography and conventional fluorescence imaging together with multiphoton microscopic tomography were implemented to study the malignant tumor morphology and physiology. Tumor structure and angiogenesis were successfully imaged by photoacoustic tomography and conventional fluorescence imaging. The important malignant tumor cellular parameters such as oxygen saturation and αvβ3 integrin concentration were measured in living small animals (rodents) using the novel photoacoustic tomography technique. By implementing multiphoton microscopy using Cy3.5 NHS ester contrast agent, tumor amino acid metabolism was successfully studied in cell culture. This method will at least give you a relative concentration map of amino acid in cells. Non invasive in vivo imaging can be achieved by modifying the current multiphoton imaging setup. A new method for studying amino acid and glucose metabolisms of tumor cells using multiphoton imaging was developed

High-resolution spectroscopic photoacoustic tomography for non-invasive functional imaging of small-animal brains in vivo

Based on the multiwavelength laser-based photoacoustic tomography, noninvasive imaging of cerebral blood oxygenation and blood volume in small-animal brains in vivo was realized. The high sensitivity of this technique is based on the spectroscopic differences between oxy- and deoxy-hemoglobins whereas its spatial resolution is diffraction-limited by the photoacoustic signals. The point-by-point distributions of hemoglobin oxygen saturation and total concentration of hemoglobin in the cerebral cortical venous vessels, altered by systemic physiological modulations including hyperoxia and hypoxia, were visualized successfully through the intact skin and skull. This technique can potentially accelerate the progress in neuroscience and provide important new insights into cerebrovascular physiology and brain function

Deep penetrating photoacoustic tomography in biological tissues

Photoacoustic tomography (PAT) in a circular scanning configuration was developed to image the deeply embedded optical heterogeneity in biological tissues. Based on the intrinsic contrast between blood and chicken breast muscle, an embedded blood object that was 5 cm deep in the tissue was detected using pulsed laser light at a wavelength of 1064 nm. Compared with detectors for flat active surfaces, cylindrically focused ultrasonic transducers can reduce the interference generated from the off-plane photoacoustic sources and make the image in the scanning plane clearer. While the optical penetration was optimized with near-infrared laser pulses of 800 nm in wavelength, the optical contrast was enhanced by indocyanine green (ICG) whose absorption peak matched the laser wavelength. This optimized PAT was able to image fine objects embedded at a depth of up to 5.2-cm, which is 6.2 times the 1/e optical penetration depth, in chicken breast muscle, at a resolution of < ~750 microns with a sensitivity of <7 pmol of ICG in blood. The resolution was found to deteriorate slowly with increasing imaging depth

High-resolution photoacoustic tomography

Optical contrast is sensitive to physiological parameters, such as the oxygen saturation and total concentration of hemoglobin, in biological tissues. Photoacoustic tomography is based on the high optical contrast yet utilizing the high ultrasonic resolution. Our work in this emerging area of research will be summarized in this invited talk. In this technology, a diffraction-based inverse-source problem is solved in the image reconstruction, for which we developed the rigorous reconstruction theory. We implemented a prototype and accomplished non-invasive transdermal and transcranial functional imaging of small-animal brains in vivo. Changes in the cerebral blood oxygenation and blood volume of a rat, as a result of the alternation from hyperoxia to hypoxia, were imaged successfully

High-resolution ultrasound-aided biophotonic imaging

Optical contrast is sensitive to functional parameters, including the oxygen saturation and total concentration of hemoglobin, in biological tissues. However, due to the overwhelming scattering encountered by light in tissues, traditional optical modalities cannot provide satisfactory spatial resolution beyond the ballistic (a few hundred microns) and quasiballistic (1-2 mm) regimes. Photoacoustic tomography is based on the high optical contrast yet utilizing the high ultrasonic resolution. Our work in this emerging area of research will be summarized in this invited talk. In this technology, a diffraction-based inverse-source problem is solved in the image reconstruction, for which we developed the rigorous reconstruction theory. We implemented a prototype and accomplished noninvasive transdermal and transcranial functional imaging of small-animal brains in vivo. Change in the cerebral blood oxygenation of a rat, as a result of the alternation from hyperoxia to hypoxia, was imaged successfully

Functional photoacoustic tomography for non-invasive imaging of cerebral blood oxygenation and blood volume in rat brain in vivo

Based on the multi-wavelength laser-based photoacoustic tomography, non-invasive in vivo imaging of functional parameters, including the hemoglobin oxygen saturation and the total concentration of hemoglobin, in small-animal brains was realized. The high sensitivity of this technique is based on the spectroscopic differences between oxy- and deoxy-hemoglobin while its spatial resolution is bandwidth-limited by the photoacoustic signals rather than by the optical diffusion as in optical imaging. The point-by-point distributions of blood oxygenation and blood volume in the cerebral cortical venous vessels, altered by systemic physiological modulations including hyperoxia, normoxia and hypoxia, were visualized successfully through the intact skin and skull. This technique, with its prominent intrinsic advantages, can potentially accelerate the progress in neuroscience and provide important new insights into cerebrovascular physiology and brain function that are of great significance to the neuroscience community

Noninvasive imaging of hemoglobin concentration and oxygenation in the rat brain using high-resolution photoacoustic tomography

Simultaneous transcranial imaging of two functional parameters, the total concentration of hemoglobin and the hemoglobin oxygen saturation, in the rat brain in vivo is realized noninvasively using laser-based photoacoustic tomography (PAT). As in optical diffusion spectroscopy, PAT can assess the optical absorption of endogenous chromophores, e.g., oxygenated and deoxygenated hemoglobins, at multiple optical wavelengths. However, PAT can provide high spatial resolution because its resolution is diffraction-limited by photoacoustic signals rather than by optical diffusion. Laser pulses at two wavelengths are used sequentially to acquire photoacoustic images of the vasculature in the cerebral cortex of a rat brain through the intact skin and skull. The distributions of blood volume and blood oxygenation in the cerebral cortical venous vessels, altered by systemic physiological modulations including hyperoxia, normoxia, and hypoxia, are visualized successfully with satisfactory spatial resolution. This technique, with its prominent sensitivity to endogenous contrast, can potentially contribute to the understanding of the interrelationship between neural, hemodynamic, and metabolic activities in the brain

In vivo functional photoacoustic imaging of brain tumor vasculature

We present a study of the functional photoacoustic imaging of tumor hypoxia in mice in vivo. Based on spectroscopic photoacoustic tomography that detects the optical absorption of oxy- and deoxy-hemoglobins, the blood oxygen saturation and the vascularization of brain tumors were visualized. U87 glioblastoma tumor cells were inoculated intracranially at a 3-mm depth from the surface of the nude mouse head seven days before the experiment. Increased blood content and hypoxia inside the tumor vasculature were detected through the intact skin and skull. This technique will be useful for future studies on tumor metabolic activities in the brain and hypoxia-related therapeutic resistance

In vivo imaging and characterization of hypoxia-induced neovascularization and tumor invasion

Hypoxia is a critical event in tumor progression and angiogenesis. Hypoxia can be detected noninvasively by a novel spectroscopic photoacoustic tomography technology (SPAT) and this finding is supported by our molecular biology investigation aimed to elucidate the etiopathogenesis of SPAT detected hypoxia and angiogenesis. The present study provides an integrated approach to define oxygen status (hypoxia) of intracranial tumor xenografts using spectroscopic photoacoustic tomography. Brain tumors can be identified based on their distorted vascular architecture and oxygen saturation (SO2) images. Noninvasive in vivo tumor oxygenation imaging using SPAT is based on the spectroscopic absorption differences between oxyhemoglobin (O2Hb) and deoxyhemoblobin (HHb). Sprague-Dawley rats inoculated intracranially with ENU1564, a carcinogen-induced rat mammary adenocarcinoma cell line, were imaged with SPAT three weeks post inoculation. Proteins important for tumor angiogenesis and invasion were detected in hypoxic brain foci identified by SPAT and were elevated compared with control brain. Immunohistochemistry, Western blotting, and semi-quantitative RT-PCR showed that HIF-1 α, VEGF-A, and VEGFR2 (Flk-1) protein and mRNA expression levels were significantly higher (P<0.05) in brain tumor tissues compared to normal brain. Gelatin zymography and RT-PCR demonstrated the upregulation of MMP-9 in tumor foci compared with brain control. Together these results suggest the critical role of hypoxia in driving tumor angiogenesis and invasion through upregulation of target genes important for these functions. Moreover this report validates our hypothesis that a novel noninvasive technology (SPAT) developed in our laboratory is suitable for detection of tumors, hypoxia, and angiogenesis