Bruce J. Tromberg - US grants

Optical techniques are used extensively in medicine to address a broad variety of disorders. Their impact has expanded from the traditional areas of dermatology and ophthalmology to include gynecology, oncology and cardiology, and, more recently dentistry, radiology, and anesthesiology. Consequently, the relationship between tissue optical properties and photo-medicine is tantamount to the importance of pharmacokinetics in clinical management. More specifically, rapid, accurate determination of tissue optical properties is essential to predicting photon dosimetry during therapeutic procedures, and interpreting the information content of back-scattered or transmitted light during diagnostic studies. Accordingly, we propose the development of frequency-domain photon migration (FDPM), a new optical method for non-invasive, quantitative assessment of tissue absorption and scattering. Since the underlying principle behind FDPM involves the propagation of diffuse photon density waves, this proposal addresses fundamental aspects of density-wave behavior in biological tissues. Our approach incorporates a balanced, interactive mix of theory and experiment. The conceptual framework is based on application of the diffusion approximation to light transport in turbid media. Relatively simple analytical solutions to the photon diffusion equation will be developed for a variety of tissue-like sample geometries. The validity of these equations will be tested in real and simulated tissues, and our experimental results will be used to highlight model limitations. The conceptual framework will be applied in a similar manner to fully describe experimental constraints. Modifications to the instrumentation, conceptual framework, and mathematical structure will, by design, occur in rapid fashion. For example, current photon density wave theory suggests that typical tissue measurements require bandwidths higher than the 250 MHz available with our existing instrument. We therefore propose expanding our system to approximately 500 MHz during the first grant year. High-bandwidth FDPM results will be compared to optical property values derived from conventional methods. Ultra-structural analyses will be performed on tissues in order to identify biological determinants of tissue optical properties. In this manner, a general model for the influence of instrumental parameters, optical properties, and geometrical boundaries on photon density waves will be developed. We expect this fundamental information will have a vital impact on nearly all aspects of photomedicine, including: 1) defining the utility, future applications and design considerations for optical diagnostic devices, 2) characterizing the limitations of current tissue light- transport models and 3) accounting for the numerous disparities between literature-reported tissue optical property values.

DESCRIPTION (provided by applicant): The Laser Microbeam and Medical Program (LAMMP) is a NIH Biomedical Technology Resource Center dedicated to the use of lasers and optics in biology and medicine. LAMMP is located within the Beckman Laser Institute (BLI), an interdisciplinary biomedical research, teaching, and clinical facility at the University of California, Irvine. Overall resource objectives are to promote a well-balanced Center with activities in technological research and development, collaborative research, service, training, and dissemination. In this sixth renewal application of LAMMP, we continue to emphasize our unique capabilities to facilitate "translational" research by rapidly moving basic science and technology discoveries from "blackboard to benchtop to bedside". This is accomplished by combining state of the art optical technologies with specialized resource facilities for cell and tissue engineering, histopathology, pre-clinical animal models, and clinical care. LAMMP provides both Microscopy and Microbeam Technologies (MMT) for high-resolution functional imaging and manipulation of living cells and tissues and Medical Translational Technologies (MTT) for non- and minimally-invasive monitoring, treating, and imaging pre-clinical animal models and human subjects. In addition, we propose to establish a new core, Virtual Photonics Technologies (VPT) for developing computational models and methods that advance the performance of biophotonic technologies, and enhance the information content derived from optical measurements. Seven Technology Research and Development projects are proposed that will result in the fabrication of several new instruments as well as the creation of multi-functional software. These projects build on our longstanding expertise in light-tissue interaction models and Biophotonics technologies, including laser microbeams, non-linear microscopy, optical coherence tomography, spatial/temporal modulation methods, and diffuse optics. LAMMP cores contain complementary technologies that are capable of quantitatively characterizing, imaging, and perturbing structure and biochemical function in cells and tissues with scalable resolution and depth sensitivity ranging from micrometers to centimeters. Collaborative studies are proposed that advance these technologies so they become widely-available, enabling methods in Biology and Medicine. Throughout the grant we emphasize the relevance of LAMMP technologies to Medicine in areas such as cancer, cardiovascular disease, metabolic syndrome, and neurologic function, as well as fundamental biological process, such as mechano-transduction, wound repair, angiogenesis, fibrosis, and cell death.

technology /technique development; lasers; education; biomedical resource;

technology /technique development; animal tissue; lasers; model design /development; biomedical resource; biological products;

microscopy; education; biomedical resource;

health care; technology /technique development; lasers; biomedical equipment development; Mammalia; bioengineering /biomedical engineering; biomedical resource; behavioral /social science research tag;

technology /technique development; animal tissue; microscopy; lasers; model design /development; genetics; biomedical resource; bioengineering /biomedical engineering;

Frequency-Domain Photon Migration (FDPM) is a non-invasive optical technique which utilizes near-infrared light to monitor physiology in bulk tissues. Optical properties derived from FDPM measurements can be used to construct low-resolution functional images and, consequently, provide a relatively low-cost adjunct to many conventional diagnostic tools. Considerable interest has been generated in applying FDPM to tumor imaging; however, relatively little is known about in vivo tissue optical properties and their relationship to physiology. Since optical properties (absorption and scattering parameters) are the fundamental determinants of contrast in FDPM-derived images, we will employ the LAMMP 1 GHz FDPM instrument to characterize optical properties in various types of normal and malignant tissues in vivo. We believe this information is essential for realistically estimating the feasibility of tumor optical imaging.

T lymphocytes (T cells) contact with antigen-presenting B cells initiates an activation cascade which includes an increase in T-cell intracellular calcium and leads to T-cell proliferation and differentiation. Although T-cell/B-cell physical contact is required for an immune response, little is known about the patterns of cellular interaction and their relation to activation. Novel optical techniques at the LAMMP facility provide insights into these puzzles at the single-cell level. In this project, we study biophysical requirements for T-cell activation using an optical trap to control the orientation of T-cell/B-cell pairs and fluorescent microscopy to measure subsequent T-cell intracellular calcium level ([Ca2+]i) response. Our specific aim is to develop an optical trapping-based high resolution fluorescence imaging system (CATS) to study1) Spatial requirements for T-cell activation and relationships between stimulus intensity and response pattern (response percentage, response latency, calcium signal pattern, shape and motility changes), 2) The role of adhesion/co-stimulatory molecules, such as CD45 molecule, in T cell activation, 3) The role of actin cytoskeleton in T cell activation.

The objective of the proposed research is to expand the current capabilities of the two-photon microscope (TPM) system constructed in the LAMMP facility to image cellular activity within tissues, especially cutaneous wounds, to depths of at least 500 (m. One of the main advantages the TPM has over a confocal microscope is that the system can be used to look at any specific excitation in the region 350-450nm due to the tunability of the excitation laser. Another advantage of this TPM is the unique capability to obtain fluorescent images from two different wavelength regions e.g. red and green, simultaneously. We propose to extend this capability by integration of a cooled CCD spectrometer which will allow for spectral measurements in the range 400-800nm to pixel size resolution. The specific aims of this work are 1) Optimization of the TPM for depth and resolution using Monte Carlo simulations and imaging through phantoms of turbid media to determine the principal factors in image degradation, 2) Image cells and tissues in vitro, especially those involved in wound healing, using exogenous localized photosensitizers (PS) and endogenous fluorophores (auto fluorescence) to provide contrast, 3) Determination of PDT dose for relevant cells using the two-photon system, 4) Image cells and tissues in vivo using animal models to demonstrate depth imaging and the monitoring of wound repair, 5) Obtain real time images of FRET signals and determine excitation wavelengths that optimize energy transfer.

Development of novel methods to allow the rapid execution of Monte Carlo computations distributed amongst several computational engines.

T-cell contact with antigen-presenting cells (APC) initiates an activation cascade which includes an increase in T-cell intracellular calcium and leads to T-cell proliferation and differentiation. Although T-cell/APC physical contact is required for an immune response, little is known about the patterns of cellular interaction and their relation to activation. We have combined fluorescence spectroscopy and imaging with optical manipulation to investigate the physical properties of T-cell activation. We study cell-cell contact requirements for T-cell activation using optical tweezers to control the orientation of T-cell/APC pairs and fluorescence microscopy to measure the subsequent T-cell intracellular calcium level ([Ca2+]i) response. T cells which are presented with antigen at the leading edge have a higher probability of responding and a shorter latency of response than those contacting APCs or antibody-coated beads with their trailing end. Alterations in anti body dens ity and bead size are used to determine the spatial requirements for T cell activation and the minimum number of receptors which must be engaged in order to transmit a positive signal. Results show that T cell responses (response percentage, latency and [Ca2+]i pattern) depend on both antibody density on bead and bead size. ~340 TCRs are required to be engaged for intracellular calcium level increase in T cell activation.

We will focus on the generalization of current optical models to allow for the accurate processing of FDPM data in a broader number of clinical situations. Specifically, we are interested in making FDPM measurements to probe layered tissue structures as well as assess the properties of small tissue volumes. In both these applications, we require FDPM measurements to be made at small source-detector (S-D) separations. However, it is well known that such measurements, when processed using algorithms based on standard optical diffusion theory (SODT), can lead to the determination of inaccurate optical values. Our goals are to generalize SODT to provide governing equations which accommodate spatially distributed collimated spaces and to solve these new equations for steady and amplitude modulated collimated point source located within an infinite medium. We will compare these results to solutions derived using SODT and experiment. We will also solve the new equati ons for steady and amplitude modulated collimated sources illuminating the surface of an infinite medium. Compare results with SODT and experiment. Finally, we will develop a theoretical framework to address cases where FDPM measurements contain significant contributions from both minimally scattered and fully diffuse photons.

We will focus on the generalization of current optical models to allow for the accurate processing of FDPM data in a broader number of clinical situations. Specifically, we are interested in making FDPM measurements to probe layered tissue structures as well as assess the properties of small tissue volumes. In both these applications, we require FDPM measurements to be made at small source-detector (S-D) separations. However, it is well known that such measurements, when processed using algorithms based on standard optical diffusion theory (SODT), can lead to the determination of inaccurate optical values. Our goals are to generalize SODT to provide governing equations which accommodate spatially distributed collimated spaces and to solve these new equations for steady and amplitude modulated collimated point source located within an infinite medium. We will compare these results to solutions derived using SODT and experiment. We will also solve the new equati ons for s teady and amplitude modulated collimated sources illuminating the surface of an infinite medium. Compare results with SODT and experiment. Finally, we will develop a theoretical framework to address cases where FDPM measurements contain significant contributions from both minimally scattered and fully diffuse photons.

Frequency domain photon migration (FDPM) was used to non-invasively quantify optical properties (absorption and scattering) of cervical tissues during photodynamic therapy (PDT). The aims of this study were 1) to quantitatively determine near-infrared optical properties of normal cervical tissues and high-grade squamous intraepithelial lesions (HSIL), 2) to assess the feasibility of differentiating normal cervical tissues from HSIL on the basis of these optical properties, and 3) to determine how cervical tissue optical properties change following PDT of HSIL in vivo. Eleven patients were scheduled for photodynamic therapy of histologically proven high-grade cervical squamous intraepithelial lesion with ALA (5-aminolevulinic acid). FDPM measurements were performed on normal cervical tissues and HSIL prior to ALA application onto the ecto-cervix, ninety minutes after ALA administration, and three minutes after completion of PDT light delivery. FDPM measurements of no rmal cerv ical tissues were compared to HSIL using two-tailed analysis of variance (ANOVA). The results of this study clearly demonstrate that FDPM is 1) sensitive to PDT induced changes in cervical optical properties (absorption changes up to 20%), 2) absorption and scattering are significantly reduced in dysplastic cervical tissue vs. normal (reduction in scattering ranges from 6-13% and absorption from 8-16%), and 3) dysplastic processes induce wavelength dependent alterations in absorption and scattering. These results suggest that near-infrared FDPM may have an important role in non-invasive detection of cervical dysplasia.

In the past year, using the two-photon microscope (TPM) system constructed in the LAMMP facility, we were able to image both collagen and NADH autofluorescence and Protoporphyrin IX localization (exogenous fluorescence) within a RAFT tissue model. Images of the RAFT tissue, composed of collagen, macrophages and fibroblasts, were obtained with submicron resolution to a depth of 200 mm, the working distance of the water immersion objective used. Selective uptake of Protoporphyrin IX by specific cellular components of the matrix was also visualized at depth utilizing the TPM system.

Type I collagen is the main component in the extracellular matrix (ECM) of the lungs. Several important diseases such as asthma and interstitial pulmonary fibrosis involve detrimental collagen production and ECM remodeling. In addition, there are several reports in the literature of important interactions between the epithelium and the fibroblast in modulating the ECM. We studied the ECM of a fibroblast-imbedded collagen gel designed as an in vitro model of the interstitial connective tissue of the lung. The model system was a 3-D co-culture consisting of human lung fibroblasts (CCD-18 lu), denatured type I collagen, fetal bovine serum (FBS), sodium hydroxide, fibroblasts, media, and a monolayer of human alveolar epithelial cells (A549). The gels were fixed at 0, 3, and 6 day intervals, and observed by immunohistochemical staining, transmission electron microscopy (TEM), and two-photon microscopy (TPM). Two cases were studied: 1) gels cultured in media supplemented with 10% serum, and 2) serum-free culture. In both cases, gels cultured without the monolayer served as a control. Preliminary analysis of live cultures shows the presence of fibers/bundles displaying TPM-induced autofluorescence signatures consistent with collagen and/or elastin. In medi a supplemented with 10% serum, matrix fibers were observed from Day 3 and increased in number through Day 6. Similarly, in serum-free culture the number of detectable fibers increased from Day 3 through Day 6. However, the total number of bundles per day was less than that of the gels in media with serum. We conclude that human lung fibroblasts remodel the ECM in vitro, and that factors present in serum (i.e., TGF-b1) enhance the remodeling process.

Novel techniques are being developed to allow for direct predictions of tissue biochemical composition from diffuse optical measurements. The specific chemometric methods used include principal component analysis and partial least squares. The promise of such methods is that it may allow quantitative extraction of chromophore concentrations in complex geometries and does not rely on having an underlying model for optical transport.

DESCRIPTION: The long-term goal of the proposed work is to develop a new, minimally-invasive treatment of benign uterine disease based on Photodynamic Therapy (PDT). Eventually, uterine PDT may have a significant impact on women's health care and its associated costs because it will provide an inexpensive, outpatient alternative to conventional surgical and medical techniques for treating dysfunctional uterine bleeding (DUB) and improving adenomyosis (AM)-related symptoms. During the past six years we have completed a number of animal, human and modeling studies that have helped us understand the complexity inherent to achieving irreversible endometrial ablation (EA) during PDT. We now propose a major new effort which builds on this foundation and further characterizes how light and drug dosimetry parameters affect clinical outcome. Specific aims include: 1) constructing, characterizing and optimizing performance of a novel, minimally-invasive intrauterine light probe (IULP); 2) completing Phase I/II and beginning Phase III human clinical trials with ALA; 3) testing a limited number of new sensitizers in animal models; and 4) evaluating pharmacokinetics, safety and efficacy of a limited number of new compounds in humans.

Diffuse Optical Imaging (DOI) is a non-invasive optical technique that employs near-infrared (NIR) light to quantitatively characterize the optical properties of thick tissues. Although NIR methods were first applied to breast diaphanography nearly 80 years ago, quantitative DOI methods employing time- or frequency-domain photon migration technologies have only recently been used for breast imaging (i.e. since the mid-1990s). In this research program we propose the formation of a flexible, multi-institutional "Network" composed of partnerships between leading, Academic, Industry and Government laboratories that have demonstrated expertise in Breast Cancer Research. Our broad goal is to advance new technology, Multi-Dimensional Diffuse Optical Imaging (MDDOI), that will dramatically improve breast cancer detection, clinical management, and quality of life for breast cancer patients. MD-DOI employs broadband technology both in spectral (approximately 650-1000 nm) and temporal (approximately IGHz) domains in order to separate absorption from scattering and quantify multiple molecular probes based on absorption or fluorescence contrast. Additional dimensionality is provided by integrating and co-registering MD-DOI functional information with Magnetic Resonance Imaging (MRI) and X-Ray mammography. Factors affecting critical MD-DOI issues, such as intrinsic and extrinsic conlrast mechanisms, quantitation of biochemical components, image formation/visualization, and multi-modality co-registration will be explored and defined in 3 research projects and delivered as usable translational technologies by three research resource cores. Based on these fmdings, a standardized MD-DOI platform will be developed that can be used as a stand-alone device or in conjunction with MRI and mammography. This technology will be tested, validated, and duplicated for translational use in multiple clinical sites: The University of Pennsylvania, The Massachusetts General Hospital (MGH), Dartmouth University, and The University of California, (Irvine and San Francisco). Clinical studies in each test site will involve coordination through their respective breast care centers housed in five NCI Comprehensive Cancer Centers. Pre-clinlcal animal studies will be performed using specially designed MD-DOI technology optimized for animal models and optical/MR/molecular imaging agents. The proposed Network will stimulate the development, validation, and standardization of new technologies, procedures, and analysis tools; as well as the formation of important multi-institutional collaborative relationships and commercial partners. This broad-based, multi-disciplinary effort will provide the community with new insight regarding the origins of breast disease and practical approaches for addressing several key challenges in breast cancer clinical management, including: detecting early disease, distinguishing between malignant and benign lesions, and understanding the impact of therapies (e.g. hormone replacement therapy (HRT) and neoadjuvant chemotherapy).

Absorption; Body Tissues; Breast; Breast Cancer Detection; Breast Tissue; Breast cancer screening; CRISP; Chemotherapy-Hormones/Steroids; Clinical; Clinical Research; Clinical Study; Complement; Complement Proteins; Computer Retrieval of Information on Scientific Projects Database; Data; Dependence; Devices; Diagnosis, Ultrasound; Echography; Echotomography; Electromagnetic, Laser; Endocrine Gland Secretion; Frequencies (time pattern); Frequency; Funding; Goals; Grant; Hand; Hemoglobin; Hormones; Hydrogen Oxide; Institution; Investigators; Lasers; Light; MMG; Mammary Gland Parenchyma; Mammary Gland Tissue; Mammogram; Mammography; Measures; Medical Imaging, Ultrasound; Menopausal Status; Menstrual cycle; NIH; National Institutes of Health; National Institutes of Health (U.S.); O element; O2 element; Operation; Operative Procedures; Operative Surgical Procedures; Optics; Oxygen; Pathology Report; Patients; Photons; Photoradiation; Physiologic; Physiological; Process of absorption; Property; Property, LOINC Axis 2; Radiation, Laser; Research; Research Personnel; Research Resources; Researchers; Resolution; Resources; Screening procedure; Source; Surgical; Surgical Interventions; Surgical Procedure; Therapeutic Hormone; Tissues; Ultrasonic Imaging; Ultrasonogram; Ultrasonography; Ultrasound Test; Ultrasound, Medical; United States National Institutes of Health; Variant; Variation; Water; absorption; base; diagnostic ultrasound; in vivo; insight; mammary cancer detection; migration; screening; screenings; sonogram; sonography; sound measurement; surgery; ultrasound; ultrasound imaging; ultrasound scanning; volunteer

Abstract Not Provided.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In this study, we propose to use spectroscopic and polarimetric techniques to push the limits of optical intrinsic signal imaging (ISI) towards measuring the fast response of an illuminated cortex to a stimulus input. For this study, fast signals are defined as those that take place on timescales shorter than the hemodynamic response, which occurs on the order of seconds. Thus we propose to image light reflectance changes in the cortex that are due to electrical signals in the activated neurons (~10s of milliseconds) and to the transient increase in deoxygenated hemoglobin known as the initial dip (~100s of milliseconds). Such fast signals are crucial to understand because they are the least understood part of the brain response to a stimulus and are a direct measure of neural activity.

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mammary gland; nephelometry; breast neoplasm /cancer diagnosis; diagnosis design /evaluation; optics; portable biomedical equipment; patient monitoring device; rapid diagnosis; mammography; clinical research; infrared spectrometry; human subject;

Abstract Not Provided.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. In the past year, using the multi-photon microscope (MPM) system constructed in the LAMMP facility, we were able to image both collagen and NADH autofluorescence collagen second hormonic generation (SHG) and Protoporphyrin IX localization (exogenous fluorescence) within a RAFT tissue model. Images of the RAFT tissue, composed of collagen, macrophages and fibroblasts, were obtained with submicron resolution to a depth of 200 mm, the working distance of the water immersion objective used. Selective uptake of Protoporphyrin IX by specific cellular components of the matrix was also visualized at depth utilizing the TPM system.

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This work extends single location measurements to mapping of optical properties. The spatial variation of the optical properties are expected to give important information about tissue physiology. The method proposed here is to illuminate tissue with structured light over a large area. Fourier analysis and filtering of the structured light image allows for subsurface imaging and the determination of the optical properties. Experiments on various heterogeneous tissue phantoms are under investigation. Tissue measurements will subsequently be performed.

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The overall objective of this research project is to investigate the utility of photodynamic therapy (PDT) for the treatment of malignant brain tumors (gliomas). The project is a close and integrated multi disciplinary collaborative effort of various institutions in he United States and Norway. The participating parties are: The Department of neurosurgery, Rikshospital, the Laser microbeam and medical program (LAMMP) facility at the Beckman Laser Institute and Medical Clinic (BLI), University of California, Irvine (UCI), California USA the Department of health physics at the University of Nevada, Las Vegas (UNLV), Nevada USA. and the Departments of surgical oncology, and pathology the Norwegian Radium Hospital.Oslo Norway. The aim of previous investigations was to determine the utility of photodynamic therapy in combination with ionizing radiation or concurrent hyperthermia in the treatment of glioblastoma multiforme. The effects of light dose, gamma radiation dose, temperature and photosensitizer type on human glioma spheroids have been investigated in detail. The project also investigated the PDT effect of ALA-ester and repetitive PDT treatments during the past few years with several publications resulted from the study.

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DESCRIPTION (provided by applicant): The 2002 Gordon Research Conference on Lasers in Medicine and biology will be held from July 14-19, 2002 at Kimball Union Academy in Meriden, New Hampshire. The intent of the conference is to gather investigators from academia, governmental agencies, national laboratories and industry to examine future applications of lasers and optical sciences in medicine and surgery. Particular emphasis is placed on technological advances that are close to clinical application and/or commercial development. Sessions include: Optical Coherence Tomography; Nanotechnologies for Optical Sensing, Imaging, and Manipulation; Biological Applications of Micro-optical Devices; Combined Imaging Methodologies; Fluorescence Methods for Biomedical Diagnostics; Advances in Optical Microscopy; Intra-vital Imaging and Microscopy; Wavefront Sensing and Adaptive Optics in Vision Correction; and Photodynamic Therapy. Seven of the nine sessions include at least one talk with substantial applications in the area of cancer diagnostics. An important focus of the Gordon Conference is to bring students and post-doctoral fellows in biomedical engineering together with academic engineers and scientists, clinicians and members of industry to discuss and define the most important questions and problems to be solved in this field. Ample time between sessions has been schedule to foster informal discussion of new ideas emerging from the conference. In addition to traditional scientific presentations, we have organized two special symposia to facilitate this discussion process. The first symposium, "Career Paths in Biomedical Engineering", will feature short presentations from clinicians and engineers in academia, industry and government to provide career advice to graduate students and post-doctoral fellows. The second symposium, a special industrial forum, will highlight new technologic developments on the cusp of clinical viability. Industrial participants will each make a brief presentation followed by questions and answers. Both symposia will conclude with a general discussion followed by a reception. Finally, a special issue of the Journal of Biomedical Optics will be devoted to Optical Detection: Clinical and Industrial Advances. Conference attendees presenting talks or posters will be invited to submit their work for this special issue.

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bioimaging /biomedical imaging; imaging /visualization /scanning

bioimaging /biomedical imaging; imaging /visualization /scanning

bioimaging /biomedical imaging; imaging /visualization /scanning

bioimaging /biomedical imaging; imaging /visualization /scanning

bioimaging /biomedical imaging; imaging /visualization /scanning

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We have developed a new method for solving photon transport problems that combines accuracy and increased speed when compared with conventional Monte Carlo simulations. This ?condensed history? method adapts a strategy routinely used to solve charged transport problems for radiation dosimetry, problems that would be prohibitively costly to solve conventionally. This method promises to be especially useful for tissue problems calling for computational models intermediate between classical diffusion theory and rigorous radiative transport. Gains in computational efficiency by factors of 4-5 over conventional simulation can be expected in many instances.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Visiting researchers from various institutions are provided with hands-on and didactic trainign in LAMMP technologies. Following completion of their training period these individuals return to their home institutions to introduce LAMMP technologies into their laboratories.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Surgical removal of brain tumors is the most common initial treatment received by brain tumor patients. Surgical resection can benefit the patients in several ways: for example, it relieves the mass effect of tumor on neurological tissue and allows histological diagnosis of the tumor, which directly affects the direction of follow-up therapeutic strategy (3). Many studies have demonstrated that aggressive surgical resection enhances the survival length and quality of life for brain tumor patients. Therefore, the goal of brain tumor resection procedures is to maximize tumor removal with minimal neurological damage. To achieve this goal accurate intraoperative identification of brain tumor margins during craniotomy is required. This study aims at the development of a means for visualizing brain tumor margins using optical techniques such as Modulated Imaging.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The Laser Microbeam and Medical Program sponsors a seminar series on a bimonthly basis involving invited speakers within the biological and biomedical optics communities. The seminars are generally an hour long with time for questions and discussion following the talk. December 6, 2006 Greg Kronmal (Ambion, Inc-The RNA Company An Applied Biosystems Business Austin, Texas) "RNAi Basics and siRNA Experimental Design " December 8, 2006 Ryan Hill (Research Group of Prof. Jason B. Shear Department of Chemistry &Biochemistry University of Texas at Austin ) "Enzyme-Nanoparticle Functionalization" December 12, 2006 Dr. Moshe Levi, MD (Professor of Medicine, Physiology and Biophysics Vice Chair of Medicine for Research Head Division of Renal Diseases and Hypertension University of Colorado at Denver and Health Sciences Center ) "Role of nuclear receptors (FXR and SREBP) in regulation of renal lipid metabolism and diabetic kidney disease" December 15, 2006 Dr. Don C. Lamb, PhD (University of Illinois at Urbana-Champaign) "Sensitive Fluorescence Methodologies and their Application to Biological Systems" January 11, 2007 Dr. Nosang V. Myung (Department of Chemical and Environmental Engineering and Center for Nanoscale Science and Engineering University of California-Riverside) "Nanosensor Array" January 18, 2007 Dr. Michael T. Longaker (Department of Surgery, Stanford University) "Skeletal Tissue Engineering" January 25, 2007 Dr. Bruce C. Wheeler (University of Illinois at Urbana-Champaign 1304 West Springfield Avenue Urbana, IL 61801) "Brain on a Chip: Engineering Form and Function in Cultured Neuronal Network" February 2, 2007 Dr. Chen-Yuan Dong (Department of Physics, National Taiwan University) "Developing Intravital Multiphoton Microscopy for Biomedical Research" February 13, 2007 Dr. Joe Fu-Jiou Lo (Biomedical Engineering University of Southern California) "Micro Optics for Biophotonics Detection--MOEMS design, implementation, and medical application." February 13, 2007 Dr. Karen L. Christman, PhD (NIH Postdoctoral Fellow University of California, Los Angeles) "Polymers in Medicine: From Tissue Engineering to Bionanotechnology" February 20, 2007 Dr. Ralf Wessel, PhD (Washington University in St. Louis Associate Professor of Physics ) "Probing Brains with Electrodes and Computation" February 22, 2007 Dr. Pedram Mohseni, PhD (Case Western Reserve University Electrical Engineering and Computer Science Department ) "Single-Chip Wireless Microsystems for Recording Neuroelectrical and Neurochemical Activity" February 27, 2007 Dr. Julia Lyubovitsky, PhD (Beckman Laser Institute) "Developing optical tools to look at biological systems" March 6, 2007 Dr. Paul Sajda, PhD (Columbia University Biomedical Engineering ) "Single-trial Neuroimaging: Identifying Neural Correlates of Trial-to-Trial Behavioral Variability" March 9, 2007 Dr. Maksim Bazhenov, PhD (The Salk Institute for Biological Studies) "Oscillatory synchronization and information coding in the olfactory system" March 13, 2007 Dr. Thomas Lu, PhD (Department of Anatomy and Neurobiology University of California, Irvine) "Temporal Processing of Time-Varying Sounds in the Auditory Cortex" March 15, 2007 Dr. Michael Lin, PhD (University of California, San Diego) "Engineering Drug-controllable Reporters of Protein History in Cells and Organisms" March 23, 2007 Dr. Vincent Wallace (TeraView Ltd, Cambridge, UK) "Development of an intra-operative THz imaging probe for breast conserving surgery" March 26, 2007 Dr. David Sampson (Optical+Biomedical Engineering Laboratory, School of Electrical, Electronic &Computer Engineering, The University of Western Australia, Perth, Australia) "Deducing tissue microstructure without resolving it" April 11, 2007 Dr. Dror Seliktar, PhD (Department of Biomedical Engineering Technion [unreadable][unreadable]" Israel Institute of Technology) "Novel Biosynthetic Hydrogel Biomaterials Engineered for Wound Healing and Tissue Regeneration" April 19, 2007 Dr. Brian Andrews (Chief of Rheumatology, UC Irvine) "The Structure of Cartilage and Synovium in Normal Joints, Rheumatoid Arthritis and Osteoarthritis" April 19, 2007 Dr. John M. Boone, PhD (Professor of Radiology and Biomedical Engineering University of California, Davis) "Breast CT: A Tool for Breast Cancer Screening, Diagnosis and Treatment" April 19, 2007 James R. Mansfield, M.Sc. and Dr. Richard M. Levenson (CRi) "Optical imaging of multiplexed molecular markers in vivo and ex vivo" April 26, 2007 Dr. Utkarsh Sharma, PhD (Kiara Technologies, Inc.) "Fiber Lasers and All-fiber Optic Devices for Applications in Sensing and Medical Imaging" May 1, 2007 Dr. Ed Leiter and Dr. Adam Kennedy (From the Jackson Laboratory) "Mouse Models for Metabolic Syndrome" May 2, 2007 Dr. Miguel A. L. Nicolelis, MD, PhD (Anne W. Deane Professor of Neuroscience Depts. of Neurobiology, Biomedical Engineering, and Psychological and Brain Sciences Co-Director, Duke Center for Neuroengineering) "Computing with Neural Ensembles" May 3, 2007 Dr. Darrell Irvine, PhD (Department of Materials Science &Engineering Massachusetts Institute of Technology ) "Engineering Immunity: Using Materials to Prod and Probe the Immune System" May 17, 2007 Dr. E. Duco Jansen, PhD (Associate Professor of Biomedical Engineering Vanderbilt University ) "Optical Stimulation of Neural Tissue" May 31, 2007 Dr. Nik Kollias (Principal Research Fellow, Johnson &Johnson Consumer and Personal Products Co., Skillman, NJ 08558) "Skin Aging (1-80 yrs) across ethnicities: Measurements and Questions" May 31, 2007 Dr. Franz Hillenkamp, PhD (Institute for Medical Physics and Biophysics University of Muenster ) "Analysis of Nucleic Acids by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry" July 10, 2007 Dr. Veneranda Guadalupe Garces-Chavez (University of St. Andrews, Scotland ) "Optical Manipulation: Novel techniques for improved photoporation, optical sorting and Raman spectroscopy" July 19, 2007 Dr. Claus-Dieter Ohl (Assistant Professor Division of Physics and Applied Physics School of Physical and Mathematical Sciences Nanyang Technological University Singapore) "Inertial and stable bubble oscillations for microfluidic applications " July 26, 2007 Dr Gal Markel (Sheba Cancer Research Center, Sheba Medical Center, Israel) "Novel immune evasion mechanism employed by melanoma" August 21, 2007 Dr. Uzi Efron (Dept of Electrical Engineering Holon Institute of Technology ) "Studies &Development Effort of a Low Vision Goggle: Recent Progress" August 24, 2007 Dr. Steven L. Jacques, PhD (Biomedical Engineering and Dermatology Oregon Health &Science University, Portland, Oregon ) "Global versus microscopic optical scattering properties of tissues, studied using reflectance-mode confocal microscopy and optical coherence tomography." September 24, 2007 Dr. Sung Sik Thomas Aquinas Hur, PhD (Beckman Laser Institute University of California, Irvine) "Roles of 3D Traction Forces in Migration and Focal Adhesion Dynamics of Bovine Aortic Endothelial Cells" September 26, 2007 Dr. Alvin T. Yeh, PhD (Assistant Professor Department of Biomedical Engineering Texas A&M University ) "Systemic Approaches for Intravital Assessment of Tissue Response, Growth and Remodeling" October 2, 2007 Dr. Elliot Hui (MIT) "Cellular Scale Tissue Engineering Through MEMS" October 11, 2007 Dr. Elliot Botvinick (Assistant Professor University of California, Irvine) "Introduction to the Mechanobiology Lab " October 18, 2007 Dr. Andrew K. Dunn (Biomedical Engineering Department University of Texas at Austin ) "High Resolution Optical Imaging of Brain Function" October 26, 2007 Dr. Gerard L. Cot[unreadable], PhD, P.E. (Charles H. &Bettye Barclay Professor Head, Department of Biomedical Engineering Texas A&M University ) "Optically-Based Biomedical Sensing Approaches" November 7, 2007 Dr. Peter Wang (University of Illinois, Urbana-Champaign) "Seeing is Believing: Molecular Imaging, NanoBiotechnology, and Mechano Biology in Live Cells" November 9, 2007 Dr. Matt Glucksberg (Professor, Northwestern University Robert R. McCormick School of Engineering and Applied Sciences Biomedical Engineering Chair, Department of Biomedical Engineering ) "Biomedical Engineering Design and Global Health" November 15, 2007 Dr. Aladin Boriek, PhD (Associate Professor of Medicine and Physiology Baylor College of Medicine Houston, Texas) "Engineering Approach to the Ventilatory Pump" November 29, 2007 Dr. Charles Limoli (Associate Professor Radiation Oncology University of California, Irvine) "Redox changes regulating the stress response of multipotent neural cells "

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Exchange of slides for a course entitled "Photomedicine" given at the master level.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Neoadjuvant chemotherapy can achieve tumor downstaging and allow breast conservation in some patients. Response to chemotherapy is assessed clinically. Although this has been shown to be associated with improved disease-free survival, it is not clear whether clinical assessment of the primary tumor accurately reflects pathologic response. The UCSF Breast Center (N. Hylton and L. Esserman) are using MRI to assess changes in tumor size and distribution in response to neoadjuvant chemotherapy. Preliminary results indicate that tumor changes measured by MRI are better than clinical exam and are meaningful predictors of survival. The MRI signal origin is related to changes in blood volume fraction, perfusion, and spatial extent of the tumor site. These are precisely the functional parameters that the hand-held FDPM optical probe can provide in a single, non-invasive measurement. As a result, we will initiate a pilot study that examines 10 UCSF patients. Each patient will receive baseline MRI scans at UCSF, followed by optical scans at UCI. Subsequent MRI scans will be performed following each cycle of chemotherapy (AC, Adriamycin/Cyclophosphamide). In order to minimize the number of trips to Irvine, a second and final optical scan will be completed following the 4th AC cycle, just prior to surgery. Because optical and MRI provide complementary functional information, we believe these results will provide valuable insight into the mechanisms these therapies as well as help establish practical criteria for predicting drug efficacy

Adhesion Plaques; Affect; Biological; Biological Models; CRISP; Cell Adhesion; Cell Communication; Cell Communication and Signaling; Cell Differentiation; Cell Differentiation process; Cell Interaction; Cell Locomotion; Cell Migration; Cell Movement; Cell Signaling; Cell-Matrix Adherens Junctions; Cell-to-Cell Interaction; Cells; Cellular Adhesion; Cellular Matrix; Cellular Migration; Collagen; Complement; Complement Proteins; Computer Retrieval of Information on Scientific Projects Database; Cytoskeletal System; Cytoskeleton; Disease; Disorder; Doppler Effect; Doppler OCT; Doppler Shift; Embryo Development; Embryogenesis; Embryonic Development; Event; Fluorescence; Focal Adhesions; Focal Contacts; Funding; Gel; Generations; Grant; INFLM; Image; Inflammation; Institution; Intracellular Communication and Signaling; Investigators; Maps; Measures; Mechanical Stimulation; Metastasis; Metastasize; Metastatic Neoplasm; Metastatic Tumor; Model System; Models, Biologic; Motility; Motility, Cellular; Motion; NIH; National Institutes of Health; National Institutes of Health (U.S.); Neoplasm Metastasis; OCT Tomography; Optical Coherence Tomography; Physiologic; Physiological; Procedures; Process; Proteins; Research; Research Personnel; Research Resources; Researchers; Resources; Role; Secondary Neoplasm; Secondary Tumor; Signal Transduction; Signal Transduction Systems; Signaling; Simulate; Source; Spectroscopy; Spectrum Analyses; Spectrum Analysis; System; System, LOINC Axis 4; Tensile Strength; Time; Tissue Model; Tomography, Optical Coherence; Tumor Cell Migration; United States National Institutes of Health; Wound Healing; Wound Repair; biological signal transduction; cancer metastasis; cell motility; disease/disorder; gene product; imaging; intracellular skeleton; multi-photon; polymerization; second harmonic; social role; tissue repair; two-photon

Absorption; Acute; Apoplexy; Biological; Blood Volume; Blood flow; Body Tissues; Brain; Brain Injury, Chronic; CRISP; Cerebral Stroke; Cerebrovascular Apoplexy; Cerebrovascular Circulation; Cerebrovascular Stroke; Cerebrovascular accident; Cerebrum; Chronic Brain Injury; Collaborations; Computer Retrieval of Information on Scientific Projects Database; Computer Simulation; Computerized Models; Deep; Depth; Diffusion; Electromagnetic, Laser; Encephalon; Encephalons; Epilepsy; Epileptic Seizures; Epileptics; Frequencies (time pattern); Frequency; Functional Imaging; Funding; Grant; Hemoglobin; Illumination; Image; Institution; Intermediary Metabolism; Investigators; Lasers; Light; Lighting; Longitudinal Studies; METBL; Maps; Mathematical Model Simulation; Mathematical Models and Simulations; Measurement; Measures; Metabolic Processes; Metabolism; Methods; Metric; Modeling; Models, Computer; NIH; National Institutes of Health; National Institutes of Health (U.S.); Nervous System, Brain; Neuroprotectants; Neuroprotective Agents; Neuroprotective Drugs; O element; O2 element; Optics; Oxygen; Photoradiation; Physiologic Imaging; Process of absorption; Property; Property, LOINC Axis 2; Radiation, Laser; Relative; Relative (related person); Research; Research Personnel; Research Resources; Researchers; Resources; Sampling; Seizure Disorder; Simulation, Computer based; Source; Stroke; Structure; System; System, LOINC Axis 4; Technology; Tissues; Trauma; United States National Institutes of Health; Vascular Accident, Brain; absorption; base; brain attack; cerebral blood flow; cerebral circulation; cerebral vascular accident; cerebrocirculation; cerebrovascular; computational modeling; computational models; computational simulation; computer based models; computerized modeling; computerized simulation; design; designing; epilepsia; epileptiform; epileptogenic; imaging; in silico; in vivo; insight; interest; long-term study; stroke; virtual; virtual simulation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Understanding such fundamental cell biological processes as migration, polarization and cell division requires the ability to noninvasively observe the dynamic changes in the structure of the involved subcellular compartments. Routinely used modes of observing these processes often require invasive imaging methods that rely on the introduction of dyes or toxins or fixation. In turn, introduction of these compounds can yield artifacts as well as impede sequential imaging. This study proposes to use an integrated multiphoton microscopy and optical coherence tomography (MPM/OCT) system as a means of observing subcellular compartments without the application of exogenous sources of contrast. The previous development of an MPM/OCT platform has permitted simultaneous observation of co-registered fluorescence and scattering contrast in cells and tissues. High contrast of scattering in the OCT modality has shown clear structures which are like actin filopodia. Both auto-fluorescence and scattering are intrinsic contrasts in cells and tissues. Furthermore, MPM/OCT can provide high-resolution and 3-dimensional images, which means cells can be observed in their natural environment of 3-dimensional extracellular matrix. Data from the proposed experiments will specifically characterize the biological basis of those structures that provide OCT contrast. It may provide the basis for a novel method of studying cytoskeletal or membranous dynamics in unlabelled cells in a more natural 3-dimensioal environment.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. John Frangioni's lab is developing wide field imaging technology for use in the surgical field that will enable quantitative imaging. Eventually Modulated Imaging will be incorporated into this approach. There can be several centimeters of variation in height of the target tissue over the width of the surgical field. In addition, tissues in the field are likely to have some shape/curvature. In order for imaging to be quantitative, we need to employ correction factors that will essentially "flatten" the surgical field. This is a series of phantom measurements aimed to develop/validate algorithms needed for correcting curved surfaces observed during in vivo measurements.

A multi-center translational study will be conducted in order to evaluate the ability of a bedside optical technology to monitor neoadjuvant chemotherapy response in breast cancer patients with locally-advanced disease. The technology, Diffuse Optical Spectroscopic Imaging (DOSI), will be combined with MRI and biomarkers that provide anatomic and molecular correlates, respectively. The proposed studies build on the infrastructure established by the NCI Network for Translational Research in Optical Imaging (NTROI), which has allowed us to validate and standardize DOSI-MRI co-registration methods, data analysis, histopathology, and protocol design. Five clinical sites with identical DOSI instruments and MRI protocols will participate: University of California, Irvine, University of California, San Francisco, University of Pennsylvania, Dartmouth, and Harvard/MGH. A 6th site, Siemens Corporate Research, will provide data and informatics support using specially developed software. The proposed NTR will be integrated into 2 well-established NCI programs supported by ACRIN and I-SPY in order to leverage DOSI with standard MRI and tissue biomarker protocols. Developmental studies will explore blood biomarkers, as well as social and economic factors affecting the acceptance of DOSI for breast cancer chemotherapy management. The resulting NTR-ACRIN-ISPY partnership will facilitate the first multi-center, multi-modality study designed to validate the role of optics in monitoring and predicting response to cancer chemotherapies. Our long-term goal is to provide oncologists with quantitative, non-invasive, standardized optical endpoints that can be used to rapidly optimize therapy for individual breast cancer patients.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In this project, NIR fluorophores are quantified in solution. The excitation and emission spectra as well as the quantum yield are measured in solution using a fluorimeter housed at the LFD.

DESCRIPTION (provided by applicant): There is considerable interest in developing new quantitative imaging methods to monitor and predict breast cancer response to neoadjuvant chemotherapy (NAC), both prior to and as early as possible during the course of treatment. Diffuse optical spectroscopic imaging (DOSI) allows patients to be followed from baseline through treatment and surgery with a cost-effective, bedside, handheld scanning probe. In this competing renewal application, we propose to further advance the development of a standardized DOSI technology platform by introducing a new, portable, miniature device (m-DOSI) that has significantly reduced cost and size with equivalent performance compared to existing instruments. m-DOSI scanners will be constructed and delivered to all collaborating clinical sites and up to 150 pre-surgical NAC patients will be evaluated in multi- center studies. Our goal is to establish quantitative DOSI functional endpoints of NAC response that predict patient clinical outcome as measured by pathologic complete response (pCR). In order to optimize the power of DOSI predictions and ease of integration into the clinical work flow, we propose to validate three measurement time-points and DOSI endpoints first discovered in our current funding cycle: 1) Tissue oxygen saturation prior to chemotherapy infusion, 2) Tissue oxyhemoglobin concentration one day following the first infusion, and 3) a Tissue optical index (TOI) at the midpoint of NAC prior to switching drug regimens. Tumor hemodynamics will also be investigated during the first week of therapy. NAC midpoint response is currently undergoing evaluation as part of our American College of Radiology Imaging Network 60-patient multi-center clinical trial (ACRIN-6691) that completed enrollment in March 2013, two months prior to the originally anticipated June 2013 end date. We hypothesize that the combination of pre-therapy and one-day post- infusion measurements with the ACRIN mid-point assessment will provide a more practical and improved approach for managing patients. In order to develop new insight into achieving pCR in the challenging triple negative (TN) breast cancer population we will stratify response by molecular subtype and adjust our multi- endpoint predictive model for the TN subgroup. Finally, we will continue to optimize and improve m-DOSI functionality, standardize clinical measurement and analysis procedures, and evaluate whether m-DOSI can be used with equivalent overall performance by different operators. Our long-term goal is to identify the right combination of quantitative clinical endpoints for informing medical decisions o chemotherapy regimen, duration, and timing of surgery. Ultimately this work is expected to lead to a bedside optical imaging technology that can be used to improve patient outcome by maximizing therapeutic response, minimizing unnecessary toxicity, and optimizing clinical decision-making.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This project is to image glioma angiogenesis in subcutaneous and intracranial tumor models using spatially modulated imaging.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Main Objective: DOSI will be used to evaluate the patient's response to chemotherapy. The primary aim of this clinical trial is to determine whether the baseline to mid-therapy changes in the DOSI measurement of the quantitative tumor tissue optical index (TOI) can predict final complete pathologic response in breast cancer patients undergoing pre-surgical neoadjuvant chemotherapy. The secondary aims investigate the correlation between additional DOSI quantitative measurements of tumor biochemical composition obtained at other timepoints, the full range of pathologic response (i.e. complete, partial, and non-response) and any corresponding imaging measurements. DOSI and standard of care imaging (e.g. serial mammograms, serial ultrasounds, etc) and/or all MRI (e.g., SOC imaging or MRI imaging from co-enrollment in ACRIN 6657 or other studies) will be collected. ACRIN #6691 is the first optical imaging trial sponsored by ACRIN.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The main goal of this project is to develop a robust method of non-linear microscopy of human skin imaging. This will include the excitation/emission parameters optimization, depth of imaging improvement (i.e. optical clearing of skin, objective-detector sensitivity) and motion artifacts prevention. The imaging goals are two-fold. First, the more cellular topical skin layers (stratum corneum and epidermis that form fist 50-75 [unreadable]m of skin) are bearing the main brunt of skin carcinogenesis: melanocytes are localized within the basal epidermal layer, epidermis is responsible for basal and squamous cell carcinoma as well as benign lesions (actinic keratosis, sebhorreic keratosis, etc) genesis. This is a first barrier protecting the body from environmental factors such as microorganisms, moisture, UV, etc. The underlaying dermis (2-4 mm thickness depending on the body part) bulk is made of connective tissue, a ECM of elastic and collagen fibers, cellular component of which is responsible for wound healing and immune response among others. Human skin reflects age, gender, race, health status. Imaging skin with microscopic resolution and spectral selectivity can provide a powerful tool in understanding important biological and biomechanical processes of wound healing, carcinogenesis, aging, and environmental response.

A multi-center translational study will be conducted in order to evaluate the ability of a bedside optical technology to monitor neoadjuvant chemotherapy response in breast cancer patients with locally-advanced disease. The technology, Diffuse Optical Spectroscopic Imaging (DOSI), will be combined with MRI and biomarkers that provide anatomic and molecular correlates, respectively. The proposed studies build on the infrastructure established by the NCI Network for Translational Research in Optical Imaging (NTROI), which has allowed us to validate and standardize DOSI-MRI co-registration methods, data analysis, histopathology, and protocol design. Five clinical sites with identical DOSI instruments and MRI protocols will participate: University of California, Irvine, University of California, San Francisco, University of Pennsylvania, Dartmouth, and Harvard/MGH. A 6th site, Siemens Corporate Research, will provide data and informatics support using specially developed software. The proposed NTR will be integrated into 2 well-established NCI programs supported by ACRIN and I-SPY in order to leverage DOSI with standard MRI and tissue biomarker protocols. Developmental studies will explore blood biomarkers, as well as social and economic factors affecting the acceptance of DOSI for breast cancer chemotherapy management. The resulting NTR-ACRIN-ISPY partnership will facilitate the first multi-center, multi-modality study designed to validate the role of optics in monitoring and predicting response to cancer chemotherapies. Our long-term goal is to provide oncologists with quantitative, non-invasive, standardized optical endpoints that can be used to rapidly optimize therapy for individual breast cancer patients.

This Integrative Graduate Education and Research Traineeship (IGERT) award initiates a novel model for interdisciplinary graduate training in biophotonics across the biomedical sciences, physical sciences, and engineering. Biophotonics technologies provide powerful capabilities to probe and manipulate biological components and processes. Their utilization in the life sciences and medicine represents an estimated annual economic impact of $50 billion. This program aims to produce the next generation of biophotonics leaders to make transformative advances in the development and application of new tools for biological and medical discovery and maintain global U.S. leadership in biotechnology, pharmaceutical and medical device industries.
Intellectual Merit: This IGERT award creates a hands-on training program that integrates physics, chemistry, engineering, and life-science principles across spatial and temporal scales. The interaction of, and collaboration between, biomedical scientists, physical scientists and engineers throughout the graduate traineeship will drive advances in biophotonics technologies, computational methods, and molecular probes to solve important problems in bio-molecular, cellular, tissue, and whole organismal systems.
Broader Impacts: The BEST IGERT project will promote dissemination of an innovative education framework aimed towards a diverse cadre of scientists and engineers. Moreover, IGERT faculty and trainees will engage vigorously in a spectrum of outreach, dissemination, recruitment, retention, and career development activities that leverages the commitment of multiple units within UC-Irvine, industry in Southern California, and a nationwide network of faculty contacts, including those at minority serving institutions, to inform the public and broaden participation by students from underrepresented groups.

IGERT is an NSF-wide program intended to meet the challenges of educating U.S. Ph.D. scientists and engineers with the interdisciplinary background, deep knowledge in a chosen discipline, and the technical, professional, and personal skills needed for the career demands of the future. The program is intended to establish new models for graduate education and training in a fertile environment for collaborative research that transcends traditional disciplinary boundaries, and to engage students in understanding the processes by which research is translated to innovations for societal benefit.

DESCRIPTION (provided by applicant): Spatial Frequency Domain Imaging (SFDI) is a model-based, wide-field, non-contact method for measuring the absorption, scattering, and fluorescence properties of biological tissue. Optical properties are determined in each pixel simultaneously, by measuring the attenuation (or fluorescence) of sinusoidal patterns of light projected onto the sample at varying spatial frequencies and phases. Images are demodulated by processing 3 phase-shifted views of the sample. The mean interrogation depth at a given wavelength is controlled by the spatial frequency of projection, and frequency-dependent differences in path length are used to calculate tissue optical properties using computational models. Because 3 specific phases are required for each projected frequency, care must be taken to perfectly sequence all projections and camera triggers. While each of these processes is fairly rapid, together they can slow the acquisition to a fraction of the camera frame rate. In order to overcome this limitation and facilitate real-time SFDI, we will develop new methods using frequency synthesis - multiple frequencies synthesized into customized projection patterns. These patterns will be optimized for speed and frequency-dependent information content in order to facilitate rapid and accurate optical property measurements, probe buried objects, and perform tomography. When properly selected, frequency synthesized projections can potentially decrease the minimum acquisition time to the frame rate of the camera, allowing real-time SFDI and SFD tomography. The ability to project custom patterns not only allows us to generate multi-frequency components, it also adds the ability to change their orientation. This allows us to explore a new mode of contrast based on probing tissue structures that are aligned with the direction of the projected pattern. This is due to the fact that many tissue types, including bone, muscle, skin, and white matter in the brain, have orientated internal structures such that the degree of optical scattering depends on the direction of light propagation. The scattering direction of these oriented tissues is determined by their microscopic structure and obeys a diffusion equation. We will derive accurate solutions to the anisotropic diffusion equation in the spatial frequency domain. In an ordered medium, the attenuation of sinusoidal patterns depends on the relative orientation of the spatial frequency pattern and scatterer direction. Thus, by projecting multiple spatial frequencies in different directions and measuring the attenuation, we will be able to image the spatially varying scattering orientation over a large field of view. We expect that the combination of spatial frequency synthesis and orientation control will lead to new methods for quantitative, real-time imaging and tomography in thick tissues, as well as the characterization of exciting new contrast mechanisms based on an optical diffusion tensor. PUBLIC HEALTH RELEVANCE: These studies will allow us to acquire sufficient data to develop and validate a new optical technology for real-time, quantitative imaging and depth sectioning of tissues based on relatively simple, cost-effective imaging cameras and patterned light projection techniques. The technology has potential to replace conventional camera-based imaging methods by allowing quantitative viewing of functional tissue attributes beneath the surface, where disease typically begins. Our approach is expected to lead to new, bedside medical imaging methods for detecting disease, monitoring therapy response, and guiding surgical procedures.

DESCRIPTION (provided by applicant): Hemodynamic activity plays an important role in brain function and disease. The connection between neuronal and vascular activity in the brain is poorly understood. This in part stems from the lack of quantitative methods to localize hemodynamic changes in three-dimensions with sufficient temporal and spatial resolution to monitor the progression of the hemodynamic response to a stimulus. Intrinsic Signal Optical Imaging (ISOI) is a camera-based method for imaging cortical activity. ISOI allows researchers to image the hemodynamic changes that accompany neuronal activity by measuring changes in intensity of light remitted from an illuminated brain. Because it is capable of imaging large cortical regions with high spatial and temporal resolution, ISOI has become an important tool for studying the functional architecture and plasticity of the cortex. However, ISOI has two major limitations that have hampered our ability to decipher the complex relationship between hemodynamics and neuronal activity. First, it is often difficult to determine whether measured signals are due to changes in hemoglobin concentration or alterations in hemoglobin oxygenation state. Second, ISOI only provides two-dimensional images, and thus cannot localize hemodynamic activity in depth. We have recently acquired preliminary results using multi-spectral ISOI that demonstrate tomographic localization of hemodynamic activity in rat cortex during whisker stimulation. This is possible because near- infrared light penetrates much more deeply into the brain than visible light. By using many wavelengths of light spanning both the visible and near-infrared (NIR), and applying tomographic imaging methods, we have succeeded in forming three-dimensional tomographic images of hemodynamic activity in the rat somatosensory cortex. In addition, we have also been able to quantify dynamic changes in hemoglobin concentration and oxygenation state by employing spatial frequency domain imaging (SFDI) techniques in conjunction with multi-spectral image analysis. The purpose of this R21 proposal is to validate and refine this exciting new finding. Over the course of the two year funding period, we will incorporate snapshot hyperspectral imaging into our established optical brain imaging methods - ISOI and SFDI. We will optimize our tomographic image reconstruction methods to work in the rat cortex using dynamic tissue simulating phantoms, and validate our approach using our established protocol for whisker stimulation in the adult rat. PUBLIC HEALTH RELEVANCE: Deciphering the connection between hemodynamic and neuronal activity is crucial for advancing our understanding of normal brain function, gaining insight into brain pathologies, and developing more effective neuro-therapies. However, current brain functional imaging methods are limited in their ability to localize and quantify hemodynamic activity in the brain. The goal of this work is to develop and validate a new technique based on principles of hyperspectral spatial frequency domain imaging to meet this need for high- resolution three-dimensional functional tomography of cerebral hemodynamics.

This Major Research Instrumentation award supports the acquisition of an ultrafast multimodal spectroscopy system. This system enables advanced time-resolved spectroscopy experiments, a key capability in material and molecular research. The system will be placed in the Laser Spectroscopy Facility (LSF), housed in the Department of Chemistry of the University of California Irvine. Traditionally, ultrafast lasers and spectroscopy have not been part of user facilities because former ultrafast laser technologies were too complex and unreliable to support many users. Consequently, the user community of ultrafast technologies has remained small, and the unique research capabilities of ultrafast optical techniques have remained inaccessible to an audience interested in such capabilities. In this Program, a state-of-the-art ultrafast laser system is made accessible through a three-pronged model based on optimized facility conditions, strong user support and a partnership with industry. This model addresses past limitations to make ultrafast laser technology and its research capabilities accessible to a much broader community of researchers. In making the technology available to a much larger user base, the impact of ultrafast spectroscopy is significantly amplified. The Program ensures broad exposure and dissemination of ultrafast spectroscopy capabilities. On the UCI campus alone, the Laser Spectroscopy Facility (LSF) in which the system will be housed supports more than 300 users, and serves 13 departments on campus. By leveraging strong connections with institutions and companies neighboring UCI, a large community of researchers will have access to the ultrafast spectroscopy capabilities offered through the facility. The impact of the requested technology is further fortified by a strong user training program, established channels of dissemination, and an outreach program designed around the ultrafast spectroscopy instrument

The most fundamental processes in matter evolve on ultrafast time scales. Examples include the making and breaking of chemical bonds, conformational motions of molecules, evolution of optical excitations, and electron transfer processes. These phenomena form the mechanistic basis of scientific challenges in chemistry, biology and materials science: designing efficient catalysts, understanding nature's solution to light harvesting materials, and optimizing the efficiency of solar cells, to name a few. Ultrafast laser technologies have proven indispensable for meeting these challenges, as they provide direct recordings of such fast fundamental processes. Consequently, the demand for ultrafast spectroscopy is growing, as an increasingly expanding pool of chemists, chemical engineers, physicists, and biologists come to rely on this technology. The goal of this Program is to bring the unique capabilities of ultrafast laser science to a broad community of researchers. This goal is achieved by implementing a three-pronged model: 1) Acquisitions of a commercial ultrafast laser light source and nonlinear optical spectrometer, with unprecedented performance, versatility, stability, and simplicity of operation. The system is made available through a staffed user facility at the University of California, Irvine (UCI); 2) A partnership with Newport Optics, the supplier of the instrument. The partnership establishes a push-pull mechanism between technology and application for impacting a broad community beyond the users at UCI; 3) The Ultrafast Consultation Board (UCB), a team of experimental and theoretical ultrafast spectroscopists who provide general assistance, technical guidance, mentoring and help with the interpretation of data. By providing depth to the analysis of acquired spectroscopic data, the UCB bolsters the scientific impact of measurements made by non-experts, expands the user base, and magnifies the breadth of research topics supported by this Program.

? DESCRIPTION (provided by applicant): Summary: Multiphoton microscopy (MPM) is a nonlinear laser scanning microscopy technique that features high three-dimensional resolution, fast imaging capabilities and label-free molecular contrast. Several endogenous tissue components can be visualized, including collagen (through second-harmonic generation, SHG), reduced nicotinamide adenine dinucleotide (NADH), flavin adenosine dinucleotide (FAD), keratin, melanin and elastin fibers (through two-photon excited fluorescence, TPEF). The ability to generate high- resolution images of tissue structure and composition without the need for exogenous labels makes MPM imaging particularly well-suited for characterizing superficial tissues in vivo. The purpose of this clinical imaging proposal is to evaluate the ability of in viv multiphoton microscopy to provide quantitative optical endpoints with sufficiently high predictive power to reliably distinguish between pigmented lesions in three groups: common nevi, atypical nevi and melanoma. The framework is based on our preliminary results obtained from a 15-lesion (14 patient) study where we identified three optical biomarkers related to TPEF and SHG signals and correlated these in vivo molecular features with conventional ex vivo histopathologic criteria. MPM biomarkers were combined to obtain a quantitative, 9-point numerical index (multi-photon melanoma index, MMI) that distinguished between common nevi (MMI = 0-1), atypical nevi (MMI = 1-4) and melanoma (MMI = 5-8) (p<0.05). We now propose a powered prospective clinical trial that follows on these promising results in order to determine whether the MMI, or similar MPM-derived index, can be reliably used in a clinical setting. We expect our results will provide a validated decision-making endpoint to increase clinical diagnosis accuracy of common nevi, atypical nevi and melanoma. In addition, our effort to correlate in vivo MPM-derived contrast with conventional histopathology is expected to lead to new insight regarding the origins of melanoma appearance and progression. Our long-term goal is to identify the right combination of quantitative clinical endpoints that would improve clinical diagnoses, guide effective treatment, and eliminate unnecessary biopsies while increasing identification of lesions requiring removal.

PROJECT SUMMARY Multiphoton microscopy (MPM) can provide sub-micron resolution images of living tissues in their native environment with label-free molecular contrast from multiple modalities, including second harmonic generation (SHG) and two-photon excited fluorescence (TPEF). Several endogenous tissue components can be visualized, including collagen (from SHG) and reduced nicotinamide adenine dinucleotide (NADH), flavin adenosine dinucleotide (FAD), keratin, melanin and elastin fibers (from TPEF). We have advanced label-free MPM technologies in skin clinical/translational studies for characterizing keratinocyte metabolism, diagnosing melanoma, understanding melanocyte biology, detecting basal cell carcinoma, quantifying skin pigmentation, and assessing the effects of cutaneous laser therapy. Many of these studies have been completed using a commercial multi-photon microscope for clinical skin imaging that has limitations in terms of field-of-view (FOV), speed, footprint, and cost. In order to address these barriers to clinical adoption, we propose to build a ?next-generation? clinical multiphoton microscope that integrates advanced benchtop technologies into a compact, practical, and cost-effective bedside device. This new instrument will have comparable FOV, resolution, and scanning features to standard-of-care reflectance confocal microscopes (RCM), yet provide unique structural and metabolic contrast from multiple modalities (TPEF and SHG) that can only be achieved with MPM. We will establish the clinical safety of this device in a light dose escalation study that assesses DNA and cellular damage, and establish key performance benchmarks in a 12-patient clinical study of healthy volunteers across a range of skin types. In addition, we will conduct pilot studies of wound re-epithelialization and melanocyte migration in the context of vitiligo micro-grafts, a clinical procedure where pigmented skin is transplanted into skin affected by vitiligo, which is devoid of melanocytes. Melanocytes migrating out of engrafted skin and keratinocytes turning over within engrafted skin can be visualized by measuring the TPEF of cellular melanin and co-factors (NADH, FAD+) in and around the grafts, effectively identifying different cell populations involved in wound healing. Our broad, long term goal is to develop ev-MPM as a practical approach for rapid, in vivo characterization of cellular morphologic and metabolic imaging endpoints in patients. These can be used to understand and optimize wound healing and provide a practical beside platform for detecting, diagnosing, and optimizing therapeutic response in skin diseases.

PROJECT SUMMARY/ABSTRACT The broad objective of the Imaging Core is to use cutting edge optical imaging technologies to non-invasively visualize the macroscopic, microscopic, and molecular processes that underlie skin biology and disease. The Core builds on technologies and expertise in the Beckman Laser Institute and Medical Clinic (BLI) and the Laboratory for Fluorescence Dynamics (LFD), longstanding UC Irvine centers that have pioneered the development and application of optics and photonics technologies for biology and medicine. BLI and LFD will create a new skin imaging suite housing specific, state-of-the-art, in vivo optical imaging platforms to P30 skin biology investigators, including: Multiphoton Microscopy, MPM; Fluorescence Lifetime Imaging Microscopy, FLIM; Coherent Raman Scattering, CRS; Spatial Frequency Domain Imaging, SFDI; Laser Speckle Imaging, LSI; and Optical Coherence Tomography (OCT); as well as computational methods and models for understanding light propagation in skin (i.e. skin optics). These technologies are used to visualize fluorescent molecular probes and reporters as well as provide label-free contrast from intrinsic tissue signals in cells, ECM, and vasculature. By employing these multi-modal, multi-scale technologies, the imaging core will facilitate studies that can lead to powerful new skin biology insights that can be translated to humans. This is accomplished by integrating imaging datasets with the Genomics and Systems biology cores to develop models that can then be tested in biological experiments, including in animals. In addition to providing cutting edge technologies for research, a key component of the Imaging Core will be to provide resources for training, dissemination, and collaboration that drive and support interdisciplinary activities. Our goal is to bring together technology developers, modeling experts, biologists, and clinicians both within UCI and the skin research community at large to build teams that can share knowledge and apply new tools to solve longstanding problems. This will be accomplished by creating a combination of hands-on training, didactic training, and dissemination activities that provide skin biology researchers with the knowledge, resources, and support needed to integrate state-of the-art optical imaging into skin biology research. Finally, our technologies will continue to be optimized, with feedback from the community, to better understand skin biology and pathophysiology, with the ultimate goal of developing new approaches that improve outcomes for patients suffering from cutaneous disease.