Our NICHD collaborators continue to develop theoretical models of light propagation in tissue, which account for scattering, absorption, and polarization state of light, and that predict the spatial profile of the surface intensity of re-emitted and transmitted light. These models allow for both re-emission of the introduced light and emission of fluorescent light generated by an embedded chromophore, either inherent to or introduced into the tissue. Measurement of a series of surface intensity profiles allows reconstruction of the three-dimensional structure of the tissue using inverse analytical techniques. Likewise, polarization-based models allow interpretation of the change of polarization state with scattering changes associated with tissue. Polarization sensitive imaging offers the potential of distinguishing hidden structures developed below the skin surface, such as the collagen network, that can be used to follow the transition from normal to diseased tissue state. Pearson correlation data analysis techniques enable enhancement of images and allow characterization of subsurface structures of biological tissue. One such clinical application being pursued by our NICHD collaborators is colposcopy to assess changes in the cervix that may result from disease or other factors. In partnership with our collaborators we designed both the optical configuration and mechanical layout of this instrument, fabricated, and tested an adaption to a commercial colposcope that permits these polarization sensitive measurements to be captured and at the same time allows the clinician simultaneous visual and photographic or video assessment of the cervix. This instrument is currently being evaluated in the clinic. In the current year, based on feedback from the clinical experience, we have made several modifications to the instrument to make it more user-friendly. Additionally, a mechanical adaptations have both increased the range and precision of the cameras focal position, which has improved the quality of the captured images and the enhanced subsequent image analysis, and provided a positioning capability that is more stable and user friendly. Key aspects of this instrumentation are the basis of an ongoing patent application. In a related on-going project to monitor the vasculature of Karposi's sarcoma, a multi-spectral imaging modality is being evaluated to assess changes in morphology due to scattering and simultaneously to use spectral information to assess changes in vasculature function - such as angiogenesis and necrosis - associated with the different treatment modalities being pursued in the clinic. The current instrumentation is based on sequential change of filtration that is time consuming, subject to sequential access, and limited in its ability to provide for variable intensity of illumination. A fast random-access near-infra-red illumination system reduces the time taken to perform the necessary measurements and allows for a significant improvement for the patient. Through millisecond resolution, random access to different wavelengths of light (600nm to 1100nm), modification of the bandwidth and intensity of illumination, exposure times from sub-millisecond to """"""""infinite and real time analysis provides more complete clinical data and allows for follow-up assessment of clinical condition. Details of this work have resulted in a publication and a submission to PLOS ONE ENTITLED: """"""""Evaluation of non-invasive multispectral imaging as a tool for measuring the effect of systemic therapy in Kaposi sarcoma"""""""" Kainerstorfer JM, Polizzotto Mn, Uldrick TS, Rahman R, Hassan M, Laley N, Ardeshirpour Y, Wyvill KM, Aleman K, Smith PD, Yarchoan R, Gandjbakhche AH. In collaboration with both NICHD and CIT, a portable instrument was designed that assesses the severity of sub-dural hematomas following head injury that should enable inexpensive, portable, on-site screening of suspected traumatic brain injury. In concept, a single near ir light source and dual separated detectors are mounted in a hand-held device that can be scanned across the skull using motion as a signalfor detecting changes in blood volume in the dural regions of the head, thus indicating the presence of brain injury. In the current year, a prototype proof of principle device and a tissue-like head phantom with a simulated sub-dural hematoma were developed and evaluated. These demonstrated the viability of such an approach and produced results with excellent localization of inclusions and quantitative assessment. Details of this work have resulted in a publication. In other collaborations with the NICHD group, we are pursuing building a small, portable, and wearable system that uses near infrared light for brain imaging. We envision that the necessary sources, detectors, and associated electronic circuits for processing, transmitting and receiving data can be mounted within a helmet. The optical source couples light into the brain and the resulting light after transmission through the diffusive brain tissue is collected by the receivers, and the processed data will enable an evaluation of the hemodynamic response. To enable this aspect of the project, we have recently ordered prototype dual-wavelength (780nm and 850nm) optrodes that we will integrate into the package worn by the patient. It is estimated that the final unit within the helmet will be less than 4cm x 4cm. We continue our interest in the use of near infrared upconverters for use in these studies (we are also pursuing the use of these as luminescent labels - see """"""""Instrumentation"""""""" annual report). Identification of deeper structures within the tissue is possible by extending to the near infra-red region of the spectrum and by the use of novel infrared compounds to act as probes localized at desired sites within the tissue.

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Riley, Jason D; Amyot, Franck; Pohida, Tom et al. (2012) A hematoma detector-a practical application of instrumental motion as signal in near infra-red imaging. Biomed Opt Express 3:192-205
Kainerstorfer, Jana M; Riley, Jason D; Ehler, Martin et al. (2011) Quantitative principal component model for skin chromophore mapping using multi-spectral images and spatial priors. Biomed Opt Express 2:1040-58