We describe the development of a rapid, noncontact imaging method, modulated imaging (MI), for quantitative, wide-field characterization of optical absorption and scattering properties of turbid media. MI utilizes principles of frequency-domain sampling and model-based analysis of the spatial modulation transfer function (s-MTF). We present and compare analytic diffusion and probabilistic Monte Carlo models of diffuse reflectance in the spatial frequency domain. Next, we perform MI measurements on tissue-simulating phantoms exhibiting a wide range of l* values (0.5 mm to 3 mm) and (μs′/μa) ratios (8 to 500), reporting an overall accuracy of approximately 6% and 3% in absorption and reduced scattering parameters, respectively. Sampling of only two spatial frequencies, achieved with only three camera images, is found to be sufficient for accurate determination of the optical properties. We then perform MI measurements in an in vivo tissue system, demonstrating spatial mapping of the absorption and scattering optical contrast in a human forearm and dynamic measurements of a forearm during venous occlusion. Last, metrics of spatial resolution are assessed through both simulations and measurements of spatially heterogeneous phantoms.

Experiments performed on turbid phantoms demonstrate that spatially modulated illumination facilitates quantitative wide-field optical property mapping and tomographic imaging in turbid media.

Abstract. We present a review of short-wave infrared

Absorption and reduced scattering coefficients of in-vivo human skin provide critical information on non-invasive skin diagnoses for aesthetic and clinical purposes. To date, very few in-vivo skin optical properties have been reported. Previously, we reported absorption and scattering properties of in-vivo skin in the wavelength range from 650 to 1000nm using the diffusing probe in the “modified two-layer geometry”. In this study, we determine the spectra of skin optical properties continuously in the range from 500 to 1000nm. It was found that the concentration of chromophores, such as oxy-hemoglobin, deoxy-hemoglobin, and melanin, calculated based on the absorption spectra of eighteen subjects at wavelengths above and below 600nm were distinct because of the inherent difference in the interrogation region. The scattering power, which is related to the average scatterer’s size, demonstrates a clear contrast between skin phototypes, skin sites, and wavelengths. We also applied venous occlusion on forearms and found that the concentrations of oxy- and deoxy-hemoglobin as assessed at wavelengths above and below 600nm were different. Our results suggest that diffuse reflectance techniques with the visible and near infrared light sources can be employed to investigate the hemodynamics and optical properties of upper dermis and lower dermis.

We describe a non-contact profile correction technique for quantitative, wide-field optical measurement of tissue absorption (µ a ) and reduced scattering (µ s ') coefficients, based on geometric correction of the sample's Lambertian (diffuse) reflectance intensity. Since the projection of structured light onto an object is the basis for both phase-shifting profilometry and modulated imaging, we were able to develop a single instrument capable of performing both techniques. In so doing, the surface of the 3-dimensional object could be acquired and used to extract the object's optical properties. The optical properties of flat polydimethylsiloxane (silicone) phantoms with homogenous tissue-like optical properties were extracted, with and without profilometry correction, after vertical translation and tilting of the phantoms at various angles. Objects having a complex shape, including a hemispheric silicone phantom and human fingers, were acquired and similarly processed, with vascular constriction of a finger being readily detectable through changes in its optical properties. Using profilometry correction, the accuracy of extracted absorption and reduced scattering coefficients improved from 2-to 10-fold for surfaces having height variations as much as 3 cm and tilt angles as high as 40°. These data lay the foundation for employing structured light for quantitative imaging during surgery.

Abstract. Oxygenation measurements are widely used in patient care. However, most clinically available instruments currently consist of contact probes that only provide global monitoring of the patient (e.g., pulse oximetry probes) or local monitoring of small areas (e.g., spectroscopy-based probes). Visualization of oxygenation over large areas of tissue, without a priori knowledge of the location of defects, has the potential to improve patient management in many surgical and critical care applications. In this study, we present a clinically compatible multispectral spatial frequency domain imaging (SFDI) system optimized for surgical oxygenation imaging. This system was used to image tissue oxygenation over a large area (16×12 cm) and was validated during preclinical studies by comparing results obtained with an FDA-approved clinical oxygenation probe. Skin flap, bowel, and liver vascular occlusion experiments were performed on Yorkshire pigs and demonstrated that over the course of the experiment, relative changes in oxygen saturation measured using SFDI had an accuracy within 10% of those made using the FDA-approved device. Finally, the new SFDI system was translated to the clinic in a firstin-human pilot study that imaged skin flap oxygenation during reconstructive breast surgery. Overall, this study lays the foundation for clinical translation of endogenous contrast imaging using SFDI. C 2011 Society of Photo-Optical Instrumentation Engineers (SPIE).

In vivo and ex vivo tissue autofluorescence (endogenous fluorescence) have been employed to investigate the presence of markers that could be used to detect tissue abnormalities and/or malignancies. We present a study of the autofluorescence of normal skin and tumor in vivo, conducted on 18 patients diagnosed with nonmelanoma skin cancers (NMSC). We observed that both in basal cell carcinomas (BCC) and squamous cell carcinomas (SCC) the endogenous fluorescence due to tryptophan residues was more intense in tumor than in normal tissue, probably due to epidermal thickening and/or hyperproliferation. Conversely, the fluorescence intensity associated with dermal collagen crosslinks was generally lower in tumors than in the surrounding normal tissue, probably because of degradation or erosion of the connective tissue due to enzymes released by the tumor. The decrease of collagen fluorescence in the connective tissue adjacent to the tumor loci was validated by fluorescence imaging on fresh-frozen tissue sections obtained from 33 NMSC excised specimens. Our results suggest that endogenous fluorescence of NMSC, excited in the UV region of the spectrum, has characteristic features that are different from normal tissue and may be exploited for noninvasive diagnostics and for the detection of tumor margins.

Clinical examination alone is not always sufficient to determine which burn wounds will heal spontaneously and which will require surgical intervention for optimal outcome. We present a review of optical modalities currently in clinical use and under development to assist burn surgeons in assessing burn wound severity, including conventional histology/ light microscopy, laser Doppler imaging, indocyanine green videoangiography, near-infrared spectroscopy and spectral imaging, in vivo capillary microscopy, orthogonal polarization spectral imaging, reflectance-mode confocal microscopy, laser speckle imaging, spatial frequency domain imaging, photoacoustic microscopy, and polarization-sensitive optical coherence tomography.