Dual-mode endomicroscopy for detection of epithelial dysplasia in the mouth: a descriptive pilot study.

Dual-mode endomicroscopy is a diagnostic tool for early cancer detection. It combines the high-resolution nuclear tissue contrast of fluorescence endomicroscopy with quantified depth-dependent epithelial backscattering as obtained by diffuse optical microscopy. In an in vivo pilot imaging study of 27 oral lesions from 21 patients, we demonstrate the complementary diagnostic value of both modalities and show correlations between grade of epithelial dysplasia and relative depth-dependent shifts in light backscattering. When combined, the two modalities provide diagnostic sensitivity to both moderate and severe epithelial dysplasia in vivo.

Fluorescence confocal endomicroscopy of the cervix: pilot study on the potential and limitations for clinical implementation.

Current diagnostic capabilities and limitations of fluorescence endomicroscopy in the cervix are assessed by qualitative and quantitative image analysis. Four cervical tissue types are investigated: normal columnar epithelium, normal and precancerous squamous epithelium, and stromal tissue. This study focuses on the perceived variability within and the subtle differences between the four tissue groups in the context of endomicroscopic in vivo pathology. Conclusions are drawn on the general ability to distinguish and diagnose tissue types, on the need for imaging depth control to enhance differentiation, and on the possible risks for diagnostic misinterpretations.

Diffuse optical microscopy for quantification of depth-dependent epithelial backscattering in the cervix.

A fiber optic imaging approach is presented using structured illumination for quantification of almost pure epithelial backscattering. We employ multiple spatially modulated projection patterns and camera-based reflectance capture to image depth-dependent epithelial scattering. The potential diagnostic value of our approach is investigated on cervical ex vivo tissue specimens. Our study indicates a strong backscattering increase in the upper part of the cervical epithelium caused by dysplastic microstructural changes. Quantization of relative depth-dependent backscattering is confirmed as a potentially useful diagnostic feature for detection of precancerous lesions in cervical squamous epithelium.

Quantifying phase function influence in subdiffusively backscattered light.

Light backscattering at short source-detector separations is considerably influenced by the scattering phase function of a turbid medium. We seek to more precisely relate a medium's subdiffusive backscattering to the angular scattering characteristics of its microstructure. First, we demonstrate the inability of the scattering asymmetry g1 = < cos θ > to predict phase function influence on backscattering and reveal ambiguities related to the established phase function parameter γ. Through the use of high-order similarity relations, we introduce a new parameter that more accurately relates a scattering phase function to its subdiffusive backscattering intensity. Using extensive analytical forward calculations based on solutions to the radiative transfer equation in the spatial domain and spatial frequency domain, we demonstrate the superiority of our empirically derived quantifier σ over the established parameter γ.

Detecting structural information of scatterers using spatial frequency domain imaging.

We demonstrate optical phantom experiments on the phase function parameter γ using spatial frequency domain imaging. The incorporation of two different types of scattering particles allows for control of the optical phantoms' microscopic scattering properties. By laterally structuring areas with either TiO2 or Al2O3 scattering particles, we were able to obtain almost pure subdiffusive scattering contrast in a single optical phantom. Optical parameter mapping was then achieved using an analytical radiative transfer model revealing the microscopic structural contrast on a macroscopic field of view. As part of our study, we explain several correction and referencing techniques for high spatial frequency analysis and experimentally study the sampling depth of the subdiffusive parameter γ.

Optical phantoms with adjustable subdiffusive scattering parameters.

A new epoxy-resin-based optical phantom system with adjustable subdiffusive scattering parameters is presented along with measurements of the intrinsic absorption, scattering, fluorescence, and refractive index of the matrix material. Both an aluminium oxide powder and a titanium dioxide dispersion were used as scattering agents and we present measurements of their scattering and reduced scattering coefficients. A method is theoretically described for a mixture of both scattering agents to obtain continuously adjustable anisotropy values g between 0.65 and 0.9 and values of the phase function parameter γ in the range of 1.4 to 2.2. Furthermore, we show absorption spectra for a set of pigments that can be added to achieve particular absorption characteristics. By additional analysis of the aging, a fully characterized phantom system is obtained with the novelty of g and γ parameter adjustment.

Model-based analysis on the influence of spatial frequency selection in spatial frequency domain imaging.

Frequency variation in spatial frequency domain imaging is a powerful tool for adjusting the penetration depth of the imaging signal and the parameter sensitivity toward absorption and diffusive and subdiffusive scattering. Through our computational analysis, using an analytical solution of the radiative transfer equation, we add quantitation to this tool by linking the different spatial frequency regimes to their relative information content and to their absolute depth sensitivity. Special focus is placed on high spatial frequencies by analysis of the phase function parameter γ and its significance and ambiguity in describing subdiffusive scattering.

Surface layering properties of Intralipid phantoms.

Intralipid has become an extensively studied and widely used reference and calibration phantom for diffuse optical imaging technologies. In this study we call attention to the layering properties of Intralipid emulsions, which are commonly assumed to have homogeneous optical properties. By measurement of spatial frequency domain reflectance in combination with an analytical solution of the radiative transfer equation for two-layered media, we make quantitative investigations on the formation of a surface layer on different dilutions of Intralipid. Our findings are verified by an independent spatially resolved reflectance setup giving evidence of a time dependent, thin and highly scattering surface layer on top of Intralipid-water emulsions. This layer should be considered when using Intralipid as an optical calibration or reference phantom.

Sources of errors in spatial frequency domain imaging of scattering media.

Knowledge of the impact of potential sources of error in spatial frequency domain imaging (SFDI) is essential for the quantitative characterization of absorption and scattering in tissue and other turbid media. We theoretically investigate the error in the derived absorption and scattering parameter, subject to typical experimental and theoretical sources of errors. This provides a guideline to properly assess the significance of various parameters related to the measurement and the theoretical evaluation of spatial frequency domain reflectance data. At the same time, this study serves as a reference to estimate the overall precision of derived optical parameters of semi-infinite scattering media using SFDI.

Extending the 3ω method: thermal conductivity characterization of thin films.

A lock-in technique for measurement of thermal conductivity and volumetric heat capacity of thin films is presented. The technique is based on the 3ω approach using electrical generation and detection of oscillatory heat along a thin metal strip. Thin films are deposited onto the backside of commercial silicon nitride membranes, forming a bilayer geometry with distinct thermal parameters. Stepwise comparison to an adapted heat diffusion model delivers these parameters for both layers. Highest sensitivity is found for metallic thin films.