A major drawback in polarization gating of light backscattered from tissue is that surface reflections dominate the image. An optically flat plate and matching fluid applied to the tissue surface, combined with off-axis detection, were previously used to address this problem. This approach is often inappropriate or inconvenient for practical use and more importantly can affect the tissue's optical properties. A method is demonstrated that combines images obtained with linearly and circularly polarized light to produce a polarization-gated image that is free from surface reflections and does not require optically flat plates or matching fluid.
The scattering of polarized light from a two layer scattering medium is investigated using Monte Carlo simulations. First order and normalized second order moments are used to analyze the spatial properties of the emerging light in different polarization states. Linearly and circularly polarized illumination is used to probe different depths. Absorption and layer thickness are varied and it is demonstrated that the determination of these values is aided by the inclusion of polarization information. The lateral and depth localization of light by polarization subtraction is also quantified. Potential applications of these techniques are burn depth and melanoma thickness measurements.
Light scattering provides a problem in optical spectroscopy as the relationship between attenuation and absorption in the presence of scattering is nonlinear. Three simple methods of reducing the effects of scattering and hence returning to an approximately linear relationship are considered in this paper, namely, extracting light that has maintained its original polarization state through subtraction of orthogonal polarization states, use of an added absorber, and spatial filtering. These can all be applied relatively easily to conventional spectrophotometers. However, there is an inevitable trade-off between the accuracy of the measurement and the signal-to-noise ratio as scattered light is rejected from the detector. It is demonstrated that polarization subtraction is the most efficient technique at selecting weakly scattered photons from a scattered light background as it enables the relationship between attenuation and absorption coefficient to become more linear while maintaining a higher number of detected photons. In practical use, the drawback of polarization subtraction over added absorber and spatial filtering methods is that a large dc background light level is maintained, which contributes to a higher shot noise. This means that when the scattering coefficient is high (micros > or = 7 mm(-1)) the added absorber method offers better performance for shot noise limited detection.
Scattering can result in erroneous determination of the concentrations of constituent absorbers in spectrophotometry. This is due to the relationship between attenuation and absorption coefficient becoming nonlinear; hence, the use of the Lambert-Beer law becomes invalid. It has previously been shown that application of polarization techniques can reduce these effects, resulting in a more linear relationship. Here we quantify the impact of this improvement on measurement of the ratio of concentrations for two general absorbing species and show that measurement using polarization-maintaining light is more accurate. This is performed using a generalized version of theory previously dependent on selection of isosbestic wavelengths. For the absorbing species and geometries considered here, the mean error on the estimation of absorber concentration ratio is 18.2% for the case of detection without polarization discrimination. When polarization-maintaining light is extracted, mean errors of 1.2% and 5.1% are achieved for linear and circular polarizations, respectively. The improvement provided by the polarization techniques is observed regardless of the illuminating wavelengths but is achieved at the expense of a reduced signal-to-noise ratio. Taking this into account, for the detection scheme considered with a detector well capacity of 4 x 10(5) electrons the improvement provided by linear polarization-maintaining light is reduced to a factor of 3.6 and for circular polarizations a factor of 2.2.
Surface reflections often present a problem in the polarization difference imaging of tissue. The technique described involves illumination in a single polarization state and detection in the orthogonal polarization state. Synchronously rotating both the illumination and orthogonal detection states provides an image free from surface reflections that is sensitive to the polarization properties of the underlying tissue. Results on tissue phantoms demonstrate that polarization sensitive measurements of a test target can be made up to a depth of 12 mean free paths within a scattering medium. Preliminary images of bovine tendon are also demonstrated.
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