Measurement of absorption in scattering media using objective laser speckle: application to blood oximetry

Multi-spectral imaging enables non-invasive sensing of chemical concentrations in biological tissue based on measurement of optical absorption, but invariably in the presence of high levels of scatter. Absorption is normally inferred from measurement of contrast of biological features, such as the vasculature, and so accuracy is degraded by the poorly characterized modulation-transfer function of the imaging optics and overlying tissue. We report how experimental characterization of the spectral variation of the tissue point-spread function and associated objective speckle pattern can be used to characterize the absorption spectrum and chromophore concentration, with a particular emphasis on determination of the ratio of oxygenated to deoxygenated hemoglobin within blood. Absorption measurements are determined purely by the geometry of the experiment, without degradation due to optical aberrations and associated light scatter. The technique offers enhanced and low-cost determination of in vitro or in vivo chromophore characterizations, including blood-gas analysis.