Laser Speckle Orthogonal Contrast Imaging Calibration by Replicating Red Blood Cell Scattering Statistics with a Moving Reference Diffuser

Recent studies have proposed improving Laser Speckle Contrast Imaging (LSCI) by using polarimetric filtering to isolate multiply scattered photons from moving red blood cells (RBCs), an approach referred to as Laser Speckle Orthogonal Contrast Imaging (LSOCI). This reliance on multiple scattering enables the development of a calibration method based on a moving reference sample, chosen to generate dynamic circular Gaussian speckle fields that replicate the statistical properties of RBC scattering in both intensity and the distribution of polarization states. Assuming that multiply scattered photons from both RBCs and the reference sample exhibit a homogeneous distribution of polarization states over the Poincaré sphere, the proposed calibration links in vivo speckle contrast reduction bijectively to an equivalent speed of the reference sample. We demonstrate that this equivalent-velocity metric yields consistent in vivo measurements across distinct instruments despite the use of different laser spectral widths, thereby providing a standardized and transferable means to quantify microcirculatory activity.