Several variables may affect the local contrast values in laser speckle contrast imaging (LSCI), irrespective of relative motion. It has been suggested that the optical properties of the moving fluid and surrounding tissues can affect LSCI values. However, a detailed study of this has yet to be presented. In this work, we examined the combined effects of the reduced scattering and absorption coefficients on LSCI. This study employs fluid phantoms with different optical properties that were developed to mimic whole blood with varying hematocrit levels. These flow phantoms were imaged with an LSCI system developed for this study. The only variable parameter was the optical properties of the flowing fluid. A negative linear relationship was seen between the changes in contrast and changes in reduced scattering coefficient, absorption coefficient, and total attenuation coefficient. The change in contrast observed due to an increase in the scattering coefficient was greater than what was observed with an increase in the absorption coefficient. The results indicate that optical properties affect contrast values and that they should be considered in the interpretation of LSCI data.
Unlike laser Doppler flowmetry, there has yet to be presented a clear description of the physical variables that laser speckle contrast imaging (LSCI) is sensitive to. Herein, we present a theoretical basis for demonstrating that LSCI is sensitive to total flux and, in particular, the summation of diffusive flux and advective flux. We view LSCI from the perspective of mass transport and briefly derive the diffusion with drift equation in terms of an LSCI experiment. This equation reveals the relative sensitivity of LSCI to both diffusive flux and advective flux and, thereby, to both concentration and the ordered velocity of the scattering particles. We demonstrate this dependence through a short series of flow experiments that yield relationships between the calculated speckle contrast and the concentration of the scatterers (manifesting as changes in scattering coefficient), between speckle contrast and the velocity of the scattering fluid, and ultimately between speckle contrast and advective flux. Finally, we argue that the diffusion with drift equation can be used to support both Lorentzian and Gaussian correlation models that relate observed contrast to the movement of the scattering particles and that a weighted linear combination of these two models is likely the most appropriate model for relating speckle contrast to particle motion.
Before laser speckle contrast imaging (LSCI) can be used reliably and quantitatively in a clinical setting, there are several theoretical and practical issues that still must be addressed. In order to address some of these issues, an electro-optical system that utilizes a nematic liquid crystal spatial light modulator (SLM) to mimic LSCI experiments was assembled. The focus of this paper is to address the issue of how incident intensity affects LSCI results. Using the SLM-based system, we systematically adjusted incident intensity on the SLM and assessed the resulting first- and second-order statistics of the imaged speckle to explain the corresponding spatial contrast values in both frozen and time-integrated speckle patterns. The SLM-based system was used to generate speckle patterns with a controlled minimum speckle size, probability intensity distribution, and temporal decorrelation behavior. By eliminating many experimental parameters, this system is capable of serving as a useful intermediary tool between computer simulation and physical experimentation for further developing LSCI as a quantitative imaging modality.
Laser speckle contrast imaging (LSCI) is a non- or minimally- invasive modality for observing relative blood flow or perfusion. Recently, there has been an effort to use LSCI for truly quantitative blood flow measurements. However, this effort has been hampered not only by real theoretical issues, but also by challenges associated with numerous experimental parameters that can potentially impact the calculated contrast values. In this work, we present our efforts at using a nematic liquid crystal, phase-only, spatial light modulator (SLM) to mimic LSCI experiments with precisely controlled experimental parameters. This approach permits the rapid experimental investigation of numerous factors including: The effects of different flow models on LSCI contrast values; the effects of multiple decorrelation times in the same depth of field; the effects of ‘static’ scatterers; and the effects of camera settings relative to speckle decorrelation times, just to name a few. We have found that an SLM is a useful tool for the experimental investigation of LSCI that eliminates many of the experimental variables associated with typical flow model experiments or in vivo experimentation. LSCI experiments with SLMs are a useful intermediary between computer simulations and physical flow models.
Laser Speckle contrast imaging (LSCI) is a non-invasive or minimally invasive method for visualizing blood flow and perfusion in biological tissues. In LSCI the motion of scattering particles results in a reduction in global and regional speckle contrast. A variety of parameters can affect the calculated contrast values in LSCI techniques, including the optical properties of the fluid and surrounding tissue. In typical LSCI where the motion of blood is of interests, optical properties are influenced by hematocrit levels. In this work we considered the combined effects of both the scattering and absorption coefficients on LSCI measurements on a flow phantom. Fluid phantoms consisting of various concentrations of neutrally buoyant ~10 micron microspheres and India ink mixed with DI water were formulated to mimic the optical properties of whole blood with various levels of hematocrit. In these flow studies, it was found that an increase in μa and/or μs led to a decrease in contrast values when all other experimental parameters were held constant. The observed reduction in contrast due to optical property changes could easily be confused with a contrast reduction due to increased flow velocity. These results suggest that optical properties need to be considered when using LSCI to make flow estimates.
The dynamic behavior of phase singularities, or optical vortices, in the pseudo-phase representation of dynamic speckle patterns is investigated. Sequences of band-limited, dynamic speckle patterns with predetermined Gaussian decorrelation behavior were generated, and the pseudo-phase realizations of the individual speckle patterns were calculated via a two-dimensional Hilbert transform algorithm. Singular points in the pseudo-phase representation are identified by calculating the local topological charge as determined by convolution of the pseudo-phase representations with a series of 2×2 nabla filters. The spatial locations of the phase singularities are tracked over all frames of the speckle sequences, and recorded in three-dimensional space (x,y,f), where f is frame number in the sequence. The behavior of the phase singularities traces 'vortex trails' which are representative of the speckle dynamics. Slowly decorrelating speckle patterns results in long, relatively straight vortex trails, while rapidly decorrelating speckle patterns results in tortuous, relatively short vortex trails. Optical vortex analysis such as described herein can be used as a descriptor of biological activity, flow, and motion.