Impact of changes in tissue optical properties on near-infrared diffuse correlation spectroscopy measures of skeletal muscle blood flow

Near-infrared diffuse correlation spectroscopy (DCS) is increasingly used to study relative changes in skeletal muscle blood flow. However, most diffuse correlation spectrometers assume that tissue optical properties-such as absorption (μ_a) and reduced scattering (μ'_s) coefficients-remain constant during physiological provocations, which is untrue for skeletal muscle. Here, we interrogate how changes in tissue μ_a and μ'_s affect DCS calculations of blood flow index (BFI). We recalculated BFI using raw autocorrelation curves and μ_a/μ'_s values recorded during a reactive hyperemia protocol in 16 healthy young individuals. First, we show that incorrectly assuming baseline μ_a and μ'_s substantially affects peak BFI and BFI slope when expressed in absolute terms (cm²/s, P < 0.01), but these differences are abolished when expressed in relative terms (% baseline). Next, to evaluate the impact of physiologic changes in μ_a and μ'_s, we compared peak BFI and BFI slope when μ_a and μ'_s were held constant throughout the reactive hyperemia protocol versus integrated from a 3-s rolling average. Regardless of approach, group means for peak BFI and BFI slope did not differ. Group means for peak BFI and BFI slope were also similar following ad absurdum analyses, where we simulated supraphysiologic changes in μ_a/μ'_s. In both cases, however, we identified individual cases where peak BFI and BFI slope were indeed affected, with this result being driven by relative changes in μ_a over μ'_s. Overall, these results provide support for past reports in which μ_a/μ'_s were held constant but also advocate for real-time incorporation of μ_a and μ'_s moving forward.NEW & NOTEWORTHY We investigated how changes in tissue optical properties affect near-infrared diffuse correlation spectroscopy (NIR-DCS)-derived indices of skeletal muscle blood flow (BFI) during physiological provocation. Although accounting for changes in tissue optical properties has little impact on BFI on a group level, individual BFI calculations are indeed impacted by changes in tissue optical properties. NIR-DCS calculations of BFI should therefore account for real-time, physiologically induced changes in tissue optical properties whenever possible.