Laser speckle contrast imaging of blood flow from anesthetized mice: correcting drifts in measurements due to breathing movements

Abstract

Background: Laser speckle contrast imaging allows non-invasive assessment of cutaneous blood flow. Although the technique is attractive to measure a quantity related to the skin blood flow (SBF) in anesthetized animal models, movements from breathing can mask the SBF signal. As a consequence, the measurement is overestimated because a variable amount of a DC component due to the breathing movements is added to the SBF signal. Objective: To evaluate a method for estimating the background level of the SBF signal, rejecting artefacts from breathing. Methods: A baseline correction method used for accurate DNA sequencing was evaluated, based on estimating the background level of a signal in small temporal sliding-windows. The method was applied to evaluate a mouse model of hindlimb ischemia. SBF signals from hindlimbs of anesthetized C57BL/6 mice (n=13) were registered. The mean SBF (Fi and Fc from ischemic and control hindlimbs) were computed from the registers and from the corresponding estimated background levels (Fib and Fcb from ischemic and control hindlimbs). Results: The mean values of the percentages (a measure of ischemia) MI = (Fi/Fc).100 and MIb = (Fib/Fcb).100 were computed to be 30±4% and 23±3% respectively (mean ± SE). Evidences of statistical differences between both, ischemic and control hindlimbs, were obtained (p<0.05, paired student-t). The mean error [(MI-MIb)/MIb].100 obtained was 45±14% (mean±SE). Conclusion: The recovery of a corrupted SBF signal by breathing artefacts is feasible, allowing more accurate measurements.