Reproducibility of measuring cerebral blood flow by laser-Doppler flowmetry in mice

Laser-Doppler flowmetry has been widely used to trace hemodynamic changes in experimental stroke research. The purpose of the present study was to evaluate the day-to-day test-retest reproducibility of measuring cerebral blood flow by LDF in awake mice. The flux indicating cerebral blood flow (CBF), red blood cell (RBC) velocity, and RBC concentration were measured with LDF via cranial windows for the bilateral somatosensory cortex in awake mice. LDF measurements were performed three times, at baseline, 1 hour after, and 7 days after the baseline measurement. Moreover, breathing rate (BR) and partial pressure of transcutaneous CO₂ (PtCO₂) were measured simultaneously with LDF measurement. Intraclass correlation coefficient (ICC) and within-subject coefficient of variation (CVw) were calculated. CBF, RBC velocity, and RBC concentration showed good day-to-day test-retest reproducibility (ICC: 0.61 - 0.95, CVw: 8.3% - 15.4%). BR and PtCO₂ in awake mice were stable during the course of the experiments. The evaluation of cerebral microcirculation using LDF appears to be applicable to long-term studies.