The purpose of this study was to investigate red blood cell (RBC) behavior during an increase in local cerebral blood flow (LCBF). We measured changes in RBC behavior by using laser-Doppler flowmetry (LDF) in &agr;-chloralose-anesthetized rats. An increase in LCBF was carried out by approximately 2.5 and 4.0% CO₂ inhalation and activation of the somatosensory cortex. The activation of the cortex was induced by electrical stimulation of the hind paw with 1.5-mA pulses (0.1 ms) applied at frequencies of 0.2, 1, 5, and 10 Hz for a 5 s duration. The increases in LCBF and RBC velocity during both CO₂ inhalations were larger than that in RBC concentration (p < 0.05). LCBF and RBC velocity during 4.0% CO₂ inhalation were larger than those during 2.5% CO₂ inhalation (p < 0.05), though there was no significant difference in RBC concentration between the two conditions, suggesting a limitation of capillary volume. During somatosensory stimulation, the evoked LCBF increased with increasing stimulus frequency up to 5 Hz and decreased at 10 Hz. The responses of RBC concentration at 0.2 and 10 Hz were greater than those of RBC velocity (p < 0.05), but no significant differences in response magnitude were found at 1 and 5 Hz between RBC concentration and RBC velocity. These results suggest that the increase in LCBF during neuronal activity is different from that of controlling the LCBF as induced by CO₂, and that the regulation of RBC concentration and RBC velocity is controlled by independent mechanisms.