Liver regeneration is delayed in mice with a defective TLR4 (C3H/HeJ) but is normal in TLR4 knockouts (TLR4(-/-) ). Here, we investigated the possible involvement of structural and hemodynamic changes in vivo in the underlying mechanism. In LPS-sensitive (C3H/HeN, C57BL/6) and LPS-insensitive (C3H/HeJ, TLR4(-/-) ) mice, a 70% partial hepatectomy (PH) was performed under inhalational anesthesia. At days 3 and 7 after PH, the hepatic microcirculation was interrogated using intravital microscopy. Delayed liver regeneration was confirmed in C3H/HeJ, but not in C3H/HeN, C57BL/6 (WT) or TLR4(-/-) mice by liver weight-to-body-weight ratio, percentage of PCNA-positive cells and mitotic index data. At day 3 after PH, sinusoidal RBC velocity increased by 100% in C3H/HeN but only by 40% in C3H/HeJ mice. Estimated sinusoidal blood flow was significantly higher at day 7 after PH in C3H/HeN than in C3H/HeJ mice. The hepatic cord width was significantly larger in C3H/HeN than in C3H/HeJ mice at day 3 and it was significantly larger in TLR4(-/-) than in C57BL/6 WT mice at day 7 after PH. Hepatocyte nucleus density and functional sinusoidal density was significantly reduced at days 3 and 7 after PH in all mouse strains compared to their zero-time controls. Functional sinusoidal density was significantly lower in C3H/HeJ compared to C3H/HeN mice at day 7 after PH. The present study indicates that altered sinusoidal blood flow and velocity in C3H/HeJ mice may contribute to the observed delay in regenerative response in these mice. In addition, restoration of normal liver architecture may be delayed in TLR4(-/-) mice. This article is protected by copyright. All rights reserved.