In vivo two-photon excited fluorescence microscopy reveals cardiac- and respiration-dependent pulsatile flow in cortical blood vessels in mice.

T.P. Santisakultarm, N.R. Cornelius, N. Nishimura, A.I. Schafer, R.T. Silver, P.C. Doerschuk, W.L. Olbricht, C.B. Schaffer
Am. J Physiol. Heart Circ. Physiol. Vol. 302, No. 7, pp. H1367-H1377

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Subtle alterations in cerebral blood flow can impact the health and function of brain cells and are linked to cognitive decline and dementia. To understand hemodynamics in the three-dimensional vascular network of the cerebral cortex, we applied two-photon excited fluorescence microscopy to measure the motion of red blood cells (RBCs) in individual microvessels throughout the vascular hierarchy in anesthetized mice. To resolve heartbeat- and respiration-dependent flow dynamics we simultaneously recorded the electrocardiogram and respiratory waveform. We found that centerline RBC speed decreased with decreasing vessel diameter in arterioles, slowed further through the capillary bed, then increased with increasing vessel diameter in venules. RBC flow was pulsatile in nearly all cortical vessels, including capillaries and venules. Heartbeat-induced speed modulation decreased through the vascular network, while the delay between heartbeat and the time of maximum speed increased. Capillary tube hematocrit was 0.21 and did not vary with centerline RBC speed or topological position. Spatial RBC flow profiles in surface vessels were blunted compared to a parabola, and could be measured at vascular junctions. Finally, we observed a transient decrease in RBC speed in surface vessels prior to inspiration. In conclusion, we have developed an approach to study detailed characteristics of RBC flow in the three-dimensional cortical vasculature, including quantification of fluctuations in centerline RBC speed due to cardiac and respiratory rhythms and flow profile measurements. These methods and the quantitative data on basal cerebral hemodynamics open the door to studies of normal and disease state cerebral microcirculation.

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