Abstract
Two-photon laser scanning microscopy is widely used to measure blood hemodynamics in brain blood vessels. Still, the algorithms used so far to extract red blood cell (RBC) size and velocity from line-scan acquisitions have ignored the extent to which scanning speed influences the measurements. Here, we used a theoretical approach that takes into account the velocity and direction of both scanning mirrors and RBCs during acquisition to provide an algorithm that measures the real RBC size and velocity. We validate our approach in brain vessels of anesthetized mice, and demonstrate that it corrects online measurement errors that can reach several 10s of percent as well as data previously acquired. To conclude, our analysis allows unbiased comparisons of blood hemodynamic parameters from brain capillaries and large vessels in control and pathological animal models.
Highlights
Blood flow mapping is widely used to image brain activity in physiological or pathological conditions because of the tight coupling that links neuronal activation and functional hyperemia (Iadecola, 2017)
red blood cell (RBC) undergo major deformation depending on blood flow dynamics within microvessels, in particular when they pass through capillaries that are smaller than their diameter (Suzuki et al, 1996; Zharov et al, 2006)
We first investigated the relationship between the size of a RBC shadow, the RBC real size and the scanning velocity
Summary
Blood flow mapping is widely used to image brain activity in physiological or pathological conditions because of the tight coupling that links neuronal activation and functional hyperemia (Iadecola, 2017). RBCs undergo major deformation depending on blood flow dynamics within microvessels, in particular when they pass through capillaries that are smaller than their diameter (Suzuki et al, 1996; Zharov et al, 2006). This deformability is impaired in many pathological conditions as hereditary disorders (for example spherocytosis, elliptocytosis, ovalocytosis, and stomatocytosis), diabetes, hypercholesterolemia (reviewed by Tomaiuolo, 2014), or during infection by plasmodium (Tiburcio et al, 2012). Accurate measurement of RBC shape and velocity with laser scanning microscopy
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