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Item A simple automated method for continuous fieldwise measurement of microvascular hemodynamics(Elsevier, 2019-05) Clendenon, Sherry G.; Fu, Xiao; Von Hoene, Robert A.; Clendenon, Jeffrey L.; Sluka, James P.; Winfree, Seth; Mang, Henry; Martinez, Michelle; Filson, Adele J.; Klaunig, James E.; Glazier, James A.; Dunn, Kenneth W.; Medicine, School of MedicineMicrovascular perfusion dynamics are vital to physiological function and are frequently dysregulated in injury and disease. Typically studies measure microvascular flow in a few selected vascular segments over limited time, failing to capture spatial and temporal variability. To quantify microvascular flow in a more complete and unbiased way we developed STAFF (Spatial Temporal Analysis of Fieldwise Flow), a macro for FIJI open-source image analysis software. Using high-speed microvascular flow movies, STAFF generates kymographs for every time interval for every vascular segment, calculates flow velocities from red blood cell shadow angles, and outputs the data as color-coded velocity map movies and spreadsheets. In untreated mice, analyses demonstrated profound variation even between adjacent sinusoids over seconds. In acetaminophen-treated mice we detected flow reduction localized to pericentral regions. STAFF is a powerful new tool capable of providing novel insights by enabling measurement of the complex spatiotemporal dynamics of microvascular flow.