Clendenon, Sherry G.Fu, XiaoVon Hoene, Robert A.Clendenon, Jeffrey L.Sluka, James P.Winfree, SethMang, HenryMartinez, MichelleFilson, Adele J.Klaunig, James E.Glazier, James A.Dunn, Kenneth W.2020-07-212020-07-212019-05Clendenon, S. G., Fu, X., Von Hoene, R. A., Clendenon, J. L., Sluka, J. P., Winfree, S., Mang, H., Martinez, M., Filson, A. J., Klaunig, J. E., Glazier, J. A., & Dunn, K. W. (2019). A simple automated method for continuous fieldwise measurement of microvascular hemodynamics. Microvascular research, 123, 7–13. https://doi.org/10.1016/j.mvr.2018.11.010https://hdl.handle.net/1805/23309Microvascular 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.en-USPublisher PolicyRed blood cell velocityIntravital microscopyHemodynamicsMicrovascularCapillaryArticle