Using quantitative intravital multiphoton microscopy to dissect hepatic transport in rats

•Hepatic transport can be characterized using microscopy of fluorescent transporter substrates.•Multiphoton microscopy can resolve hepatic transport at subcellular resolution in vivo.•Quantitative intravital microscopy can quantify disruptions in hepatic transport in vivo. Hepatic solute transport is a complex process whose disruption is associated with liver disease and drug-induced liver injury. Intravital multiphoton fluorescence excitation microscopy provides the spatial and temporal resolution necessary to characterize hepatic transport at the level of individual hepatocytes in vivo and thus to identify the mechanisms and cellular consequences of cholestasis. Here we present an overview of the use of fluorescence microscopy for studies of hepatic transport in living animals, and describe how we have combined methods of intravital microscopy and digital image analysis to dissect the effects of drugs and pathological conditions on the function of hepatic transporters in vivo.