Abstract
Sorting and trafficking of proteins to their target membranes is crucial for the function of epithelia as fluid transporting entities. A powerful tool to study membrane trafficking is total internal reflection fluorescence (TIRF) microscopy at which only a ∼100 nm thin layer at the glass buffer interface is illuminated. In particular it allows for monitoring motion and membrane fusion of vesicles carrying membrane proteins with high temporal resolution in living cells. However, TIRF microscopy was limited so far to study membrane trafficking at the basolateral membrane at the sites where cells are attached to the glass cover slip.To overcome this limitation we developed a microfluidic biochip which allows for approaching the apical membrane of polarized cells towards a glass cover slip in a controlled way. The chip was applied to visualize fluorescently tagged aquaporins at the apical membrane of Madin-Darby canine kidney (MDCK) cells by TIRF microscopy.
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