Other Required Materials:
Visualize bile canaliculi structure and MRP2 transporter function in Emulate Organ-Chips
Carboxy-DCFDA (5-(and-6)-Carboxy-2′,7′-Dichlorofluorescein Diacetate) is a reagent that passively diffuses into cells. It is colorless / nonfluorescdent and is cleaved by intracellular esterases to yield a fluoresecent fluirophore, 5-(and-6)-carboxy-2′,7′ –dichlorofluorescein (CDFDA). This reagent can be used to visualize the formation of bile canaliculi in polarized hepatocytes which is indicative of MRP-2 trasnporter activity. Carboxy-DCFDA is a substrate of multidrug resistance-associated protein 2 (MRP2), a hepatocyte efflux transporter, thus it can be used to visualize MRP2-mediated canalicular uptake and efflux, as well as to label the bile canaliculi structures.
See Protocol EP155 Live Staining of Cells.
Recommended reagent dilution and incubation time
Dilute 5 mM stock CDFDA solution 1:500 in culture medium. Incubate in the dark at 37°C for 30 minutes.
Image of CDFDA (green) and nuclei staining (blue) indicating hepatocyte polarization, establishment of bile canaliculi structures, and active MRP2 transporters in hepatocytes in the human Liver-Chip (top channel).
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