Visualizing Bile Acid and MDR1 Transport

The bile acid conjugate CLF has been used for visualisation of bile acid transportation across liver tissue^{3 (link),5 (link),6 ,25 (link)}, in particular as regulated by MRP2 whereas the activities of MDR1 was assessed by rhodamine 123 efflux^{8 (link),9 (link),19 (link)}. Independently, cysts were washed and incubated separately with 5 μM CLF (BD, Japan) and 100 μM rhodamine 123 (BD) in the free serum-culture medium, at 37 °C within 30 min. Subsequently, cysts were rinsed twice with fresh culture medium. The fluorescence dye accumulation was then recorded using a confocal microscope (Olympus, Japan) employing FITC (495–519 nm) for CLF and rhodamine green (502–527 nm) emission spectra.

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Rizki-Safitri A., Shinohara M., Miura Y., Danoy M., Tanaka M., Miyajima A, & Sakai Y. (2018). Efficient functional cyst formation of biliary epithelial cells using microwells for potential bile duct organisation in vitro. Scientific Reports, 8, 11086.

Publication 2018

rhodamine 123 confocal microscope

Bile acid Confocal microscope Culture medium Cysts Fitc Fluorescence dye Liver Rhodamine Rhodamine 123 Serum

Corresponding Organization : The University of Tokyo

Other organizations : Waseda University, Laboratory for Integrated Micro-Mechatronic Systems

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Variable analysis

independent variables

Concentration of CLF (5 μM)
Concentration of rhodamine 123 (100 μM)

dependent variables

Fluorescence dye accumulation

control variables

Incubation time (30 min)
Incubation temperature (37 °C)
Culture medium (free serum-culture medium)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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