File:Fetal-Calcium-Regulates-Branching-Morphogenesis-in-the-Developing-Human-and-Mouse-Lung-Involvement-pone.0080294.s002.ogv

English: Time-lapse video of DiSBAC2(3) loaded lung epithelium. A lung explant from an E12.5 C57BL/6 mouse was cultured for 48 h before being loaded with DiSBAC2(3) through intra-luminal injection. DiSBAC2(3) fluorescence was visualized using a Cell Map IC confocal laser-scanning microscope with a water-immersion BX50WI and a UM Plan Fl 10x/0.30 W objective. Lungs were secured using a slice anchor in a 35 mm cell culture dish in 5 mL of solution containing in (mM): 135 NaCl, 5 KCl, 1.23 MgCl2, 1.0 mM CaCl2, 5 HEPES, 10 glucose, pH 7.4. DiSBAC2(3) was excited at 532 nm and images were collected using a 560 nm long pass filter, by the direct method. Images were taken at 6s intervals over a ten minute time period at room temperature and after 5 minutes, the KCl concentration in the extracellular solution was increased to 50 mM.