BSC1 (monkey kidney epithelial) cells stably expressing rat brain clathrin light chain-EGFP (kindly provided by T. Kirchhausen, Harvard Medical School, Boston, MA) were cultured and prepared as specified in Supplementary Note 13 online. For live cell imaging, BSC1 cells were plated on glass coverslips, and through-the-objective TIR-FM was performed on a Nikon TE2000U inverted microscope using a 100X/1.45NA oil-immersion objective. Images were captured at 0.5 Hz with 200ms exposure time using a Hamamatsu Orca II-ERG.
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Live-cell Imaging of Clathrin-mediated Endocytosis
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Other organizations : Scripps Research Institute, University of Toronto
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Variable analysis
independent variables
- Stable expression of rat brain clathrin light chain-EGFP in BSC1 (monkey kidney epithelial) cells
- Dynamics of clathrin light chain-EGFP in live BSC1 cells
- BSC1 (monkey kidney epithelial) cells
- Glass coverslips for cell plating
- Nikon TE2000U inverted microscope with 100X/1.45NA oil-immersion objective for live cell imaging
- TIR-FM (through-the-objective total internal reflection fluorescence microscopy) for imaging
- Hamamatsu Orca II-ERG camera for image capture at 0.5 Hz with 200ms exposure time
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