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Chapters
- 00:04Title
- 01:48Sample Preparation
- 02:38TIRF Imaging: Alignment of the Laser Line and Optimization of TIRF Illumination
- 04:05TIRF Imaging: Capture of the Time Series
- 05:46Number & Brightness (N&B): Quality Check of the Time Series
- 06:34Number & Brightness (N&B): Computation of the B-values in Selected Region-of-interest (ROI)
- 08:19Results: N&B Analysis and Kinetics of FGFR1 Oligomerization
- 09:42Conclusion
We describe an imaging approach for the determination of the average oligomeric state of mEGFP-tagged-receptor oligomers induced by ligand binding in the plasma membrane of living cells. The protocol is based on Total Internal Reflection Fluorescence (TIRF) microscopy combined with Number and Brightness (N&B) analysis.
