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通过总内部反射荧光显微镜与数量和亮度分析相结合,活细胞中细胞表面受体的显微化动力学
JoVE Journal
Biology
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JoVE Journal Biology
Oligomerization Dynamics of Cell Surface Receptors in Living Cells by Total Internal Reflection Fluorescence Microscopy Combined with Number and Brightness Analysis
DOI:

10:43 min

November 06, 2019

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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

Summary

Automatic Translation

我们描述了一种成像方法,用于测定由活细胞血浆膜中的配体结合诱导的mEGFP标记受体寡聚体的平均寡聚状态。该协议基于总内部反射荧光 (TIRF) 显微镜与数字和亮度 (N&B) 分析相结合。

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