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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
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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
Please note that all translations are automatically generated.
Click here for the English version.
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
DOI:
10.3791/60398-v
•
10:43 min
•
November 06, 2019
•
Moreno Zamai
,
Antonio Trullo
,
Elvira Arza
,
Ugo Cavallaro
,
Valeria R. Caiolfa
2
1
Unit of Microscopy and Dynamic Imaging
,
Centro Nacional de Investigaciones Cardiovasculares (CNIC), Madrid, Spain
,
2
Centro di Imaging Sperimentale
,
Ospedale San Raffaele, Milan, Italy
,
3
Unit of Gynecological Oncology Research
,
European Institute of Oncology IRCCS, Milan, Italy
Chapters
00:04
Title
01:48
Sample Preparation
02:38
TIRF Imaging: Alignment of the Laser Line and Optimization of TIRF Illumination
04:05
TIRF Imaging: Capture of the Time Series
05:46
Number & Brightness (N&B): Quality Check of the Time Series
06:34
Number & Brightness (N&B): Computation of the B-values in Selected Region-of-interest (ROI)
08:19
Results: N&B Analysis and Kinetics of FGFR1 Oligomerization
09:42
Conclusion
Summary
Automatic Translation
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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。
Tags
Oligomerization
Cell Surface Receptors
TIRF Microscopy
Number And Brightness Analysis
Receptor Clustering
FGFR1
Fluorescence Fluctuation
Live Cell Imaging
Protein Oligomerization
Cell Signaling
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