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活体哺乳动物细胞器中单膜蛋白的多色定位显微术
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Biologia
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JoVE Journal Biologia
Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
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活体哺乳动物细胞器中单膜蛋白的多色定位显微术

DOI:
10.3791/57690-v

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11:06 min

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June 30, 2018

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, , , ,
1School of Biology,University of Osnabrück, 2Center of Cellular Nanoanalytics, Integrated Bioimaging Facility,University of Osnabrück, 3Department of Biology,WWU Münster

Capítulos

  • 00:04Título
  • 00:54Physical Alignment of Image Splitter Generated Images
  • 02:25Alignment of Spectrally Separated Channels by Software Performing Spatial Transformation
  • 04:34Single Molecule Imaging of Mitochondrial Membrane Proteins
  • 08:32Results: Multi-color Localization of Mammalian Mitochondrial PINK1
  • 10:15Conclusion

Summary

Tadução automática

Tadução automática

在这里, 我们提出了一个协议的多颜色本地化的单膜蛋白在细胞器的活体细胞。要附加显影, 使用自贴标蛋白。蛋白质, 位于同一细胞器的不同的细胞膜车厢, 可以本地化的精度为 18 nm。

Tags

Keywords: Multi-color Localization MicroscopySingle Membrane ProteinsOrganellesLive Mammalian CellsProtein DynamicsSpatiotemporal OrganizationMitochondrial Membrane ProteinsTransfectionSample PreparationMicroscope SetupFluorescent BeadsObjective LensImmersion OilTransmission LightLaserFluorescence ChannelsImage SplitterTIRF MicroscopyTransformation MatrixDual-color ImagesCalibrationUnit ConversionLocalization ManagerPoint Spread Function (PSF)

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