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Image Processing Protocol for the Analysis of the Diffusion and Cluster Size of Membrane Receptors by Fluorescence Microscopy
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JoVE 杂志 免疫与感染
Image Processing Protocol for the Analysis of the Diffusion and Cluster Size of Membrane Receptors by Fluorescence Microscopy

Image Processing Protocol for the Analysis of the Diffusion and Cluster Size of Membrane Receptors by Fluorescence Microscopy

DOI:
10.3791/59314-v

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12:15 min

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April 09, 2019

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, , , , , ,
1Department of Macromolecular Structures, Centro Nacional de Biotecnología,Campus Univ. Autónoma de Madrid, 2Campus Urb. Montepríncipe s/n,Univ. San Pablo CEU, 3Department of Immunology and Oncology, Centro Nacional de Biotecnología,Campus Univ. Autónoma de Madrid, 4Department of Cell Signaling,Centro Andaluz de Biología Molecular y Medicina Regenerativa (CSIC), 5Meyer Cancer Center, 6Department of Anatomy and Cell Biology,McGill Univ.

Chapters

  • 00:04Title
  • 01:37Preparation of Biological Samples and Image Acquisition
  • 02:57Selection of Images and Creation of Masks
  • 05:45Tracking Particles
  • 08:09Calculation of the Diffusion Coefficients and Classification of Trajectories
  • 09:10Calculation of Cluster Size
  • 09:50Results: Analysis of Dynamic Fluorescence Particle Characteristics
  • 11:24Conclusion

Summary

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Here, we present a protocol for single particle tracking image analysis that allows quantitative evaluation of diffusion coefficients, types of motion and cluster sizes of single particles detected by fluorescence microscopy.

Tags

Image ProcessingSingle-molecule TrackingLateral DiffusionSpot SizeMembrane ReceptorsFluorescence MicroscopyTIRF MicroscopyJurkat CellsChemokine ReceptorCXCL12

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