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Studying Organelle Dynamics in B Cells During Immune Synapse Formation
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Chapters
- 00:00Title
- 01:16B Cell Activation with Antigen-coated Beads
- 02:48B Cell Activation on Antigen-coated Coverslips
- 03:51Immunofluoresence and Image Analysis
- 04:30Analysis of the Distribution of Organelles in B Cells Activated with Ag-coated Beads
- 07:42Analysis of Cell Spreading and Distribution of Organelles in B Cells Activated on Ag-coated Coverslips
- 09:29Isolation of Centrosome-enriched Fractions from Resting and Activated B Cells
- 11:23Results
- 15:07Conclusion
Herein we describe two approaches to characterize cell polarization events in B lymphocytes during the formation of an IS. The first, involves quantification of organelle recruitment and cytoskeleton rearrangements at the synaptic membrane. The second is a biochemical approach, to characterize changes in composition of the centrosome, which undergoes polarization to the immune synapse.
