Journal
/
Immunology and Infection
/
Visualizing the Actin and Microtubule Cytoskeletons at the B-cell Immune Synapse Using Stimulated Emission Depletion (STED) Microscopy
JoVE Journal
Immunology and Infection
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Immunology and Infection
Visualizing the Actin and Microtubule Cytoskeletons at the B-cell Immune Synapse Using Stimulated Emission Depletion (STED) Microscopy

Visualizing the Actin and Microtubule Cytoskeletons at the B-cell Immune Synapse Using Stimulated Emission Depletion (STED) Microscopy

DOI:
10.3791/57028-v

•

11:00 min

•

April 09, 2018

•

, ,
1Department of Microbiology and Immunology,University of British Columbia

Chapters

  • 00:05Title
  • 01:18GFP-Fusion Protein Expression and Anti-IgG Antibody Glass Coverslip Coating
  • 03:14Cell Spreading, Fixing, and Immunostaining
  • 05:44Stimulated Emission Depletion Microscope Imaging
  • 08:02Results: Representative STED Imaging
  • 09:50Conclusion

Summary

Automatic Translation

Automatic Translation

We present a protocol for using STED microscopy to simultaneously image actin structures, microtubules, and microtubule plus-end binding proteins in B cells that have spread on coverslips coated with antibodies to the B-cell receptor, a model for the initial phase of immune synapse formation.

Tags

Keywords: ActinMicrotubuleCytoskeletonB-cellImmune SynapseSTED MicroscopyCytoskeleton OrganizationCytoskeleton ReorganizationImmune Cell ActivationFluorophoresA20 B-lymphoma CellsTransfectionGoat Anti-mouse Immunoglobulin G AntibodyCoverslips

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/57028?article_type=v

Seven Steps to Stellate Cells
check_url/57028?article_type=v

Measuring Bacterial Load and Immune Responses in Mice Infected with Listeria monocytogenes
check_url/57028?article_type=v

An Introduction to Parasitic Wasps of Drosophila and the Antiparasite Immune Response
check_url/57028?article_type=v

Intravital Imaging of the Mouse Thymus using 2-Photon Microscopy
check_url/57028?article_type=v

Investigating the Effects of Probiotics on Pneumococcal Colonization Using an In Vitro Adherence Assay
check_url/57028?article_type=v

Visualization of the Immunological Synapse by Dual Color Time-gated Stimulated Emission Depletion (STED) Nanoscopy
check_url/57028?article_type=v

In Vitro Analysis of Myd88-mediated Cellular Immune Response to West Nile Virus Mutant Strain Infection
check_url/57028?article_type=v

Qualitative and Quantitative Analysis of the Immune Synapse in the Human System Using Imaging Flow Cytometry
check_url/57028?article_type=v

Visualizing Macrophage Extracellular Traps Using Confocal Microscopy
check_url/57028?article_type=v

Co-staining Blood Vessels and Nerve Fibers in Adipose Tissue

Read Article