Visualization and Quantitative Analysis of the Actin Cytoskeleton Upon B Cell Activation
| Autor: | Vid Šuštar, Marika Vainio, Pieta K. Mattila |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Cell signaling Phalloidin macromolecular substances Actin cytoskeleton Immunological synapse Cell biology 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology 0302 clinical medicine chemistry Live cell imaging Fluorescence microscope Cytoskeleton 030217 neurology & neurosurgery Actin |
| Zdroj: | Methods in Molecular Biology ISBN: 9781493974733 |
| DOI: | 10.1007/978-1-4939-7474-0_18 |
| Popis: | The formation of the immunological synapse upon B cell activation critically depends on the rearrangement of the submembranous actin cytoskeleton. Polymerization of actin monomers into filaments provides the force required for B cell spreading on the antigen-presenting cell (APC). Interestingly, the actin network also participates in cellular signaling at multiple levels. Fluorescence microscopy plays a critical role in furthering our understanding of the various functions of the cytoskeleton, and has become an important tool in the studies on B cell activation. The actin cytoskeleton can be tracked in live cells with various fluorescent probes binding to actin, or in fixed cells typically with phalloidin staining. Here, we present the usage of TIRF microscopy and an image analysis workflow for studying the overall density and organization of the actin network upon B cell spreading on antigen-coated glass, a widely used model system for the formation of the immunological synapse. |
| Databáze: | OpenAIRE |
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