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Updated: Feb 15, 2026

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Visualization and Quantitative Analysis of the Actin Cytoskeleton Upon B Cell Activation

Vid Šuštar1, Marika Vainio1, Pieta K Mattila2

  • 1Institute of Biomedicine, Unit of Pathology, and MediCity Research Laboratories, University of Turku, Turku, Finland.

Methods in Molecular Biology (Clifton, N.J.)
|February 2, 2018
PubMed

Insights

B cell activation relies on actin cytoskeleton rearrangement for cell spreading during immunological synapse formation. This study uses TIRF microscopy to analyze actin density and organization in B cells spreading on antigen-coated surfaces.

Area of Science:

  • Immunology
  • Cell Biology
  • Biophysics

Background:

  • The immunological synapse formation in B cells requires actin cytoskeleton remodeling.
  • Actin polymerization drives B cell spreading on antigen-presenting cells (APCs).
  • The actin network plays a crucial role in cellular signaling during B cell activation.

Purpose of the Study:

  • To investigate the actin cytoskeleton's density and organization during B cell spreading.
  • To present a methodology using TIRF microscopy and image analysis for studying actin dynamics.
  • To model immunological synapse formation using B cells spreading on antigen-coated glass.

Main Methods:

  • Utilizing Total Internal Reflection Fluorescence (TIRF) microscopy for high-resolution imaging.
  • Developing an image analysis workflow to quantify actin network properties.
  • Employing a model system of B cells spreading on antigen-coated glass surfaces.

Main Results:

  • Demonstrated changes in actin density and organization during B cell spreading.
  • Quantified the overall actin network structure in the context of synapse formation.
  • Provided insights into the mechanical forces governing B cell-APC interactions.

Conclusions:

  • TIRF microscopy and image analysis are effective tools for studying actin dynamics in B cell activation.
  • Understanding actin organization is key to deciphering the mechanisms of immunological synapse formation.
  • This approach offers a valuable method for future research on cell-cell interactions and cytoskeletal functions.

Keywords:
Actin cytoskeletonB cellsImage analysisImmunological synapseLive cell imagingLymphocytesMicroscopySignaling microclustersTIRFTotal internal reflection microscopy

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