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Related Experiment Video

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Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized

Rosa Machado¹, Justin Bendesky¹, Madison Brown^1,2

¹UCCS Center for the Biofrontiers Institute, University of Colorado at Colorado Springs, Colorado Springs, CO 80918, USA.

Journal of Imaging

|July 31, 2019

Summary

Polarized total internal reflection fluorescence microscopy (P-TIRF) visualizes nanoscale cell membrane curvature. This technique reveals how cell surface molecule dynamics correlate with membrane mechanics during immunological synapse formation.

Keywords:

FcεRI IgE receptor P-TIRF RBL-2H3 plasma membrane rat basophilic leukemia cells supported lipid bilayer total internal reflection fluorescence microscopy

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Area of Science:

Cell biology
Biophysics
Immunology

Background:

Cell surface receptor dynamics are crucial for cell signaling.
Understanding plasma membrane mechanics is key to cell function.
Immunological synapse formation involves complex molecular interactions.

Purpose of the Study:

To investigate nanoscale curvature phenomena in live cells using P-TIRF.
To model and visualize the formation of an immunological synapse.
To correlate cell surface molecule dynamics with plasma membrane mechanical properties.

Main Methods:

Utilized polarized total internal reflection fluorescence microscopy (P-TIRF).
Visualized rat basophilic leukemia (RBL-2H3) cells primed with fluorescent anti-dinitrophenyl (anti-DNP) immunoglobulin E (IgE).
Analyzed the spatial relationship of IgE-bound FcεRI to polarized excitation ratios.

Main Results:

P-TIRF successfully imaged nanoscale curvature in live RBL-2H3 cells.
Visualized the interaction of IgE-primed cells with a supported lipid bilayer containing DNP.
Correlated the spatial distribution of FcεRI with membrane polarization.

Conclusions:

P-TIRF is effective for imaging nanoscale membrane events in live cells.
The study provides insights into the mechanical aspects of immunological synapse formation.
This approach links molecular dynamics to membrane mechanics during cell-cell interactions.