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

Journal of Imaging
Rosa MachadoGuy M. Hagen

Abstract

Total internal reflection fluorescence microscopy with polarized excitation (P-TIRF) can be used to image nanoscale curvature phenomena in live cells. We used P-TIRF to visualize rat basophilic leukemia cells (RBL-2H3 cells) primed with fluorescent anti-dinitrophenyl (anti-DNP) immunoglobulin E (IgE) coming into contact with a supported lipid bilayer containing mobile, monovalent DNP, modeling an immunological synapse. The spatial relationship of the IgE-bound high affinity IgE receptor (FcεRI) to the ratio image of P-polarized excitation and S-polarized excitation was analyzed. These studies help correlate the dynamics of cell surface molecules with the mechanical properties of the plasma membrane during synapse formation.

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Citations

Sep 5, 2019·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Mirsana P Ebrahimkutty, Milos Galic
Jun 8, 2021·ACS Photonics·Olha MelnykRobert E Camley

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Methods Mentioned

BETA
fluorescence recovery after photobleaching
super-resolution microscopy

Software Mentioned

micromanager
TIRF
Arduino
MATLAB
DIPImage

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