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Article

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

1
UCCS Center for the Biofrontiers Institute, University of Colorado at Colorado Springs, Colorado Springs, CO 80918, USA
2
Department of Physics and Energy Science, University of Colorado at Colorado Springs, Colorado Springs, CO 80918, USA
*
Authors to whom correspondence should be addressed.
Received: 15 April 2019 / Revised: 28 June 2019 / Accepted: 28 June 2019 / Published: 4 July 2019
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. View Full-Text
Keywords: total internal reflection fluorescence microscopy; P-TIRF; rat basophilic leukemia cells; RBL-2H3; IgE receptor; FcεRI; plasma membrane; supported lipid bilayer total internal reflection fluorescence microscopy; P-TIRF; rat basophilic leukemia cells; RBL-2H3; IgE receptor; FcεRI; plasma membrane; supported lipid bilayer
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MDPI and ACS Style

Machado, R.; Bendesky, J.; Brown, M.; Spendier, K.; Hagen, G.M. Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized Excitation. J. Imaging 2019, 5, 63. https://0-doi-org.brum.beds.ac.uk/10.3390/jimaging5070063

AMA Style

Machado R, Bendesky J, Brown M, Spendier K, Hagen GM. Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized Excitation. Journal of Imaging. 2019; 5(7):63. https://0-doi-org.brum.beds.ac.uk/10.3390/jimaging5070063

Chicago/Turabian Style

Machado, Rosa; Bendesky, Justin; Brown, Madison; Spendier, Kathrin; Hagen, Guy M. 2019. "Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized Excitation" J. Imaging 5, no. 7: 63. https://0-doi-org.brum.beds.ac.uk/10.3390/jimaging5070063

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