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

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...

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Detalles Bibliográficos
Autores principales: Machado, Rosa, Bendesky, Justin, Brown, Madison, Spendier, Kathrin, Hagen, Guy M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6663088/
https://www.ncbi.nlm.nih.gov/pubmed/31360699
http://dx.doi.org/10.3390/jimaging5070063
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author Machado, Rosa
Bendesky, Justin
Brown, Madison
Spendier, Kathrin
Hagen, Guy M.
author_facet Machado, Rosa
Bendesky, Justin
Brown, Madison
Spendier, Kathrin
Hagen, Guy M.
author_sort Machado, Rosa
collection PubMed
description 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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spelling pubmed-66630882019-07-29 Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized Excitation Machado, Rosa Bendesky, Justin Brown, Madison Spendier, Kathrin Hagen, Guy M. J Imaging Article 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. MDPI 2019-07-04 /pmc/articles/PMC6663088/ /pubmed/31360699 http://dx.doi.org/10.3390/jimaging5070063 Text en © 2019 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Article
Machado, Rosa
Bendesky, Justin
Brown, Madison
Spendier, Kathrin
Hagen, Guy M.
Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized Excitation
title Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized Excitation
title_full Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized Excitation
title_fullStr Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized Excitation
title_full_unstemmed Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized Excitation
title_short Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized Excitation
title_sort imaging membrane curvature inside a fcεri-centric synapse in rbl-2h3 cells using tirf microscopy with polarized excitation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6663088/
https://www.ncbi.nlm.nih.gov/pubmed/31360699
http://dx.doi.org/10.3390/jimaging5070063
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