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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Main Authors:	Rosa Machado, Justin Bendesky, Madison Brown, Kathrin Spendier, Guy M. Hagen
Format:	Article
Language:	English
Published:	MDPI AG 2019-07-01
Series:	Journal of Imaging
Subjects:	total internal reflection fluorescence microscopy P-TIRF rat basophilic leukemia cells RBL-2H3 IgE receptor FcεRI plasma membrane supported lipid bilayer
Online Access:	https://www.mdpi.com/2313-433X/5/7/63

id	doaj-05542c59290d4531b7a92ea2f0e56986
record_format	Article
spelling	doaj-05542c59290d4531b7a92ea2f0e569862020-11-24T20:44:10ZengMDPI AGJournal of Imaging2313-433X2019-07-01576310.3390/jimaging5070063jimaging5070063Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized ExcitationRosa Machado0Justin Bendesky1Madison Brown2Kathrin Spendier3Guy M. Hagen4UCCS Center for the Biofrontiers Institute, University of Colorado at Colorado Springs, Colorado Springs, CO 80918, USAUCCS Center for the Biofrontiers Institute, University of Colorado at Colorado Springs, Colorado Springs, CO 80918, USAUCCS Center for the Biofrontiers Institute, University of Colorado at Colorado Springs, Colorado Springs, CO 80918, USAUCCS Center for the Biofrontiers Institute, University of Colorado at Colorado Springs, Colorado Springs, CO 80918, USAUCCS Center for the Biofrontiers Institute, University of Colorado at Colorado Springs, Colorado Springs, CO 80918, USATotal 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.https://www.mdpi.com/2313-433X/5/7/63total internal reflection fluorescence microscopyP-TIRFrat basophilic leukemia cellsRBL-2H3IgE receptorFcεRIplasma membranesupported lipid bilayer
collection	DOAJ
language	English
format	Article
sources	DOAJ
author	Rosa Machado Justin Bendesky Madison Brown Kathrin Spendier Guy M. Hagen
spellingShingle	Rosa Machado Justin Bendesky Madison Brown Kathrin Spendier Guy M. Hagen Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized Excitation Journal of Imaging total internal reflection fluorescence microscopy P-TIRF rat basophilic leukemia cells RBL-2H3 IgE receptor FcεRI plasma membrane supported lipid bilayer
author_facet	Rosa Machado Justin Bendesky Madison Brown Kathrin Spendier Guy M. Hagen
author_sort	Rosa Machado
title	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_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_sort	imaging membrane curvature inside a fcεri-centric synapse in rbl-2h3 cells using tirf microscopy with polarized excitation
publisher	MDPI AG
series	Journal of Imaging
issn	2313-433X
publishDate	2019-07-01
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.
topic	total internal reflection fluorescence microscopy P-TIRF rat basophilic leukemia cells RBL-2H3 IgE receptor FcεRI plasma membrane supported lipid bilayer
url	https://www.mdpi.com/2313-433X/5/7/63
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Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized Excitation

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