Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized Excitation.
FcεRI
IgE receptor
P-TIRF
RBL-2H3
plasma membrane
rat basophilic leukemia cells
supported lipid bilayer
total internal reflection fluorescence microscopy
Journal
Journal of imaging
ISSN: 2313-433X
Titre abrégé: J Imaging
Pays: Switzerland
ID NLM: 101698819
Informations de publication
Date de publication:
07 2019
07 2019
Historique:
entrez:
31
7
2019
pubmed:
31
7
2019
medline:
31
7
2019
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 31360699
doi: 10.3390/jimaging5070063
pmc: PMC6663088
mid: NIHMS1041488
pii: jimaging5070063
pii:
doi:
Types de publication
Journal Article
Langues
eng
Subventions
Organisme : NIGMS NIH HHS
ID : R15 GM128166
Pays : United States
Déclaration de conflit d'intérêts
Conflicts of Interest: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the National Science Foundation.
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