scholarly journals Imaging Membrane Curvature inside a Model Immunological Synapse in RBL-2H3 Cells using TIRF Microscopy with Polarized Excitation

2019 ◽  
Author(s):  
Rosa Machado ◽  
Justin Bendesky ◽  
Madison Brown ◽  
Kathrin Spendier ◽  
Guy M. Hagen
Keyword(s):  
Total Internal Reflection ◽  
Synapse Formation ◽  
Immunological Synapse ◽  
Membrane Curvature ◽  
Leukemia Cells ◽  
Live Cells ◽  
Supported Lipid Bilayer ◽  
Surface Molecules ◽  
Rat Basophilic Leukemia Cells ◽  
Basophilic Leukemia

AbstractTotal 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) coming into contact with a supported lipid bilayer, modeling an immunological synapse. These studies help correlate the dynamics of cell surface molecules with the mechanical properties of the plasma membrane during synapse formation.

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

2019 ◽  
Vol 5 (7) ◽  
pp. 63 ◽  
Author(s):  
Machado ◽  
Bendesky ◽  
Brown ◽  
Spendier ◽  
Hagen
Keyword(s):  
Synapse Formation ◽  
Immunological Synapse ◽  
Immunoglobulin E ◽  
Spatial Relationship ◽  
Membrane Curvature ◽  
Live Cells ◽  
Ratio Image ◽  
Rat Basophilic Leukemia Cells ◽  
High Affinity Ige Receptor ◽  
Relationship Of

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.

The distribution of antigen on the surface of RBL-2H3 rat basophilic leukemia cells observed by back-scattered electron imaging

1986 ◽  
Vol 44 ◽  
pp. 274-275
Author(s):  
R.F. Stump ◽  
J.R. Pfeiffer ◽  
JC. Seagrave ◽  
D. Huskisson ◽  
J.M. Oliver
Keyword(s):  
Cell Surface ◽  
Dynamic Properties ◽  
Leukemia Cells ◽  
Antigen Binding ◽  
Scattered Electron ◽  
Ige Receptor ◽  
Receptor Complexes ◽  
Rat Basophilic Leukemia Cells ◽  
Electron Imaging ◽  
Basophilic Leukemia

In RBL-2H3 rat basophilic leukemia cells, antigen binding to cell surface IgE-receptor complexes stimulates the release of inflammatory mediators and initiates a series of membrane and cytoskeletal events including a transformation of the cell surface from a microvillous to a lamellar topography. It is likely that dynamic properties of the IgE receptor contribute to the activation of these responses. Fewtrell and Metzger have established that limited crosslinking of IgE-receptor complexes is essential to trigger secretion. In addition, Baird and colleagues have reported that antigen binding causes a rapid immobilization of IgE-receptor complexes, and we have demonstrated an apparent increase with time in the affinity of IgE-receptor complexes for antigen.

Immunoglobulin E mediated membrane conductance changes in rat basophilic leukemia cells

1988 ◽  
Vol 66 (3) ◽  
pp. 328-331 ◽  
Author(s):  
Carlos Barajas-López ◽  
Jan D. Huizinga
Keyword(s):  
Immunoglobulin E ◽  
Calcium Influx ◽  
Membrane Conductance ◽  
Membrane Resistance ◽  
Leukemia Cells ◽  
Rat Basophilic Leukemia Cells ◽  
Conductance Changes ◽  
Electrophysiological Effects ◽  
Basophilic Leukemia ◽  
Stimulation Of

Electrophysiological effects of anaphylactic stimulation of rat basophilic leukemia cells (RBL-2H3) were studied using conventional microelectrodes. Stimulation of passively sensitized cells by anti-immunoglobulin E resulted in hyperpolarization followed by depolarization. These changes in membrane polarization were associated with a decrease in input membrane resistance. No effect of anaphylactic stimulation was seen in Ca2+-free solution or when Ca2+ influx was blocked by Co2+, but it was mimicked by the Ca2+ ionophore A-23187. This suggests that the changes in ionic conductances were associated with calcium influx. These results support the hypothesis that membrane conductance changes are involved in the stimulus-secretion process of the RBL-2H3 cells.

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