Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized Excitation
Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized Excitation
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DOI：
10.3390/jimaging5070063
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发表时间：
2019-07-01
影响因子：
3.2
通讯作者：
Hagen, Guy M.
中科院分区：
文献类型：
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作者：
Machado, Rosa;Bendesky, Justin;Hagen, Guy M.
偏振激发全内反射荧光显微镜（P-TIRF）可用于成像活细胞中的纳米级曲率现象。我们使用P-TIRF可视化大鼠嗜碱性白血病细胞（RBL-2 H3细胞）与荧光抗二硝基苯基（抗-DNP）免疫球蛋白E（IgE）接触的支持脂质双层含有移动的，单价DNP，模拟免疫突触。分析了IgE结合的高亲和力IgE受体（Fc ε RI）与P极化激发和S极化激发的比值图像的空间关系。这些研究有助于将突触形成过程中细胞表面分子的动力学与质膜的机械性质联系起来。
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 (Fc epsilon RI) 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.