利用偏振激发的全内反射荧光显微镜成像RBL-2H3细胞中以FcεRI为中心的突触内的膜曲率。

Machado Rosa, Bendesky Justin, Brown Madison, Spendier Kathrin, Hagen Guy M

UCCS Center for the Biofrontiers Institute, University of Colorado at Colorado Springs, Colorado Springs, CO 80918, USA.

Department of Physics and Energy Science, University of Colorado at Colorado Springs, Colorado Springs, CO 80918, USA.

J Imaging. 2019 Jul;5(7). doi: 10.3390/jimaging5070063. Epub 2019 Jul 4.

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.

采用偏振激发的全内反射荧光显微镜（P-TIRF）可用于对活细胞中的纳米级曲率现象进行成像。我们利用P-TIRF来观察预先用荧光抗二硝基苯基（抗DNP）免疫球蛋白E（IgE）致敏的大鼠嗜碱性白血病细胞（RBL-2H3细胞）与含有可移动单价DNP的支持脂质双层接触的情况，以此模拟免疫突触。分析了与IgE结合的高亲和力IgE受体（FcεRI）与P偏振激发和S偏振激发的比率图像之间的空间关系。这些研究有助于将细胞表面分子的动力学与突触形成过程中质膜的力学特性联系起来。