Super-Resolution Imaging and Quantitative Analysis of Membrane Protein/Lipid Raft Clustering Mediated by Cell-Surface Self-Assembly of Hybrid Nanoconjugates.
Super-Resolution Imaging and Quantitative Analysis of Membrane Protein/Lipid Raft Clustering Mediated by Cell-Surface Self-Assembly of Hybrid Nanoconjugates.
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DOI：
10.1002/cbic.201500278
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发表时间：
2015-08-17
期刊：
影响因子：
--
通讯作者：
Kopeček J
中科院分区：
文献类型：
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作者：
Hartley JM;Chu TW;Peterson EM;Zhang R;Yang J;Harris J;Kopeček J
采用超分辨成像技术对杂化纳米结合物处理后细胞质膜的组织变化进行定量研究。该纳米偶联物在恶性B细胞表面交联化CD20，从而诱导细胞凋亡。使用对相关分析对超分辨率图像进行分析，以确定簇的大小并计算簇中的平均分子数量。通过用胆固醇螯合剂和肌动蛋白去稳定剂对细胞进行预处理以防止脂筏的形成，研究了脂筏的作用。脂筏簇的大小与纳米结合物处理后的细胞凋亡诱导有关。在杂化纳米结合物处理的细胞中，脂筏簇的半径为200 nm。超分辨率图像提供了精确的分子位置坐标，可用于确定结合的连接物的密度、簇大小和每个簇的分子数量。
Super-resolution imaging was used to quantify organizational changes in the plasma membrane after treatment with hybrid nanoconjugates. The nanoconjugates crosslinked CD20 on the surface of malignant B cells thereby inducing apoptosis. Super-resolution images were analyzed using pair-correlation analysis to determine cluster size and to count the average number of molecules in the clusters. The role of lipid rafts was investigated by pre-treating cells with a cholesterol chelator and actin destabilizer to prevent lipid raft formation. Lipid raft cluster size correlated with apoptosis induction after treatment with the nanoconjugates. Lipid raft clusters had radii ~200 nm in cells treated with the hybrid nanoconjugates. Super-resolution images provided precise molecule location coordinates that could be used to determine density of bound conjugates, cluster size, and number of molecules per cluster.