PDF(Pubmed)
Abstract：
Preventing nonspecific binding is essential for sensitive surface-based quantitative single-molecule microscopy. Here we report a much-simplified RainX-F127 (RF-127) surface with improved passivation. This surface achieves up to 100-fold less nonspecific binding from protein aggregates compared to commonly used polyethylene glycol (PEG) surfaces. The method is compatible with common single-molecule techniques including single-molecule pull-down (SiMPull), super-resolution imaging, antibody-binding screening and single exosome visualization. This method is also able to specifically detect alpha-synuclein (α-syn) and tau aggregates from a wide range of biofluids including human serum, brain extracts, cerebrospinal fluid (CSF) and saliva. The simplicity of this method further allows the functionalization of microplates for robot-assisted high-throughput single-molecule experiments. Overall, this simple but improved surface offers a versatile platform for quantitative single-molecule microscopy without the need for specialized equipment or personnel.
摘要：
防止非特异性结合对于敏感的基于表面的定量单分子显微镜至关重要。在这里，我们报告了一个简化的RainX-F127（RF-127）表面，具有改进的钝化。与通常使用的聚乙二醇（PEG）表面相比，该表面与蛋白质聚集体的非特异性结合减少了100倍。该方法与常见的单分子技术兼容，包括单分子下拉（SiMPull），超分辨率成像，抗体结合筛选和单个外泌体可视化。该方法还能够从包括人血清在内的各种生物流体中特异性检测α-突触核蛋白（α-syn）和tau聚集体，大脑提取物，脑脊液（CSF）和唾液。该方法的简单性进一步允许用于机器人辅助的高通量单分子实验的微孔板的功能化。总的来说,这个简单但改进的表面为定量单分子显微镜提供了一个通用的平台，而不需要专门的设备或人员。
