%0 Journal Article
%T Improved Imaging Surface for Quantitative Single-Molecule Microscopy.
%A Zhang YP
%A Lobanova E
%A Dworkin A
%A Furlepa M
%A Yang WS
%A Burke M
%A Meng JX
%A Potter N
%A Sala RL
%A Kahanawita L
%A Layburn F
%A Scherman OA
%A Williams-Gray CH
%A Klenerman D
%J ACS Appl Mater Interfaces
%V 16
%N 28
%D 2024 Jul 17
%M 38979642
%F 10.383
%R 10.1021/acsami.4c06512
%X 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.