{Reference Type}: Journal Article
{Title}: Improved Imaging Surface for Quantitative Single-Molecule Microscopy.
{Author}: Zhang YP;Lobanova E;Dworkin A;Furlepa M;Yang WS;Burke M;Meng JX;Potter N;Sala RL;Kahanawita L;Layburn F;Scherman OA;Williams-Gray CH;Klenerman D;
{Journal}: ACS Appl Mater Interfaces
{Volume}: 16
{Issue}: 28
{Year}: 2024 Jul 17
{Factor}: 10.383
{DOI}: 10.1021/acsami.4c06512
{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.