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Characterization and optimization of paper SERS substrate were performed in detail, in which morphologies and distribution of silver nanoparticles (AgNPs) on the paper substrate pretreated with different concentrations of NaCl and the subsequent soaking with colloidal AgNPs for different period of time were evaluated. Our results show that both NaCl concentration and soaking time with AgNPs have a significant influence on SERS enhancement, showing that an optimal EF of 2.27 × 107 was achieved when the paper substrate was treated with 20 mM NaCl and one-hour soak with AgNPs. Moreover, seminal plasma (SP) was specifically selected to evaluate the performance of paper-based SERS substrate for potential clinical detection and diagnosis. The optimization of the paper SERS substrate demonstrates potential applications in reliable on-site detection of SP and clinical diagnosis of fertility-related diseases as well

Evaluation and Optimization of Paper-Based SERS Substrate for Potential Label-Free Raman Analysis of Seminal Plasma