Plasmon-Amplified Endogenous Fluorescence Nanospectroscopic Sensor Based on Inherent Elastic Scattering for Ultratrace Ratiometric Detection of Capsaicinoids.

Endogenous fluorescence imaging techniques are key for modern single-molecule quantification without the use of additional labeling probes. However, the drawback of weak fluorescence signal is the primary challenge in meeting the ever-increasing demands of single-molecule detection. Here, we report a simple and reliable method that provides up to ∼100-fold uniform fluorescence enhancement of endogenous fluorescence of the capsaicinoid molecule. The method is based on a single nanoparticle plasmon-amplified endogenous fluorescence nanospectroscopic sensor (PAEFS). This work demonstrated the applicability of PAEFS in refining sensitivity at the single-molecule level by showing ultralow limits of detection (106 times lower than previous reports) of fluorescence-based capsaicinoids with a wide range of linear response (18 zM to 85 pM). Spectrally overlapped capsaicinoid analogues were quantified ratiometrically to detect the analogue percentages in real samples. The novel endogenous fluorescence enhancement approach presented here represents a universal sensor for enhanced detection of single molecules using existing techniques without altering the original molecular features or using add-on labeling probes.