Real-Time Fluorescence Imaging of His-Tag-Driven Conjugation of mCherry Proteins to Silver Nanowires

Abstract

In this work, we aimed to apply fluorescence microscopy to image protein conjugation to Ni-NTA modified silver nanowires in real time via the His-tag attachment. First, a set of experiments was designed and performed for the mixtures of proteins and silver nanowires in order to demonstrate plasmon enhancement of mCherry protein fluorescence as well as the ability to image fluorescence of single molecules. The results indicated strong enhancement of single-protein fluorescence emission upon coupling with silver nanowires. This conclusion was supported by a decrease in the fluorescence decay time of mCherry proteins. Real-time imaging was carried out for a structure created by dropping protein solution onto a glass substrate with functionalized silver nanowires. We observed specific attachment of mCherry proteins to the nanowires, with the recognition time being much longer than in the case of streptavidin–biotin conjugation. This result indicated that it is possible to design a universal and efficient real-time sensing platform with plasmonically active functionalized silver nanowires.

Keywords

• silver nanowires
• surface functionalizatio
• conjugation
• fluorescence imaging
• photoactive protein
• biosensing