Resolution enhancement of single-molecule microscopy by using surface plasmon-coupled emission

Jian-Zong Lai^1*, Chun-Yu Lin², Gerald Abrigo¹, Fan-Ching Chien¹

¹Department of Optics and Photonics, National Central University, Taoyuan, Taiwan
²College of Photonics, National Yang Ming Chiao Tung University, Tainan, Taiwan

* Presenter:Jian-Zong Lai, email:zenus109226067@g.ncu.edu.tw

Surface plasmon-coupled emission (SPCE) substrates designed for the enhancement of fluorescence intensity of blinking fluorophores in single-molecule localization microscopy (SMLM). The investigation on the various thicknesses of metal layers was performed and optimized the thickness of the silver metal layer to provide the highest fluorescence enhancement of 4.8-fold and highly directional fluorescence. The combination of both techniques allows imaging super-resolution images in nanoscale-level resolution, and the use of excitation field created by SPCE substrate is beneficial for rejection of background in the detection of fluorophores, which led to results in high resolution as the reveal of the real position of fluorophores. Moreover, the ability to enhancement of fluorescence intensity by SPCE substrate potentially enabled in increasing the image resolution, improving the temporal resolution, and reducing illumination power density. Low illumination power density allows the low effect of phototoxicity and photodamage of a living cell, and thus maximizes the observation time.

Keywords: surface plasmon-coupled emission, single-molecule localization microscopy