Recently, far-field optical imaging with a resolution significantly beyond diffraction limit has attracted tremendous attention allowing for high resolution imaging in living objects. Various methods have been proposed that are divided in to two basic approaches; deterministic super-resolution like STED or RESOLFT and stochastic super-resolution like PALM or STORM. We propose to achieve super-resolution in far-field fluorescence imaging by the use of controllable (on-demand) bursts of pulses that can change the fluorescence signal of long-lived component over one order of magnitude. We demonstrate that two beads, one labeled with a long-lived dye and another with a short-lived dye, separated by a distance lower than 100 nm can be easily resolved in a single experiment. The proposed method can be used to separate two biological structures in a cell by targeting them with two antibodies labeled with long-lived and short-lived fluorophores.
In this report, we describe a plasmonic platform with silver fractals for metal enhanced fluorescence (MEF) measurements. When a dye containing surface was brought into contact with silver fractals, a significantly enhanced fluorescence signal from the dye was observed. Fluorescence enhancement was studied with the N-methyl-azadioxatriangulenium chloride salt (Me-ADOTA.Cl) in PVA films made from 0.2 % PVA (w/v) solution spin-coated on a clean glass coverslip. The Plasmonic Platforms (PP) was assembled by pressing together silver fractals on one glass slide and a separate glass coverslip spin-coated with a uniform Me-ADOTA.Cl in PVA film. In addition, we also tested the ADOTA labeled human serum albumin (HSA) deposited on a glass slide for potential PP bioassay applications. Using the new PP, we could achieve more than 20-fold fluorescence enhancement (bright spots) accompanied by decrease in fluorescence lifetime. The experimental results were used to calculate the extinction (excitation) enhancement factor (GA) and fluorescence radiative rate enhancements factor (GF). No change in emission spectrum was observed for a dye with and without contact with fractals. Our studies indicate that this type of PP can be a convenient approach for constructing assays utilizing metal enhanced fluorescence (MEF) without the need for depositing the material directly on metal structures platforms.