Fluorescent proteins are indispensable for selective, quantitative visualization of localization, dynamics, and interactions of key molecular constituents of live cells. Incorporation of fluorescent proteins into an optical cavity can lead to a significant increase in fluorescence signal levels due to stimulated emission and light amplification in the cavity, forming a laser with biological gain medium. Utilization of lasing emission from fluorescent biological molecules can then greatly enhance the performance of fluorescence-based biosensors benefiting from the high sensitivity of non-linear lasing processes to small perturbations in the cavity and the gain medium. Here we study optofluidic biolasers that exploit active liquid optical resonators formed by surface-supported aqueous microdroplets containing purified yellow fluorescent protein or a suspension of live E. coli bacterial cells expressing the fluorescent protein. We first demonstrate lasing in fluorescent protein solutions at concentrations as low as 49 μM. Subsequently, we show that a single fluorescent bacterial cell of micrometre size confined in a droplet-based cavity can serve as a laser gain medium. Aqueous droplet microcavities allow the maintenance of the bacterial cells under conditions compatible with unimpeded growth. Therefore, our results also suggest a direct route to microscopic sources of laser light with self-regenerating gain media.
We introduce a portable, all-liquid microlaser based on optically pumped dye-doped emulsion microdroplets held in a single beam optical trap. We show high stability of the laser emission spectra during prolonged optical manipulation of the droplets within an immiscible host liquid. We investigate the effects of droplet size and dye concentration on the spectral position of lasing wavelength and show how these parameters can be used for the emission wavelength tuning. We also study shifting of the average lasing wavelength to the blue side of the spectrum due to dye photobleaching. The presented optically manipulated fluidic microlasers are disposable and can be easily combined with microfluidic chip technology. This makes them especially attractive for on-chip applications in chemical and biological analysis and sensing.
Incorporating fluorescence resonance energy transfer (FRET) into a laser cavity can increase the sensitivity of FRET-based biochemical sensors due to the nonlinear dependence of the lasing output on the FRET parameters. Here, we carry out a comprehensive theoretical analysis of optofluidic FRET lasers based on a Fabry-Pérot microcavity using a rate equation model. We compare conceptually distinct cases of donor and acceptor molecules diffusing freely in bulk solution versus molecules connected by a fixed-length linker and show that the latter arrangement is especially well-suited for sensing of lowconcentration analytes. By comparing FRET lasing-based sensors with conventional FRET sensors, we show that for optimal pump fluence and FRET-pair concentration, FRET lasing can lead to more than 100-fold enhancement in detection sensitivities of conformational changes in the Förster radius range. We also show that for optimal experimental conditions, donor and acceptor emission intensities become over 20-fold more sensitive to FRETpair concentration changes in the presence of FRET lasing. We study the dependence of the sensitivity enhancement on the cavity Q-factor. We show that the highest enhancements can be obtained for Q-factors between 10 4 − 10 6 , and enhancement values decrease for Q-factors above 10 6 due to the radiative energy transfer in the cavity.
We introduce tunable optofluidic microlasers based on active optical resonant cavities formed by optically stretched, dye-doped emulsion droplets confined in a dual-beam optical trap. To achieve tunable dye lasing, optically pumped droplets of oil dispersed in water are stretched by light in the dual-beam trap. Subsequently, resonant path lengths of whispering gallery modes (WGMs) propagating in the droplet are modified, leading to shifts in the microlaser emission wavelengths. Using this technique, we present all-optical, almost reversible spectral tuning of the lasing WGMs and show that the direction of tuning depends on the position of the pump beam focus on the droplet. In addition, we study the effects of temperature changes on the spectral position of lasing WGMs and demonstrate that droplet heating leads to red-tuning of the droplet lasing wavelength.
We demonstrate optofluidic microlasers using highly efficient non-radiative Förster resonance energy transfer (FRET) for pumping of gain medium placed within liquid microdroplets situated on a superhydrophobic surface. Microdroplets generated from a mixture of ethylene glycol, glycerol, and water and stained with the FRET donor-acceptor dye pair Rhodamine 6G-Rhodamine 700 serve as active optical resonant cavities hosting high-quality whispering gallery modes. Upon direct optical pumping of the donor with a pulsed laser, lasing is observed in the emission band of the acceptor as a result of efficient FRET coupling between the acceptor and donor molecules. FRET lasing is characterized for different acceptor and donor concentrations, and threshold pump fluences of acceptor lasing as low as 6.3 mJ cm −2 are demonstrated. We also verify the dominance of the non-radiative FRET over cavity-assisted radiative energy transfer for the range of parameters studied in the experiments.
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