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Optics Express

Nanosecond control and optical pulse shaping by stimulated emission depletion in a liquid crystal

Maruša Vitek, Igor Muševič

Published Jun 19, 2015• 12 citations
Publisher

Abstract

We study anisotropic Stimulated Emission Depletion (STED) from dye molecules, which are collectively ordered in a host liquid crystal. Due to the ordering of fluorescent emitters, the STED efficiency depends on the polarization of the depletion beam and time-delay of the STED pulse. The depletion efficiency is highest at lower temperatures in the highly ordered smectic-A phase and deteriorates in the higher temperature nematic and isotropic phases. We demonstrate by temporal tuning of STED that it is possible to generate an arbitrary sequence of nanosecond fluorescent pulses with variable width and variable delay. Our results show that the STED mechanism in principle allows for very fast (GHz) and efficient control of light by light, which could in the future be used for all-optical control of the flow of light in photonic microdevices based on liquid crystals. Using STED anisotropy and time-control, new modalities of STED imaging in liquid crystals could be developed.

DOI
10.1364/oe.23.016921

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Scientific Reports•Sep 6, 2017•74 citations
We study anisotropic Stimulated Emission Depletion (STED) from dye molecules, which are collectively ordered in a host liquid crystal. Due to the ordering of fluorescent emitters, the STED efficiency depends on the polarization of the depletion beam and time-delay of the STED pulse. The depletion efficiency is highest at lower temperatures in the highly ordered smectic-A phase and deteriorates in the higher temperature nematic and isotropic phases. We demonstrate by temporal tuning of STED that it is possible to generate an arbitrary sequence of nanosecond fluorescent pulses with variable width and variable delay. Our results show that the STED mechanism in principle allows for very fast (GHz) and efficient control of light by light, which could in the future be used for all-optical control of the flow of light in photonic microdevices based on liquid crystals. Using STED anisotropy and time-control, new modalities of STED imaging in liquid crystals could be developed.
The imaging technology of stimulated emission depletion (STED) utilizes the nonlinearity relationship between the fluorescence saturation and the excited state stimulated depletion. It implements three-dimensional (3D) imaging and breaks the diffraction barrier of far-field light microscopy by restricting fluorescent molecules at a sub-diffraction spot. In order to improve the resolution which attained by this technology, the computer simulation on temporal behavior of population probabilities of the sample was made in this paper, and the optimized parameters such as intensity, duration and delay time of the STED pulse were given.
The feasibility of manipulating the single molecule absorption-emission cycle using picosecond stimulated emission depletion (STED) is investigated using a stochastic computer simulation. In the simulation the molecule is subjected to repeated excitation and depletion events using time delayed pairs of excitation (PUMP) and depletion (DUMP) pulses derived from a high repetition rate pulsed laser system. The model is used to demonstrate that a significant and even substantial reduction in the occurrence of 'dark states' in the fluorescence emission can be achieved using stimulated emission depletion. Variation in the PUMP-DUMP window allows precise control of the fluorescence yield with substantial increases in the fluorescence intensity observed at early PUMP-DUMP delays.
Stimulated emission depletion (STED) is the key optical technology enabling super-resolution microscopy below the diffraction limit. Here, we demonstrate that modulation of STED in the time domain, combined with properly designed lock-in detection, can radically enhance the contrast of fluorescent images of strongly autofluorescent biotissues. In our experiments, the temporally modulated STED technique, implemented with low-intensity continuous-wave laser sources, is shown to provide an efficient all-optical suppression of a broadband fluorescent background, allowing the contrast of fluorescent images of mammal brain tissues tagged with nitrogen-vacancy diamond to be increased by five orders of magnitude.
As one of the most important realizations of stimulated emission depletion (STED) microscopy, the continuous-wave (CW) STED system, constructed by using CW lasers as the excitation and STED beams, has been investigated and developed for nearly a decade. However, a theoretical model of the suppression factors in CW STED has not been well established. In this investigation, the factors that affect the spatial resolution of a CW STED system are theoretically and numerically studied. The full-width-at-half-maximum (FWHM) of a CW STED with a doughnut-shaped STED beam is also reanalyzed. It is found that the suppression function is dominated by the ratio of the local STED and excitation beam intensities. In addition, the FWHM is highly sensitive to both the fluorescence rate (inverse of fluoresce lifetime) and the quenching rate, but insensitive to the rate of vibrational relaxation. For comparison, the suppression function in picosecond STED is only determined by the distribution of the STED beam intensity scaled with the saturation intensity. Our model is highly consistent with published experimental data for evaluating the spatial resolution. This investigation is important in guiding the development of new CW STED systems.
In stimulated emission depletion (STED) nanoscopy the wavelength of the STED beam is usually tuned towards the red tail of the emission maximum of the fluorophore. Shifting the STED wavelength closer to the emission peak, i.e. towards the blue region, favorably increases the stimulated emission cross-section. However, this blue-shifting also increases the probability to excite fluorophores that have remained in their ground state, compromising the image contrast. Here we present a method to exploit the higher STED efficiency of blue-shifted STED beams while maintaining the contrast in the image. The method is exemplified by imaging immunolabeled features in mammalian cells with an up to 3-fold increased STED efficiency compared to that encountered in standard STED nanoscopy implementations.
In STED (stimulated emission depletion) nanoscopy, the resolution and signal are limited by the fluorophore de-excitation efficiency and photobleaching. Here, we investigated their dependence on the pulse duration and power of the applied STED light for the popular 750 nm wavelength. In experiments with red- and orange-emitting dyes, the pulse duration was varied from the sub-picosecond range up to continuous-wave conditions, with average powers up to 200 mW at 80 MHz repetition rate, i.e. peak powers up to 1 kW and pulse energies up to 2.5 nJ. We demonstrate the dependence of bleaching on pulse duration, which dictates the optimal parameters of how to deliver the photons required for transient fluorophore silencing. Measurements with the dye ATTO647N reveal that the bleaching of excited molecules scales with peak power with a single effective order ~1.4. This motivates peak power reduction while maintaining the number of STED-light photons, in line with the superior resolution commonly achieved for nanosecond STED pulses. Other dyes (ATTO590, STAR580, STAR635P) exhibit two distinctive bleaching regimes for constant pulse energy, one with strong dependence on peak power, one nearly independent. We interpret the results within a photobleaching model that guides quantitative predictions of resolution and bleaching.