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Dispersion management is critical in many optical applications, whether to reduce impairments in fiber optic communication or chirp pulse amplification, or to create time stretch instruments for single-shot continuous recording of fast phenomena. The most common solutions for achieving large dispersion with low loss include dispersion compensation fiber, fiber Bragg grating, and diffraction grating pairs. Such dispersive elements have finite operational bandwidth, limited total dispersion, or insufficient power handling. In this Letter, we demonstrate a new, to the best of our knowledge, implementation of the chromo-modal dispersion device based on a silicon waveguide slab that addresses these limitations. The device provides extremely large dispersion with a widely tunable spectrum. We also propose a new time-stretch spectrometer where the absorption cell simultaneously provides spectrum-to-time mapping for fast single-shot spectroscopy.
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Microalgal biofuels and biomass have ecofriendly advantages as feedstocks. Improved understanding and utilization of microalgae require large-scale analysis of the morphological and metabolic heterogeneity within populations. Here, with Euglena gracilis as a model microalgal species, we evaluate how fluorescence- and brightfield-derived-image-based descriptors vary during environmental stress at the single-cell level. This is achieved with a new multiparameter fluorescence-imaging cytometric technique that allows the assaying of thousands of cells per experiment. We track morphological changes, including the intensity and distribution of intracellular lipid droplets, and pigment autofluorescence. The combined fluorescence-morphological analysis identifies new metrics not accessible with traditional flow cytometry, including the lipid-to-cell-area ratio (LCAR), which shows promise as an indicator of oil productivity per biomass. Single-cell metrics of lipid productivity were highly correlated ( R2 > 0.90, p < 0.005) with bulk oil extraction. Such chemomorphological atlases of algal species can help optimize growth conditions and selection approaches for large-scale biomass production.
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Euglena gracilis/citologia , Euglena gracilis/metabolismo , Citometria de Fluxo , Imagem Óptica , Análise de Célula Única/métodos , Espaço Intracelular/metabolismoRESUMO
Improving the efficiency, cell survival, and throughput of methods to modify and control the genetic expression of cells is of great benefit to biology and medicine. We investigate, both computationally and experimentally, a nanostructured substrate made of tipless pyramids for plasmonic-induced transfection. By optimizing the geometrical parameters for an excitation wavelength of 800 nm, we demonstrate a 100-fold intensity enhancement of the electric near field at the cell-substrate contact area, while the low absorption typical for gold is maintained. We demonstrate that such a substrate can induce transient poration of cells by a purely optically induced process.
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Ouro/química , Lasers , Nanoestruturas/química , Transfecção/métodos , Sobrevivência Celular , Células HeLa , Humanos , Nanoestruturas/ultraestrutura , Óptica e Fotônica , Ressonância de Plasmônio de SuperfícieRESUMO
The angular light scattering profile of microscopic particles significantly depends on their morphological parameters, such as size and shape. This dependency is widely used in state-of-the-art flow cytometry methods for particle classification. We introduce a new spectrally encoded angular light scattering method, with potential application in scanning flow cytometry. We show that a one-to-one wavelength-to-angle mapping enables the measurement of the angular dependence of scattered light from microscopic particles over a wide dynamic range. Improvement in dynamic range is obtained by equalizing the angular dependence of scattering via wavelength equalization. Continuous angular spectrum is obtained without mechanical scanning enabling single-shot measurement. Using this information, particle morphology can be determined with improved accuracy. We derive and experimentally verify an analytic wavelength-to-angle mapping model, facilitating rapid data processing. As a proof of concept, we demonstrate the method's capability of distinguishing differently sized polystyrene beads. The combination of this technique with time-stretch dispersive Fourier transform offers real-time and high-throughput (high frame rate) measurements and renders the method suitable for integration in standard flow cytometers.
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High-throughput real-time optical integrators are of great importance for applications that require ultrafast optical information processing, such as real-time phase reconstruction of ultrashort optical pulses. In many of these applications, integration of wide optical bandwidth signals is required. Unfortunately, conventional all-optical integrators based on passive devices are usually sensitive to the wavelength and bandwidth of the optical carrier. Here, we propose and demonstrate a passive all-optical intensity integrator whose operation is independent of the optical signal wavelength and bandwidth. The integrator is implemented based on modal dispersion in a multimode waveguide. By controlling the launch conditions of the input beam, the device produces a rectangular temporal impulse response. Consequently, a temporal intensity integration of an arbitrary optical waveform input is performed within the rectangular time window. The key advantage of this device is that the integration operation can be performed independent of the input signal wavelength and optical carrier bandwidth. This is preferred in many applications where optical signals of different wavelengths are involved. Moreover, thanks to the use of a relatively short length of multimode waveguide, lower system latency is achieved compared to the systems using long dispersive fibers. To illustrate the versatility of the optical integrator, we demonstrate temporal intensity integration of optical waveforms with different wavelengths and optical carrier bandwidths. Finally, we use this device to perform high-throughput, single-shot, real-time optical phase reconstruction of phase-modulated signals at telecommunications bit rates.
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Dispositivos Ópticos , Processamento de Sinais Assistido por Computador/instrumentação , Telecomunicações/instrumentação , Sistemas Computacionais , Desenho Assistido por Computador , Desenho de Equipamento , Análise de Falha de EquipamentoRESUMO
Fast and selective isolation of single cells with unique spatial and morphological traits remains a technical challenge. Here, we address this by establishing high-speed image-enabled cell sorting (ICS), which records multicolor fluorescence images and sorts cells based on measurements from image data at speeds up to 15,000 events per second. We show that ICS quantifies cell morphology and localization of labeled proteins and increases the resolution of cell cycle analyses by separating mitotic stages. We combine ICS with CRISPR-pooled screens to identify regulators of the nuclear factor κB (NF-κB) pathway, enabling the completion of genome-wide image-based screens in about 9 hours of run time. By assessing complex cellular phenotypes, ICS substantially expands the phenotypic space accessible to cell-sorting applications and pooled genetic screening.
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Citometria de Fluxo , Imagem Óptica , Transporte Ativo do Núcleo Celular , Animais , Sistemas CRISPR-Cas , Núcleo Celular/metabolismo , Forma Celular , Técnicas Genéticas , Genoma , Genoma Humano , Humanos , Microscopia de Fluorescência , Mitose , NF-kappa B/metabolismo , Organelas/ultraestrutura , Fenótipo , Fator de Transcrição RelA/metabolismoRESUMO
The ability to control chromatic dispersion is paramount in applications where the optical pulsewidth is critical, such as chirped pulse amplification and fiber optic communications. Typically, devices used to generate large amounts (>100 ps/nm) of chromatic dispersion are based on diffraction gratings, chirped fiber Bragg gratings, or dispersion compensating fiber. Unfortunately, these dispersive elements suffer from one or more of the following restrictions: (i) limited operational bandwidth, (ii) limited total dispersion, (iii) low peak power handling, or (iv) large spatial footprint. Here, we introduce a new type of tunable dispersive device, which overcomes these limitations by leveraging the large modal dispersion of a multimode waveguide in combination with the angular dispersion of diffraction gratings to create chromatic dispersion. We characterize the device's dispersion, and demonstrate its ability to stretch a sub-picosecond optical pulse to nearly 2 nanoseconds in 20 meters of multimode optical fiber. Using this device, we also demonstrate single-shot, time-wavelength atomic absorption spectroscopy at a repetition rate of 90.8 MHz.
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Tecnologia de Fibra Óptica/instrumentação , Refratometria/instrumentação , Ressonância de Plasmônio de Superfície/instrumentação , Cor , Desenho de Equipamento , Análise de Falha de EquipamentoRESUMO
We present a method for improving femtomole-level trace detection (10(9) molecules) using large-area surface-enhanced Raman scattering (SERS) substrates. Using multiphoton-induced exposure of a commercial photoresist, we physically limit the available molecular adsorption sites to only the electromagnetic "hot spots" on the substrate. This process prevents molecules from adsorbing to sites of weak SERS enhancement, while permitting adsorption to sites of extraordinary SERS enhancement. For a randomly adsorbed submonolayer of benzenethiol molecules the average Raman scattering cross section of the processed sample is 27 times larger than that of an unprocessed SERS substrate.
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Frequency domain fluorescence lifetime imaging is a powerful technique that enables the observation of subtle changes in the molecular environment of a fluorescent probe. This technique works by measuring the phase delay between the optical emission and excitation of fluorophores as a function of modulation frequency. However, high-resolution measurements are time consuming, as the excitation modulation frequency must be swept, and faster low-resolution measurements at a single frequency are prone to large errors. Here, we present a low cost optical system for applications in real-time confocal lifetime imaging, which measures the phase vs. frequency spectrum without sweeping. Deemed Lifetime Imaging using Frequency-multiplexed Excitation (LIFE), this technique uses a digitally-synthesized radio frequency comb to drive an acousto-optic deflector, operated in a cat's-eye configuration, to produce a single laser excitation beam modulated at multiple beat frequencies. We demonstrate simultaneous fluorescence lifetime measurements at 10 frequencies over a bandwidth of 48 MHz, enabling high speed frequency domain lifetime analysis of single- and multi-component sample mixtures.
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Porous silicon nanoparticles (PSiNPs) are attractive carriers for targeted drug delivery in nanomedicine. For in vivo applications, the biodegradation property of PSiNPs provides a pathway for their safe clearance from the body. Particles sizes of 80-120 nm are of particular interest as they are important for cellular applications, such as drug delivery for cancer therapy, because these nanoparticles can take advantage of the enhanced permeability and retention effect to deliver drug preferentially to tumors with leaky vasculature, yet large enough to avoid renal clearance. However, the biodegradability rate of such particles is often too fast, which limits particle half-life and potentially reduces their in vivo delivery efficiency. In this work, we focus on the degradation of nanoscale particles and study the effect of both thermal oxidation and silica coating on the stability of PSiNPs in phosphate buffered saline solution (a close mimic of a basic biological fluid). Using thermal oxidation, the half-life of PSiNPs can be varied from 10 min up to 3 h. Using silica coating, the half-life can be extended further to 8 h. The particles produced using both these techniques can be functionalized using standard silica surface chemistries developed for applications in drug delivery.
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Nanopartículas/química , Silício/química , Biodegradação Ambiental , Cinética , Nanopartículas/ultraestrutura , Oxirredução , Porosidade , Dióxido de Silício/química , TemperaturaRESUMO
We present a new type of surface-enhanced Raman scattering (SERS) substrate that exhibits extremely large and uniform cross-section enhancements over a macroscopic (greater than 25 mm2) area. The substrates are fabricated using a femtosecond laser nanostructuring process, followed by thermal deposition of silver. SERS signals from adsorbed molecules show a spatially uniform enhancement factor of approximately 10(7). Spectroscopic characterization of these substrates suggests their potential for use in few or single-molecule Raman spectroscopy.