Nanospeckle Illumination Microscopy (NanoSIM) utilizes plasmonic nanoisland structures to enable super-resolution surface imaging of live cells. By analyzing the intensity fluctuations of plasmonic nanospeckles, we achieved three-fold improved spatial resolution and the ability to identify multiple cellular structures. Experimental results demonstrate the potential of NanoSIM as an effective and versatile tool for investigating dynamic cellular processes within live cell membranes of HeLa cells, providing crucial insights into complex cellular interactions.
Metallic nanostructures have the potential to be used in a variety of applications related to sensing and imaging biological molecules due to their ability to enhance the way molecules absorb and emit light. However, the interaction between metallic nanostructure and molecules can give rise to difficulty with determining precise molecular positions and orientations and therefore pose major challenges in the field of super-resolution imaging. In this work, we used axially defocused imaging to analyze the interaction between a single fluorescent molecule and a metallic nanostructure. In addition, a pattern matching algorithm was used to analyze the images, explore the interaction between the molecule and the nanostructure and thereby determine the lateral position. The accuracy was found to improve while the degree was dependent on the dipolar orientation and the distance between dipole and nanostructure. This approach has the potential to improve the reliability of using metallic nanostructures for imaging and sensing in the future and opens up new possibilities for various imaging and sensing methods.
Surface plasmon, collective electron oscillation induced by light absorption in noble metals, has received renewed attention that opens a new area of photonics research in what is known as thermoplasmonics. As thermoplasmonics develops, opto-thermal response measurement of a single nanostructure becomes essential. In this study, we propose a collection-type near-field scanning optical microscopy (NSOM) that can simultaneously measure light absorption and near-field enhancement on a single nanostructure. We analyzed light absorption from optically induced thermal expansion while measuring a near-field coupled with the NSOM tip. We have observed discrepancy and nonlinearity of angular spectrum between light absorption and near-field enhancement on gold thin films and compared with simulation results based on iterative opto-thermal analysis. We were able to determine the cause of the axial shift on the NSOM and the mechanisms by which the discrepancy may ariss. The proposed technique can also acquire optical characteristics of a single disk in a periodic array of gold nanodisks, and even measure the gaps between the disks. Furthermore, we expect the proposed technology to be extended to measuring near-field thermal characteristics of more complicated structure such as metamaterials.
Various plasmonic nanostructure-based substrates are used to detect biological signals beyond the diffraction limit with a high signal-to-noise ratio. These approaches take advantage of excitation of localized surface plasmon to acquire high-frequency biological signals while preserving photon energy. Numerous techniques, including focused ion beam, electronbeam lithography, and reactive ion etching, have been used to fabricate plasmonic substrates. However, these fabrication techniques are time and resource-consuming. In contrast, disordered nanostructure-based substrates have attracted interests due to the easy fabrication steps and potential cost savings. Metallic nanoisland substrates, for instance, can be mass-produced using thin film deposition and annealing without lithographic process. In this work, we have investigated nanospeckle illumination microscopy (NanoSIM) using disordered near-field speckle illumination generated by nanoisland substrate. Selectively activated fluorescence wide-field images were obtained by nanospeckle illumination generated on the nanoisland substrate. Super-resolved fluorescence images were reconstructed by an optimization algorithm based on blind structured illumination microscopy. Experimental studies of various biological targets including HeLa cell membranes were performed to demonstrate the performance of NanoSIM. Using NanoSIM, we were able to improve spatial resolution of ganglioside distribution in HeLa cells targeted by CT-B by more than threefold compared to the diffraction-limited images. Note that the accessibility of super-resolution imaging techniques can be enhanced by the nanospeckle illumination of disordered metallic nanoislands. These results may be used in imaging and sensing systems that work with detecting biological signals beyond diffraction limits in various applications.
We have investigated the feasibility of disordered plasmonic nanocomposites for super-resolution imaging. Annealing-based nanocomposite substrate has a great potential in biomedical optical and sensing technology because it can be mass-produced without difficult manufacturing processes. We introduce a new approach for wide-field super-resolution fluorescence imaging based on the nanocomposite island substrates, which we call nanospeckle illumination microscopy (NanoSIM). Near-field speckle patterns produced on disordered nanoisland substrates can help reconstruction of high-resolution fluorescence images with appropriate basis images. We have acquired basis images using azimuthal scanning illumination (ASI). Each ASI produces nonuniform nanoscale near-field speckles which can excite fluorescent dyes within a subdiffraction-limited area. While exploiting the random nature of plasmonic nanocomposite, NanoSIM does not require any specific polarization state to be maintained for ASI. We have tested NanoSIM to obtain super-resolved mages of molecules on the HeLa cell membrane. The full-width-at-half maximum was shown to improve by more than three times over the diffraction-limit with 360 basis images. Reconstructed images of gangliosides distribution on the HeLa cell suggest that fewer basis images may produce almost the same resolution with a shorter computation time. The optical resolution and sensitivity of disordered plasmonic substrate can be further enhanced by controlling the geometrical features of nanoislands structure.
Beyond structured illumination microscopy (SIM) which uses diffraction-limited light illumination, specially designed nanostructures such as metallic nanoantenna arrays generating localized surface plasmon have been developed to expand the frequency information without increasing photon energy. In this study, disordered temperature-annealed nanocomposite islands were used to create random distribution of nanospeckles because nanoisland substrates can be mass-produced in a large observation area by thin film deposition and annealing process. In our nanospeckle illumination microscopy (NanoSIM) system, azimuthal scanning illumination (ASI) on nanoislands creates a randomly localized nearfield distribution that induces an arbitrary number of fluorescence images. By the difficulty of obtaining structured illumination patterns of random nanostructures, images were reconstructed using a modified blind-SIM algorithm which fits well with the ASI system. A 100 nm fluorescent nanobead experiment confirms that NanoSIM provides resolution enhancement of spatial information in good agreement with the results obtained from AFM images. We emphasize that using random nanospeckles of disordered nanocomposite islands can provide highly accessible super-resolution. The results can be applied to imaging and sensing techniques, such as switching-based multi-channel microscopy.
In this work, we explore a way to design meta-plasmonic structure-based biosensors using machine learning methods. Plasmonic biosensing is a label-free detection method that is widely used to measure various biomolecular interactions. One of the main challenges has been how to improve the sensitivity and detection limit to detect very small molecules at low concentrations. Here, metamaterial was employed to address these issues using machine learning for the design. Transfer matrix algorithm was used to calculate optical characteristics of meta-plasmonic structure to generate training data. The multilayer perceptron was then applied to predict the optical characteristics of the meta-plasmonic structure. The performance was compared with conventional interpolation methods. Multilayer perceptron was shown to achieve mean squared error lower by about 1.5 times. Autoencoder and t-Stochastic Neighbor Embedding were also used to cluster the optical characteristics. Structural parameters which provide resonance in reflection can be found through clustering of optical characteristics. It was shown that meta-plasmonic structure improves sensitivity by more than ten times over conventional plasmonic biosensors. We expect that machine learning methods can be further extended to other biosensing modalities.
We have investigated the plasmonic effect of a gold thin film on the optical properties under a range of combinations of incident wavelengths, incident angles and polarization states, while assuming various film thicknesses. Theoretical calculation was performed with rigorous coupled-wave analysis based on the temperature-dependent Drude-Lorentz dispersion model. The calculation method considers the effects of absorption, which is converted to heat in a gold thin film and can affect material parameters such as permittivity. Experimentally, light absorption and field enhancement factor were directly measured using near-field scanning optical microscopy. We have also measured the near-field distribution and thermal effects in the gold thin film. Absorption and field enhancement experimentally measured using three incident wavelengths of 488, 532, and 721 nm for a thin gold film with thicknesses 20, 50, and 70 nm showed good agreement with calculated data. Also observed was the disparity between the incident angles that correspond to maximum absorption and highest field enhancement. The results can help understand the thermal effects on optical properties of plasmonic nanostructures for applications in biological imaging and sensing techniques.
We have investigated the effect of metal induced energy transfer from gold nanoaperture to fluorescence lifetime. We changed the orientation and position of dipole inside various diameter of nanohole and calculated relative lifetime by COMSOL Multiphysics and MATLAB. For the experiment, confocal microscopy setup was customized with blue laser at 479 nm and the nanoaperture was fabricated by both focused ion beam and e-beam lithography. Also, HPTS fluorescent dye in PVA solution was deposited on the nanoapertures by spin coating method instead of water droplets to avoid the effect from the film side and improve the contrast of the lifetime image. While the diameter of nanoapertures changed from 70 to 200 nm with 50 and 100 nm height, the maximum value of fluorescence lifetime tended to be proportional to the diameter and height due to energy transfer between dipole and metal. The maximum lifetime was 1.8 ns at 200 nm diameter which corresponded with calculation results. However, the lifetime difference between two heights was not linear tendency. The lifetime reduction factor was of about almost 10 for a 70 nm nanohole, and 3 for a 200 nm nanohole which was a maximum of 3 in the case of droplet. It is possible to distinguish the size of the nanohole using the lifetime reduction even at a scale below the diffraction limit and applied to single biomolecule detection.
We have studied how the light-matter colocalization effect, which is the overlap effect between target and electric field distribution, affects the sensitivity of nanogap-based surface plasmon resonance (SPR) sensors. The SPR characteristics of the nanogap structure were calculated using a rigorous coupled-wave analysis program with DNA immobilization and hybridization cases. The colocalized shift (COS) and its relative value, relative optical signature (ROS), were defined to explain the capability change according to the gap size (g) along with the momentum matching condition of the nanogap. The maximum sensitivity, defined as the COS value compared to the changed refractive index, was 514.8 deg / (μm * RIU), which appeared at a relatively large interval (g = 17nm). When increasing the nanogap size, the ROS and reflectance also increased. However, the trend reversed, and the decrease in ROS reached a negative range at a 9 nm gap or more. This ROS is due to damping in the multiple localized surface plasmon polariton mode, and the strong field confinement inside the nanogap induces a negative shift. The zero-shift, the most avoidable when designing an SPR sensor, and near-field distribution with plasmon mode, is observed across different gap size and period pairs. The effect of colocalization inside the nanogap, as identified through various metrics, promises a highly sensitive detection potential for biological applications.
In this report, we describe improvement of image resolution in surface plasmon resonance microscopy (SPRM) which suffers from poor quality due to severe surface plasmon (SP) propagation. Our approach takes two-channel momentum sampling by switched light incidence followed by minimum filtering to implement spatially switched SPRM (ssSPRM). The performance evaluated with periodic wires in comparison with conventional SPRM and bright-field microscopy shows that the effect of SP propagation can be circumvented and the effective decay length of SPRM is calculated to increase by only 7% compared to that of bright-field images.
We analyze the characteristics of plasmonics-based enhancement of a wire-grid polarizer (WGP) by rigorous coupledwave analysis (RCWA). We consider blazed WGP (bWGP) for improvement of polarimetric performance based on plasmonic momentum-matching in the metal/dielectric interface. The analysis used a model of triangular wire-grids approximated with five graded layers of identical thickness. We have compared the performance to that of a conventional WGP (cWGP) with a corresponding lamellar grating shape profile. As a performance measure, we calculated transmittance (TR) and extinction ratio (ER). It was found that TR in both cases tends to decrease monotonically with a longer period (Λ). The maximum TR of bWGPs is lower than cWGPs. On the other hand, maximum ER of bWGPs is much higher than that of cWGPs, particularly at a longer period, with an extinction peak peaked at Λ = 800 nm. For cWGPs, an extinction peak is observed at Λ = 200 nm with comparable enhancement (~42 dB). We have also computed relative TR (RTR) and relative ER (RER) for assessment of performance relative to cWGP. RTR decreases slowly in a manner similar to TR, however, RER increases exponentially with a longer wire-grid period. The results suggest that strong localization of near-fields observed with bWGPs can be used to improve polarimetric performance of a WGP.
Plasmonic optical trapping allows trapping and manipulation of micro- and even nanometer-sized particles using localized and enhanced electric fields by plasmon resonance in metallic nanostructure. We consider an optical conveyor belt consisting of an array of nanodisks acting as optical tweezers with different sizes to implement a system to trap and manipulate particles through a laser-induced gradient force. An electric field induced and localized at each optical resonator is sensitive to the wavelength and polarization. The maximum electric field is enhanced at resonant wavelength depending on the shape and size of the plasmonic nanostructure used for light localization. By changing the light wavelength and polarization, the position of localized light induced in the disk can be determined and nanoparticles can be moved to a desired location through the variation of resonance conditions without any mechanical forces.
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