Hari P. Paudel

Field of view advantage of conjugate adaptive optics in microscopy applications

The imaging performance of an optical microscope can be degraded by sample-induced aberrations. A general strategy to undo the effect of these aberrations is to apply wavefront correction with a deformable mirror (DM). In most cases the DM is placed conjugate to the microscope pupil, called pupil adaptive optics (AO). When the aberrations are spatially variant an alternative configuration involves placing the DM conjugate to the main source of aberrations, called conjugate AO. We provide a theoretical and experimental comparison of both configurations for the simplified case where spatially variant aberrations are produced by a well-defined phase screen. We pay particular attention to the resulting correction field of view (FOV). Conjugate AO is found to provide a significant FOV advantage. While this result is well known in the astronomical community, our goal here is to recast it specifically for the optical microscopy community.

Light-Controlled Plasmon Switching Using Hybrid Metal-Semiconductor Nanostructures

We present a proof of concept for the dynamic control over the plasmon resonance frequencies in a hybrid metal-semiconductor nanoshell structure with Ag core and TiO(2) coating. Our method relies on the temporary change of the dielectric function ε of TiO(2) achieved through temporarily generated electron-hole pairs by means of a pump laser pulse. This change in ε leads to a blue shift of the Ag surface plasmon frequency. We choose TiO(2) as the environment of the Ag core because the band gap energy of TiO(2) is larger than the Ag surface plasmon energy of our nanoparticles, which allows the surface plasmon being excited without generating electron-hole pairs in the environment at the same time. We calculate the magnitude of the plasmon resonance shift as a function of electron-hole pair density and obtain shifts up to 126 nm at wavelengths around 460 nm. Using our results, we develop the model of a light-controlled surface plasmon polariton switch.

Adaptive optics in multiphoton microscopy: comparison of two, three and four photon fluorescence

We demonstrate adaptive optics system based on nonlinear feedback from 3- and 4-photon fluorescence. The system is based on femtosecond pulses created by soliton self-frequency shift of a 1550-nm fiber-based femtosecond laser together with micro-electro-mechanical system (MEMS) phase spatial light modulator (SLM). We perturb the 1020-segment SLM using an orthogonal Walsh sequence basis set with a modified version of three-point phase shifting interferometry. We show the improvement after aberrations correction in 3-photon signal from fluorescent beads. In addition, we compare the improvement obtained in the same adaptive optical system for 2-, 3- and 4-photon fluorescence using dye pool. We show that signal improvement resulting from aberration correction grows exponentially as a function of the order of nonlinearity.

Focusing polychromatic light through strongly scattering media

We demonstrate feedback-optimized focusing of spatially coherent polychromatic light after transmission through strongly scattering media, and describe the relationship between optimized focus intensity and initial far-field speckle contrast. Optimization is performed using a MEMS spatial light modulator with camera-based or spectrometer-based feedback. We observe that the spectral bandwidth of the optimized focus depends on characteristics of the feedback signal. We interpret this dependence as a modification in the number of independent frequency components, or spectral correlations, transmitted by the sample, and introduce a simple model for polychromatic focus enhancement that is corroborated by experiment with calibrated samples.

Geometry dependence of field enhancement in 2D metallic photonic crystals

Geometry dependence of surface plasmon resonance of 2D metallic photonic crystals (PCs) was assessed using rigorous 3D finite difference time domain analysis. PCs of noble metallic rectangular and cylindrical nanopillars in square and triangular lattices on thick noble metal film were simulated for maximum field enhancement. It was found that the period, size and thickness of the nanopillars can be tuned to excite of surface plasmons at desired wavelengths in visible and near-infrared ranges. Maximum electric field enhancement near the nanopillars was found to be greater than 10X. The detail analysis of PCs tuned for 750 nm wavelength showed that thickness of nanopillars was the most sensitive parameter for field enhancement, and triangular lattice PCs had the wider enhancement bandwidth than square lattice PCs. Results showed that these PCs are sensitive with incident angle (theta) but not with polarization angle (phi).

Plasmon resonance modes in two-dimensional arrays of metallic nanopillars

Surface plasmon polaritons (SPPs) and localized plasmon resonance modes in two-dimensional arrays of silver nanopillars on silver surface were analyzed using the three-dimensional finite-difference time domain method. What we believe to be a new type of plasmon resonance modes at oblique incident angle for p-polarized light was observed in two-dimensional arrays of silver nanopillars in a square lattice. This resonance mode is associated with two SPP-like electric field patterns along the metal surface. We found that this resonance mode is localized and excited by the transverse polarization mode of nanopillars. Using Poynting vector plots, it was observed that the plasmon resonances in arrays of nanopillars are always associated with large energy cyclones near the nanopillars leading to light absorption.

Intravital imaging of osteocytes in mouse calvaria using third harmonic generation microscopy

Osteocytes are the most abundant cell in the bone, and have multiple functions including mechanosensing and regulation of bone remodeling activities. Since osteocytes are embedded in the bone matrix, their inaccessibility makes in vivo studies problematic. Therefore, a non-invasive technique with high spatial resolution is desired. The purpose of this study is to investigate the use of third harmonic generation (THG) microscopy as a noninvasive technique for high-resolution imaging of the lacunar-canalicular network (LCN) in live mice. By performing THG imaging in combination with two- and three-photon fluorescence microscopy, we show that THG signal is produced from the bone-interstitial fluid boundary of the lacuna, while the interstitial fluid-osteocyte cell boundary shows a weaker THG signal. Canaliculi are also readily visualized by THG imaging, with canaliculi oriented at small angles relative to the optical axis exhibiting stronger signal intensity compared to those oriented perpendicular to the optical axis (parallel to the image plane). By measuring forward- versus epi-detected THG signals in thinned versus thick bone samples ex vivo, we found that the epi-collected THG from the LCN of intact bone contains a superposition of backward-directed and backscattered forward-THG. As an example of a biological application, THG was used as a label-free imaging technique to study structural variations in the LCN of live mice deficient in both histone deacetylase 4 and 5 (HDAC4, HDAC5). Three-dimensional analyses were performed and revealed statistically significant differences between the HDAC4/5 double knockout and wild type mice in the number of osteocytes per volume and the number of canaliculi per lacunar surface area. These changes in osteocyte density and dendritic projections occurred without differences in lacunar size. This study demonstrates that THG microscopy imaging of the LCN in live mice enables quantitative analysis of osteocytes in animal models without the use of dyes or physical sectioning.

Axial range of conjugate adaptive optics in two-photon microscopy

We describe an adaptive optics technique for two-photon microscopy in which the deformable mirror used for aberration compensation is positioned in a plane conjugate to the plane of the aberration. We demonstrate in a proof-of-principle experiment that this technique yields a large field of view advantage in comparison to standard pupil-conjugate adaptive optics. Further, we show that the extended field of view in conjugate AO is maintained over a relatively large axial translation of the deformable mirror with respect to the conjugate plane. We conclude with a discussion of limitations and prospects for the conjugate AO technique in two-photon biological microscopy.

Design, fabrication, and characterization of a plasmonic upconversion enhancer and its prospects for photovoltaics

Abstract. The design, fabrication, and characterization of an upconversion-luminescence enhancer based on a two-dimensional plasmonic crystal are described. Full-wave finite-difference time domain analysis was used for optimizing the geometrical parameters of the plasmonic crystal for maximum plasmon activity, as signified by minimum light reflection. The optimum design produced >20× enhancement in the average electromagnetic field intensity within a one-micron-thick dielectric film over the plasmonic crystal. The optimized plasmonic upconverter was fabricated and used to enhance the upconversion efficiency of sodium yttrium fluoride: 3% erbium, 17% ytterbium nanocrystals dispersed in a poly(methylmethcrylate) matrix. A thin film of the upconversion layer, 105 nm in thickness, was spin-coated on the surface of the plasmonic crystal, as well as on the surfaces of planar gold and bare glass, which were used as reference samples. Compared to the sample with a planar gold back reflector, the plasmonic crystal showed an enhancement of 3.3× for upconversion of 980-nm photons to 655-nm photons. The upconversion enhancement was 25.9× compared to the same coating on bare glass. An absorption model was developed to assess the viability of plasmonically enhanced upconversion for photovoltaic applications.

A 3D topological insulator quantum dot for optically controlled quantum memory and quantum computing

We present the model of a quantum dot (QD) consisting of a spherical core-bulk heterostructure made of 3D topological insulator (TI) materials, such as PbTe/Pb