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We demonstrate a novel method for second harmonic generation-detected circular dichroism (CD) imaging based on the use of phase-locked, temporally delayed femtosecond laser pulses. The polarization state of the fundamental wave was controllably changed over 2μ rad by using a birefringent delay line, which provided attosecond inter-pulse delays for orthogonal phase-locked replicas; the achievable phase stability was 14 as. By introducing either a positive or negative delay of ~667 as, we induced a ±μ/2 phase shift between the orthogonally polarized pulses, resulting in left circularly polarized or right circularly polarized light. CD imaging performance using the pulse sequence was compared to results obtained for plasmonic nanoantennas using a rotating quarter-wave plate. The pulse sequence is expected to simplify polarization-resolved optical imaging by reducing experimental artifacts and decreasing image acquisition times. This method can be easily extended to other CD spectroscopy measurements. [ABSTRACT FROM AUTHOR]

The use of plasmon amplification of nonlinear optical wave-mixing signals to generate optical images in which the position of the scattering point source can be determined with nanometer accuracy is described. Solid gold nanosphere dimers were used as a model system for the nonlinear medium, which converted the Ti:sapphire fundamental to its second harmonic frequency. Matching the fundamental wave energy to the localized surface plasmon resonance of the electromagnetically coupled nanospheres was critical for achieving the high localization accuracy. Our technique, named Nonlinear Optical Localization using Electromagnetic Surface fields (NOLES) imaging, routinely yielded nonlinear optical images with 1-nm localization accuracy at rates ≥2 fps and can also be used as a photo-switching localization contrast method. This high level of accuracy in pinpointing the signal point source position exceeded that made possible using conventional diffraction-limited far-field methods by 160×. The NOLES technique, with its high temporal resolution and spatial accuracy that far surpass the performance typical of fluorescence-based imaging, will be relevant for imaging dynamic chemical, biological, and material environments. [ABSTRACT FROM AUTHOR]

TheNonlinear Optical Localization using Electromagnetic Surfacefields (NOLES) imaging technique was used to generate optical imagesin which the position of a chiral object could be determined withnanometer precision. Asymmetric gold bowtie nanostructures were usedas a model system with 2D chirality. The bowties functioned as a chiralnonlinear medium that converted the fundamental of a Ti:sapphire laserto its second harmonic frequency. The bowties consisted of two lithographicallyprepared equilateral triangles (base = 75 nm, height = 85 nm, thickness= 25 nm) separated by a 20 nm gap. Asymmetric bowties were formedby lateral displacement of one triangle by 10 nm, yielding C2point group symmetry. The chirality of thebowtie nanostructures was confirmed via nonzero second-harmonic generationcircular dichroism (SHG-CDR) ratios, which came from single-particleSHG measurements. The SHG-CDR ratios were validated using numericalfinite difference time domain simulations that quantified the relativemagnitudes of gap-localized electromagnetic fields at the harmonicfrequency resulting from excitation by left and right circularly (LCPand RCP) and linearly polarized fundamental waves. The relative electricdipolar and magnetic dipolar contributions to the SHG responses weredetermined using single-particle continuous polarization variation(CPV) SHG measurements. The spatial localization precision obtainablefor individual chiral nanostructures was determined by statisticalanalysis of the SHG image point spread function. Our results demonstratedthat both the chiral image contrast, which resulted from LCP and RCPexcitation, and the corresponding localization precision was dependentupon the relative magnetic dipole/electric dipole ratio (G/F). A localization precision of 1.13 ± 0.13nm and left-to-right image enhancements of 400% were obtained forbowties with the highest G/Fratiosusing 5 s frame exposure times. The polarization dependence and magneticdipole amplification confirmed here demonstrate that the NOLES imagingtechnique is a powerful method for studying chiral specimens withhigh spatial precision. [ABSTRACT FROM AUTHOR]

Here we introduce a fiber amplifier and a diamond Raman laser that output high powers (6.5 W, 1.3 W) at valuable wavelengths (1060 nm, 1250 nm) for two-photon excitation of red-shifted fluorophores. These custom excitation sources are both simple to construct and cost-efficient in comparison to similar custom and commercial alternatives. Furthermore, they operate at a repetition rate (80 MHz) that allows fast image acquisition using resonant scanners. With our system we demonstrate compatibility with fast resonant scanning, the ability to acquire neuronal images, and the capability to image vasculature at deep locations (>1 mm) within the mouse cerebral cortex., Competing Interests: The authors declare no conflicts of interest., (© 2022 Optica Publishing Group under the terms of the Optica Open Access Publishing Agreement.)

Deep in vivo imaging of vasculature requires small, bright, and photostable fluorophores suitable for multiphoton microscopy (MPM). Although semiconducting polymer dots (pdots) are an emerging class of highly fluorescent contrast agents with favorable advantages for the next generation of in vivo imaging, their use for deep MPM has never before been demonstrated. Herein, we characterize the multiphoton properties of three pdot variants and perform deep in vivo MPM imaging of cortical rodent microvasculature. We find pdot brightness exceeds conventional fluorophores, including quantum dots, and their broad multiphoton absorption spectrum permits imaging at wavelengths better-suited for biological imaging and confers compatibility with a range of longer excitation wavelengths. This results in substantial improvements in signal-to-background ratio (>3.5-fold) and greater cortical imaging depths (z = 1,300 µm). Ultimately, pdots are a versatile tool for MPM due to their extraordinary brightness and broad absorption, enabling interrogation of deep structures in vivo ., Competing Interests: J. Yu and D. T. Chiu have financial interest in Lamprogen, which has licensed the pdot technology from the University of Washington.

Optical manipulation of colloidal nanoparticles and molecules is significant in numerous fields. Opto-thermoelectric nanotweezers exploiting multiple coupling among light, heat, and electric fields enables the low-power optical trapping of nanoparticles on a plasmonic substrate. However, the management of light-to-heat conversion for the versatile and precise manipulation of nanoparticles is still elusive. Herein, we explore the opto-thermoelectric trapping at plasmonic antennas that serve as optothermal nanoradiators to achieve the low-power (∼0.08 mW/μm 2 ) and deterministic manipulation of nanoparticles. Specifically, precise optical manipulation of nanoparticles is achieved via optical control of the subwavelength thermal hot spots. We employ a femtosecond laser beam to further improve the heat localization and the precise trapping of single ∼30 nm semiconductor quantum dots at the antennas where the plasmon-exciton coupling can be tuned. With its low-power, precise, and versatile particle control, the opto-thermoelectric manipulation can have applications in photonics, life sciences, and colloidal sciences.

We perform high-resolution, non-invasive, in vivo deep-tissue imaging of the mouse neocortex using multiphoton microscopy with a high repetition rate optical parametric amplifier laser source tunable between λ =1,100 and 1,400 nm. By combining the high repetition rate (511 kHz) and high pulse energy (400 nJ) of our amplifier laser system, we demonstrate imaging of vasculature labeled with Texas Red and Indocyanine Green, and neurons expressing tdTomato and yellow fluorescent protein. We measure the blood flow speed of a single capillary at a depth of 1.2 mm, and image vasculature to a depth of 1.53 mm with fine axial steps (5 μ m) and reasonable acquisition times. The high image quality enabled analysis of vascular morphology at depths to 1.45 mm.

Two-color multiphoton microscopy through wavelength mixing of synchronized lasers has been shown to increase the spectral window of excitable fluorophores without the need for wavelength tuning. However, most currently available dual output laser sources rely on the costly and complicated optical parametric generation approach. In this report, we detail a relatively simple and low cost diamond Raman laser pumped by a ytterbium fiber amplifier emitting at 1055 nm, which generates a first Stokes emission centered at 1240 nm with a pulse width of 100 fs. The two excitation wavelengths of 1055 and 1240 nm, along with the effective two-color excitation wavelength of 1140 nm, provide an almost complete coverage of fluorophores excitable within the range of 1000-1300 nm. When compared with 1055 nm excitation, two-color excitation at 1140 nm offers a 90% increase in signal for many far-red emitting fluorescent proteins (for example, tdKatushka2). We demonstrate multicolor imaging of tdKa-tushka2 and Hoechst 33342 via simultaneous two-color two-photon, and two-color three-photon microscopy in engineered 3D multicellular spheroids. We further discuss potential benefits and applications for two-color three-photon excitation. In addition, we show that this laser system is capable of in vivo imaging in mouse cortex to nearly 1 mm in depth with two-color excitation., Competing Interests: Conflict Of Interest: The authors declare no conflict of interest.

We report plasmon-mediated two-photon photoluminescence (TPPL)-detected circular dichroism (CD) from colloidal metal nanoparticle assemblies. Two classes of solid gold nanosphere (SGN) dimers--heterodimers and homodimers--were examined using polarization-resolved TPPL, second harmonic generation (SHG), and one-photon photoluminescence (OPPL). Unambiguous CD was detected in both the TPPL and SHG signals, and the magnitudes of the CD responses in these measurements showed agreement for individual nanostructures. Heterodimers gave larger CD responses (average TPPL-CDR = 0.62 ± 0.33; average SHG-CDR = 0.51 ± 0.21) than homodimers (average TPPL-CDR = 0.19 ± 0.04; average SHG-CDR = 0.18 ± 0.06). OPPL-CD was not detected for either structure. Analysis of dimer emission properties suggested the CD responses were determined by properties of the one-photon-resonant mode excited by the laser. Average TPPL signals were (4.3 ± 0.6)× larger than those for SHG. Because signal amplitude is a primary determinant for spatial accuracies and precisions obtained from optical microscopy, CD contrast generated from plasmon-mediated TPPL, which we report for the first time, can extend the suite of super-resolution imaging techniques.

Superatom state-resolved dynamics of the Au25(SC8H9)18(-) monolayer-protected cluster (MPC) were examined using femtosecond two-dimensional electronic spectroscopy (2DES). The electronic ground state of the Au25(SC8H9)18(-) MPC is described by an eight-electron P-like superatom orbital. Hot electron relaxation (200 ± 15 fs) within the superatom D manifold of lowest-unoccupied molecular orbitals was resolved from hot hole relaxation (290 ± 20 fs) in the superatom P states by using 2DES in a partially collinear pump-probe geometry. Electronic relaxation dynamics mediated by specific superatom states were distinguished by examining the time-dependent cross-peak amplitudes for specific excitation and detection photon energy combinations. Quantification of the time-dependent amplitudes and energy positions of cross peaks in the 2.21/1.85 eV (excitation/detection) region confirmed that an apparent energetic blue shift observed for transient bleach signals results from rapid hot electron relaxation in the superatom D states. The combination of structurally precise MPCs and state-resolved 2DES can be used to examine directly the influence of nanoscale structural modifications on electronic carrier dynamics, which are critical for developing nanocluster-based photonic devices.

The characteristic feature of noble metal nanoparticles is the localized surface plasmon resonance (LSPR). Plasmon-supporting nanoparticles can function as transducers because of the LSPR's ability to amplify electromagnetic fields and its sensitivity to changes in the surrounding dielectric. The performance of these materials in transducer applications is inherently related to nanoparticle structure. This Perspective describes the use of femtosecond laser-based spectroscopies to elucidate the nanoscale structure-property interplay. First, femtosecond time-resolved transient extinction measurements that probe the LSPR following nanoparticle photoexcitation are described. These measurements illustrate how nanostructure dimensions influence sensitivity to changes in the interfacial dielectric. The combination of single-particle nonlinear optical (NLO) measurements and electron microscopy is also used to describe the symmetry of plasmon surface fields in nanoparticle assemblies. In particular, the use of continuous polarization variation-detected second-harmonic generation to describe electric and magnetic dipolar contributions to NLO properties is discussed.