Andrii Rogov

Simultaneous Multiharmonic Imaging of Nanoparticles in Tissues for Increased Selectivity

We investigate the use of bismuth ferrite (BFO) nanoparticles for tumor tissue labeling in combination with infrared multiphoton excitation at 1250 nm. We report the efficient and simultaneous generation of second- and third-harmonic signals by the nanoparticles. On this basis, we set up a novel imaging protocol based on the co-localization of the two harmonic signals and demonstrate its benefits in terms of increased selectivity against endogenous background sources in tissue samples. Finally, we discuss the use of BFO nanoparticles as mapping reference structures for correlative light–electron microscopy.

Harmonic nanoparticles: noncentrosymmetric metal oxides for nonlinear optics

response, often referred to as harmonicnanoparticles (HNPs). HNPs feature a series of properties which distinguish them from otherphotonics nanoprobes (quantum dots, up-conversion nanoparticles, noble metal particles). HNPsemission is inherently nonlinear and based on the efficient generation of harmonics as opposed tofluorescence or surface plasmon scattering. In addition, the fully coherent signal emitted byHNPs together with their polarization sensitive response and absence of resonant interactionmake them appealing for several applications ranging from multi-photon (infrared) microscopyand holography, to cell tracking and sensing.Keywords: multiphoton microscopy, second harmonic generation, third harmonic generation,nonlinear optics, nanophotonics, perovskites(Some figures may appear in colour only in the online journal)1. IntroductionSince the advent of ultrafast lasers in the early nineties andrelated availability of high peak power pulses at compara-tively low energies, the investigation of nonlinear opticalresponse have become a widespread activity resulting ininnovative applications. In particular, in the field of imagingthis has led to the introduction of multi-photon microscopy byW Webb [1]. This technique, as compared to one-photonexcited fluorescence microscopy, features increased imagingdepth, no out-of-focus bleaching, and spatial resolutioncomparable to confocal techniques without compromise insensitivity. The latest technological advancements in this fieldare related to its extension towards longer wavelengths,facilitated by the timely introduction of novel tunable sourcesin the 0.7–1.3 μm spectral range and new optical componentswith optimized performances in this region (microscopeobjectives, acousto-optic modulators, ...). Although these newinfrared tunable devices are supposedly more adapted forimaging through thick (living) samples because of reducedscattering [2–4], the available choice of imaging targets(fluorescent molecules, dyes) with two-photon absorptionbands >1200nm is very limited.Nanoparticle-based labelling, which is sometimes pre-ferred over other approaches for its superior photo-stability,brightness, and chemical selectivity usually requires two-photon interactions in the 700–900nm range, with a fewnotable exceptions [5]. However most optically active labelsmay suffer from bleaching and blinking and their use inbiological research might be prevented because of low bio-compatibility. For these reasons, since 2006, several researchgroups worldwide have proposed a complementary approach,

Nonlinear optical and magnetic properties of BiFeO3 harmonic nanoparticles

Second Harmonic Generation (SHG) from BiFeO3 nanocrystals is investigated for the first time to determine their potential as biomarkers for multiphoton imaging. Nanocrystals are produced by an auto-combustion method with 2-amino-2-hydroxymethyl-propane-1,3-diol as a fuel. Stable colloidal suspensions with mean particle diameters in the range 100–120 nm are then obtained after wet-milling and sonication steps. SHG properties are determined using two complementary experimental techniques, Hyper Rayleigh Scattering and nonlinear polarization microscopy. BiFeO3 shows a very high second harmonic efficiency with an averaged 〈d〉 coefficient of 79 ± 12 pm/V. From the nonlinear polarization response of individual nanocrystals, relative values of the independent dij coefficients are also determined and compared with recent theoretical and experimental studies. Additionally, the particles show a moderate magnetic response, which is attributed to γ-Fe2O3 impurities. A combination of high nonlinear optical efficiency and magnetic response within the same particle is of great interest for future bio-imaging and diagnostic applications.

Cellular uptake and biocompatibility of bismuth ferrite harmonic advanced nanoparticles

UNLABELLED Bismuth Ferrite (BFO) nanoparticles (BFO-NP) display interesting optical (nonlinear response) and magnetic properties which make them amenable for bio-oriented diagnostic applications as intra- and extra membrane contrast agents. Due to the relatively recent availability of this material in well dispersed nanometric form, its biocompatibility was not known to date. In this study, we present a thorough assessment of the effects of in vitro exposure of human adenocarcinoma (A549), lung squamous carcinoma (NCI-H520), and acute monocytic leukemia (THP-1) cell lines to uncoated and poly(ethylene glycol)-coated BFO-NP in the form of cytotoxicity, haemolytic response and biocompatibility. Our results support the attractiveness of the functional-BFO towards biomedical applications focused on advanced diagnostic imaging. FROM THE CLINICAL EDITOR Bismuth Ferrite nanoparticles (BFO-NP) have been recently successfully introduced as photodynamic tools and imaging probes. However, how these nanoparticles interact with various cells at the cellular level remains poorly understood. In this study, the authors performed in vitro experiments to assess the effects of uncoated and PEG-coated BFO-NP in the form of cytotoxicity, haemolytic response and biocompatibility.

Multi-Order Investigation of the Nonlinear Susceptibility Tensors of Individual Nanoparticles

We use Hyper Rayleigh Scattering and polarization resolved multiphoton microscopy to investigate simultaneously the second and third-order nonlinear response of Potassium Niobate and Bismuth Ferrite harmonic nanoparticles. We first derive the second-to-third harmonic intensity ratio for colloidal ensembles and estimate the average third-order efficiency of these two materials. Successively, we explore the orientation dependent tensorial response of individual nanoparticles fixed on a substrate. The multi-order polarization resolved emission curves are globally fitted with an analytical model to retrieve individual elements of susceptibility tensors.

Preparation from a revisited wet chemical route of phase-pure, monocrystalline and SHG-efficient BiFeO 3 nanoparticles for harmonic bio-imaging

We present two new synthetic routes for bismuth ferrite harmonic nanoparticles (BiFeO3 HNPs). Both phase-pure and mixed phase BiFeO3 materials were produced after improvement of the solvent evaporation and sol-gel combustion routes. Metal nitrates with a series of dicarboxylic acids (tartronic, tartaric and mucic) were used to promote crystallization. We found that the longer the carbon backbone with a hydroxyl group attached to each carbon, the lower the annealing temperature. We also demonstrate that nanocrystals more readily formed at a given temperature by adding glycerol but to the detriment of phase purity, whereas addition of NaCl in excess with mucic acid promotes the formation of phase-pure, monocrystalline nanoparticles. This effect was possibly associated with a better dispersion of the primary amorphous precursors and formation of intermediate complexes. The nanoparticles have been characterized by XRD, TEM, ζ-potential, photon correlation spectroscopy, two-photon microscopy and Hyper-Rayleigh Scattering measurements. The improved crystallization leads to BiFeO3 HNPs without defect-induced luminescence and with a very high averaged second harmonic efficiency (220 pm/V), almost triple the efficiency previously reported. This development of simple, scalable synthesis routes which yield phase-pure and, crucially, monocrystalline BiFeO3 HNPs demonstrates a significant advance in engineering the properties of nanocrystals for bio-imaging and diagnostics applications.

Nonlinear optical properties of silicon carbide (SiC) nanoparticles by carbothermal reduction

SiC nanoparticles by carbothermal reduction show promising properties in terms of second harmonic and multiphoton excited luminescence. In particular, we estimate a nonlinear efficiency < d < = 17 pm/V, as obtained by Hyper Rayleigh Scattering. We also present results of cell labelling to demonstrate the potential use of SiC nanoparticles for nonlinear bioimaging by simultaneous detection of second harmonic and luminescence.

Harmonic nanoparticles for nonlinar bio-imaging and detection

In this contribution we present the motivations underlying the introduction of harmonic nanoparticles, i.e. second harmonic contrast agents for nonlinear microscopy. Their properties will be discussed in the light of various biological applications including imaging of stem cells and rare event detection in physiological media.

Spinning-disc confocal microscopy in the second near-infrared window (NIR-II)

Fluorescence microscopy in the second near-infrared optical window (NIR-II, 1000–1350 nm) has become a technique of choice for non-invasive in vivo imaging. The deep penetration of NIR light in living tissue, as well as negligible tissue autofluorescence within this optical range, offers increased resolution and contrast with even greater penetration depths. Here, we present a custom-built spinning-disc confocal laser microscope (SDCLM) that is specific to imaging in the NIR-II. The SDCLM achieves a lateral resolution of 0.5 ± 0.1 µm and an axial resolution of 0.6 ± 0.1 µm, showing a ~17% and ~45% enhancement in lateral and axial resolution, respectively, compared to the corresponding wide-field configuration. We furthermore showcase several applications that demonstrate the use of the SDCLM for in situ, spatiotemporal tracking of NIR particles and bioanalytes within both synthetic and biological systems.

Tailoring single-cycle electromagnetic pulses in the 2–9 THz frequency range using DAST/SiO 2 multilayer structures pumped at Ti:sapphire wavelength

We present a numerical parametric study of single-cycle electromagnetic pulse generation in a DAST/SiO₂multilayer structure via collinear optical rectification of 800 nm femtosecond laser pulses. It is shown that modifications of the thicknesses of the DAST and SiO₂layers allow tuning of the average frequency of the generated THz pulses in the frequency range from 3 to 6 THz. The laser-to-THz energy conversion efficiency in the proposed structures is compared with that in a bulk DAST crystal and a quasi-phase-matching periodically poled DAST crystal and shows significant enhancement.