Can T. Xu

High-Resolution Fluorescence Diffuse Optical Tomography Developed with Nonlinear Upconverting Nanoparticles

Fluorescence diffuse optical tomography (FDOT) is an emerging biomedical imaging technique that can be used to localize and quantify deeply situated fluorescent molecules within tissues. However, the potential of this technique is currently limited by its poor spatial resolution. In this work, we demonstrate that the current resolution limit of FDOT can be breached by exploiting the nonlinear power-dependent optical emission property of upconverting nanoparticles doped with rare-earth elements. The rare-earth-doped core-shell nanoparticles, NaYF(4):Yb(3+)/Tm(3+)@NaYF(4) of hexagonal phase, are synthesized through a stoichiometric method, and optical characterization shows that the upconverting emission of the nanoparticles in tissues depends quadratically on the power of excitation. In addition, quantum-yield measurements of the emission from the synthesized nanoparticles are performed over a large range of excitation intensities, for both core and core-shell particles. The measurements show that the quantum yield of the 800 nm emission band of core-shell upconverting nanoparticles is 3.5% under an excitation intensity of 78 W/cm(2). The FDOT reconstruction experiments are carried out in a controlled environment using liquid tissue phantoms. The experiments show that the spatial resolution of the FDOT reconstruction images can be significantly improved by the use of the synthesized upconverting nanoparticles and break the current spatial resolution limits of FDOT images obtained from using conventional linear fluorophores as contrast agents.

Autofluorescence insensitive imaging using upconverting nanocrystals in scattering media

Autofluorescence is a nuisance in the field of fluorescence imaging and tomography of exogenous molecular markers in tissue, degrading the quality of the collected data. In this letter, we report autofluorescence insensitive imaging using highly efficient upconverting nanocrystals (NaYF4:Yb3+∕Tm3+) in a tissue phantom illuminated with near-infrared radiation of 85mW∕cm2. It was found that imaging with such nanocrystals leads to an exceptionally high contrast compared to traditional downconverting fluorophores due to the absence of autofluorescence. Upconverting nanocrystals may be envisaged as important biological markers for tissue imaging purposes.

Fluorescence diffuse optical tomography using upconverting nanoparticles

Fluorescence diffuse optical tomography (FDOT) can provide important information in biomedical studies. In this ill-posed problem, suppression of background tissue autofluorescence is of utmost importance. We report a method for autofluorescence-insensitive FDOT using nonlinear upconverting nanoparticles (NaYF4:Yb3+/Tm3+) in a tissue phantom under excitation intensities well below tissue-damage thresholds. Even with the intrinsic autofluorescence from the phantom only, the reconstruction of the nanoparticles is of much better quality than the reconstruction of a Stokes-shifting dye. In addition, the nonlinear power dependence leads to more confined reconstructions and may increase the resolution in FDOT.

Disordered, strongly scattering porous materials as miniature multipass gas cells.

We investigate the interaction of light and gas in strongly scattering nano- and macroporous media. Manufacturing and structural characterization of ZrO(2), Al(2)O(3) and TiO(2) ceramics with different pore sizes, measurements of optical properties using photon time-of-flight spectroscopy, and high-resolution laser spectroscopy of O(2) at 760 nm are reported. We show that extreme light scattering can be utilized to realize miniature spectroscopic gas cells. Path length enhancement factors up to 750 are reached (5.4 m path through gas for light transmitted through a 7 mm ZrO(2) with 49% porosity and 115 nm pores).

Balancing power density based quantum yield characterization of upconverting nanoparticles for arbitrary excitation intensities.

Upconverting nanoparticles (UCNPs) have recently shown great potential as contrast agents in biological applications. In developing different UCNPs, the characterization of their quantum yield (QY) is a crucial issue, as the typically drastic decrease in QY for low excitation power densities can either impose a severe limitation or provide an opportunity in many applications. The power density dependence of the QY is governed by the competition between the energy transfer upconversion (ETU) rate and the linear decay rate in the depopulation of the intermediate state of the involved activator in the upconversion process. Here we show that the QYs of Yb(3+) sensitized two-photon upconversion emissions can be well characterized by the balancing power density, at which the ETU rate and the linear decay rate have equal contributions, and its corresponding QY. The results in this paper provide a method to fully describe the QY of upconverting nanoparticles for arbitrary excitation power densities, and is a fast and simple approach for assessing the applicability of UCNPs from the perspective of energy conversion.

Deep tissue optical imaging of upconverting nanoparticles enabled by exploiting higher intrinsic quantum yield through use of millisecond single pulse excitation with high peak power

We have accomplished deep tissue optical imaging of upconverting nanoparticles at 800 nm, using millisecond single pulse excitation with high peak power. This is achieved by carefully choosing the pulse parameters, derived from time-resolved rate-equation analysis, which result in higher intrinsic quantum yield that is utilized by upconverting nanoparticles for generating this near infrared upconversion emission. The pulsed excitation approach thus promises previously unreachable imaging depths and shorter data acquisition times compared with continuous wave excitation, while simultaneously keeping the possible thermal side-effects of the excitation light moderate. These key results facilitate means to break through the general shallow depth limit of upconverting-nanoparticle-based fluorescence techniques, necessary for a range of biomedical applications, including diffuse optical imaging, photodynamic therapy and remote activation of biomolecules in deep tissues.

Fluorescence Diffuse Optical Tomography using Upconverting Nanoparticles

In the fluorescence diffuse optical tomography (FDOT) problem, suppressing background is of utmost importance. We demonstrate autofluorescence-insensitive FDOT using upconverting nanoparticles and methods to exploit the nonlinearity to obtain reconstructions of higher resolutions.

Use of nonlinear upconverting nanoparticles provides increased spatial resolution in fluorescence diffuse imaging

Fluorescence diffuse imaging (FDI) suffers from limited spatial resolution. In this Letter, we report a scanning imaging approach to increase the resolution of FDI using nonlinear fluorophores. The resolution of a linear fluorophore was compared with nonlinear upconverting nanoparticles (NaYF(4):Yb(3+)/Tm(3+)) in a tissue phantom. A resolution improvement of a factor of 1.3 was found experimentally. Simulations suggested a maximum resolution improvement of a factor of 1.45. Usage of nonlinear fluorophores is a promising method for increasing the spatial resolution in FDI.

Multibeam fluorescence diffuse optical tomography using upconverting nanoparticles

Fluorescence diffuse optical tomography (FDOT) is a biomedical imaging modality that can be used for localization and quantification of fluorescent molecules inside turbid media. In this ill-posed problem, the reconstruction quality is directly determined by the amount and quality of the information obtained from the boundary measurements. Regularly, more information can be obtained by increasing the number of excitation positions in an FDOT system. However, the maximum number of excitation positions is limited by the finite size of the excitation beam. In the present work, we demonstrate a method in FDOT to exploit the unique nonlinear power dependence of upconverting nanoparticles to further increase the amount of information in a raster-scanning setup by including excitation with two beams simultaneously. We show that the additional information can be used to obtain more accurate reconstructions.

Drug quantification in turbid media by fluorescence imaging combined with light-absorption correction using white Monte Carlo simulations

Accurate quantification of photosensitizers is in many cases a critical issue in photodynamic therapy. As a noninvasive and sensitive tool, fluorescence imaging has attracted particular interest for quantification in pre-clinical research. However, due to the absorption of excitation and emission light by turbid media, such as biological tissue, the detected fluorescence signal does not have a simple and unique dependence on the fluorophore concentration for different tissues, but depends in a complex way on other parameters as well. For this reason, little has been done on drug quantification in vivo by the fluorescence imaging technique. In this paper we present a novel approach to compensate for the light absorption in homogeneous turbid media both for the excitation and emission light, utilizing time-resolved fluorescence white Monte Carlo simulations combined with the Beer-Lambert law. This method shows that the corrected fluorescence intensity is almost proportional to the absolute fluorophore concentration. The results on controllable tissue phantoms and murine tissues are presented and show good correlations between the evaluated fluorescence intensities after the light-absorption correction and absolute fluorophore concentrations. These results suggest that the technique potentially provides the means to quantify the fluorophore concentration from fluorescence images.