Upconversion FRET quantitation: the role of donor photoexcitation mode and compositional architecture on the decay and intensity based responses.

Lanthanide-doped colloidal nanoparticles capable of photon upconversion (UC) offer long luminescence lifetimes, narrowband absorption and emission spectra, and efficient anti-Stokes emission. These features are highly advantageous for Förster Resonance Energy Transfer (FRET) based detection. Upconverting nanoparticles (UCNPs) as donors may solve the existing problems of molecular FRET systems, such as photobleaching and limitations in quantitative analysis, but these new labels also bring new challenges. Here we have studied the impact of the core-shell compositional architecture of upconverting nanoparticle donors and the mode of photoexcitation on the performance of UC-FRET from UCNPs to Rose Bengal (RB) molecular acceptor. We have quantitatively compared luminescence rise and decay kinetics of Er3+ emission using core-only NaYF4: 20% Yb, 2% Er and core-shell NaYF4: 20% Yb @ NaYF4: 20% Yb, 5% Er donor UCNPs under three photoexcitation schemes: (1) direct short-pulse photoexcitation of Er3+ at 520 nm; indirect photoexcitation of Er3+ through Yb3+ sensitizer with (2) 980 nm short (5-7 ns) or (3) 980 nm long (4 ms) laser pulses. The donor luminescence kinetics and steady-state emission spectra differed between the UCNP architectures and excitation schemes. Aiming for highly sensitive kinetic upconversion FRET-based biomolecular assays, the experimental results underline the complexity of the excitation and energy-migration mechanisms affecting the Er3+ donor responses and suggest ways to optimize the photoexcitation scheme and the architecture of the UCNPs used as luminescent donors.

Quantitative Comparison of the Light-to-Heat Conversion Efficiency in Nanomaterials Suitable for Photothermal Therapy.

Functional colloidal nanoparticles capable of converting between various energy types are finding an increasing number of applications. One of the relevant examples concerns light-to-heat-converting colloidal nanoparticles that may be useful for localized photothermal therapy of cancers. Unfortunately, quantitative comparison and ranking of nanoheaters are not straightforward as materials of different compositions and structures have different photophysical and chemical properties and may interact differently with the biological environment. In terms of photophysical properties, the most relevant information to rank these nanoheaters is the light-to-heat conversion efficiency, which, along with information on the absorption capacity of the material, can be used to directly compare materials. In this work, we evaluate the light-to-heat conversion properties of 17 different nanoheaters belonging to different groups (plasmonic, semiconductor, lanthanide-doped nanocrystals, carbon nanocrystals, and metal oxides). We conclude that the light-to-heat conversion efficiency alone is not meaningful enough as many materials have similar conversion efficiencies─in the range of 80-99%─while they significantly differ in their extinction coefficient. We therefore constructed their qualitative ranking based on the external conversion efficiency, which takes into account the conventionally defined light-to-heat conversion efficiency and its absorption capacity. This ranking demonstrated the differences between the samples more meaningfully. Among the studied systems, the top-ranking materials were black porous silicon and CuS nanocrystals. These results allow us to select the most favorable materials for photo-based theranostics and set a new standard in the characterization of nanoheaters.

Engineering the Compositional Architecture of Core-Shell Upconverting Lanthanide-Doped Nanoparticles for Optimal Luminescent Donor in Resonance Energy Transfer: The Effects of Energy Migration and Storage.

Förster Resonance Energy Transfer (FRET) between single molecule donor (D) and acceptor (A) is well understood from a fundamental perspective and is widely applied in biology, biotechnology, medical diagnostics, and bio-imaging. Lanthanide doped upconverting nanoparticles (UCNPs) have demonstrated their suitability as alternative donor species. Nevertheless, while they solve most disadvantageous features of organic donor molecules, such as photo-bleaching, spectral cross-excitation, and emission bleed-through, the fundamental understanding and practical realizations of bioassays with UCNP donors remain challenging. Among others, the interaction between many donor ions (in donor UCNP) and many acceptors anchored on the NP surface and the upconversion itself within UCNPs, complicate the decay-based analysis of D-A interaction. In this work, the assessment of designed virtual core-shell NP (VNP) models leads to the new designs of UCNPs, such as …@Er, Yb@Er, Yb@YbEr, which are experimentally evaluated as donor NPs and compared to the simulations. Moreover, the luminescence rise and decay kinetics in UCNP donors upon RET is discussed in newly proposed disparity measurements. The presented studies help to understand the role of energy-transfer and energy migration between lanthanide ion dopants and how the architecture of core-shell UCNPs affects their performance as FRET donors to organic acceptor dyes.

Standardization of Methodology of Light-to-Heat Conversion Efficiency Determination for Colloidal Nanoheaters.

Localized photothermal therapy (PTT) has been demonstrated to be a promising method of combating cancer, that additionally synergistically enhances other treatment modalities such as photodynamic therapy or chemotherapy. PTT exploits nanoparticles (called nanoheaters), that upon proper biofunctionalization may target cancerous tissues, and under light stimulation may convert the energy of photons to heat, leading to local overheating and treatment of cancerous cells. Despite extensive work, there is, however, no agreement on how to accurately and quantitatively compare light-to-heat conversion efficiency (ηQ) and rank the nanoheating performances of various groups of nanomaterials. This disagreement is highly problematic because the obtained ηQ values, measured with various methods, differ significantly for similar nanomaterials. In this work, we experimentally review existing optical setups, methods, and physical models used to evaluate ηQ. In order to draw a binding conclusion, we cross-check and critically evaluate the same Au@SiO2 sample in various experimental conditions. This critical study let us additionally compare and understand the influence of the other experimental factors, such as stirring, data recording and analysis, and assumptions on the effective mass of the system, in order to determine ηQ in a most straightforward and reproducible way. Our goal is therefore to contribute to the understanding, standardization, and reliable evaluation of ηQ measurements, aiming to accurately rank various nanoheater platforms.

Fluorescent electrospun PMMA microfiber mats with embedded NaYF4: Yb/Er upconverting nanoparticles.

Functional upconverting nanoparticles (UCNPs) can offer new possibilities in fluorescent applications as they exhibit desired characteristic properties like large shift between the fluorescent emission signal and the infrared excitation wavelength, multi- and narrow-band absorption and emission in visible and near infrared - Vis/NIR, together with excellent photostability and low toxicity as opposed to semiconducting quantum dots. The upconversion luminescence emission or quenching characteristics of UCNPs can be altered upon exposure to physical or chemical environmental factors providing thus a functionality that can be utilized for sensing or imaging. Furthermore their functionalization with suitable indicator dyes or recognition elements can extend the range of luminescence response and ratiometric sensing to specific analytes. Synergistically, electrospun nano- and microfibers offering large surface area can enhance the functionality of UCNPs by retaining the fluorescence efficiency and improving the overall responsivity due to dramatically increased surface. For the optimization of this hybrid material system the controllable incorporation of UCNPs is required especially at increased concentration conditions needed for high brightness. Herein, we report the fabrication, morphological and optical characterization of electrospun polymer-based nanocomposite fibers, consisting of poly(methyl methacrylate) (PMMA) and upconverting lanthanide doped nanoparticles of the type NaYF4 : 20% Yb3+/2% Er3+ @ NaYF4. Morphological studies regarding the uniformity and aggregation effects of the UCNP inclusion within the fibers have been implemented followed by upconversion emission characterization by pulsed near-infrared excitation. The study and optimization of such nanocomposite fibrous systems could provide useful insights for the development of efficient upconverting electrospun fiber mats for a number of imaging and sensing applications.