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Inorganic nanocrystals with linear and nonlinear luminescence in the ultraviolet, visible, near infrared and short-wave infrared like semiconductor quantum dots and spectrally shifting lanthanide-based nanophosphors have meanwhile found applications in the life and material sciences ranging from optical reporters for bioimaging and sensing over security barcodes to solid state lighting and photovoltaics. These nanomaterials commonly have increasingly sophisticated core/shell particle architectures with shells of different chemical composition and thickness to minimize radiationless deactivation at the particle surface that is usually the main energy loss mechanism [1]. For lanthanide-based spectral shifters, particularly for very small nanoparticles, also surface coatings are needed which protect near-surface lanthanide ions from luminescence quenching by high energy vibrators like O-H groups and prevent the disintegration of these nanoparticles under high dilution conditions. [2,3,4]. The identification of optimum particle structures requires quantitative spectroscopic studies focusing on the key performance parameter photoluminescence quantum yield [5,6], ideally flanked by single particle studies to assess spectroscopic inhomogeneities on a particle-to-particle level for typical preparation methods [7,8], Moreover, in the case of upconversion nanoparticles with a multi-photonic and hence, excitation power density (P)-dependent luminescence, quantitative luminescence studies over a broad P range are required to identify particle architectures that are best suited for applications in fluorescence assays up to fluorescence microscopy. Here, we present methods to quantify the photoluminescence of these different types of emitters in the vis/NIR/SWIR and as function of P and demonstrate the importance of such measurements for a profound mechanistic understanding of the nonradiative deactivation pathways in semiconductor and upconversion nanocrystals of different size and particle architecture in different environments.
The surface chemistry of nanomaterials controls their interaction with the environment and biological species and their fate and is hence also relevant for their potential toxicity. This has meanwhile led to an increasing interest in validated and preferably standardized methods for the determination and quantification of surface functionalities on nanomaterials and initiated different standardization projects within ISO/TC 229 and IEC/TC 113 as well as interlaboratory comparisons (ILCs) of different analytical methods for the quantification of surface coatings by OECD. Here we present the results of a first ILC on the quantification of the amount of amino functionalities on differently sized inorganic nanoparticles done by division Biophotonics and the National Research Council of Canada (NRC) and the PWI 19257 on the Characterization and Quantification of Surface Functional Groups and Coatings on Nanoobjects approved by ISO/TC 229 (WG2) in fall 2022 that will result in a VAMAS study on this topic organized by division Biophotonics. Key words: nanoparticles, surface analysis, surface functional groups, quantification, optical assay, qNMR, VAMAS, standardization, ICL, quality assurance, reference material.
Inorganic nanocrystals with linear and nonlinear luminescence in the ultraviolet, visible, near infrared and short-wave infrared like semiconductor quantum dots and spectrally shifting lanthanide-based nanophosphors have meanwhile found many applications in the life and material sciences. This includes optical reporters for bioimaging and sensing, security and authentication barcodes, solid state lighting, converter materials, and photovoltaics. The identification of optimum particle structures requires quantitative spectroscopic studies under application-relevant conditions, focusing on the key performance parameter photoluminescence quantum yield, ideally flanked by single particle studies to assess spectroscopic inhomogeneities on a particle-to-particle level for typical preparation methods. In this context, methods to quantify the photoluminescence of these different nanoscale emitters are shown and utilized as a basis for a profound mechanistic understanding of the nonradiative deactivation pathways in semiconductor and upconversion nanocrystals of different size and particle architecture in different environments. Exemplary for the application potential of such nanomaterials, in addition, the design of optical sensors from different nanomaterials and functional organic dyes is briefly summarized.
In the focus of division Biophotonics are the design, preparation, analytical and spectroscopic characterization, and application of molecular and nanoscale
functional materials, particularly materials with a photoluminescence in the visible, near infrared (NIR) and short-wave infrared (SWIR). This includes optical reporters for bioimaging and sensing, security and authentication barcodes, and materials for solid state lighting, energy conversion, and photovoltaics. For the identification of optimum particle structures quantitative spectroscopic studies are performed under application-relevant conditions, focusing on the key performance parameter photoluminescence quantum yield. In addition, simple, cost-efficient, and standardizable strategies for quantifying functional groups on the surface of nano- and microparticles are developed, here with a focus on optical assays and electrochemical titration methods, cross-validated by more advanced methods such as quantitative NMR. In addition, reference materials and reference products are developed for optical methods, particularly luminescence techniques, and for analytical methods utilized for the characterization of nanomaterials.
•Overview of expertise of division Biophotonics
•Functional molecular and nanoscale luminescent reporters, probes, and
sensors including design principles available from division Biophotonics
•Examples for pH and O2 responsive molecular probes and particle sensors
•Surface group quantification Analytical methods and cleavable probes
•Concepts & standards for the validation and traceability of optical measurements
pH and oxygen are amongst the most important and frequently measured analytes in the life and material sciences, indicating, e.g., diseases and corrosion processes. This includes the optical monitoring of pH in living cells for studying cellular internalization pathways, such as phagocytosis, endocytosis, and receptor ligand internalization with the aid of molecular and nanoscale fluorescent sensors. Nanoparticle (NP)-based sensors, that are labeled or stained with a multitude of sensor dyes, have several advantages as compare to conventional molecular probes like enhanced brightness, i.e., amplified signals, ease of designing ratiometric systems by combining analyte sensitive and inert reference dyes, and increased photostability. Moreover, this can enable the use of hydrophobic dyes in aqueous environments. Versatile templates and carriers for the fabrication of nanosensors by the staining and/or labelling with different fluorophores and sensor molecules or surface functionalized NP like silica (SiO2-NP) and polystyrene (PS-NP) particles provide. Here we present the design of a versatile platform of color emissive nanosensors and stimuli-responsive microparticles for the measurement of pH, oxygen, and other targets utilizing both types of matrices and sets of spectrally distinguishable sensor and reference dyes and their characterization and demonstrate the applicability of representative sensor particle for cellular studies.
Inorganic nanocrystals with linear and nonlinear luminescence in the ultraviolet, visible, near infrared and shortwave infrared like semiconductor quantum dots and spectrally shifting lanthanide-based nanophosphors have meanwhile found applications in the life and material sciences ranging from optical reporters for bioimaging and sensing over security barcodes to solid state lighting and photovoltaics. These nanomaterials commonly have increasingly sophisticated core/shell particle architectures with shells of different chemical composition and thickness to minimize radiationless deactivation at the particle surface that is usually the main energy loss mechanism [1]. For lanthanide-based spectral shifters, particularly for very small nanoparticles, also surface coatings are needed which protect near-surface lanthanide ions from luminescence quenching by high energy vibrators like O-H groups and prevent the disintegration of these nanoparticles under high dilution conditions. [2,3,4]. The identification of optimum particle structures requires quantitative spectroscopic studies focusing on the key performance parameter photoluminescence quantum yield [5,6], ideally flanked by single particle studies to assess spectroscopic inhomogeneities on a particle-to-particle level for typical preparation methods [7], Moreover, in the case of upconversion nanoparticles with a multi-photonic and hence, excitation power density (P)-dependent luminescence, quantitative luminescence studies over a broad P range are required to identify particle architectures that are best suited for applications in fluorescence assays up to fluorescence microscopy. Here, we present methods to quantify the photoluminescence of these different types of emitters in the vis/NIR/SWIR and as function of Pand demonstrate the importance of such measurements for a profound mechanistic understanding of the nonradiative deactivation pathways in semiconductor and upconversion nanocrystals of different size and particle architecture in different environments.
Boron neutron capture therapy (BNCT) relies on the activation of 10B by thermal neutrons, which results in small highly energetic particle emission inducing cancer cells damage. However, in order to overcome the limits of the currently used BNCT agents, it is necessary to design new systems, which can specifically accumulate and deliver a sufficient amount of 10B in tumors. In this study, we designed a 10B-BSH-containing aza-BODIPY (aza-SWIR-BSH). It enabled the efficient vectorization of clinically used 10B-BSH to the tumor, resulting in higher therapeutic activity than the 10B-BSH alone.
Accurate and quantitative photoluminescence measurements are mandatory for the comparison of different emitter classes and the rational design of the next generation of molecular and nanoscale reporters as well as for most applications relying on their luminescence features in the life and material sciences and nanobiophotonics. In the following, procedures for the determination of the spectroscopic key parameter photoluminescence quantum yield, i.e., the number of emitted per absorbed photons, in the UV/vis/NIR/SWIR are presented including pitfalls and achievable uncertainties and material-specific effects related to certain emitter classes are addressed. Special emphasis is dedicated to luminescent nanocrystals.
Lanthanide-based upconversion nanoparticles (UCNPs) like hexagonal 𝛽-NaYF4 UCNPs doped with Yb3+ and Er3+, which efficiently convert 976 nm light to ultraviolet, visible, and near infrared photons, offer new strategies for luminescence-based sensing, barcoding, and Imaging. Their upconversion (UC) luminescence (UCL) features like UCL intensity, quantum yield, relative spectral distribution / UCL luminescence color, and luminescence decay kinetics are, however, strongly influenced by particle size, dopant ion concentration, particle architecture, surface chemistry including presence and thickness of surface passivation and shielding shells, microenvironment/presence of quenchers with high energy vibrations, and excitation power density (P).
We present here a comprehensive study of the influence of excitation power density on the UCL features of different types of UCNPs, focusing on Yb3+ and Er3+ co-doped NaYF4 core-only and core-shell nanostructures with different sizes and doping ion concentration, which underlines the importance of P-dependent optimum dopant concentrations for UCNP performance and the potential of P-tuning of UCL.