Ingenieurwissenschaften und zugeordnete Tätigkeiten
Filtern
Dokumenttyp
- Posterpräsentation (25) (entfernen)
Referierte Publikation
- nein (25)
Schlagworte
- Fluorescence (10)
- Dye (5)
- Nanoparticle (4)
- Assay (3)
- Flow cytometry (3)
- Iron oxide nanoparticles (3)
- Nano (3)
- Nanomaterial (3)
- Quality assurance (3)
- Quantum dot (3)
Organisationseinheit der BAM
- 1.2 Biophotonik (25) (entfernen)
SWIR luminescent nanomaterials – key chemical parameters for bright probes for in vivo bioimaging
(2024)
A current challenge for studying physio-pathological phenomena and diseaserelated processes in living organisms with non-invasive optical bioimaging is the development of bright optical reporters that enable deep tissue penetration, a high detection sensitivity, and a high spatial and temporal resolution. The focus of this project are nanomaterials, which absorb and emit in the shortwave infrared (SWIR) between ~900–2500 nm where scattering, absorption, and autofluorescence of the tissue are strongly reduced compared to the visible and NIR.
Surface functionalized silica nanoparticles (SiO2-NP) gained great interest in the life and material sciences, as they can be used e.g. as drug carriers, fluorescent sensors, and multimodal labels in bioanalytical assays and imaging applications. They are highly stable, are easily produced and modified on a large scale at low cost and can be labeled or stained with a multitude of sensor dyes. These dye modified particle conjugates have several advantages as compared to conventional molecular probes like enhanced brightness, ease of designing ratiometric systems by combining analyte-sensitive and inert reference dyes, and increased photostability. Moreover, stained nanoparticles can enable the use of hydrophobic dyes in aqueous environments.
Here we present our work on multicolored sensors for the measurement of pH, oxygen and saccharides utilizing amorphous SiO2 NPs.
Engineered nanoparticles (NPs) with various chemical compositions and surface functionalities are routinely fabricated for industrial applications such as medical diagnostics, drug delivery, sensing, catalysis, energy conversion and storage, opto-electronics, and information storage which improve the quality of life and European prosperity. NP function, performance, interaction with biological species, and environmental fate are largely determined by their surface functionalities. Standardized repeatable surface characterization methods are therefore vital for quality control of NPs, and to meet increasing concerns regarding their safety. Therefore, industry, regulatory agencies, and policymakers need validated traceable measurement methods and reference materials. This calls for fit-for-purpose, validated, and standardized methods, and reference data and materials on the surface chemistry of engineered NPs. Here, we present a concept for the development of such standardized measurement protocols utilizing method cross-validation and interlaboratory comparisons (ILCs) with emphasis on both advanced measurement methods such as quantitative Nuclear Magnetic Resonance (qNMR), X-ray photoelectron spectroscopy (XPS) and secondary ion mass spectrometry (SIMS) and cost-efficient, non-surface specific methods like optical assays and electrochemical titration methods.
Engineered and tailored nanomaterials (NM) are of great interest in the life and material sciences, as they can be used, e.g., as drug carriers, barcodes, fluorescent sensors, and multimodal labels in bioanalytical assays and imaging applications. Their performance and safety depend not only on their particle size, size distribution, and morphology, but also on their surface chemistry, i.e., the total number of surface functional groups (FG) and the number of FG accessible for subsequent functionalization with ligands or biomolecules, which in turn determines surface charge, colloidal stability, biocompatibility, and toxicity. It also underlines the importance of validated analytical methods that provide accurate information on these application-relevant physicochemical properties with a known uncertainty. In the case of FG quantification, this calls for robust, fast, inexpensive, and reliable methods which allow for the characterization of a broad variety of NM differing in size, chemical composition, and optical properties.
Methods
Aiming at the development of simple, versatile, and multimodal tools for the quantification of bioanalytically relevant FG such as amine, carboxy, thiol, and aldehyde functionalities, we investigated and compared various analytical methods commonly used for FG quantification. This includes electrochemical titration methods, dye-based optical assays, and other instrumental analytical techniques such as nuclear magnetic resonance (NMR), mass spectrometry (MS), and thermal analysis methods.
Results
Here, we will present examples for different types of NMs and FGs including results from a currently running interlaboratory comparison (ILC) with the National Research Council of Canada (NRC) to pave the road for method standardization.
Innovative aspects
• Surface analysis
• Performance and safety of nanomaterials
• Standardization
A series of aroyl-S,N-ketene acetal based bichromophores is readily synthesized by Buchwald-Hartwig amination and Ullmann reaction in moderate to good yields. The aminated aroyl-S,N-ketene acetals are emissive in the solid state and in the aggregate, but not in solution, thus, they are AIEgens (aggregation induced emission chromogens). Aggregation is induced by fractional alternation of the solvent mixture, here by increasing the water fraction of ethanol/water mixtures. For most derivatives, the emission upon induced aggregation stems solely from the aroyl-S,N-ketene acetal chromophore, regardless whether excitation occurs at the absorption maximum of the triarylamine or the aroyl-S,N-ketene acetal. Therefore, a pronounced energy transfer from the triarylamine donor to the aroyl-S,N-ketene acetal acceptor can be inferred. The color of the emission can be controlled by choosing the para-aroyl substituent. A partial energy transfer could also be observed for some bichromophores, leading to aggregation-induced dual emission (AIDE). In addition, four examples of aminated diaroyl-S,N-ketene acetals were added to the compound library. The electron-withdrawing properties of the additional aroyl group provide a bathochromic shift of the emission band of the aroyl-S,N-ketene acetal. These bichromophores also show AIDE and in one case even aggregation-induced white light emission as a result of additive color mixing.
Tailor-made nanoparticles are of increasing interest in e.g. catalysis, as sensor materials, analytical assays, or can have superior photophysical properties. A major issue concerning the preparation of high-quality and functional nanoparticles is a good control of particle size, shape, polydispersity, and composition.
Small Angle X-ray Scattering (SAXS) is a non-destructive method for the analysis of nanostructures in a wide variety of materials. This method allows determining averaged structural parameters on a length scale from just above atomic sizes up to several 100 nanometers such as sizes, size distributions, volume fractions, and inner surface sizes. Moreover, anomalous Small Angle X-ray Scattering (ASAXS) exploits the anomalous dispersion of the scattering amplitudes near the X-ray absorption edges of the elements contained in the sample. These element sensitive contrast variations can be used to analyse average composition fluctuations on the nm scale. Two kinds of nanoparticles are chosen here to elaborate the advantages of ASAXS in the analysis of complex materials.
A facile and efficient methodology is developed for the thermal synthesis of size-tunable, stable, and uniform bimetallic NiCu core–shell nanoparticles (NPs) for various application in catalysis. Their diameter can be tuned in a range from 6 nm to 30 nm and the Ni:Cu ratio is adjustable in a wide range from 1:1 to 30:1. The NPs are structurally characterized by a method combination of transmission electron microscopy, anomalous small-angle X-ray scattering (ASAXS), X-ray absorption fine structure, and X-ray photoelectron spectroscopy. Here, we focus on the ASAXS method and its ability to analyses nanostructure parts and their compositions at once. As a result, a NiCu alloyed core surrounded by a Ni enriched shell and an outer NiO shell was found.
Semiconductor nanocrystals (quantum dots, QDs) are well known for their superior photophysical properties and enabled advancements in several key technologies of the 21st century and numerous technological applications like in photovoltaics, LED displays, photocatalysis, and biosensing. To achieve high photoluminescence quantum yields (PLQY) and enhanced photostability the QD core needs to be passivated by a second semiconductor, which possess a larger band gap to confine the charges within the QD core. An important parameter is thereby the lattice mismatch between the core and shell. To avoid strong lattice strain, which would alter the photophysical properties, an intermediary shell can be used as a lattice adapter between the core and the outer shell leading to core/shell/shell systems. These systems have shown to possess high PLQYs combined with a strong long-term stability and can be found in modern QLED displays. ASAXS was used here to better understand the core/shell/shell structure of InP/ZnSe/ZnS QDs to enable a correlation between their structural and photophysical properties.
Photoluminescence Quantum Yields of Luminescent Nanocrystals and Particles in the UV/vis/NIR/SWIR
(2023)
The rational design of functional luminescent materials such as semiconductor quantum dots and lanthanide-based upconversion nanoparticles, all photophysical and mechanistic studies, and the comparison of different emitters require accurate and quantitative photoluminescence measurements. Particularly the reliable determination of the key performance parameter photoluminescence quantum yield (f), the number of emitted per absorbed photons, and the brightness are of special importance for luminescence applications in the life and material sciences and nano(bio)photonics.[1] In this context, examples for absolute measurements of the photoluminescence quantum yields of UV/vis/NIR/SWIR emissive semiconductor quantum dots and rods, made from different materials, and spectrally shifting lanthanide upconversion nanocrystals with different surface chemistries in transparent matrices are presented including excitation wavelength and power density dependent studies utilizing integration sphere spectroscopy.[2,3] In addition, procedures for the absolute determination of the photoluminescence quantum yields of scattering dispersions of larger size quantum rods and differently sized inorganic particles have been developed as well as procedures for the characterization of solid luminescent nanomaterials such as different perovskites and YAG:Cer converter materials.[4] Thereby, challenges and pitfalls of f measurements in different wavelength regions including the SWIR and material-specific effects related to certain emitter classes are addressed, achievable uncertainties are quantified, and relative and absolute measurements of photoluminescence quantum yield measurements are compared to underline limitations of the former approach. Finally, a set of novel UV/vis/NIR quantum yield standards is presented including their certification with a complete uncertainty budget.[5]
Current challenges and objectives for non-invasive optical bioimaging are deep tissue penetration, high detection sensitivity, high spatial and temporal resolution, and fast data acquisition. A promising spectral window to tackle these challenges is the short-wave infrared (SWIR) ranging from 900 nm to 1700 nm where scattering, absorption, and autofluorescence of biological components are strongly reduced compared to the visible/NIR. At present, the best performing SWIR contrast agents are based on nanomaterials containing toxic heavy-metal ions like cadmium or lead, which raises great concerns for biological applications. Promising heavy-metal free nanoscale candidates are gold nanoclusters (AuNCs) and Ag2S nanoparticles (NPs). The photoluminescence (PL) of both types of nanomaterials is very sensitive to their size, composition of their surface ligand shell, and element composition, which provides an elegant handle to fine-tune their absorption and emission features and boost thereby the size of the signals recorded in bioimaging studies.
Aiming for the development of SWIR contrast agents with optimum performance, we dived deeper into the photophysical processes occurring in these nanomaterials, thereby exploring in depth how the environment, surface ligand composition, and the incorporation of transition metals influence the optical properties of AuNCs and Ag2S NPs. We observed a strong enhancement of the SWIR emission of AuNCs upon exposure to different local environments (in solution, polymer, and in the solid state). Addition of metal ions such as Zn2+ to Ag2S based NPs led to a strong PL enhancement, yielding PL quantum yields of about 10% and thus making them highly suitable for non-invasive deep imaging of vascular networks and 3D fluid flow mapping.
In recent years, chromium (III) complexes have received a lot of attention as novel near-infrared (NIR) emitters. This interest was triggered by the report on the first molecular ruby Cr(ddpd)2(BF4)3 with a high photoluminescence quantum yield of 13.7% of its near infrared (NIR) emission band and a long luminescence lifetime of 1.122 ms at room temperature. Meanwhile, the influence of triplet oxygen, temperature, and pressure on the optical properties of different molecular rubies have been assessed. These features make these molecular rubies promising candidates for multi-analyte optical sensing applications and the generation of singlet oxygen for photocatalysis and photodynamic therapy. However, in an oxygen-containing environment, the photoluminescence quantum yields and luminescence lifetimes of these chromium(III) complexes show only very small values. This hampers their application as NIR luminescence labels. This application, that cannot be tackled by conventional deoxygenating approaches, requires suitable strategies to protect the luminescence of the chromium(III) complexes from oxygen quenching. Typical approaches to reduce the oxygen sensitivity of long-lived luminophores include the encapsulation into an oxygen-shielding matrix or less commonly employed, by tuning the bulkiness of the ligands for oxygen-sensitive coordination compounds. An elegant approach to reduce the undesired luminescence quenching by triplet oxygen explored by us presents the incorporation of these chromium(III) complexes into amorphous, non-porous silica nanoparticles, that can be simply surface functionalized, e.g., with targeting ligands and/or other sensor molecules. This can enable the use of such chromium(III) complexes as reporters for bioanalytical assays and bioimaging without the need to introduce reactive groups into the ligands and can pave the road to lifetime tuning.
In this work, as first proof-of-concept experiments, a set of chromium (III) complexes constituting of different ligands and counter anions, were embedded into the core of silica nanoparticles. As an alternative synthesis strategy, selected complexes were incorporated into a silica shell formed around the core of self-made silica nanoparticles. Subsequently, the optical properties of the resulting luminescent silica nanoparticles were spectroscopically assessed by steady state and time-resolved luminescence spectroscopy. First results of time-resolved luminescence measurements of the Cr(ddpd)2(PF6)3 complex incorporated into 25nm large silica nanoparticles dispersed in aerated water in comparison to the decay kinetics obtained for this complex in acetonitrile in air showed an increase in lifetime from 46 µs to 1147 µs. This confirming our design concept of nanoscale NIR emissive Cr(III) reporters.
At present, the field of research on nanostructures is actively developing, which is due to their unique physico-chemical properties compared to bulk materials. Many research activities are focused on obtaining nanocomposites, which combine various types of nanostructures with different properties and function. For example, the development of magneto-luminescent nanocomposites makes it possible to use their luminescence for optical imaging, and their magnetic properties for magnetic targeted delivery and as agents of hyperthermia and magnetic resonance imaging.
My master studies as part of the project Goszadanie 2019-1080 at ITMO were focused on the investigation of nanocomposites, consisting of semiconductor quantum dots (QDs) as luminescent component and superparamagnetic iron oxide nanoparticles (SPIONs) as magnetic one, in solution and during their incubation with HeLa cells. The spectrally resolved analysis of the QD photoluminescence (PL) kinetics of the free QDs and the QDs incorporated in these nanocomposites undergoing energy transfer processes allowed for (1) understanding the reasons for the quenching of QD luminescence in cells, (2) evaluating the average distance between the QDs and, based on this, concluding the degree of QD aggregation in cells, and (3) drawing conclusions about the QD-quencher composites integrity in cells. Overall, the analysis of the PL kinetics confirmed that QDs and SPIONs remain bound in the obtained nanocomposites during incubation with cells.
To ensure the successful advancement of nanomaterials in biomedicine and the transition from their laboratory preparation and studies to their use in different applications and in industry, it is crucial to develop reliable measurement methods and reference materials candidates for the characterization of functional nanomaterials and assessing the quality of the obtained nanostructures. My recently started project at BAM, which is part of the EU metrology project MeTrINo, will be devoted to this topic. There we will focus on the development of methodologies for the synthesis and characterization of iron oxide nanoparticles, already used in biomedicine, and multi-element lanthanide-based nanoparticles with attractive upconversion luminescence, as reference materials with high monodispersity and reproducibility. Also, these nanoparticles will be functionalized with organic dyes for optical imaging and, probably, the study of the energy transfer phenomena.