The metallic-associated adverse local tissue reactions (ALTR) and events accompanying worn-broken implant materials are still poorly understood on the subcellular and molecular level. Current immunohistochemical techniques lack spatial resolution and chemical sensitivity to investigate causal relations between material and biological response on submicron and even nanoscale. In our study, new insights of titanium alloy debris-tissue interaction were revealed by the implementation of label-free high-resolution correlative microscopy approaches. We have successfully characterized its chemical and biological impact on the periprosthetic tissue obtained at revision surgery of a fractured titanium-alloy modular neck of a patient with hip osteoarthritis. We applied a combination of photon, electron and ion beam micro-spectroscopy techniques, including hybrid optical fluorescence and reflectance micro-spectroscopy, scanning electron microscopy (SEM), Energy-dispersive X-ray Spectroscopy (EDS), helium ion microscopy (HIM) and micro-particle-induced X-ray emission (micro-PIXE). Micron-sized wear debris were found as the main cause of the tissue oxidative stress exhibited through lipopigments accumulation in the nearby lysosome. This may explain the indications of chronic inflammation from prior histologic examination. Furthermore, insights on extensive fretting and corrosion of the debris on nm scale and a quantitative measure of significant Al and V release into the tissue together with hydroxyapatite-like layer formation particularly bound to the regions with the highest Al content were revealed. The functional and structural information obtained at molecular and subcellular level contributes to a better understanding of the macroscopic inflammatory processes observed in the tissue level. The established label-free correlative microscopy approach can efficiently be adopted to study any other clinical cases related to ALTR.

Correlated light and electron microscopy (CLEM) has become essential in life sciences due to advancements in imaging resolution, sensitivity, and sample preservation. In nanotoxicology─specifically, studying the health effects of particulate matter exposure─CLEM can enable molecular-level structural as well as functional analysis of nanoparticle interactions with lung tissue, which is key for the understanding of modes of action. In our study, we implement an integrated high-resolution fluorescence lifetime imaging microscopy (FLIM) and hyperspectral fluorescence imaging (fHSI), scanning electron microscopy (SEM), ultrahigh resolution helium ion microscopy (HIM) and synchrotron micro X-ray fluorescence (SR μXRF), to characterize the nanobio interface and to better elucidate the modes of action of lung epithelial cells response to known inflammatory titanium dioxide nanotubes (TiO₂ NTs). Morpho-functional assessment uncovered several mechanisms associated with extensive DNA, essential minerals, and iron accumulation, cellular surface immobilization, and the localized formation of fibrous structures, all confirming immunomodulatory responses. These findings advance our understanding of the early cellular processes leading to inflammation development after lung epithelium exposure to these high-aspect-ratio nanoparticles. Our high-resolution experimental approach, exploiting light, ion, and electron sources, provides a robust framework for future research into nanoparticle toxicity and its impact on human health.

Color centers in hexagonal boron nitride (hBN) have been emerging as a multifunctional platform for various optical applications including quantum information processing, quantum computing and imaging. Simultaneously, due to its biocompatibility and biodegradability hBN is a promising material for biomedical applications. In this work, we demonstrate single-photon emission from hBN color centers embedded inside live cells and their application to cellular barcoding. The generation and internalization of multiple color centers into cells was performed via simple and scalable procedure while keeping the cells unharmed. The emission from live cells was observed as multiple diffraction-limited spots, which exhibited excellent single-photon characteristics with high single-photon purity of 0.1 and superb emission stability without photobleaching or spectral shifts over several hours. Due to different emission wavelengths and peak widths of the color centers, they were employed as barcodes. We term them Quantum Photonic Barcodes (QPBs). Each QPB can exist in one out of 470 possible distinguishable states and a combination of a few QPBs per cell can be used to uniquely tag virtually an unlimited number of cells. The barcodes developed here offer some excellent properties, including ease of production by a single-step procedure, biocompatibility and biodegradability, emission stability, no photobleaching, small size and a huge number of unique barcodes. This work provides a basis for the use of hBN color centers for robust barcoding of cells and due to the single photon emission, presented concepts could in future be extended to quantum-limited sensing and super-resolution imaging.

Knowing the temperature distribution within the conducting walls of various multilayer-type materials is crucial for a better understanding of heat-transfer processes. This applies to many engineering fields, good examples being photovoltaics and microelectronics. In this work we present a novel fluorescence technique that makes possible the non-invasive imaging of local temperature distributions within a transparent, temperature-sensitive, co-doped Er:GPF1Yb0.5Er glass-ceramic with micrometer spatial resolution. The thermal imaging was performed with a high-resolution fluorescence microscopy system, measuring different focal planes along the z-axis. This ultimately enabled a precise axial reconstruction of the temperature distribution across a 500-µm-thick glass-ceramic sample. The experimental measurements showed good agreement with computer-modeled heat simulations and suggest that the technique could be adopted for the spatial analyses of local thermal processes within optically transparent materials. For instance, the technique could be used to measure the temperature distribution of intermediate, transparent layers of novel ultra-high-efficiency solar cells at the micron and sub-micron levels.