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A variety of luminescent dyes including the most common indicators for optical oxygen sensors were investigated in regard to their stability and photophysical properties in the presence of nitrogen dioxide. The dyes were immobilized in polystyrene and subjected to NO 2 concentrations from 40 to 5500 ppm. The majority of dyes show fast degradation of optical properties due to the reaction with NO 2 . The class of phosphorescent metalloporphyrins shows the highest resistance against nitrogen dioxide. Among them, palladium(II) and platinum(II) complexes of octasubstituted sulfonylated benzoporphyrins are identified as the most stable dyes with almost no decomposition in the presence of NO 2 . The phosphorescence of these dyes is reversibly quenched by nitrogen dioxide. Immobilized in various polymeric matrices, the sulfonylated Pt(II) benzoporphyrin demonstrates about one order of magnitude more efficient quenching by NO 2 than by molecular oxygen. Our study demonstrates that virtually all commercially available and reported optical oxygen sensors are likely to show either irreversible decomposition in the presence of nitrogen dioxide or reversible luminescence quenching. They should be used with extreme caution if NO 2 is present in relatively high concentrations or it may be generated from other species such as nitric oxide. As an important consequence of nearly anoxic systems, production of nitrogen dioxide or nitric oxide may be therefore erroneously interpreted as an increase in oxygen concentration.

Multicellular spheroids are important tools for studying tissue and cancer physiology in 3D and are frequently used in tissue engineering as tissue assembling units for biofabrication. While the main power of the spheroid model is in mimicking physical-chemical gradients at the tissue microscale, the real physiological environment (including dynamics of metabolic activity, oxygenation, cell death, and proliferation) inside the spheroids is generally ignored. At the same time, the effects of the growth medium composition and the formation method on the resulting spheroid phenotype are well documented. Thus, characterization and standardization of spheroid phenotype are required to ensure the reproducibility and transparency of the research results. The analysis of average spheroid oxygenation and the value of O2 gradients in three dimensions (3D) can be a simple and universal way for spheroid phenotype characterization, pointing at their metabolic activity, overall viability, and potential to recapitulate in vivo tissue microenvironment. The visualization of 3D oxygenation can be easily combined with multiparametric analysis of additional physiological parameters (such as cell death, proliferation, and cell composition) and applied for continuous oxygenation monitoring and/or end-point measurements. The loading of the O2 probe is performed during the stage of spheroid formation and is compatible with various protocols of spheroid generation. The protocol includes a high-throughput method of spheroid generation with introduced red and near-infrared emitting ratiometric fluorescent O2 nanosensors and the description of multi-parameter assessment of spheroid oxygenation and cell death before and after bioprinting. The experimental examples show comparative O2 gradients analysis in homo- and hetero-cellular spheroids as well as spheroid-based bioprinted constructs. The protocol is compatible with a conventional fluorescence microscope having multiple fluorescence filters and a light-emitting diode as a light source.

Reaction of [NiCl 2 (PnH) 4 ] (1) (PnH=6-tert-butyl-pyridazine-3-thione) with NiCl 2 affords the binuclear paddlewheel (PW) complex [Ni 2 (Pn) 4 ] (2). Diamagnetic complex 2 is the first example of a PW complex capable of reversibly binding and releasing NH 3 . The NH 3 ligand in [Ni 2 (Pn) 4 (NH 3 )] (2⋅NH 3 ) enforces major spectroscopic and magnetic susceptibility changes, thus displaying vapochromic properties (λ max (2)=532 nm, λ max (2⋅NH 3 )=518 nm) and magnetochemical switching (2: S=0; 2⋅NH 3 : S=1). Upon repeated adsorption/desorption cycles of NH 3 the PW core remains intact. Compound 2 can be embedded into thin polyurethane films (2 P ) under retention of its sensing abilities. Therefore, 2 qualifies as reversible optical probe for ammonia. The magnetochemical switching of 2 and 2⋅NH 3 was studied in detail by SQUID measurements showing that in 2⋅NH 3 , solely the Ni atom coordinated the NH 3 molecule is responsible for the paramagnetic behavior., (© 2021 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.)

Aprotic alkali metal-O 2 batteries face two major obstacles to their chemistry occurring efficiently, the insulating nature of the formed alkali superoxides/peroxides and parasitic reactions that are caused by the highly reactive singlet oxygen ( 1 O 2 ). Redox mediators are recognized to be key for improving rechargeability. However, it is unclear how they affect 1 O 2 formation, which hinders strategies for their improvement. Here we clarify the mechanism of mediated peroxide and superoxide oxidation and thus explain how redox mediators either enhance or suppress 1 O 2 formation. We show that charging commences with peroxide oxidation to a superoxide intermediate and that redox potentials above ~3.5 V versus Li/Li + drive 1 O 2 evolution from superoxide oxidation, while disproportionation always generates some 1 O 2 . We find that 1 O 2 suppression requires oxidation to be faster than the generation of 1 O 2 from disproportionation. Oxidation rates decrease with growing driving force following Marcus inverted-region behaviour, establishing a region of maximum rate.

Metallocorroles involving 5d transition metals are currently of interest as near-IR phosphors and as photosensitizers for oxygen sensing and photodynamic therapy. Their syntheses, however, are often bedeviled by capricious and low-yielding protocols. Against this backdrop, we describe rhenium-imido corroles, a new class of 5d metallocorroles, synthesized simply and in respectable (∼30%) yields via the interaction of a free-base corrole, Re 2 (CO) 10 , K 2 CO 3 , and aniline in 1,2,4-trichlorobenzene at ∼190 °C in a sealed vial under strict anaerobic conditions. The generality of the method was shown by the synthesis of six derivatives, including those derived from meso -tris(pentafluorophenyl)corrole, H 3 [TPFPC], and five different meso -tris( p -X-phenyl)corroles, H 3 [T p XPC], where X = CF 3 , F, H, CH 3 , OCH 3 . Single-crystal X-ray structures obtained for two of the complexes, Re[T p FPC](NPh) and Re[T p CF 3 PC](NPh), revealed relatively unstrained equatorial Re-N distances of ∼2.00 Å, a ∼ 0.7-Å displacement of the Re from the mean plane of the corrole nitrogens, and an Re-N imido distance of ∼1.72 Å. Details of the corrole skeletal bond distances, diamagnetic 1 H NMR spectra, relatively substituent-independent Soret maxima, and electrochemical HOMO-LUMO gaps of ∼2.2 V all indicated an innocent corrole macrocycle. Surprisingly, unlike several other classes of 5d metallocorroles, the Re-imido complexes proved nonemissive in solution at room temperature and also failed to sensitize singlet oxygen formation, indicating rapid radiationless deactivation of the triplet state, presumably via the rapidly rotating axial phenyl group. By analogy with other metal-oxo and -imido corroles, we remain hopeful that the Re-imido group will prove amenable to further elaboration and thereby contribute to the development of a somewhat challenging area of coordination chemistry.

A set of rhenium(V)-oxo meso -triarylcorroles bearing ester and carboxylic acid functionalities were synthesized with a view to determining their potential for photodynamic therapy. Toward this end, we measured their near-IR phosphorescence and their ability to sensitize singlet oxygen formation. The two esters studied, Re V O 5,10,15-tris( meta -carbomethoxyphenyl)corrole and Re V O 5,10,15-tris( para -carbomethoxyphenyl)corrole, were found to exhibit phosphorescence quantum yields of around 1% and fairly long phosphorescence lifetimes of about 60 μs in toluene. The corresponding carboxylic acids, which were examined in ethanolic/aqueous media, in contrast, showed much lower phosphorescence quantum yields on the order of 0.01% and somewhat shorter phosphorescent lifetimes. The quantum yields for singlet oxygen formation, on the other hand, turned out to be equally high (0.72 ± 0.02) for the esters and corresponding carboxylic acids. For the two carboxylic acids, we also carried out photocytotoxicity measurements on rat bladder cancer cells (AY27) and human colon carcinoma cells (WiDr). Cell viability measurements (MTT assays) indicated 50% cell death (LD 50 ) for AY27 cells upon 5 min of blue light exposure with the meta carboxylic acid and upon 7 min of exposure with the para carboxylic acid; complete cell death resulted after 20 min for both compounds. The WiDr cells proved less sensitive, and LD 50 values were reached after 8 and 12 min illumination with the meta and para carboxylic acids, respectively., Competing Interests: The authors declare no competing financial interest., (Copyright © 2020 American Chemical Society.)

A new luminescent indicator is presented that enables simultaneous measurement of oxygen and temperature at a single wavelength. The indicator, an alkylsulfone-substituted Zn(II)- meso -tetraphenyltetrabenzoporphyrin, emits prompt and thermally activated delayed fluorescence (TADF). TADF is sensitive toward oxygen and temperature and is referenced against prompt fluorescence (PF) that is not affected by oxygen. The information on both parameters is accessed from the decay time of TADF and the temperature-dependent ratio of TADF and PF. Sensor foils, made from poly(styrene- co -acrylonitrile) and the indicator dye, enable temperature-compensated trace oxygen sensing (0.002-6 hPa pO 2 ) at ambient conditions. Compared to the previously reported dual sensors based on two emitters, the new sensor significantly simplifies the experimental setup and eliminates risks of different leaching or photobleaching rates by utilizing only one indicator dye and operating at a single wavelength.

Thermally activated delayed fluorescence (TADF) is a highly temperature-dependent process and can be used in optical thermometry. TADF-based optical thermometers reported so far show fairly high-temperature sensitivity but have poor brightness and significant oxygen cross-talk. A new class of TADF emitters, Zn(II) Schiff base complexes, possess excellent brightness and high temperature sensitivity of the decay time at ambient temperature (4.1%/K change of TADF lifetime at 25 °C), enabling a resolution better than 0.03 °C. Oxygen cross-sensitivity is eliminated by covering the sensing layer (luminophore in polystyrene) with an off-stoichiometry thiol-ene polymer as an oxygen-consuming layer, and a poly(vinylidene chloride- co -acrylonitrile) layer as an oxygen barrier. The material is stable after more than 2 months of storage at ambient air, which enables long-term temperature monitoring., Competing Interests: The authors declare no competing financial interest., (Copyright © 2020 American Chemical Society.)

Emulsion templated norbornadiene is cured via ring-opening metathesis polymerization yielding macroporous poly(norbornadiene) foams of 76% porosity exhibiting appealing stiffness combined with considerable ductility. The foams are readily oxidized in the presence of air at room temperature exhibiting an oxygen uptake capacity of more than 300 mg O 2 g -1 foam. In closed volumes of air, a final oxygen level of a maximum of 0.0005 vol%, that is, 5 ppm (v) can be achieved after several hours at room temperature. The synergism of the porous morphology and the chemical nature of the polymer allows for the first example of an organic oxygen scavenger material with properties distinctly surpassing the state-of-the art in the field., (© 2020 The Authors. Published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Biomeshes based on decellularized bovine pericardium (DBP) are widely used in reconstructive surgery due to their wide availability and the attractive biomechanical properties. However, their efficacy in clinical applications is often affected by the uncontrolled immunogenicity and proteolytic degradation. To address this issue, we present here in vivo multiparametric imaging analysis of epoxy crosslinked DBPs to reveal their fate after implantation. We first analyzed the structure of the crosslinked DBP using scanning electron microscopy and evaluated proteolytic stability and cytotoxicity. Next, using combination of fluorescence and hypoxia imaging, X-ray computed microtomography and histology techniques we studied the fate of DBPs after subcutaneous implantation in animals. Our approach revealed high resistance to biodegradation, gradual remodeling of a surrounding tissue forming the connective tissue capsule and calcification of crosslinked DBPs. These changes were concomitant to the development of hypoxia in the samples within 3 weeks after implantation and subsequent induction of angiogenesis and vascularization. Collectively, presented approach provides new insights on the transplantation of the epoxy crosslinked biomeshes, the risks associated with its applications in soft-tissue reconstruction and can be transferred to studies of other types of implants., (Copyright © 2020 Elagin, Kuznetsova, Grebenik, Zolotov, Istranov, Zharikova, Istranova, Polozova, Reunov, Kurkov, Shekhter, Gafarova, Asadchikov, Borisov, Dmitriev, Zagaynova and Timashev.)

Presented herein is a first photophysical investigation of a series of rhenium(v)-oxo tris(p-X-phenyl)corroles, Re[TpXPC](O), where X = CF3, F, H, CH3, and OCH3. The complexes all exhibit near-IR phosphorescence in anoxic toluene at room temperature, with emission maxima at 780 ± 10 nm, phosphorescence decay times of 56-74 μs and quantum yields of 1.07-1.52%, the highest reported to date for phosphorescent corrole derivatives. We have also demonstrated that the dyes may serve as indicators in optical oxygen sensors and as sensitizers in triplet-triplet-annihilation-based upconversion. Although the ReO corroles are not superior to benchmark benzoporphyrin complexes that are currently used in these applications, they may prove useful in certain areas such as optical multianalyte sensing owing to improved flexibility in the available spectral properties of the dyes. The high thermal and photochemical stability of the complexes also bodes well for their deployment as new, phosphorescent sensitizers in photodynamic therapy.

Singlet oxygen ( 1 O 2 ) causes a major fraction of the parasitic chemistry during the cycling of non-aqueous alkali metal-O 2 batteries and also contributes to interfacial reactivity of transition-metal oxide intercalation compounds. We introduce DABCOnium, the mono alkylated form of 1,4-diazabicyclo[2.2.2]octane (DABCO), as an efficient 1 O 2 quencher with an unusually high oxidative stability of ca. 4.2 V vs. Li/Li + . Previous quenchers are strongly Lewis basic amines with too low oxidative stability. DABCOnium is an ionic liquid, non-volatile, highly soluble in the electrolyte, stable against superoxide and peroxide, and compatible with lithium metal. The electrochemical stability covers the required range for metal-O 2 batteries and greatly reduces 1 O 2 related parasitic chemistry as demonstrated for the Li-O 2 cell., (© 2019 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.)

A series of stable Pt(IV) corrole complexes with the general formula Pt IV [T p XPC]( m / p -C 6 H 4 CN)(py), where T p XPC 3- is the trianion of a tris( p -X-phenyl)corrole and X = CF 3 , H, and CH 3 , has been synthesized, affording key physicochemical data on a rare and elusive class of metallocorroles. Single-crystal X-ray structures of two of the complexes revealed very short equatorial Pt-N distances of 1.94-1.97 Å, an axial Pt-C distance of ∼2.03 Å, and an axial Pt-N distance of ∼2.22 Å. The complexes exhibit Soret maxima at ∼430 nm, which are essentially independent of the meso -aryl para substituents, and strong Q bands with the most intense peak at 595-599 nm. The substituent-independent Soret maxima are consistent with an innocent Pt IV -corrole 3- description for the complexes. The low reduction potentials (-1.45 ± 0.08 V vs saturated calomel reference electrode) also support a highly stable Pt(IV) ground state as opposed to a noninnocent corrole •2- description. The reductions, however, are irreversible, which suggests that they involve concomitant cleavage of the Pt-aryl bond. Unlike Pt(IV) porphyrins, two of the complexes, Pt IV [T p XPC]( m -C 6 H 4 CN)(py) (X = CF 3 and CH 3 ), were found to exhibit room-temperature near-IR phosphorescence with emission maxima at 813 and 826 nm, respectively. The quantum yield of ∼0.3% is comparable to those observed for six-coordinate Ir(III) corroles., Competing Interests: The authors declare no competing financial interest.

The imaging of real-time fluxes of K + ions in live cell with high dynamic range (5-150 mM) is of paramount importance for neuroscience and physiology of the gastrointestinal tract, kidney and other tissues. In particular, the research on high-performance deep-red fluorescent nanoparticle-based biosensors is highly anticipated. We found that BODIPY-based FI3 K + -sensitive fluoroionophore encapsulated in cationic polymer RL100 nanoparticles displays unusually strong efficiency in staining of broad spectrum of cell models, such as primary neurons and intestinal organoids. Using comparison of brightness, photostability and fluorescence lifetime imaging microscopy (FLIM) we confirmed that FI3 nanoparticles display distinctively superior intracellular staining compared to the free dye. We evaluated FI3 nanoparticles in real-time live cell imaging and found that it is highly useful for monitoring intra- and extracellular K + dynamics in cultured neurons. Proof-of-concept in vivo brain imaging confirmed applicability of the biosensor for visualization of epileptic seizures. Collectively, this data makes fluoroionophore FI3 a versatile cross-platform fluorescent biosensor, broadly compatible with diverse experimental models and that crown ether-based polymer nanoparticles can provide a new venue for design of efficient fluorescent probes., Competing Interests: Competing financial interests None declared.

Phosphors based on magnesium titanate activated with Mn 4 + ions are of great interest because, when excited with blue light, they display a strong red-emitting luminescence. They are characterized by a luminescence decay which is strongly temperature dependent in the range from -50 ∘ C to 150 ∘ C, making these materials very promising for temperature sensing in the biochemical field. In this work, the optical and thermal properties of the luminescence of Mg 2 TiO 4 are investigated for different Mn 4 + doping concentrations. The potential of this material for temperature sensing is demonstrated by fabricating a fiber optic temperature microsensor and by comparing its performance against a standard resistance thermometer. The response of the fiber optic sensor is exceptionally fast, with a response time below 1 s in the liquid phase and below 1.1 s in the gas phase., Competing Interests: The authors declare no conflict of interest.

Aprotic sodium-O 2 batteries require the reversible formation/dissolution of sodium superoxide (NaO 2 ) on cycling. Poor cycle life has been associated with parasitic chemistry caused by the reactivity of electrolyte and electrode with NaO 2 , a strong nucleophile and base. Its reactivity can, however, not consistently explain the side reactions and irreversibility. Herein we show that singlet oxygen ( 1 O 2 ) forms at all stages of cycling and that it is a main driver for parasitic chemistry. It was detected in- and ex-situ via a 1 O 2 trap that selectively and rapidly forms a stable adduct with 1 O 2 . The 1 O 2 formation mechanism involves proton-mediated superoxide disproportionation on discharge, rest, and charge below ca. 3.3 V, and direct electrochemical 1 O 2 evolution above ca. 3.3 V. Trace water, which is needed for high capacities also drives parasitic chemistry. Controlling the highly reactive singlet oxygen is thus crucial for achieving highly reversible cell operation., (© 2017 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.)

High brightness, chemical and photostability, tunable characteristics, and spectral and surface properties are important attributes for nanoparticle probes designed for live cell imaging. We describe a class of nanoparticles for high-resolution imaging of O2 that consists of a substituted conjugated polymer (polyfluorene or poly(fluorene-alt-benzothiadiazole)) acting as a FRET antenna and a fluorescent reference with covalently bound phosphorescent metalloporphyrin (PtTFPP, PtTPTBPF). The nanoparticles prepared from such copolymers by precipitation method display stability, enhanced (>5-10 times) brightness under one- and two-photon excitation, compatibility with ratiometric and lifetime-based imaging modes, and low toxicity for cells. Their cell-staining properties can be modulated with positively and negatively charged groups grafted to the backbone. The "zwitter-ionic" nanoparticles show high cell-staining efficiency, while their cell entry mechanisms differ for the different 3D models. When injected in the bloodstream, the cationic and anionic nanoparticles show similar distribution in vivo. These features and tunable properties make the conjugated polymer based phosphorescent nanoparticles a versatile tool for quantitative O2 imaging with a broad range of cell and 3D tissue models.

Optical oxygen sensing is of broad interest in many areas of research, such as medicine, food processing, and micro- and marine biology. The operation principle of optical oxygen sensors is well established and these sensors are routinely employed in lab and field experiments. Ultratrace oxygen sensors, which enable measurements in the sub-nanomolar region (dissolved oxygen), are becoming increasingly important. Such sensors prominently exhibit phenomena that complicate calibration and measurements. However, these phenomena are not constrained to ultratrace sensors; rather, these effects are inherent to the way optical oxygen sensors work and may influence any optical oxygen measurement when certain conditions are met. This scenario is especially true for applications that deal with high-excitation light intensities, such as microscopy and microfluidic applications. Herein, we present various effects that we could observe in our studies with ultratrace oxygen sensors and discuss the reasons for their appearance, the mechanism by which they influence measurements, and how to best reduce their impact. The phenomena discussed are oxygen photoconsumption in the sensor material; depletion of the dye ground state by high-excitation photon-flux values, which can compromise both intensity and ratiometric-based measurements; triplet-triplet annihilation; and singlet-oxygen accumulation, which affects measurements at very low oxygen concentrations., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A new measurement system enables combination of continuous glucose monitoring (CGM) and insulin infusion. A sensor system comprising an optical glucose biosensor and an optical oxygen sensor is integrated into the insulin infusion catheter of an insulin pump. Both sensors rely on near infrared (NIR) phosphorescent porphyrin dyes, wherefore the signals can be read out transcutaneous and non-invasively with a custom-built phase fluorometer measurement module. The spectral properties of the indicator dyes and the optical setup of the measurement module were optimized to enable independent read-out in two channels. Dynamic ranges from 0 mmHg to 160 mmHg oxygen and 0mg/dL to 360 mg/dL glucose (LOD 2mg/dL) are covered by the oxygen and the glucose sensor, respectively. In-vivo measurements in pigs demonstrate good correlation of reference blood glucose levels and glucose values obtained with the presented sensor system. The evaluation of the clinical accuracy of the system with Clarke Error Grid Analysis showed similar results to CGM-devices currently on the market., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Cell-permeable phosphorescent probes enable the study of cell and tissue oxygenation, bioenergetics, metabolism, and pathological states such as stroke and hypoxia. A number of such probes have been described in recent years, the majority consisting of cationic small molecule and nanoparticle structures. While these probes continue to advance, adequate staining for the study of certain cell types using live imaging techniques remains elusive; this is particularly true for neural cells. Here we introduce novel probes for the analysis of neural cells and tissues: negatively charged poly(methyl methacrylate-co-methacrylic acid)-based nanoparticles impregnated with a phosphorescent Pt(II)-tetrakis(pentafluorophenyl)porphyrin (PtPFPP) dye (this form is referred to as PA1), and with an additional reference/antennae dye poly(9,9-diheptylfluorene-alt-9,9-di-p-tolyl-9H-fluorene) (this form is referred to as PA2). PA1 and PA2 are internalised by endocytosis, result in efficient staining in primary neurons, astrocytes, and PC12 cells and multi-cellular aggregates, and allow for the monitoring of local O(2) levels on a time-resolved fluorescence plate reader and PLIM microscope. PA2 also efficiently stains rat brain slices and permits detailed O(2) imaging experiments using both one and two-photon intensity-based modes and PLIM modes. Multiplexed analysis of embryonic rat brain slices reveals age-dependent staining patterns for PA2 and a highly heterogeneous distribution of O(2) in tissues, which we relate to the localisation of specific progenitor cell populations. Overall, these anionic probes are useful for sensing O(2) levels in various cells and tissues, particularly in neural cells, and facilitate high-resolution imaging of O(2) in 3D tissue models.

New optical pH-sensors relying on 1,4-diketopyrrolo-[3,4- c ]pyrroles (DPPs) as fluorescent pH-indicators are presented. Different polymer hydrogels are useful as immobilization matrices, achieving excellent sensitivity and good brightness in the resulting sensor. The operational pH can be tuned over a wide range (pH 5-12) by selecting the fine structure of the indicator and the matrix. A ratiometric sensor in the form of nanoparticles is also presented. It is suitable for RGB camera readout, and its practical applicability for fluorescence imaging in microfluidic systems is demonstrated. The indicators are synthesized starting from the commercially available DPP pigments by a straightforward concept employing chlorosulfonation and subsequent reaction with amines. Their sensitivity derives from two distinct mechanisms. At high pH (>9), they exhibit a remarkable alteration of both absorption and fluorescence spectra due to deprotonation of the lactam nitrogen atoms. If a phenolic group is introduced, highly effective fluorescence quenching at near-neutral pH occurs due to photoinduced electron transfer (PET) involving the phenolate form.

A new intracellular O(2) (icO(2)) sensing probe is presented, which comprises a nanoparticle (NP) formulation of a cationic polymer Eudragit RL-100 and a hydrophobic phosphorescent dye Pt(II)-tetrakis(pentafluorophenyl)porphyrin (PtPFPP). Using the time-resolved fluorescence (TR-F) plate reader set-up, cell loading was investigated in detail, particularly the effects of probe concentration, loading time, serum content in the medium, cell type, density, etc. The use of a fluorescent analogue of the probe in conjunction with confocal microscopy and flow cytometry analysis, revealed that cellular uptake of the NPs is driven by nonspecific energy-dependent endocytosis and that the probe localizes inside the cell close to the nucleus. Probe calibration in biological environment was performed, which allowed conversion of measured phosphorescence lifetime signals into icO(2) concentration (μM). Its analytical performance in icO(2) sensing experiments was demonstrated by monitoring metabolic responses of mouse embryonic fibroblast cells under ambient and hypoxic macroenvironment. The NP probe was seen to generate stable and reproducible signals in different types of mammalian cells and robust responses to their metabolic stimulation, thus allowing accurate quantitative analysis. High brightness and photostability allow its use in screening experiments with cell populations on a commercial TR-F reader, and for single cell analysis on a fluorescent microscope.

A sensitive and broad-ranged optical thermometer, based on the thermally activated delayed fluorescence of fullerene C70, is presented. It consists of C70 molecularly dispersed in a polymer film. Several polymer matrices were investigated. In the absence of oxygen the fluorescence intensity increases markedly with temperature. At 25 degrees C the fluorescence intensity of C70 increases maximally by a factor of between 17 and 22, depending on the polymer, whereas at 100 degrees C the fluorescence intensity can be 79 times higher. In the absence of oxygen and for temperatures above 20 degrees C, the red fluorescence of C70 in the films is so intense that it is easily perceived by the naked eye. For the systems studied, the fluorescence intensity is very sensitive to temperature. This results in a working range from -80 to at least 140 degrees C in the case of C70 in poly(tert-butyl methacrylate) (PtBMA). Perylene was incorporated into the film as an internal reference in order to enable ratiometric measurements. The sensitivity of the lifetime of the delayed fluorescence to temperature is also high and results in an even wider working range. The performance of the C70/PtBMA film was measured against a well-known optical temperature probe, [Ru(phen)3] (phen=phenanthroline). The results show that the C70/PtBMA film is a very good system for optical temperature-sensing over a wide range of temperatures, outperforming known standards.