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Chapartegui-Arias, Ander ; Raysyan, A. ; Belenguer, A. M. ; Jaeger, Carsten ; Tchipilov, Teodor ; Prinz, Carsten ; Abad Andrade, Carlos Enrique ; Beyer, S. ; Schneider, Rudolf ; Emmerling, Franziska
Zeolitic imidazolate framework (ZIF) hybrid fluorescent nanoparticles and ZIF antibody conjugates have been synthesized, characterized, and employed in lateral-flow immunoassay (LFIA). The bright fluorescence of the conjugates and the possibility to tailor their mobility gives a huge potential for diagnostic assays. An enzyme-linked immunosorbent assay (ELISA) with horseradish peroxidase (HRP) as label, proved the integrity, stability, and dispersibility of the antibody conjugates, LC-MS/MS provided evidence that a covalent link was established between these metal-organic frameworks and lysine residues in IgG antibodies.
Riedel, Juliane ; Recknagel, Sebastian ; Sassenroth, Diana ; Mauch, Tatjana ; Buttler, Sabine ; Sommerfeld, Thomas ; Penk, Sibylle ; Koch, Matthias
Zearalenone (ZEN), an estrogenic mycotoxin produced by several species of Fusarium fungi, is a common contaminant of cereal-based food worldwide. Due to frequent occurrences associated with high levels of ZEN, maize oil is a particular source of exposure. Although a European maximumlevel for ZEN in maize oil exists according to Commission Regulation (EC) No. 1126/2007 along with a newly developed international standard method for analysis, certified reference materials (CRM) are still not available. To overcome this lack, the first CRM for the determination of ZEN in contaminated maize germ oil (ERM®-BC715) was developed in the frame of a European Reference Materials (ERM®) project according to the requirements of ISO Guide 35. The whole process of CRM development including preparation, homogeneity and stability studies, and value assignment is presented. The assignment of the certified mass fraction was based upon an in-house study using high-performance liquid chromatography isotope dilution tandem mass spectrometry. Simultaneously, to support the in-house certification study, an interlaboratory comparison study was conducted with 13 expert laboratories using different analytical methods. The certified mass fraction and expanded uncertainty (k=2) of ERM®-BC715 (362± 22) μg kg−1 ZEN are traceable to the SI. This reference material is intended for analytical quality control and contributes to the improvement of consumer protection and food safety.
Bornemann-Pfeiffer, Martin ; Wolf, Jakob ; Meyer, Klas ; Kern, S. ; Angelone, D. ; Leonov, A. ; Cronin, L. ; Emmerling, Franziska
A big problem with the chemistry literature is that it is not standardized with respect to precise operational parameters, and real time corrections are hard to make without expert knowledge. This lack of context means difficult reproducibility because many steps are ambiguous, and hence depend on tacit knowledge. Here we present the integration of online NMR into an automated chemical synthesis machine (CSM aka. “Chemputer” which is capable of small-molecule synthesis using a universal programming language) to allow automated analysis and adjustment of reactions on the fly. The system was validated and benchmarked by using Grignard reactions which were chosen due to their importance in synthesis. The system was monitored in real time using online-NMR, and spectra were measured continuously during the reactions. This shows that the synthesis being done in the Chemputer can be dynamically controlled in response to feedback optimizing the reaction conditions according to the user requirements.
Witte, F. ; Rietsch, P. ; Nirmalananthan-Budau, Nithiya ; Weigert, Florian ; Götze, J. P. ; Resch-Genger, Ute ; Eigler, S. ; Paulus, B.
The concept of aggregation-induced emission represents a means to rationalise photoluminescence of usually nonfluorescent excimers in solid-state materials. In this publication, we study the photophysical properties of selected diaminodicyanoquinone (DADQ) derivatives in the solid state using a combined approach of experiment and theory. DADQs are a class of high-dipole organic chromophores promising for applications in non-linear optics and light-harvesting devices. Among the compounds investigated, we find both aggregation-induced emission and aggregation-caused quenching effects rationalised by calculated energy transfer rates. Analysis of fluorescence spectra and lifetime measurements provide the interesting result that (at least) two emissive species seem to contribute to the photophysical properties of DADQs. The main emission peak is notably broadened in the long-wavelength limit and exhibits a blue-shifted shoulder. We employ high-level quantum-chemical methods to validate a molecular approach to a solid-state problem and show that the complex emission features of DADQs can be attributed to a combination of H-type aggregates, monomers, and crystal structure defects.
Biesen, L. ; May, L. ; Nirmalananthan-Budau, Nithiya ; Hoffmann, Katrin ; Resch-Genger, Ute ; Müller, T. J. J.
Aroyl-S,N-ketene acetal-based bichromophores can be readily synthesized in a consecutive three-component synthesis in good to excellent yields by condensation of aroyl chlorides and an N-(p-bromobenzyl) 2-methyl benzothiazolium salt followed by a Suzuki coupling, yielding a library of 31 bichromophoric fluorophores with substitution patterntunable emission properties. Varying both chromophores enables different communication pathways between the chromophores, exploiting aggregation-induced emission (AIE) and energy transfer (ET) properties, and thus, furnishing aggregation-based fluorescence switches. Possible applications range from fluorometric analysis of alcoholic beverages to pH sensors.
Nelson, G. ; Boehm, U. ; Bagley, S. ; Bajcsy, P. ; Bischof, J. ; Brown, C. M. ; Dauphin, A. ; Dobbie, I. M. ; Eriksson, J. E. ; Faklaris, O. ; Fernandez-Rodriguez, J. ; Ferrand, A. ; Gelman, L ; Gheisari, A. ; Hartmann, H. ; Kukat, C. ; Laude, A. ; Mitkovski, M. ; Munck, S. ; North, A. J. ; Rasse, T. ; Resch-Genger, Ute ; Schuetz, L. C. ; Seitz, A. ; Strambio-De-Castillia, C. ; Swedlow, J. R. ; Alexopoulos, I. ; Aumayr, K. ; Avilov, S. ; Bakker, G.-J. ; Bammann, R. R. ; Bassi, A. ; Beckert, H. ; Beer, S. ; Belyaev, Y. ; Bierwagen, J. ; Birngruber, K. A. ; Bosch, M. ; Breitlow, J. ; Cameron, L. A. ; Chalfoun, J. ; Chambers, J. J. ; Chen, C.-L. ; Conde-Sousa, E. ; Corbett, A. D. ; Cordelieres, F. P. ; Del Nery, E. ; Dietzel, R. ; Eismann, F. ; Fazeli, E. ; Felscher, A. ; Fried, H. ; Gaudreault, N. ; Goh, W. I. ; Guilbert, T. ; Hadleigh, R. ; Hemmerich, P. ; Holst, G. A. ; Itano, M. S. ; Jaffe, C. B. ; Jambor, H. K. ; Jarvis, S. C. ; Keppler, A. ; Kirchenbuechler, D. ; Kirchner, M. ; Kobayashi, N. ; Krens, G. ; Kunis, S. ; Lacoste, J. ; Marcello, M. ; Martins, G. G. ; Metcalf, D. J. ; Mitchell, C. A. ; Moore, J. ; Mueller, T. ; Nelson, M. S. ; Ogg, S. ; Onami, S. ; Palmer, A. L. ; Paul-Gilloteaux, P. ; Pimentel, J. A. ; Plantard, L. ; Podder, S. ; Rexhepaj, E. ; Royon, A. ; Saari, M. A. ; Schapman, D. ; Schoonderwoert, V. ; Schroth-Diez, B. ; Schwartz, S. ; Shaw, M. ; Spitaler, M. ; Stoeckl, M. T. ; Sudar, D. ; Teillon, J. ; Terjung, S. ; Thuenauer, R. ; Wilms, C. D. ; Wright, G. D. ; Nitschke, R.
A modern day light microscope has evolved from a tool devoted to making primarily empirical observations to what is now a sophisticated, quantitative device that is an integral part of both physical and life science research. Nowadays, microscopes are found in nearly every experimental laboratory. However, despite their prevalent use in capturing and quantifying scientific phenomena, neither a thorough understanding of the principles underlying quantitative imaging techniques nor appropriate knowledge of how to calibrate, operate and maintain microscopes can be taken for granted. This is clearly demonstrated by the well-documented and widespread difficulties that are routinely encountered in evaluating acquired data and reproducing scientific experiments. Indeed, studies have shown that more than 70% of researchers have tried and failed to repeat another scientist’s experiments, while more than half have even failed to reproduce their own experiments1. One factor behind the reproducibility crisis of experiments published in scientific journals is the frequent underreporting of imaging methods caused by a lack of awareness and/or a lack of knowledge of the applied technique2,3. Whereas quality control procedures for some methods used in biomedical research, such as genomics (e.g., DNA sequencing, RNA-seq) or cytometry, have been introduced (e.g. ENCODE4), this issue has not been tackled for optical microscopy instrumentation and images. Although many calibration standards and protocols have been published, there is a lack of awareness and agreement on common Standards and guidelines for quality assessment and reproducibility5.
In April 2020, the QUality Assessment and REProducibility for instruments and images in Light Microscopy (QUAREP-LiMi) initiative6 was formed. This initiative comprises imaging scientists from academia and industry who share a common interest in achieving a better understanding of the performance and limitations of microscopes and improved quality control (QC) in light microscopy. The ultimate goal of the QUAREP-LiMi initiative is to establish a set of common QC standards, guidelines, metadata models7,8, and tools9,10, including detailed protocols, with the ultimate aim of improving reproducible advances in scientific research.
This White Paper 1) summarizes the major obstacles identified in the field that motivated the launch of the QUAREP-LiMi initiative; 2) identifies the urgent need to address these obstacles in a grassroots manner, through a community of Stakeholders including, researchers, imaging scientists11, bioimage analysts, bioimage informatics developers, corporate partners, Funding agencies, standards organizations, scientific publishers, and observers of such; 3) outlines the current actions of the QUAREPLiMi initiative, and 4) proposes future steps that can be taken to improve the dissemination and acceptance of the proposed guidelines to manage QC.
To summarize, the principal goal of the QUAREP-LiMi initiative is to improve the overall quality and reproducibility of light microscope image data by introducing broadly accepted standard practices and accurately captured image data metrics.
We present the synthesis and characterization of a family of regioisomerically pure pH-sensitive rosamine fluorophores consisting of xanthene fluorophore cores, which determine the dyes’ photophysical properties such as excitation/emission wavelength, fluorescence quantum yield, and fluorescence lifetime, and differently substituted phenol moieties. The hydroxyl substituent of the phenol moiety introduces a pH sensitivity of the dyes’ fluorescence exploiting a photoinduced electron transfer (PET), that leads to a protonation-induced switching ON of the rosamine emission. Rational tuning of the pKa value of the rosamine fluorescence between 4 to 9 is achieved by altering the substitution pattern and degree of bromination of the phenolic subunits. Additionally, a temperature sensitivity of the fluorescence quantum yield is introduced or suppressed based upon the degree of rigidity of the xanthene scaffold.
We present the rational design, synthesis and spectroscopic characterization of a novel dual excitation, three color emitting, pH-responsive fluorescent probe consisting of two phenanthrene and one rhodamine B units linked by click chemistry. The rhodamine moiety, excitable at λEx = 315 nm and at λEx = 560 nm in its ring-opened form, provides the pH-responsive fluorophore, while the pH-insensitive phenanthrene, excited at λEx = 315 nm, serves as inert internal reference, The presence of two phenanthrene moieties enables a blue monomer and a blueish green excimer emission at 351 nm and 500 nm, respectively. Opening of the rhodamine B spirolactam ring at an acidic pH below 5.0 (pKa = 2.59 ± 0.04) switches on its emission at 580 nm. Simultaneously, the phenanthrene excimer emission decreases caused by a change in orientation of the phenanthrene units, while the monomer emission is barely affected. This sensor design enables ratiometric measurements in the low acidic pH range utilizing the intensity ratios of the rhodamine B and phenanthrene excimer emission at 580 nm and 500 nm. Alternatively, also the intensity ratios of the rhodamine B and the phenanthrene monomer emission could be exploited or the sum of the phenanthrene monomer and excimer fluorescence. To the best of our knowledge, this is the first report of ratiometric sensing utilizing such a versatile type of tricolor emissive dyad probe bearing phenanthrene moieties and showing phenanthrene monomer and excimer emission.
Reichenauer, F. ; Wang, Cui ; Förster, C. ; Boden, P. ; Ugur, N. ; Báez-Cruz, R. ; Kalmbach, J. ; Carrella, L. M. ; Rentschler, E. ; Ramanan, C. ; Niedner-Schatteburg, G. ; Gerhards, M. ; Seitz, M. ; Resch-Genger, Ute ; Heinze, K.
Gaining chemical control over the thermodynamics and kinetics of photoexcited states is paramount to an efficient and sustainable utilization of photoactive transition metal complexes in a plethora of technologies. In contrast to energies of charge Transfer states described by spatially separated orbitals, the energies of spinflip states cannot straightforwardly be predicted as Pauli Repulsion and the nephelauxetic effect play key roles. Guided by multireference quantum chemical calculations, we report a novel highly luminescent spin-flip emitter with a quantum chemically predicted blue-shifted luminescence. The spin-flip emission band of the chromium complex [Cr(bpmp)2]3+ (bpmp = 2,6-bis(2-pyridylmethyl) pyridine) shifted to higher energy from ca. 780 nm observed for known highly emissive chromium(III) complexes to 709 nm. The photoluminescence quantum yields climb to 20%, and very long excited state lifetimes in the millisecond range are achieved at room temperature in acidic D2O solution. Partial ligand deuteration increases the quantum yield to 25%. The high excited state energy of [Cr(bpmp)2]3+ and its facile reduction to [Cr(bpmp)2]2+ result in a high excited state redox potential. The ligand’s methylene bridge acts as a Brønsted acid quenching the luminescence at high pH. Combined with a pH-insensitive chromium(III) emitter, ratiometric optical pH sensing is achieved with single wavelength excitation. The photophysical and Ground state properties (quantum yield, lifetime, redox potential, and acid/base) of this spin-flip complex incorporating an earth-abundant metal surpass those of the classical precious metal [Ru(α-diimine)3]2+ charge transfer complexes, which are commonly employed in optical sensing and photo(redox) catalysis, underlining the bright future of these molecular ruby analogues.
Riedel, Soraya ; Hülagü, Deniz ; Bennet, Francesca ; Carl, Peter ; Flemig, Sabine ; Schmid, Thomas ; Schenk, J. A. ; Hodoroaba, Vasile-Dan ; Schneider, Rudolf
Electrochemical methods offer great promise in meeting the demand for user-friendly on-site devices for Monitoring important parameters. The food industry often runs own lab procedures, for example, for mycotoxin analysis, but it is a major goal to simplify analysis, linking analytical methods with smart technologies. Enzyme-linked immunosorbent assays, with photometric detection of 3,3’,5,5’-tetramethylbenzidine (TMB),form a good basis for sensitive detection. To provide a straightforward approach for the miniaturization of the detectionstep, we have studied the pitfalls of the electrochemical TMB detection. By cyclic voltammetry it was found that the TMB electrochemistry is strongly dependent on the pH and the electrode material. A stable electrode response to TMB could be achieved at pH 1 on gold electrodes. We created a smartphonebased, electrochemical, immunomagnetic assay for the detection of ochratoxin A in real samples, providing a solid basis forsensing of further analytes.