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Objective The main purpose of the present study was to assess the risk for major revision surgery after perichondrium transplantation (PT) at a minimum of 22 years postoperatively and to evaluate the influence of patient characteristics. Design Primary outcome was treatment success or failure. Failure of PT was defined as revision surgery in which the transplant was removed, such as (unicondylar) knee arthroplasty or patellectomy. The functioning of nonfailed patients was evaluated using the International Knee Documentation Committee (IKDC) score. In addition, the influence of patient characteristics was evaluated. Results Ninety knees in 88 patients, aged 16 to 55 years with symptomatic cartilage defects, were treated by PT. Eighty knees in 78 patients were eligible for analysis and 10 patients were lost to follow-up. Twenty-eight knees in 26 patients had undergone major revision surgery. Previous surgery and a longer time of symptoms prior to PT were significantly associated with an increased risk for failure of cartilage repair. Functioning of the remaining 52 patients and influence of patient characteristics was analyzed using their IKDC score. Their median IKDC score was 39.08, but a relatively young age at transplantation was associated with a higher IKDC score. Conclusions This 22-year follow-up study of PT, with objective outcome parameters next to patient-reported outcome measurements in a unique group of patients, shows that overall 66% was without major revision surgery and patient characteristics also influence long-term outcome of cartilage repair surgery.

Four years after its first report, expansion microscopy (ExM) is now being routinely applied in laboratories worldwide to achieve super-resolution imaging on conventional fluorescence microscopes. By chemically anchoring all molecules of interest to the polymer meshwork of an expandable hydrogel, their physical distance is increased by a factor of ∼4-5× upon dialysis in water, resulting in an imprint of the original sample with a lateral resolution up to 50-70 nm. To ensure a correct representation of the original spatial distribution of the molecules, it is crucial to confirm that the expansion is isotropic, preferentially in all three dimensions. To address this, we present an approach to evaluate the local expansion factor within a biological sample and in all three dimensions. We use photobleaching to introduce well-defined three-dimensional (3D) features in the cell and, by comparing the size and shape pre- and postexpansion, these features can be used as an intrinsic ruler. In addition, our method is capable of pointing out sample distortions and can be used as a quality control tool for expansion microscopy experiments in biological samples. ispartof: ACS OMEGA vol:5 issue:12 pages:6792-6799 ispartof: location:United States status: published

Methyltransferasen (MTasen) bilden eine grose Familie von Enzymen, die vielfaltige Zielstrukturen methylieren, von DNA, RNA und Proteinen bis zu niedermolekularen Verbindungen. Die meisten MTasen nutzen S-Adenosyl-l-methionin (AdoMet) als Methylquelle. In den letzten Jahren wurde intensiv an der Entwicklung von AdoMet-Analoga gearbeitet, mit dem Ziel, andere Einheiten als einfache Methylgruppen zu ubertragen. Derzeit gibt es zwei Hauptklassen von AdoMet-Analoga – doppelt aktivierte Molekule und Molekule auf Aziridinbasis – die jeweils einen anderen Ansatz zur Transalkylierung des Zielmolekuls verfolgen. Dieser Aufsatz erortert die Methoden zur Markierung und Funktionalisierung von Biomolekulen unter Verwendung von AdoMet-abhangigen MTasen und AdoMet-Analoga. Er behandelt die Synthesewege hin zu AdoMet-Analoga, die Stabilitat von AdoMet-Analoga in biologischen Umgebungen und ihre Anwendung in Transalkylierungen. Schlieslich werden einige Perspektiven fur die Anwendung von AdoMet-Analoga in der biologischen Forschung, Genetik oder Epigenetik und Nanotechnologie aufgezeigt.

The development of novel molecular sieves opens opportunities in the development of more sensitive analytical devices. In this paper, metal organic frameworks (MOFs), specifically ZIF-8 and ZIF-93, are grown on fiber optic based surface plasmon resonance (FO-SPR) sensors. FO-SPR has enabled sensitive sensing capabilities in biomedical settings and the addition of an MOF coating opens the way for the sensing of volatile organic compounds (VOCs) in gaseous media. FO-SPR probes were homogeneously functionalized with ZIF-8 and ZIF-93 in each case using two different precursor solutions to obtain a sequential nucleation and growth phase. The difference in MOF nucleation and growth kinetics of the two solutions was directly monitored by the FO-SPR system. The two established MOF-FO-SPR sensors were then subjected to sensing experiments with several alcohol vapors to establish their sensing capabilities. Vapors with mPa partial pressures, ppm concentrations, could successfully be detected, e.g., an LOD of 2.5 ppm for methanol detection was acquired. The difference in recognition behavior of the hydrophobic ZIF-8 and more hydrophilic ZIF-93 recognition layers can be exploited to yield qualitative information regarding the vapor composition.

The generation of cartilage from progenitor cells for the purpose of cartilage repair is often hampered by hypertrophic differentiation of the engineered cartilaginous tissue caused by endochondral ossification. Since a healthy cartilage matrix contains high amounts of Aggrecan and COMP, we hypothesized that their supplementation in the biogel used in the generation of subperiosteal cartilage mimics the composition of the cartilage extracellular matrix environment, with beneficial properties for the engineered cartilage. Supplementation of COMP or Aggrecan was studied in vitro during chondrogenic differentiation of rabbit periosteum cells and periosteum-derived chondrocytes. Low melting agarose was supplemented with bovine Aggrecan, human recombinant COMP or vehicle and was injected between the bone and periosteum at the upper medial side of the tibia of New Zealand white rabbits. Generated subperiosteal cartilage tissue was analyzed for weight, GAG and DNA content and ALP activity. Key markers of different phases of endochondral ossification were measured by RT-qPCR. For the in vitro experiments, no significant differences in chondrogenic marker expression were detected following COMP or Aggrecan supplementation, while in vivo favorable chondrogenic marker expression was detected. Gene expression levels of hypertrophic markers as well as ALP activity were significantly decreased in the Aggrecan and COMP supplemented conditions compared to controls. The wet weight and GAG content of the in vivo generated subperiosteal cartilage tissue was not significantly different between groups. Data demonstrate the potential of Aggrecan and COMP to favorably influence the subperiosteal microenvironment for the in vivo generation of cartilage for the optimization of cartilage regenerative approaches.

Single-molecule DNA mapping has the potential to serve as a powerful complement to high-throughput sequencing in metagenomic analysis. Offering longer read lengths and forgoing the need for complex library preparation and amplification, mapping stands to provide an unbiased view into the composition of complex viromes and/or microbiomes. To fully enable mapping-based metagenomics, sensitivity and specificity of DNA map analysis and identification need to be improved. Using detailed simulations and experimental data, we first demonstrate how fluorescence imaging of surface stretched, sequence specifically labeled DNA fragments can yield highly sensitive identification of targets. Second, a new analysis technique is introduced to increase specificity of the analysis, allowing even closely related species to be resolved. Third, we show how an increase in resolution improves sensitivity. Finally, we demonstrate that these methods are capable of identifying species with long genomes such as bacteria with high sensitivity. ispartof: NAR GENOMICS AND BIOINFORMATICS vol:2 issue:1 ispartof: location:England status: published

Strain-stabilized perovskites The perovskite materials used for solar cells and light-emitting diodes (which are black in color) are generally less stable at room temperature than the electronically inactive nonperovskite phases (which are yellow in color). Steele et al. show that for CsPbI 3 , strain induced in a thin film after annealing the material to 330°C and then rapidly cooling it to room temperature kinetically trapped the black phase. Grazing-incidence wide-angle x-ray scattering revealed the crystal distortions and texture formation created by interfacial strain. Science , this issue p. 679

The high-temperature, all-inorganic CsPbI

A highly reproducible and controllable deposition procedure for gold nanostructures on a titanium dioxide (TiO2) surface using femtosecond laser light has been demonstrated. This is realized by precisely focusing onto the TiO2 surface in the presence of a pure gold ion solution. The deposition is demonstrated both in dot arrays and line structures. Thanks to the multi-photon excitation, we observe that the deposition area of the nanostructures can be confined to a degree far greater than the diffraction limited focal spot. Finally, we demonstrate that catalytic activity with visible light irradiation is enhanced, proving the applicability of our new deposition technique to the catalytic field. ispartof: Nanoscale vol:9 issue:35 pages:13025-13033 ispartof: location:England status: published

Photocatalytic reactions occur at the crystal-solution interface, and hence specific crystal facet expression and surface defects can play an important role. Here we investigate the structure-related photoreduction at zinc oxide (ZnO) microparticles via integrated light and electron microscopy in combination with silver metal photodeposition. This enables a direct visualization of the photoreduction activity at specific crystallographic features. It is found that silver nanoparticle photodeposition on dumbbell-shaped crystals mainly takes place at the edges of O-terminated (0001̅) polar facets. In contrast, on ZnO microrods photodeposition is more homogeneously distributed with an increased activity at {101̅1̅} facets. Additional time-resolved measurements reveal a direct spatial link between the enhanced photoactivity and increased charge carrier lifetimes. These findings contradict previous observations based on indirect, bulk-scale experiments, assigning the highest photocatalytic activity to polar facets. The presented research demonstrates the need for advanced microscopy techniques to directly probe the location of photocatalytic activity. ispartof: JOURNAL OF PHYSICAL CHEMISTRY LETTERS vol:8 issue:2 pages:340-346 ispartof: location:United States status: published

© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim An integrated optical-electron microscope is presented that enables the in situ study of dynamic processes on photoactive materials. Here, the deposition of metallic silver nanostructures at ZnO photocatalyst particles is monitored in real time under ambient conditions by means of scanning electron microscopy. Zinc oxide crystals are immobilized on an electron transparent silicon nitride window. By passing UV light through an opposing optically transparent window, the zinc oxide is illuminated resulting in the photocatalytic formation of silver nanostructures. Both windows are part of a specially designed liquid cell filled with a dilute aqueous silver nitrate solution. Using the presented system, different electron detectors are evaluated for their ability to provide detailed images despite the interference caused by the liquid surrounding the sample. Special care has to be taken since direct silver reduction from solution induced by the electron beam interferes with the photocatalytic process. Oxygen gas, produced during the photocatalytic reaction, is also shown to complicate the imaging of the dynamic nanoscale processes in the scanning electron microscope. Nevertheless, the integrated approach allows to directly establish structure–activity relationships and to unravel optically induced processes at nanostructured materials. ispartof: PARTICLE & PARTICLE SYSTEMS CHARACTERIZATION vol:33 issue:7 pages:412-418 status: published

Optical microscopy has been a tool of choice ever since van Leeuwenhoek used Hooke's microscope to observe biological specimens. Chief among its advantages is the fact that imaging is noninvasive. In combination with the straightforward sample preparation and general convenience, optical microscopes remain essential to many aspects of modern-day research.

The impressive optoelectronic performance and low production cost of metal halide perovskites have inspired applications well beyond efficient solar cells. Herein, we widen the materials engineering options available for the efficient and selective photocatalytic oxidation of benzylic alcohols, an industrially significant reaction, using formamidinium lead bromide (FAPbBr(3)) and other perovskite-based materials. The best performance was obtained using a FAPbBr(3)/TiO2 hybrid photo catalyst under simulated solar illumination. Detailed optical studies reveal the synergetic photophysical pathways arising in FAPbBr(3)/TiO2 composites. An experimentally supported model rationalizing the large conversion enhancement over the pure constituents shows that this strategy offers new prospects for metal halide perovskites in photocatalytic applications. Research Foundation-Flanders (FWO) [G.0962.13, G0B39.15, ZW15_09 GOH6316N, G.0B49.15]; KU Leuven Research Fund [C14/15/053]; Flemish government through long term structural funding Methusalem (CASAS2) [Meth/15/04]; Hercules foundation [HER/11/14]; European Union (Horizon) Marie Sklodowska-Curie innovation program [722591, 641887]; NSFC [21773031]

Organic-inorganic halide perovskites (OIHPs) have demonstrated outstanding energy conversion efficiency in solar cells and light-emitting devices. In spite of intensive developments in both materials and devices, electronic traps and defects that significantly affect their device properties remain under-investigated. Particularly, it remains challenging to identify and to resolve traps individually at the nanoscopic scale. Here, photo-active traps (PATs) are mapped over OIHP nanocrystal morphology of different crystallinity by means of correlative optical differential super-resolution localization microscopy (Delta-SRLM) and electron microscopy. Stochastic and monolithic photoluminescence intermittency due to individual PATs is observed on monocrystalline and polycrystalline OIHP nanocrystals. Delta-SRLM reveals a heterogeneous PAT distribution across nanocrystals and determines the PAT density to be 1.3 x 10(14) and 8 x 10(13) cm(-3) for polycrystalline and for monocrystalline nanocrystals, respectively. The higher PAT density in polycrystalline nanocrystals is likely related to an increased defect density. Moreover, monocrystalline nanocrystals that are prepared in an oxygen and moisture-free environment show a similar PAT density as that prepared at ambient conditions, excluding oxygen or moisture as chief causes of PATs. Hence, it is conduded that the PATs come from inherent structural defects in the material, which suggests that the PAT density can be reduced by improving crystalline quality of the material.

Developments in microscopy have been instrumental to progress in the life sciences, and many new techniques have been introduced and led to new discoveries throughout the last century. A wide and diverse range of methodologies is now available, including electron microscopy, atomic force microscopy, magnetic resonance imaging, small-angle x-ray scattering and multiple super-resolution fluorescence techniques, and each of these methods provides valuable read-outs to meet the demands set by the samples under study. Yet, the investigation of cell development requires a multi-parametric approach to address both the structure and spatio-temporal organization of organelles, and also the transduction of chemical signals and forces involved in cell-cell interactions. Although the microscopy technologies for observing each of these characteristics are well developed, none of them can offer read-out of all characteristics simultaneously, which limits the information content of a measurement. For example, while electron microscopy is able to disclose the structural layout of cells and the macromolecular arrangement of proteins, it cannot directly follow dynamics in living cells. The latter can be achieved with fluorescence microscopy which, however, requires labelling and lacks spatial resolution. A remedy is to combine and correlate different readouts from the same specimen, which opens new avenues to understand structure-function relations in biomedical research. At the same time, such correlative approaches pose new challenges concerning sample preparation, instrument stability, region of interest retrieval, and data analysis. Because the field of correlative microscopy is relatively young, the capabilities of the various approaches have yet to be fully explored, and uncertainties remain when considering the best choice of strategy and workflow for the correlative experiment. With this in mind, the Journal of Physics D: Applied Physics presents a special roadmap on the correlative microscopy techniques, giving a comprehensive overview from various leading scientists in this field, via a collection of multiple short viewpoints. ispartof: JOURNAL OF PHYSICS D-APPLIED PHYSICS vol:51 issue:44 ispartof: location:England status: published

The methyltransferase enzymes can be applied to deliver a range of modifications to pre-determined sites on large DNA molecules with exceptional specificity and efficiency. To date, however, a limited number of modifications have been delivered in this way because of the complex chemical synthesis that is needed to produce a cofactor analogue carrying a specific function, such as a fluorophore. Here, we describe a method for the direct transfer of a series of functional compounds (seven fluorescent dyes, biotin and polyethylene glycol) to the DNA duplex. Our approach uses a functional cofactor analogue, whose final preparative step is performed alongiside the DNA modification reaction in a single pot, with no purification needed. We show that fluorophore conjugation efficiency in these mixtures is significantly improved compared to two-step labeling approaches. Our experiments highlight the remarkable malleability and selectivity of the methyltransferases tested. Additional analysis using high resolution localization of the fluorophore distribution indicates that target sites for the methyltransferase are predominantly labeled on a single strand of their palindromic site and that a small and randomly-distributed probability of off-site labeling exists. ispartof: Nucleic Acids Research vol:46 issue:11 pages:1-8 ispartof: location:England status: Published online

The performance of zeolites as solid acid catalysts is strongly influenced by the accessibility of active sites. However, synthetic zeolites typically grow as complex aggregates of small nanocrystallites rather than perfect single crystals. The structural complexity must therefore play a decisive role in zeolite catalyst applicability. Traditional tools for the characterization of heterogeneous catalysts are unable to directly relate nanometer scale structural properties to the corresponding catalytic performance. In this work, an innovative correlative super-resolution fluorescence and scanning electron microscope is applied and the appropriate analysis procedures are developed to investigate the effect of small-port H-mordenite (H-MOR) morphology on the catalytic performance, along with the effects of extensive acid leaching. These correlative measurements revealed catalytic activity at the interface between intergrown H-MOR crystallites that was assumed inaccessible, without compromising the shape selective properties. Furthermore, it was found that extensive acid leaching led to an etching of the originally accessible microporous structure, rather than the formation of an extended mesoporous structure. The associated transition of small-port to large-port H-MOR therefore did not render the full catalyst particle functional for catalysis. The applied characterization technique allows a straightforward investigation of the zeolite structure-activity relationship beyond the single-particle level. We conclude that such information will ultimately lead to an accurate understanding of the relationship between the bulk scale catalyst behavior and the nanoscale structural features, enabling a rationalization of catalyst design. ispartof: ACS Catalysis vol:7 issue:8 pages:5234-5242 ispartof: location:United States status: published

Osteoarthritis (OA) is a big burden of disease worldwide and one of the most common causes of disability in the adult population. Currently applied therapies consist of physical therapy, oral medication, intra-articular injections, and surgical interventions, with the main goal being to reduce pain and improve function and quality of life. Intra-articular (IA) administration of drugs has potential benefits in OA treatment because it minimizes systemic bioavailability and side effects associated with oral administration of drugs without compromising the therapeutic effect in the joint. However, IA drug residence time is short and there is a clinical need for a vehicle that is able to provide a sustained release long enough for IA therapy to fulfill its promise. This review summarizes the use of different polymeric systems and the incorporated drugs for IA drug delivery in the osteoarthritic joint with a primary focus on clinical needs and opportunities.

Many studies have reported on the effects of cyclooxygenase-2 (COX-2) inhibition on osteogenesis. However, far less is known about the effects of COX-2 inhibition on chondrogenic differentiation. Previous studies conducted by our group show that COX-2 inhibition influences in vitro chondrogenic differentiation. Importantly, this might have consequences on endochondral ossification processes occurring in vivo, such as bone fracture healing, growth plate development and ectopic generation of cartilage. The goal of our study was to investigate, in vivo, the effect of COX-2 inhibition by celecoxib on the cartilaginous phase of three different endochondral ossification scenarios.10 mg/kg/d celecoxib or placebo were orally administered for 25 d to skeletally-immature New Zealand White rabbits (n = 6 per group). Endochondral ossification during fracture healing of a non-critical size defect in the ulna, femoral growth plate and ectopically-induced cartilaginous tissue were examined by radiography, micro-computed tomography (mu-CT), histology and gene expression analysis.Celecoxib treatment resulted in delayed bone fracture healing, alterations in growth plate development and progression of mineralisation. In addition, chondrogenic differentiation of ectopically-induced cartilaginous tissue was severely impaired by celecoxib. In conclusion, we found that celecoxib impaired the chondrogenic phase of endochondral ossification.

A home-built fiber optic surface plasmon resonance platform (FO-SPR) was applied to directly screen PCR amplified DNA for mutations. The FO-SPR sensor was used for real-time monitoring of DNA duplex melting during high resolution temperature cycling. The signal of the DNA melting was enhanced by means of gold nanoparticle labels. This FO-SPR genetic assay allowed for detection of single-point mutations (SNP) in less than 20 min. The concept was demonstrated for the analysis of 9 different serogroups of the bacterium Legionella pneumophila, a common human pathogen responsible for atypical pneumonia. FO-SPR allowed us to detect genetic mutations inhibiting PCR, which could lead to amplification bias when molecular diagnostics are applied for L. pneumophila detection. All serogroups were found to display unique melting temperatures, indicating that mutations have accumulated in the target sequence. In a next step, clinical samples of L. pneumophila were analyzed using the FO-SPR sensor. This technology was proven to be reliable for the detection of mutations for those samples that previously displayed ambiguous qPCR quantification results. When these results were benchmarked, FO-SPR results were found to be consistent with Sanger sequencing but not with fluorescence based DNA melting. The presented results convincingly advocate the advantages of FO-SPR as a high resolution and fast genetic screening tool that can compete with the current standard techniques for SNP detection.

Affinity probe capillary electrophoresis (APCE) is potentially one of the most versatile technologies for protein diagnostics, offering an excellent balance between robustness, analysis speed and sensitivity. Combining the immunosensing and separating strength of capillary electrophoresis with the signal enhancement power of nucleic acid amplification, aptamers can further push the analytical limits of APCE to offer ultrasensitive, multiplexed detection of protein biomarkers, even when differences in electrophoretic mobility between the different aptamer-target complexes are limited. It is demonstrated how, through careful selection of experimental parameters, simultaneous detection of picomolar levels of three target proteins can be achieved even with aptamers that were initially selected under very different conditions and further taking into account that the aptamers need to be modified to allow successful PCR amplification. Aptamer-enhanced APCE offers limits of detection that are orders of magnitude lower than those that can be achieved through traditional capillary electrophoresis-based immunosensing. With recent developments in aptamer selection that for the first time realise the promise of aptamers as easily accessible, high affinity recognition molecules, it can therefore be envisioned that aptamer-enhanced APCE on parallel microfluidic platforms can be the basis for a truly high-throughput multiplexed proteomics platform, rivalling genetic screening for the first time.

Please cite this paper as: Janssen P, Brolmann H, van Kesteren P, Bongers M, Thurkow A, Heymans M, Huirne J. Perioperative outcomes using LigaSure compared with conventional bipolar instruments in laparoscopic hysterectomy: a randomised controlled trial. BJOG 2011;118:1568–1575. Objective To compare the effects of LigaSure versus the conventional bipolar technique on operating time and blood loss during laparoscopic hysterectomy. Design A randomised controlled trial. Setting Three teaching hospitals. Population Women undergoing a laparoscopic hysterectomy for benign indications. Methods 140 women undergoing a laparoscopic hysterectomy were randomised for LigaSure or conventional bipolar instruments. Main outcome measures Primary outcome was operating time from initial skin incision till detachment of the uterus. Secondary outcome measures were total operating time (from initial skin incision till final skin closure), time to dissect the adnexal ligaments, intra-operative blood loss and subjective evaluation by the surgeon of the instrument used. Results No differences in operating time (from initial skin incision till uterine detachment and initial skin incision till final skin closure) using LigaSure versus conventional bipolar instruments: 97.6 versus 91.8 minutes (P = 0.39, 95% CI – 7.6 to 19.2), and 148.1 versus 142.1 minutes (P = 0.46, 95% CI – 10.1 to 22.3), respectively. The mean blood loss using LigaSure versus conventional bipolar was 234.1 versus 273.1 ml (P = 0.46, 95% CI -39.1 to 52.7). Various subjective efficacy and instrument handling parameters were significantly different between the two instruments and between the different participating centres. Conclusions There were no significant differences in operating time and blood loss between the use of LigaSure and the use of conventional bipolar instruments during laparoscopic hysterectomy, even after correction for potential confounders. User satisfaction parameters were assessed as significantly different by surgeons of the participating centres.

The cationic dyes 9-aminoacridine (9AA) and safranine (Sf) were entrapped into silica spheres of about 0.2 μm diameter prepared by modified Stober method. The fluorescent materials are investigated by steady-state and time-resolved emission, in addition of confocal fluorescence microscopy. Silica particles containing 9-aminoacridine (SP9AA) and safranine (SPSf) or both dyes (SPSf9AA) are emissive particles. When both dyes are present in the same particle but loaded in sequential stages 9AA emission is quenched as a consequence of energy transfer from 9AA (donor) to Sf (acceptor). This result suggests that particle growing processes where the acceptor is incorporated first into the core do not prevent donor/acceptor pairs to be close due to an overlay of the concentration gradients of both dyes in a radial core-shell like distribution.

This paper is the first report of a fiber optic SPR biosensor with nanobead signal enhancement. We evaluated the system with a bioassay for the fast and accurate detection of peanut allergens in complex food matrices. Three approaches of an immunoassay to detect Ara h1 peanut allergens in chocolate candy bars were compared; a label-free assay, a secondary antibody sandwich assay and a nanobead enhanced assay. Although label-free detection is the most convenient, our results illustrate that functionalized nanobeads can offer a refined solution to improve the fiber SPR detection limit. By applying magnetite nanoparticles as a secondary label, the detection limit of the SPR bioassay for Ara h1 was improved by two orders of magnitude from 9 to 0.09 μg/mL. The super paramagnetic character of the nanoparticles ensured easy handling. The SPR fibers could be regenerated easily and one fiber could be reused for up to 35 times without loss of sensitivity. The results were benchmarked against a commercially available polyclonal ELISA kit. An excellent correlation was found between the Ara h1 concentrations obtained with the ELISA and the concentrations measured with the SPR fiber assay. In addition, with the SPR fiber we could measure the samples twice as fast as compared to the fastest ELISA protocol. Since the dipstick fiber has no need for microchannels that can become clogged, time consuming rinsing step could be avoided. The linear dynamic range of the presented sensor was between 0.1 and 2 μg/mL, which is considerably larger than the ELISA benchmark.

Ever since the inception of light microscopy, the laws of physics have seemingly thwarted every attempt to visualize the processes of life at its most fundamental, sub-cellular, level. The diffraction limit has restricted our view to length scales well above 250 nm and in doing so, severely compromised our ability to gain true insights into many biological systems. Fortunately, continuous advancements in optics, electronics and mathematics have since provided the means to once again make physics work to our advantage. Even though some of the fundamental concepts enabling super-resolution light microscopy have been known for quite some time, practically feasible implementations have long remained elusive. It should therefore not come as a surprise that the 2014 Nobel Prize in Chemistry was awarded to the scientists who, each in their own way, contributed to transforming super-resolution microscopy from a technological tour de force to a staple of the biologist's toolkit. By overcoming the diffraction barrier, light microscopy could once again be established as an indispensable tool in an age where the importance of understanding life at the molecular level cannot be overstated. This review strives to provide the aspiring life science researcher with an introduction to optical microscopy, starting from the fundamental concepts governing compound and fluorescent confocal microscopy to the current state-of-the-art of super-resolution microscopy techniques and their applications.

The conversion of dihydroxyacetone to ethyl lactate catalysed by different solid acids was investigated by means of High-Throughput Experimentation. A mesoporous amorphous aluminosilicate catalyst displaying very high selectivity and a good yield towards the lactate was identified. The correlation between the type of acid sites and the catalytic behaviour is discussed.

Organometal halide perovskites show promising features for cost-effective application in photovoltaics. The material instability remains a major obstacle to broad application because of the poorly understood degradation pathways. Here, we apply simultaneous luminescence and electron microscopy on perovskites for the first time, allowing us to monitor in situ morphology evolution and optical properties upon perovskite degradation. Interestingly, morphology, photoluminescence (PL), and cathodoluminescence of perovskite samples evolve differently upon degradation driven by electron beam (e-beam) or by light. A transversal electric current generated by a scanning electron beam leads to dramatic changes in PL and tunes the energy band gaps continuously alongside film thinning. In contrast, light-induced degradation results in material decomposition to scattered particles and shows little PL spectral shifts. The differences in degradation can be ascribed to different electric currents that drive ion migration. Moreover, solution-processed perovskite cuboids show heterogeneity in stability which is likely related to crystallinity and morphology. Our results reveal the essential role of ion migration in perovskite degradation and provide potential avenues to rationally enhance the stability of perovskite materials by reducing ion migration while improving morphology and crystallinity. It is worth noting that even moderate e-beam currents (86 pA) and acceleration voltages (10 kV) readily induce significant perovskite degradation and alter their optical properties. Therefore, attention has to be paid while characterizing such materials using scanning electron microscopy or transmission electron microscopy techniques. ispartof: Journal of Physical Chemistry Letters vol:7 issue:3 pages:561-566 ispartof: location:United States status: published

A high-throughput research platform was developed for the preparation and subsequent catalytic liquid-phase screening of ion-exchanged zeolites, for instance with regard to their use as heterogeneous catalysts. In this system aqueous solutions and other liquid as well as solid reagents are employed as starting materials and 24 samples are prepared on a library plate with a 4 × 6 layout. Volumetric dispensing of metal precursor solutions, weighing of zeolite and subsequent mixing/washing cycles of the starting materials and distributing reaction mixtures to the library plate are automatically performed by liquid and solid handlers controlled by a single common and easy-to-use programming software interface. The thus prepared materials are automatically contacted with reagent solutions, heated, stirred and sampled continuously using a modified liquid handling. The high-throughput platform is highly promising in enhancing synthesis of catalysts and their screening. In this paper the preparation of lanthanum-exchanged NaY zeolites (LaNaY) on the platform is reported, along with their use as catalyst for the conversion of renewables.

Various transition-metal-free oxides have been studied as catalysts for the epoxidation of cyclooctene with hydrogen peroxide by means of high-throughput experimentation. Different boron, aluminium, and gallium oxides were prepared according to various synthesis methods. A number of pure aluminium and gallium oxides showed very good catalytic performances, while the results obtained with boron oxides or mixed oxides were less positive. The best results were obtained with a gallium oxide catalyst, which gave an epoxide yield of 71 % and a selectivity of 99 % after reaction for 4 h at 80 degrees C. Gallium oxides had not been reported previously as active epoxidation catalysts. The use of high-throughput experimentation proved useful both for discovering new active catalysts and for identifying a number of relationships between the synthesis conditions and the catalytic properties of the transition-metal-free oxides.

Herein we report a new, one-step procedure for the metal-free ketonization of unsaturated fatty acid methyl esters and triacylglycerol mixtures with nitrous oxide (N 2 O). The conversion of various substrates can be tuned by parameters such as temperature, reaction time and N 2 O partial pressure. This ketonization chemistry offers various advantages over the classic Wacker catalytic process and a state-of-the-art two-step procedure via intermediate epoxides.

Plasma coating technologies have been demonstrated as being promising for the fabrication of bioactive and biocompatible materials, among others. Reported efforts are exclusively focused on the two-step approaches, in which the bioactive component is first immobilized on a substrate, followed by a (vacuum) plasma polymerization treatment or vice versa. However, we believe that upon minimizing the plasma energy, numerous bioactive substances such as enzymes and nucleic acids can be immobilized directly in plasmas via copolymerization with organic precursors, or by direct entrapment in the organic polymer. Therefore, a dielectric barrier discharge was employed at atmospheric pressure and ambient temperature to deposit organic coatings with reasonable growth rates at power input and frequency values as low as possible. Two promising precursors, acetylene and pyrrole, were selected out of 22 organic monomers for full physicochemical characterization. While the acetylene polymer film shows resemblance with its vacuum plasma analogue, polypyrrole coatings produced in vacuum and atmospheric plasmas differ significantly.

Supported metal nanoparticle catalysts are commonly obtained through deposition of metal precursors onto the support using incipient wetness impregnation. Typically, empirical relations between metal nanoparticle structure and catalytic performance are inferred from ensemble averaged data in combination with high-resolution electron microscopy. This approach clearly underestimates the importance of heterogeneities present in a supported metal catalyst batch. Here we show for the first time how incipient wetness impregnation leads to 10-fold variations in silver loading between individual submillimeter-sized silica support granules. This heterogeneity has a profound impact on the catalytic performance, with 100-fold variations in hydrogenation performance at the same level. In a straightforward fashion, optical microscopy interlinks single support particle level catalytic measurements to structural and compositional information. These detailed correlations reveal the optimal silver loading. A thorough consideration of catalyst heterogeneity and the impact thereof on the catalytic performance is indispensable in the development of catalysts. ispartof: ACS Catalysis vol:5 issue:11 pages:6690-6695 ispartof: location:United States status: published

Spatial restrictions around catalytic sites, provided by molecular-sized micropores, are beneficial to reaction selectivity but also inherently limit diffusion. The molecular transport can be enhanced by introducing meso- and macropores. However, the impact of this extraframework porosity on the local nanoscale reactivity is relatively unexplored. Herein we show that the area of enhanced reactivity in hierarchical zeolite, examined with super-resolution fluorescence microscopy, is spatially restricted to narrow zones around meso- and macropores, as observed with focused ion-beam-assisted scanning electron microscopy. This comparison indicates that reagent molecules efficiently reach catalytic active sites only in the micropores surrounding extraframework porosity and that extensive macroporosity does not warrant optimal reactivity distribution throughout a hierarchical porous zeolite. ispartof: ChemCatChem vol:7 issue:22 pages:3646-3650 ispartof: location:Germany status: published

© The Royal Society of Chemistry 2015. We report an investigation on the effect of stabilization agents and surface curvatures on oxidative etching of three classes of anisotropically shaped gold nanoparticles namely, rods, bipyramids and prisms. In particular, the dual role of the stabilizing agent CTAB in the etching process is explored, showing how it acts both as a source of bromine ions, accelerating etching and as a protection agent, resulting in anisotropic reshaping. ispartof: RSC Advances vol:5 issue:9 pages:6829-6833 status: published

Structural and temporal inhomogeneities can have a marked influence on the performance of inorganic and biocatalytic systems alike. While these subtle variations are hardly ever accessible through bulk or ensemble averaged activity screening, insights into the molecular mechanisms underlying these diverse phenomena are absolutely critical for the development of optimized or novel catalytic systems and processes. Fortunately, state-of-the-art fluorescence microscopy methods have allowed experimental access to this intriguing world at the nanoscale. In this tutorial review we will first provide a broad overview of key concepts and developments in the application of single molecule fluorescence spectroscopy to (bio)catalysis research. In the second part topics specific to both bio and heterogeneous catalysis will be reviewed in more detail.

Detection of allergens and micro bacteria on a fiber optic SPR sensor is performed by means of an oligonucleotide bio-recognition surface. In this work, the DNA surface is optimized by mixing with PEG monolayers to suppress unspecific and enhance specific detection. Furthermore, due to the surface characteristic of DNA functionalized gold nanoparticles the bioassay sensitivity is increased and a mean of real-time multiplexing is introduced.

Detection of single nucleotide polymorphisms (SNPs) is a crucial challenge in the development of a novel generation of diagnostic tools. Accurate detection of SNPs can prove elusive, as the impact of a single variable nucleotide on the properties of a target sequence is limited, even if this sequence consists of only a few nucleotides. New, accurate and facile strategies for the detection of point mutations are therefore absolutely necessary for the increased adoption of point-of-care molecular diagnostics. Currently, PCR and sequencing are mostly applied for diagnosing SNPs. However these methods have serious drawbacks as routine diagnostic tools because of their labour intensity and cost. Several new, more suitable methods can be applied to enable sensitive detection of mutations based on specially designed hybridization probes, mutation recognizing enzymes and thermal denaturation. Here, an overview is presented of the most recent advances in the field of fast and sensitive SNP detection assays with strong potential for integration in point-of-care tests. ispartof: The Analyst vol:139 issue:2 pages:353-370 ispartof: location:England status: published

Structural and temporal inhomogeneities can have a marked influence on the performance of inorganic and biocatalytic systems alike. While these subtle variations are hardly ever accessible through bulk or ensemble averaged activity screening, insights into the molecular mechanisms underlying these diverse phenomena are absolutely critical for the development of optimized or novel catalytic systems and processes. Fortunately, state-of-the-art fluorescence microscopy methods have allowed experimental access to this intriguing world at the nanoscale. In this tutorial review we will first provide a broad overview of key concepts and developments in the application of single molecule fluorescence spectroscopy to (bio)catalysis research. In the second part topics specific to both bio and heterogeneous catalysis will be reviewed in more detail. ispartof: Chemical Society Reviews vol:43 issue:4 pages:990-1006 ispartof: location:England status: published

Ever increasing demands in sensitivity and specificity of biosensors have recently established a trend toward the use of multivalent bioreceptors. This trend has also been introduced in the field of bacteriophage affinity peptides, where the entire phage is used as a receptor rather than the individual peptides. Although this approach is gaining in popularity due to the numerous advantages, binding kinetics of complete phage particles have never been studied in detail, notwithstanding being essential for the efficient design of such applications. In this paper we used an in house developed fiber-optic surface plasmon resonance (FO-SPR) biosensor to study the affinity and binding kinetics of phages, displaying peptide libraries. By using either peptide expression on the p3 or on the p8 coat proteins, a corresponding density of 5 up to more than 2000 peptides on a single virus particle was obtained. Binding parameters of 26 different filamentous phages, displaying peptides selective for enhanced Green Fluorescent Protein (eGFP), were characterized. This study revealed a broad affinity range of phages for the target eGFP, indicating their potential to be used for applications with different requirements in binding kinetics. Moreover, detailed analysis of koff and kon values of several selected p3 and p8 phages, using the FO-SPR biosensor, clearly showed the correlation between the binding parameters and the density at which eGFP-peptides are being expressed. Consequently, although p3 and p8-based phages both revealed exceptionally high affinities for eGFP, two p8 phages were found to have the highest affinity with dissociation constants (Kd) in the femtomolar range.

Aptamers are short, single stranded DNA/RNA oligonucleotides with the ability to fold into well-defined three-dimensional structures, which are fundamental for their highly selective target recognition, rivaling thus antibodies in terms of affinity and specificity. Thanks to these properties, they have been integrated into different micro- and nanoscale transducers, resulting in a new type of biosensors, known as `aptasensors', used in a truly new generation of diagnostic tools. Here, an overview will be given of different aptamer selection approaches, with the focus on lab-on-a-chip technology based methods, different concepts reported for integrating aptamers into micro- and nanobiosystems aiming at the development of sensitive diagnostics concluding with an outlook on the future of aptamers in theranostics.

Major advancements in molecular biology and clinical diagnostics cannot be brought about strictly through the use of genomics based methods. Improved methods for protein detection and proteomic screening are an absolute necessity to complement to wealth of information offered by novel, high-throughtput sequencing technologies. Only then will it be possible to advance insights into clinical processes and to characterize the importance of specific protein biomarkers for disease detection or the realization of `personalized medicine'. Currently however, large-scale proteomic information is still not as easily obtained as its genomic counterpart mainly because traditional, antibody-based technologies struggle to meet the stringent sensitivity- and throughput requirements that are required whereas mass-spectrometry based methods might be burdened by significant costs involved. However, recent years have seen the development of new biodetection strategies linking nucleic acids with existing antibody technology or replacing antibodies with oligonucleotide recognition elements altogether. These advancements have unlocked many new strategies to lower detection limits and dramatically increase throughput of protein detection assays. In this review, an overview of these new strategies will be given. ispartof: Sensors vol:13 issue:1 pages:898-930 ispartof: location:Switzerland status: published

The rising prevalence to food allergies in the past two decades, together with the fact that the only existing therapy is avoidance of allergen-containing food next to the implementation of anti-allergic drugs, urges the need for improved performance of current assays to detect potential allergens in food products. Therein, the focus has been on aptamer-based biosensors in recent years. In this paper we report for the first time the selection of aptamers against one of the most important peanut allergens, Ara h 1. Several Ara h1 DNA aptamers were selected after eight selection rounds using capillary electrophoresis (CE)-SELEX. The selected aptamers specifically recognized Ara h 1 and did not significantly bind with other proteins, including another peanut allergen Ara h 2. The dissociation constant of a best performing aptamer was in the nanomolar range as determined independently by three different approaches, which are surface plasmon resonance, fluorescence anisotropy, and capillary electrophoresis (353 ± 82 nM, 419 ± 63 nM, and 450 ± 60 nM, respectively). Furthermore, the selected aptamer was used for bioassay development on a home-built fiber optic surface plasmon resonance (FO-SPR) biosensor platform for detecting Ara h 1 protein in both buffer and food matrix samples demonstrating its real potential for the development of novel, more accurate aptamer-based biosensors. In conclusion, the reported aptamer holds a great potential for the detection of Ara h 1 in both the medical field and the food sector due to its high affinity and specificity for the target protein.