Nanoscale intermittent contact-scanning electrochemical microscopy

A major theme in scanning electrochemical microscopy (SECM) is a methodology for nanoscale imaging with distance control and positional feedback of the tip. We report the expansion of intermittent contact (IC)-SECM to the nanoscale, using disk-type Pt nanoelectrodes prepared using the laser-puller sealing method. The Pt was exposed using a focused ion beam milling procedure to cut the end of the electrode to a well-defined glass sheath radius, which could also be used to reshape the tips to reduce the size of the glass sheath. This produced nanoelectrodes that were slightly recessed, which was optimal for IC-SECM on the nanoscale, as it served to protect the active part of the tip. A combination of finite element method simulations, steady-state voltammetry and scanning electron microscopy for the measurement of critical dimensions, was used to estimate Pt recession depth. With this knowledge, the tip-substrate alignment could be further estimated by tip approach curve measurements. IC-SECM has been implemented by using a piezo-bender actuator for the detection of damping of the oscillation amplitude of the tip, when IC occurs, which was used as a tip-position feedback mechanism. The piezo-bender actuator improves significantly on the performance of our previous setup for IC-SECM, as the force acting on the sample due to the tip is greatly reduced, allowing studies with more delicate tips. The capability of IC-SECM is illustrated with studies of a model electrode (metal/glass) substrate

Mr-Nom: Multi-Scale Resolution of Neuronal Cells in Nissl-Stained Histological Slices Via Deliberate over-Segmentation and Merging

In comparative neuroanatomy, the characterization of brain cytoarchitecture is critical to a better understanding of brain structure and function, as it helps to distill information on the development, evolution, and distinctive features of different populations. The automatic segmentation of individual brain cells is a primary prerequisite and yet remains challenging. A new method (MR-NOM) was developed for the instance segmentation of cells in Nissl-stained histological images of the brain. MR-NOM exploits a multi-scale approach to deliberately over-segment the cells into superpixels and subsequently merge them via a classifier based on shape, structure, and intensity features. The method was tested on images of the cerebral cortex, proving successful in dealing with cells of varying characteristics that partially touch or overlap, showing better performance than two state-of-the-art methods

Enfermedad de pompe: presentación de un caso

Presentamos el caso de una niña de 6 meses de ascendencia china con enfermedad de Pompe diagnosticada a partir de una miocardiopatía hipertrófica biventricular e hipotonía global. En esta enfermedad el acúmulo de glucógeno lisosomal principalmente en el corazón y músculo esquelético ocasiona la mayoría de la sintomatología clínica. En esta paciente la hipotonía generalizada, macroglosia, debilidad en el llanto y la confirmación ecocardiográfica de una miocardiopatía hipertrófica biventricular fueron el punto inicial para la sospecha clínica de la enfermedad que, junto a la elevación de enzimas musculares y hepáticas, condujo al estudio de alfa-glucosidasa ácida (AGA) que confirmó el diagnóstico. Desde 2006 existe terapia de reemplazo enzimático que permite mejorar la sobrevida y la calidad de vida si el diagnóstico se realiza precozmente.We are presenting the case of a 7 month old infant with infantile Pompe disease, who was the first case treated by enzyme replacement therapy (ERT) at our Hospital.The patient was of chinese ancestry and was suspected to have the disease after finding biventricular hypertrophic myocardiopathy and muscle weakness and hypotonia. Other relevant features were macroglosia and increase in CK level as well as the results of the electrophysiology studies. In view of the new possibility of ERT, educational efforts must be done in order to perform early diagnosis before signs of severe cardiorespiratory failure develops, and to improve survival and quality of life of patients with this condition

Importance of mass transport and spatially heterogeneous flux processes for in situ atomic force microscopy measurements of crystal growth and dissolution kinetics

It is well-established that important information about the dissolution and growth of crystals can be obtained by the investigation of step movement on single-crystal faces via in situ AFM. However, a potential drawback of this approach for kinetic measurements is that the small region of investigation may not be representative of the overall surface. It is shown that the investigation of local processes without accounting for the processes outside the region of interest can lead to significant misinterpretation of the data collected. Taking the case of gypsum dissolution as an example, we critically analyze literature data and develop 3 different finite element method models that treat in detail the coupled mass transport–surface kinetic problem pertaining to dissolution processes in a typical AFM environment. It is shown that mass transport cannot be neglected when performing in situ AFM on macroscopic surfaces even with high-convection fluid cells. Moreover, crystal dissolution kinetics determined by AFM is mainly influenced by processes occurring in areas of the surface outside the region of interest. When this is recognized, and appropriate models are applied, step velocities due to dissolution are consistent with expectations based on macroscopic measurements, and the kinetic gap that is often apparent between nanoscale and macroscopic measurements is closed. This study provides a framework for the detailed analysis of AFM kinetic data that has wide utility and applicability

Multi-aspect testing and ranking inference to quantify dimorphism in the cytoarchitecture of cerebellum of male, female and intersex individuals: a model applied to bovine brains

The dimorphism among male, female and freemartin intersex bovines, focusing on the vermal lobules VIII and IX, was analyzed using a novel data analytics approach to quantify morphometric differences in the cytoarchitecture of digitalized sections of the cerebellum. This methodology consists of multivariate and multi-aspect testing for cytoarchitecture-ranking, based on neuronal cell complexity among populations defined by factors, such as sex, age or pathology. In this context, we computed a set of shape descriptors of the neural cell morphology, categorized them into three domains named size, regularity and density, respectively. The output and results of our methodology are multivariate in nature, allowing an in-depth analysis of the cytoarchitectonic organization and morphology of cells. Interestingly, the Purkinje neurons and the underlying granule cells revealed the same morphological pattern: female possessed larger, denser and more irregular neurons than males. In the Freemartin, Purkinje neurons showed an intermediate setting between males and females, while the granule cells were the largest, most regular and dense. This methodology could be a powerful instrument to carry out morphometric analysis providing robust bases for objective tissue screening, especially in the field of neurodegenerative pathologies

The prefrontal cortex of the bottlenose dolphin (Tursiops truncatus Montagu, 1821): a tractography study and comparison with the human

Cetaceans are well known for their remarkable cognitive abilities including self-recognition, sound imitation and decision making. In other mammals, the prefrontal cortex (PFC) takes a key role in such cognitive feats. In cetaceans, however, a PFC could up to now not be discerned based on its usual topography. Classical in vivo methods like tract tracing are legally not possible to perform in Cetacea, leaving diffusion-weighted imaging (DWI) as the most viable alternative. This is the first investigation focussed on the identification of the cetacean PFC homologue. In our study, we applied the constrained spherical deconvolution (CSD) algorithm on 3 T DWI scans of three formalin-fixed brains of bottlenose dolphins (Tursiops truncatus) and compared the obtained results to human brains, using the same methodology. We first identified fibres related to the medio-dorsal thalamic nuclei (MD) and then seeded the obtained putative PFC in the dolphin as well as the known PFC in humans. Our results outlined the dolphin PFC in areas not previously studied, in the cranio-lateral, ectolateral and opercular gyri, and furthermore demonstrated a similar connectivity pattern between the human and dolphin PFC. The antero-lateral rotation of the PFC, like in other areas, might be the result of the telescoping process which occurred in these animals during evolution

The motor cortex of the sheep: laminar organization, projections and diffusion tensor imaging of the intracranial pyramidal and extrapyramidal tracts

The laminar organization of the motor cortex of the sheep and other large domestic herbivores received scarce attention and is generally considered homologous to that of rodents and primates. Thickness of the cortex, subdivision into layers and organization are scarcely known. In the present study, we applied different modern morphological, mathematical and image-analyses techniques to the study of the motor area that controls movements of the forelimb in the sheep. The thickness of the cortex resulted comparable to that of other terrestrial Cetartiodactyls (but thicker than in marine Cetartiodactyls of similar body mass). The laminar organization showed marked development of layer 1, virtual absence of layer 4, and image analysis suggested prevalence of large irregular neural cells in the deeper layers. Diffusion tensor imaging revealed robust projections from the motor cortex to the pyramids in the brainstem, and well evident tracts descending to the tegmentum of the mesencephalon and dorsal pons. Our data contrast the general representation of the motor system of this species, considered to be predominantly based on extra-pyramidal tracts that originate from central pattern generators in the brainstem. © 2019, Springer-Verlag GmbH Germany, part of Springer Nature

Hopping intermittent contact-scanning electrochemical microscopy (HIC-SECM) as a new local dissolution kinetic probe : application to salicylic acid dissolution in aqueous solution

Dissolution kinetics of the (110) face of salicylic acid in aqueous solution is determined by hopping intermittent contact-scanning electrochemical microscopy (HIC-SECM) using a 2.5 μm diameter platinum ultramicroelectrode (UME). The method operates by translating the probe UME towards the surface at a series of positions across the crystal and inducing dissolution via the reduction of protons to hydrogen, which titrates the weak acid and promotes the dissolution reaction, but only when the UME is close to the crystal. Most importantly, as dissolution is only briefly and transiently induced at each location, the initial dissolution kinetics of an as-grown single crystal surface can be measured, rather than a surface which has undergone significant dissolution (pitting), as in other techniques. Mass transport and kinetics in the system are modelled using finite element method simulations which allows dissolution rate constants to be evaluated. It is found that the kinetics of an ‘as-grown’ crystal are much slower than for a surface that has undergone partial bulk dissolution (mimicking conventional techniques), which can be attributed to a dramatic change in surface morphology as identified by atomic force microscopy (AFM). The ‘as-grown’ (110) surface presents extended terrace structures to the solution which evidently dissolve slowly, whereas a partially dissolved surface has extensive etch features and step sites which greatly enhance dissolution kinetics. This means that crystals such as salicylic acid will show time-dependent dissolution kinetics (fluxes) that are strongly dependent on crystal history, and this needs to be taken into account to fully understand dissolution

The laminar organization of the motor cortex in monodactylous mammals: a comparative assessment based on horse, chimpanzee, and macaque

The architecture of the neocortex classically consists of six layers, based on cytological criteria and on the layout of intra/interlaminar connections. Yet, the comparison of cortical cytoarchitectonic features across different species proves overwhelmingly difficult, due to the lack of a reliable model to analyze the connection patterns of neuronal ensembles forming the different layers. We first defined a set of suitable morphometric cell features, obtained in digitized Nissl-stained sections of the motor cortex of the horse, chimpanzee, and crab-eating macaque. We then modeled them using a quite general non-parametric data representation model, showing that the assessment of neuronal cell complexity (i.e., how a given cell differs from its neighbors) can be performed using a suitable measure of statistical dispersion such as the mean absolute deviation\u2014mean absolute deviation (MAD). Along with the non-parametric combination and permutation methodology, application of MAD allowed not only to estimate, but also to compare and rank the motor cortical complexity across different species. As to the instances presented in this paper, we show that the pyramidal layers of the motor cortex of the horse are far more irregular than those of primates. This feature could be related to the different organizations of the motor system in monodactylous mammals