Molecular and functional profiling of cell diversity and identity in the lateral superior olive, an auditory brainstem center with ascending and descending projections.

The lateral superior olive (LSO), a prominent integration center in the auditory brainstem, contains a remarkably heterogeneous population of neurons. Ascending neurons, predominantly principal neurons (pLSOs), process interaural level differences for sound localization. Descending neurons (lateral olivocochlear neurons, LOCs) provide feedback into the cochlea and are thought to protect against acoustic overload. The molecular determinants of the neuronal diversity in the LSO are largely unknown. Here, we used patch-seq analysis in mice at postnatal days P10-12 to classify developing LSO neurons according to their functional and molecular profiles. Across the entire sample (n = 86 neurons), genes involved in ATP synthesis were particularly highly expressed, confirming the energy expenditure of auditory neurons. Two clusters were identified, pLSOs and LOCs. They were distinguished by 353 differentially expressed genes (DEGs), most of which were novel for the LSO. Electrophysiological analysis confirmed the transcriptomic clustering. We focused on genes affecting neuronal input-output properties and validated some of them by immunohistochemistry, electrophysiology, and pharmacology. These genes encode proteins such as osteopontin, Kv11.3, and Kvß3 (pLSO-specific), calcitonin-gene-related peptide (LOC-specific), or Kv7.2 and Kv7.3 (no DEGs). We identified 12 "Super DEGs" and 12 genes showing "Cluster similarity." Collectively, we provide fundamental and comprehensive insights into the molecular composition of individual ascending and descending neurons in the juvenile auditory brainstem and how this may relate to their specific functions, including developmental aspects.

Hydrothiolation of Triisopropylsilyl Acetylene Sulfur Pentafluoride - Charting the Chemical Space of ß-SF5 Vinyl Sulfides.

Recently, we suggested liquid and high-boiling TIPS-CC-SF5 (TASP) as a versatile reagent to access so far elusive SF5-containing building blocks by less specialized laboratories under bench-top conditions. The synthesis of non-aromatic SF5 building blocks generally requires on-site fluorination or pentafluorosulfanylation steps employing toxic and/or gaseous reagents. Herein, we underline the versatility of this reagent by reporting a benign bench-top protocol for the synthesis of Z-configured ß-pentafluorosulfanylated vinyl sulfides in good to excellent yields (up to 99 %) with exclusive (Z)-diasteroselectivity and broad functional group tolerance. This method exploits an in-situ protodesilylation-hydrothiolation sequence. This so far uncharted class of compounds was characterized by means of NMR-spectroscopy as well as SC-XRD. Furthermore, we suggest the reaction to proceed via a kinetically controlled closed-shell reaction pathway, corroborated by in-silico experiments.

Photophysical image analysis: Unsupervised probabilistic thresholding for images from electron-multiplying charge-coupled devices.

We introduce the concept photophysical image analysis (PIA) and an associated pipeline for unsupervised probabilistic image thresholding for images recorded by electron-multiplying charge-coupled device (EMCCD) cameras. We base our approach on a closed-form analytic expression for the characteristic function (Fourier-transform of the probability mass function) for the image counts recorded in an EMCCD camera, which takes into account both stochasticity in the arrival of photons at the imaging camera and subsequent noise induced by the detection system of the camera. The only assumption in our method is that the background photon arrival to the imaging system is described by a stationary Poisson process (we make no assumption about the photon statistics for the signal). We estimate the background photon statistics parameter, λbg, from an image which contains both background and signal pixels by use of a novel truncated fit procedure with an automatically determined image count threshold. Prior to this, the camera noise model parameters are estimated using a calibration step. Utilizing the estimates for the camera parameters and λbg, we then introduce a probabilistic thresholding method, where, for the first time, the fraction of misclassified pixels can be determined a priori for a general image in an unsupervised way. We use synthetic images to validate our a priori estimates and to benchmark against the Otsu method, which is a popular unsupervised non-probabilistic image thresholding method (no a priori estimates for the error rates are provided). For completeness, we lastly present a simple heuristic general-purpose segmentation method based on the thresholding results, which we apply to segmentation of synthetic images and experimental images of fluorescent beads and lung cell nuclei. Our publicly available software opens up for fully automated, unsupervised, probabilistic photophysical image analysis.

Benign synthesis of terpene-based 1,4-p-menthane diamine.

Terpenes represent a promising renewable feedstock for the substitution of fossil resources in the synthesis of renewable platform chemicals, like diamines. This work describes the synthesis and full characterization of 1,4-p-menthane diamine (1,4-PMD) obtained from α-terpinene (1). A two-step procedure using dibenzyl azodicarboxylate (DBAD) and H2 as rather benign reagents was employed under comparatively mild conditions. Both C-N bonds were formed simultaneously during a visible-light mediated Diels-Alder reaction, which was investigated in batch or flow, avoiding regioselectivity issues during the amination steps that are otherwise typical for terpene chemistry. Heterogeneously catalyzed quadruple hydrogenation of the cycloadduct (2a) yielded 1,4­PMD (3). While the intermediate cycloadduct was shown to be distillable, the target diamine can be sublimed, offering sustainable purification methods.

Visible-Light Activation of Diorganyl Bis(pyridylimino) Isoindolide Aluminum(III) Complexes and Their Organometallic Radical Reactivity.

We report on the synthesis and characterization of a series of (mostly) air-stable diorganyl bis(pyridylimino) isoindolide (BPI) aluminum complexes and their chemistry upon visible-light excitation. The redox non-innocent BPI pincer ligand allows for efficient charge transfer homolytic processes of the title compounds. This makes them a universal platform for the generation of carbon-centered radicals. The photo-induced homolytic cleavage of the Al-C bonds was investigated by means of stationary and transient UV/Vis spectroscopy, spin trapping experiments, as well as EPR and NMR spectroscopy. The experimental findings were supported by quantum chemical calculations. Reactivity studies enabled the utilization of the aluminum complexes as reactants in tin-free Giese-type reactions and carbonyl alkylations under ambient conditions, which both indicated radical-polar crossover behavior. A deeper understanding of the physical fundamentals and photochemical process was provided, furnishing in turn a new strategy to control the reactivity of bench-stable aluminum organometallics.

Comparative anatomy of the spinneret musculature in cribellate and ecribellate spiders (Araneae).

Silk production is a prominent characteristic of spiders. The silk is extruded through spigots located on the spinnerets, which are single- to multimembered paired appendages at the end of the abdomen. Most extant spiders have three pairs of spinnerets, and in between either a cribellum (spinning plate) or a colulus (defunct vestigial organ), dividing these spiders into cribellate and ecribellate species. Previous research has shown that cribellate and ecribellate spiders differ not only in the composition of their spinning apparatus but also in the movements of their spinnerets during silk spinning. The objective of this study was to determine whether the differences in spinneret movements are solely due to variations in spinneret shape or whether they are based on differences in muscular anatomy. This was accomplished by analyzing microcomputed tomography scans of the posterior abdomen of each three cribellate and ecribellate species. It was found that the number of muscles did not generally differ between cribellate and ecribellate species, but varied considerably between the species within each of these two groups. Muscle thickness, particularly of the posterior median spinneret, varied slightly between groups, with cribellate spiders exhibiting more robust muscles, possibly to aid in the combing process during cribellar thread production. Interestingly, the vestigial colulus still possesses muscles, that can be homologized with those of the cribellum. This exploration into spinneret anatomy using microcomputed tomography data reveals that despite being small appendages, the spider spinnerets are equipped with a complex musculature that enables them to perform fine-scaled maneuvers to construct different fiber-based materials.

Geometry-Dependent Elastic Flow Dynamics in Micropillar Arrays.

Regular device-scale DNA waves for high DNA concentrations and flow velocities have been shown to emerge in quadratic micropillar arrays with potentially strong relevance for a wide range of microfluidic applications. Hexagonal arrays constitute another geometry that is especially relevant for the microfluidic pulsed-field separation of DNA. Here, we report on the differences at the micro and macroscopic scales between the resulting wave patterns for these two regular array geometries and one disordered array geometry. In contrast to the large-scale regular waves visible in the quadratic array, in the hexagonal arrays, waves occur in a device-scale disordered zig-zag pattern with fluctuations on a much smaller scale. We connect the large-scale pattern to the microscopic flow and observe flow synchronization that switches between two directions for both the quadratic and hexagonal arrays. We show the importance of order using the disordered array, where steady-state stationary and highly fluctuating flow states persist in seemingly random locations across the array. We compare the flow dynamics of the arrays to that in a device with sparsely distributed pillars. Here, we observe similar vortex shedding, which is clearly observable in the quadratic and disordered arrays. However, the shedding of these vortices couples only in the flow direction and not laterally as in the dense, ordered arrays. We believe that our findings will contribute to the understanding of elastic flow dynamics in pillar arrays, helping us elucidate the fundamental principles of non-Newtonian fluid flow in complex environments as well as supporting applications in engineering involving e.g., transport, sorting, and mixing of complex fluids.

Intramolecular dearomative 1,4-addition of silyl and germyl radicals to a phenyl moiety.

Herein, we present that the radicals [Ph3PC(Me)EMes2]˙ (2Si and 2Ge) can be generated from the α-silylated and α-germylated phosphorus ylides Ph3PC(Me)E(Cl)Mes2 (1Si and 1Ge) through one-electron reduction with Jones' dimer (MesNacNacMg)2 in benzene. Although isolation of the free radicals was not possible, the products of the intramolecular addition of the radicals to a phenyl substituent of the phosphorus moiety, followed by subsequent reaction with 2Si or 2Ge to the isolated species 3Si and 3Ge, respectively, were observed. This transformation witnesses a dearomative 1,4-addition of tetryl radical species to the phenyl scaffold in a stereoselective anti-fashion.

Disruption of a three-component solution as a novel strategy for Cu and Ni extraction from vegetable oils for their determination by GF AAS.

In this work, we propose a novel approach for extracting Cu and Ni from vegetable oils (which can be expanded to other metals). The method is based on the transference of the analytes to an aqueous acid phase due to the disruption of a three-component solution. The extraction was carried out in two steps. In the first step, a three-component solution was prepared comprising the sample, 1-octanol, and HNO3 solution. Next, the homogeneous system was disrupted by adding 1.0 mL of deionized water, and two phases were formed. The aqueous extract deposited in the bottom of the flask was collected with a micropipette, and Cu and Ni were determined by graphite furnace atomic absorption spectrometry (GF AAS). The developed method presented limits of quantification (LOQ) of 0.25 and 0.17 ng g-1 for Cu and Ni, respectively, and was successfully applied in the analysis of eleven oil samples from different origins.

Hydroamination of Triisopropylsilyl Acetylene Sulfur Pentafluoride - a Bench-top Route to Pentafluorosulfanylated Enamines.

Synthetic access to a variety of aliphatic and vinylic pentafluorosulfanylated building blocks remains a major challenge in contemporary organofluorine chemistry hampering its investigation in the context of medicinal chemistry, agrochemistry and functional materials. Herein, we report a bench-top protocol to access the virtually unknown class of α-SF5 -enamines under mild reaction conditions in good to excellent yields (up to 95 %). This reaction combines the protodesilylation of the commercially available precursor TASP with the in situ hydroamination of HC≡C-SF5 . The on-site use of highly toxic gases or corrosive reagents is avoided, making access to this motif applicable to a wide chemical audience. The excellent E-diastereoselectivity of this two-step cascade reaction is suggested to be the result of the convergence of the fast Z-/E- isomerization of a vinyl anion as well as the isomerization of the iminium ion. The remarkable thermal stability of these SF5 -enamines encourages further studies of their synthetic utility.

Facilitating taxonomy and phylogenetics: An informative and cost-effective protocol integrating long amplicon PCRs and third-generation sequencing.

Phylogenetic inference has become a standard technique in integrative taxonomy and systematics, as well as in biogeography and ecology. DNA barcodes are often used for phylogenetic inference, despite being strongly limited due to their low number of informative sites. Also, because current DNA barcodes are based on a fraction of a single, fast-evolving gene, they are highly unsuitable for resolving deeper phylogenetic relationships due to saturation. In recent years, methods that analyse hundreds and thousands of loci at once have improved the resolution of the Tree of Life, but these methods require resources, experience and molecular laboratories that most taxonomists do not have. This paper introduces a PCR-based protocol that produces long amplicons of both slow- and fast-evolving unlinked mitochondrial and nuclear gene regions, which can be sequenced by the affordable and portable ONT MinION platform with low infrastructure or funding requirements. As a proof of concept, we inferred a phylogeny of a sample of 63 spider species from 20 families using our proposed protocol. The results were overall consistent with the results from approaches based on hundreds and thousands of loci, while requiring just a fraction of the cost and labour of such approaches, making our protocol accessible to taxonomists worldwide.

Dynamic evolution of locomotor performance independent of changes in extended phenotype use in spiders.

Many animals use self-built structures (extended phenotypes) to enhance body functions, such as thermoregulation, prey capture or defence. Yet, it is unclear whether the evolution of animal constructions supplements or substitutes body functions-with disparate feedbacks on trait evolution. Here, using brown spiders (Araneae: marronoid clade), we explored if the evolutionary loss and gain of silken webs as extended prey capture devices correlates with alterations in traits known to play an important role in predatory strikes-locomotor performance (sprint speed) and leg spination (expression of capture spines on front legs). We found that in this group high locomotor performance, with running speeds of over 100 body lengths per second, evolved repeatedly-both in web-building and cursorial spiders. There was no correlation with running speed, and leg spination only poorly correlated, relative to the use of extended phenotypes, indicating that web use does not reduce selective pressures on body functions involved in prey capture and defence per se. Consequently, extended prey capture devices serve as supplements rather than substitutions to body traits and may only be beneficial in conjunction with certain life-history traits, possibly explaining the rare evolution and repeated loss of trapping strategies in predatory animals.

Using symmetry to control viscoelastic waves in pillar arrays.

Solutions of macromolecules exhibit viscoelastic properties and unlike Newtonian fluids, they may break time-reversal symmetry at low Reynolds numbers resulting in elastic turbulence. Furthermore, under some conditions, instead of the chaotic turbulence, the result is large-scale waves in the form of cyclic spatial and temporal concentration variations, as has been shown for macromolecular DNA flowing in microfluidic pillar arrays. We here demonstrate how altering the symmetry of the individual pillars can be used to influence the symmetry of these waves. We control the extent of instabilities in viscoelastic flow by leveraging the effects of the symmetry of the pillars on the waves, demonstrating suppressed viscoelastic fluctuations with relevance for transport and sorting applications, or conversely opening up for enhanced viscoelasticity-mediated mixing. The onset of waves, which changes flow resistance, occurs at different Deborah numbers for flow in different directions through the array of triangular pillars, thus breaking the symmetry of the flow resistance along the device, opening up for using the occurrence of the waves to construct a fluidic diode.

Cryogenic characteristics of graphene composites-evolution from thermal conductors to thermal insulators.

The development of cryogenic semiconductor electronics and superconducting quantum computing requires composite materials that can provide both thermal conduction and thermal insulation. We demonstrated that at cryogenic temperatures, the thermal conductivity of graphene composites can be both higher and lower than that of the reference pristine epoxy, depending on the graphene filler loading and temperature. There exists a well-defined cross-over temperature-above it, the thermal conductivity of composites increases with the addition of graphene; below it, the thermal conductivity decreases with the addition of graphene. The counter-intuitive trend was explained by the specificity of heat conduction at low temperatures: graphene fillers can serve as, both, the scattering centers for phonons in the matrix material and as the conduits of heat. We offer a physical model that explains the experimental trends by the increasing effect of the thermal boundary resistance at cryogenic temperatures and the anomalous thermal percolation threshold, which becomes temperature dependent. The obtained results suggest the possibility of using graphene composites for, both, removing the heat and thermally insulating components at cryogenic temperatures-a capability important for quantum computing and cryogenically cooled conventional electronics.

The Relationship between Diurnal Measures of Tonic Alertness and Self-Reported Fatigue in Persons with Multiple Sclerosis-A Retrospective Data Analysis.

OBJECTIVE: Fatigue in multiple sclerosis (MS) is common, burdensome, and usually assessed by self-report measures. This retrospective data analysis of the twice-daily Alertness test (Test battery of Attentional Performance) examined the extent to which this assessment procedure is associated with MS-related fatigue. METHOD: Two-hundred and thirteen German inpatients (136 women) aged 18-69 years with predominantly relapsing MS (72.8%) were included. Based on reaction time (RT) differences between morning tonic alertness (8:30-11:00 a.m.) and afternoon tonic alertness (3:00-4:30 p.m.), patients were divided into an "improver," "maintainer," or "decliner" group. Multinomial logistic regression (MLR) was calculated to predict the likelihood of belonging to one of these performance groups, taking into account cognitive fatigue (Fatigue Scale of Motor and Cognition, FSMCcog), disease severity (Expanded Disability Status Scale, EDSS), depression (Center for Epidemiologic Studies Depression Scale, CES-D), gender, and tonic alertness (a.m.). RESULTS: The final MLR model (R2 = .30) included tonic alertness (a.m.) (<.001), FSMCcog (.008), EDSS (.038), CES-D (.161), and gender (.057). Using this model, correct assignment to alertness performance groups was 56.8%. Tonic alertness (p.m.) demonstrated the greatest potential for differentiation among the three performance groups (<.001). CONCLUSIONS: These results show a relationship between subjective fatigue and tonic alertness. However, other variables also contribute to this association, suggesting that the RT differences between twice-daily measures of tonic alertness is not related to increased subjective fatigue in a substantial number of pwMS, which diminishes the diagnostic value. Further studies including relevant variables such as sleepiness are urgently needed.

Geometric regularity in webs of non-orb-weaving spiders.

Geometric regularity of spider webs has been intensively studied in orb-weaving spiders, although it is not exclusive of orb weavers. Here, we document the geometrically regular, repetitive elements in the webs of the non-orb-weaving groups Leptonetidae and Telemidae for the first time. Similar to orb weavers, we found areas with regularly spaced parallel lines in the webs of Calileptoneta helferi, Sulcia sp., and cf. Pinelema sp. Furthermore, we provide a detailed account of the regular webs of Ochyrocera (Ochyroceratidae). The sections of the web with regularly disposed parallel lines are built as U-shaped modules reminiscent of orb webs. It has been suggested that the regularly spaced parallel lines in the webs of Ochyroceratidae and Psilodercidae may be produced in a single sweep of their posterior lateral spinnerets, which have regularly spaced aciniform gland spigots, perhaps involving expansion of the spinnerets. To test this hypothesis, we compared the spacing between parallel lines with the spacing between spigots, searched for expansible membranes in the spinnerets, and examined the junctions of regularly spaced lines. The distance between parallel lines was 10-20 times the distance between spigots, and we found no expansible membranes, and the intersection of parallel lines are cemented, which opposes the single sweep hypothesis. Furthermore, we found cues of viscid silk in the parallel lines of the psilodercid Althepus and broadened piriform gland spigots that may be responsible of its production. Finally, we evaluated the presence or absence of geometrically regular web elements across the spider tree of life. We found reports of regular webs in 31 spider families, including 20 families that are not orb weavers and hypothesize that the two basic aspects of regularity (parallel lines spaced at regular intervals, and radial lines spaced at regular angles) probably appeared many times in the evolution of spiders.

Short and long-range cyclic patterns in flows of DNA solutions in microfluidic obstacle arrays.

We observe regular patterns emerging across multiple length scales with high-concentration DNA solutions in microfluidic pillar arrays at low Reynolds numbers and high Deborah numbers. Interacting vortices between pillars lead to long-range order in the form of large travelling waves consisting of DNA at high concentration and extension. Waves are formed in quadratic arrays of pillars, while randomizing the position of the pillar in each unit cell of a quadratic array leads to suppression of the long-range patterns. We find that concentrations exceeding the overlap concentration of the DNA enables the waves, and exploring the behavior of the waves as a function of flow rate, buffer composition, concentration and molecular length, we identify elastic effects as central to the origin of the waves. Our work may not only help increase the low throughput that often limits sample processing in microfluidics, it may also provide a platform for further studies of the underlying viscoelastic mechanisms.

Relacionamentos ab-usivos / Abusive relationships / Relation abusive / Relación ab-usiva

O presente artigo tem como objetivo compreender os aspectos que envolvem a permanência e a passividade das vítimas de relacionamentos abusivos a partir de conceitos psicanalíticos. Para tanto, utilizou-se conceitos de Freud e Lacan, articulando a relação do sujeito com a linguagem e suas implicações na relação sujeito e Outro, que envolvem aspectos como desejo, amor e gozo. Assim, optou-se por nomear tal dinâmica relacional de relacionamento ab-usivo, entendendo que algo da ordem de uma satisfação inconsciente se atualiza nessas relações, e que neste sentido, toda relação comporta algo de ab-usivo. Considera-se fundamental que, diante de tal demanda, o analista tenha um olhar para a cena inconsciente do sujeito que se apresenta, e que a análise possibilite que algo do "um" se sustente, para que o sujeito sustente uma condição de menos assujeitamento ao seu modo de gozo.

This article aims to understand the aspects that involve the permanence and passivity of victims of abusive relationships based on psychoanalytic concepts. For this purpose, concepts from Freud and Lacan were used, articulating the subject's relationship with language and its implications in the subject's relationship with the Other, which involve aspects such as desire, love, and jouissance. Thus, it was decided to name such relational dynamics an ab-usive relationship, understanding that something of the order of an unconscious satisfaction is actualized in these relationships, and that, in this sense, every relationship involves something ab-usive. It is considered essential that, in the face of such a demand, the analyst has a look at the unconscious scene of the subject who presents himself and that the analysis allows something of the "one" to be sustained so that the subject sustains a condition of less subjection to your way of jouissance.

Cet article vise à comprendre les aspects qui impliquent la permanence et la passivité des victimes de relations abusives à partir de concepts psychanalytiques. À cette fin, nous avons utilisé des concepts de Freud et de Lacan qui articulent la relation du sujet avec le langage et ses implications dans la relation du sujet à l'Autre, qui impliquent des aspects tels que le désir, l'amour et la jouissance. Nous avons donc choisi de nommer cette dynamique relationnelle relation a-busive, considérant qu'une sorte de satisfaction inconsciente se produit dans ces relations, et qu'en ce sens, toute relation contient quelque chose d'a-busif. Il est considéré comme fondamental que, face à une telle demande, l'analyste ait un regard sur la scène inconsciente du sujet qui se présente, et que l'analyse permette de soutenir "l'un", afin que le sujet parvienne à une condition de moindre soumission à son mode de jouissance.

El presente artículo tiene como objetivo comprender los aspectos que implican la permanencia y la pasividad de las víctimas de relaciones abusivas, usando como referencia los conceptos psicoanalíticos. Se utilizaron conceptos de Freud y Lacan, articulando la relación entre el sujeto y el lenguaje y sus implicaciones en la relación entre el sujeto y el Otro, que involucran aspectos tales como el deseo, el amor y el gozo. De esta forma, se decidió denominar a tal dinámica relacional como relación ab-usiva, entendiendo que algo del orden de una satisfacción inconsciente se actualiza en este tipo de relaciones, y que en ese sentido, toda relación implica algo ab-usivo. Se considera fundamental que, ante tal demanda, el analista tenga una mirada en la escena inconsciente del sujeto que se presenta, y que el análisis permita que algo del "uno" se sustente, para que el sujeto sustente una condición de menor sujeción a su modo de gozo.

Use of dielectrophoresis for directing T cells to microwells before nanostraw transfection: modelling and experiments.

Nanostraw substrates have great potential for achieving minimally invasive cell transfection. Cells located on the nanostraw substrate are subjected to mild DC electric pulses applied across the nanostraw substrate, which open pores in the cell membrane on top of the nanostraws and drives charged cargo through these pores via electrophoresis. However, with this method, the current may leak through uncovered nanostraws, thereby decreasing the desired effect in the cell-covered nanostraws. A minimization of the number of uncovered nanostraws could be achieved by high cell coverage, but this is challenging when working with small cell populations. Nanostraw substrates of smaller area could be covered by smaller cell populations but are hard to integrate into fluidics systems. Here, we use simulations and experiments to show that this issue can be addressed by covering the nanostraw substrate with an insulating layer containing pores of similar size to cells. The pores act as traps into which cells can be guided using dielectrophoresis, ensuring a high degree of occupancy while maintaining a high cell viability, even if the total number of cells is low.

High-Throughput Separation of Long DNA in Deterministic Lateral Displacement Arrays.

Length-based separation of DNA remains as relevant today as when gel electrophoresis was introduced almost 100 years ago. While new, long-read genomics technologies have revolutionised accessibility to powerful genomic data, the preparation of samples has not proceeded at the same pace, with sample preparation often constituting a considerable bottleneck, both in time and difficulty. Microfluidics holds great potential for automated, sample-to-answer analysis via the integration of preparatory and analytical steps, but for this to be fully realised, more versatile, powerful and integrable unit operations, such as separation, are essential. We demonstrate the displacement and separation of DNA with a throughput that is one to five orders of magnitude greater than other microfluidic techniques. Using a device with a small footprint (23 mm × 0.5 mm), and with feature sizes in the micrometre range, it is considerably easier to fabricate than parallelized nano-array-based approaches. We show the separation of 48.5 kbp and 166 kbp DNA strands achieving a significantly improved throughput of 760 ng/h, compared to previous work and the separation of low concentrations of 48.5 kbp DNA molecules from a massive background of sub 10 kbp fragments. We show that the extension of DNA molecules at high flow velocities, generally believed to make the length-based separation of long DNA difficult, does not place the ultimate limitation on our method. Instead, we explore the effects of polymer rotations and intermolecular interactions at extremely high DNA concentrations and postulate that these may have both negative and positive influences on the separation depending on the detailed experimental conditions.