Virtual slit scanning microscopy

We present a novel slit scanning confocal microscope with a CCD camera image sensor and a virtual slit aperture for descanning that can be adjusted during post-processing. A very efficient data structure and mathematical criteria for aligning the virtual aperture guarantee the ease of use. We further introduce a method to reduce the anisotropic lateral resolution of slit scanning microscopes. System performance is evaluated against a spinning disk confocal microscope on identical specimens. The virtual slit scanning microscope works as the spinning disk type and outperforms on thick specimen

Widefield fluorescence microscopy with extended resolution

Widefield fluorescence microscopy is seeing dramatic improvements in resolution, reaching today 100nm in all three dimensions. This gain in resolution is achieved by dispensing with uniform Köhler illumination. Instead, non-uniform excitation light patterns with sinusoidal intensity variations in one, two, or three dimensions are applied combined with powerful image reconstruction techniques. Taking advantage of non-linear fluorophore response to the excitation field, the resolution can be further improved down to several 10nm. In this review article, we describe the image formation in the microscope and computational reconstruction of the high-resolution dataset when exciting the specimen with a harmonic light pattern conveniently generated by interfering laser beams forming standing waves. We will also discuss extensions to total internal reflection microscopy, non-linear microscopy, and three-dimensional imagin

Schneller sehen durch Regelungstechnik - Moderne Bildgebung in der Nanotechnologie

Die Nanotechnologie, also die Wissenschaft von Untersuchungen und Manipulationen im Nanometermaßstab, wird als eine der Schlüsseltechnologien des 21. Jahrhunderts angesehen. Wichtige Komponenten dieses aktuellen Forschungszweiges sind das Rastertunnelmikroskop und das Rasterkraftmikroskop, die den Blick in die Nanowelt erlauben. Dieser Artikel gibt einen einführenden Einblick in die Bildgebung der Nanowelt und versucht, den Beitrag von Systemtheorie und Regelungstechnik für die Nanotechnologie anhand der Regelung von Rasterkraftmikroskopen allgemeinverständlich zu beschreiben

Deterministic assembly of linear gold nanorod chains as a platform for nanoscale applications

We demonstrate a method to assemble gold nanorods highly deterministically into a chain formation by means of directed capillary assembly. This way we achieved straight chains consisting of end-to-end aligned gold nanorods assembled in one specific direction with well-controlled gaps of [similar]6 nm between the individual constituents. We determined the conditions for optimum quality and yield of nanorod chain assembly by investigating the influence of template dimensions and assembly temperature. In addition, we transferred the gold nanorod chains from the assembly template onto a Si/SiO2 target substrate, thus establishing a platform for a variety of nanoscale electronic and optical applications ranging from molecular electronics to optical and plasmonic devices. As a first example, electrical measurements are performed on contacted gold nanorod chains before and after their immersion in a solution of thiol end-capped oligophenylenevinylene molecules showing an increase in the conductance by three orders of magnitude, indicating molecular-mediated transport

Robust Stabilization of Elastic Joint Robots by ESP and PID Control: Theory and Experiments

This work addresses the problem of global set-point control of elastic joint robots by combining elastic structure preserving (ESP) control with non-collocated integral action. Despite the popularity and extensive research on PID control for rigid joint robots, such schemes largely evaded adoption to elastic joint robots. This is mainly due to the underactuation inherent to these systems, which impedes the direct implementation of PID schemes with non-collocated (link position) feedback. We remedy this issue by using the recently developed concept of “quasi-full actuation,” to achieve a link-side PID control structure with “delayed” integral action. The design follows the structure preserving design philosophy of ESP control and ensures global asymptotic stability and local passivity of the closed loop. A key feature of the proposed controller is the switching logic for the integral action that enables the combination of excellent positioning accuracy in free motion with compliant manipulation in contact with the environment. Its performance is evaluated on an elastic joint testbed and a compliant robot arm. The results demonstrate that elastic robots may achieve positioning accuracy comparable to rigid joint robots

A Gravity Compensation Strategy for On-ground Validation of Orbital Manipulators

The on-ground validation of orbital manipulators is a challenging task because the robot is designed for a gravity-free operational environment, but it is validated under the effect of gravity. As a consequence, joint torque limits can be easily reached in certain configurations when gravity is actively compensated by the joints. Hence, the workspace for on-ground testing is restricted. In this paper, an optimal strategy is proposed for achieving gravity compensation of an orbital manipulator arm on ground. The strategy minimizes the joint torques acting on the manipulator by solving an optimization problem and it computes the necessary forces to be tracked by an external carrier. Hence, full gravity compensation is achieved for the orbital manipulator. Experimental results validate the effectiveness of the method on the DLR CAESAR space robot, which uses a cable suspended system as external carrier to track the desired gravity compensation force, resulting from the proposed method

Single-step assembly of a gene and entire plasmid from large numbers of oligodeoxyribonucleotides.

SUMMARY Here, we describe assembly PCR as a method for the synthesis of long DNA sequences from large numbers of oligodeoxyribonucleotides (oligos). The method, which is derived from DNA shuffling [Stemmer, Nature 370 (1994a) 389-391], does not rely on DNA ligase but instead relies on DNA polymerase to build increasingly longer DNA fragments during the assembly process. A 1.1-kb fragment containing the TEM-1 [3-1actamase-encoding gene (bla) was assembled in a single reaction from a total of 56 oligos, each 40 nucleotides (ntl in length. The synthetic gene was PCR amplified and cloned in a vector containing the tetracycline-resistance gene (Tc R) as the sole selectable marker. Without relying on ampicillin (Ap) selection, 76% of the Tc R colonies were Ap R, making this approach a general method for the rapid and cost-effective synthesis of any gene. We tested the range of assembly PCR by synthesizing, in a single reaction vessel containing 134 oligos, a high-molecular-mass multimeric form of a 2.7-kb plasmid containing the bla gene, the s-fragment of the lacZ gene and the pUC origin of replication. Digestion with a unique restriction enzyme, followed by ligation and transformation in Escherichia coli, yielded the correct plasmid. Assembly PCR is well suited for several in vitro mutagenesis strategies

Multistep screening and selection of COVID‐19 convalescent plasma donors at the early stage of the SARS‐CoV‐2 pandemic: A retrospective analysis

Background and Aims The COVID-19 pandemic reached Bavaria in February 2020. Almost simultaneously, Chinese physicians published reports on the first successful treatments with plasma from COVID-19 convalescent donors. With these silver linings on the horizon, we decided to establish the manufacturing of anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibody-containing plasma from COVID-19 convalescent donors at our site. Here we describe our donor selection process, built from the ground up, which enabled us to cope with the immense resonance after our social media call for donors. Methods As a first step, we created a specific questionnaire for telephone interviews applied by trained students to filter the wave of callers interested in plasma donation. Afterward, the medical staff evaluated the hotline questionnaires and chose eligible donors to be invited for on-site donor evaluation. Data documentation was performed with MS Excel, and statistical analyses were calculated with GraphPad Prism 8. A quantitative in-house ELISA was used to detect anti-SARS-CoV-2 antibodies and determine specific titers. Results Out of 1465 calls from potential plasma donors, we could register 420 persons with a completed questionnaire. Evaluation of questionnaires identified 222 of 420 persons as eligible for donation, and 55 were directly asked for on-site donor qualification. Subsequently, as anti-SARS-CoV-2 antibody titers ≥1:800 were required, we invited 89 of 222 potential donors for an antibody screening. This procedure resulted in another 28 potential donors for an on-site evaluation. Finally, 12 donors qualified with a titer of 1:400 and 24 with ≥1:800. Conclusion Identifying suitable COVID-19 convalescent plasma donors was expected to be highly time-consuming. Implementing a screening procedure with our hotline questionnaire helped us streamline the donor selection process and reduce the workload for the staff. We propose combining the described selection process with the later introduced on-site antibody screening as an effective strategy

Identification and Characterization of Human Observational Studies in Nutritional Epidemiology on Gut Microbiomics for Joint Data Analysis

In any research field, data access and data integration are major challenges that even large, well-established consortia face. Although data sharing initiatives are increasing, joint data analyses on nutrition and microbiomics in health and disease are still scarce. We aimed to identify observational studies with data on nutrition and gut microbiome composition from the Intestinal Microbiomics (INTIMIC) Knowledge Platform following the findable, accessible, interoperable, and reusable (FAIR) principles. An adapted template from the European Nutritional Phenotype Assessment and Data Sharing Initiative (ENPADASI) consortium was used to collect microbiome-specific information and other related factors. In total, 23 studies (17 longitudinal and 6 cross-sectional) were identified from Italy (7), Germany (6), Netherlands (3), Spain (2), Belgium (1), and France (1) or multiple countries (3). Of these, 21 studies collected information on both dietary intake (24 h dietary recall, food frequency questionnaire (FFQ), or Food Records) and gut microbiome. All studies collected stool samples. The most often used sequencing platform was Illumina MiSeq, and the preferred hypervariable regions of the 16S rRNA gene were V3-V4 or V4. The combination of datasets will allow for sufficiently powered investigations to increase the knowledge and understanding of the relationship between food and gut microbiome in health and disease

Current status and recommendations for biomarkers and biobanking in neurofibromatosis

Objective: Clinically validated biomarkers for neurofibromatosis 1 (NF1), neurofibromatosis 2 (NF2), and schwannomatosis (SWN) have not been identified to date. The biomarker working group’s goals are to (1) define biomarker needs in NF1, NF2, and SWN; (2) summarize existing data on biomarkers in NF1, NF2, and SWN; (3) outline recommendations for sample collection and biomarker development; and (4) standardize sample collection and methodology protocols where possible to promote comparison between studies by publishing standard operating procedures (SOPs). Methods: The biomarker group reviewed published data on biomarkers in NF1, NF2, and SWN and on biobanking efforts outside these diseases via literature search, defined the need for biomarkers in NF, and developed recommendations in a series of consensus meetings. Results: We describe existing biomarkers in NF and report consensus recommendations for SOP and a minimal clinical dataset to accompany samples derived from patients with NF1, NF2, and SWN in decentralized biobanks. Conclusions: These recommendations are intended to provide clinicians and researchers with a common set of guidelines to collect and store biospecimens and for establishment of biobanks for NF1, NF2, and SWN