Staying on track - Keeping things running in a high-end scientific imaging core facility.

Modern life science research is a collaborative effort. Few research groups can single-handedly support the necessary equipment, expertise and personnel needed for the ever-expanding portfolio of technologies that are required across multiple disciplines in today's life science endeavours. Thus, research institutes are increasingly setting up scientific core facilities to provide access and specialised support for cutting-edge technologies. Maintaining the momentum needed to carry out leading research while ensuring high-quality daily operations is an ongoing challenge, regardless of the resources allocated to establish such facilities. Here, we outline and discuss the range of activities required to keep things running once a scientific imaging core facility has been established. These include managing a wide range of equipment and users, handling repairs and service contracts, planning for equipment upgrades, renewals, or decommissioning, and continuously upskilling while balancing innovation and consolidation.

Ångström-resolution fluorescence microscopy.

Fluorescence microscopy, with its molecular specificity, is one of the major characterization methods used in the life sciences to understand complex biological systems. Super-resolution approaches1-6 can achieve resolution in cells in the range of 15 to 20 nm, but interactions between individual biomolecules occur at length scales below 10 nm and characterization of intramolecular structure requires Ångström resolution. State-of-the-art super-resolution implementations7-14 have demonstrated spatial resolutions down to 5 nm and localization precisions of 1 nm under certain in vitro conditions. However, such resolutions do not directly translate to experiments in cells, and Ångström resolution has not been demonstrated to date. Here we introdue a DNA-barcoding method, resolution enhancement by sequential imaging (RESI), that improves the resolution of fluorescence microscopy down to the Ångström scale using off-the-shelf fluorescence microscopy hardware and reagents. By sequentially imaging sparse target subsets at moderate spatial resolutions of >15 nm, we demonstrate that single-protein resolution can be achieved for biomolecules in whole intact cells. Furthermore, we experimentally resolve the DNA backbone distance of single bases in DNA origami with Ångström resolution. We use our method in a proof-of-principle demonstration to map the molecular arrangement of the immunotherapy target CD20 in situ in untreated and drug-treated cells, which opens possibilities for assessing the molecular mechanisms of targeted immunotherapy. These observations demonstrate that, by enabling intramolecular imaging under ambient conditions in whole intact cells, RESI closes the gap between super-resolution microscopy and structural biology studies and thus delivers information key to understanding complex biological systems.

BASIN: A Semi-automatic Workflow, with Machine Learning Segmentation, for Objective Statistical Analysis of Biomedical and Biofilm Image Datasets.

Micrograph comparison remains useful in bioscience. This technology provides researchers with a quick snapshot of experimental conditions. But sometimes a two- condition comparison relies on researchers' eyes to draw conclusions. Our Bioimage Analysis, Statistic, and Comparison (BASIN) software provides an objective and reproducible comparison leveraging inferential statistics to bridge image data with other modalities. Users have access to machine learning-based object segmentation. BASIN provides several data points such as images' object counts, intensities, and areas. Hypothesis testing may also be performed. To improve BASIN's accessibility, we implemented it using R Shiny and provided both an online and offline version. We used BASIN to process 498 image pairs involving five bioscience topics. Our framework supported either direct claims or extrapolations 57% of the time. Analysis results were manually curated to determine BASIN's accuracy which was shown to be 78%. Additionally, each BASIN version's initial release shows an average 82% FAIR compliance score.

Extracting, quantifying, and comparing dynamical and biomechanical properties of living matter through single particle tracking.

A panoply of new tools for tracking single particles and molecules has led to an explosion of experimental data, leading to novel insights into physical properties of living matter governing cellular development and function, health and disease. In this Perspective, we present tools to investigate the dynamics and mechanics of living systems from the molecular to cellular scale via single-particle techniques. In particular, we focus on methods to measure, interpret, and analyse complex data sets that are associated with forces, materials properties, transport, and emergent organisation phenomena within biological and soft-matter systems. Current approaches, challenges, and existing solutions in the associated fields are outlined in order to support the growing community of researchers at the interface of physics and the life sciences. Each section focuses not only on the general physical principles and the potential for understanding living matter, but also on details of practical data extraction and analysis, discussing limitations, interpretation, and comparison across different experimental realisations and theoretical frameworks. Particularly relevant results are introduced as examples. While this Perspective describes living matter from a physical perspective, highlighting experimental and theoretical physics techniques relevant for such systems, it is also meant to serve as a solid starting point for researchers in the life sciences interested in the implementation of biophysical methods.

Testing how different narrative perspectives achieve communication objectives and goals in online natural science videos.

Communication of science through online media has become a primary means of disseminating and connecting science with a public audience. However, online media can come in many forms and stories of scientific discovery can be told by many individuals. We tested whether the relationship of a spokesperson to the science story being told (i.e., the narrative perspective) influences how people react and respond to online science media. We created five video stimuli that fell into three treatments: a scientist presenting their own research (male or female), a third-party summarizing research (male or female), and an infographic-like video with no on-screen presenter. Each of these videos presented the same fabricated science story about the discovery of a new ant species (Formicidae). We used Qualtrics to administer and obtain survey responses from 515 participants (~100 per video). Participants were randomly assigned to one of the videos and after viewing the stimulus answered questions assessing their perceptions of the video (trustworthiness and enjoyment), the spokesperson (trustworthiness and competence), scientists in general (competence and warmth), and attitudes towards the research topic and funding. Participants were also asked to recall what they had seen and heard. We determined that when participants watched a video in which a scientist presented their own research, participants perceived the spokesperson as having more expertise than a third-party presenter, and as more trustworthy and having more expertise than the no-spokesperson stimuli. Viewing a scientist presenting their own work also humanized the research, with participants more often including a person in their answer to the recall question. Overall, manipulating the narrative perspective of the source of a single online video communication effort is effective at impacting immediate objective outcomes related to spokesperson perceptions, but whether those objectives can positively influence long-term goals requires more investigation.

Causal and non-causal explanations in code biology.

In the philosophy of science, we can consider debates about the nature of non-causal explanations in general (e.g. Reutlinger, Saatsi 2018; Lange 2017) and then especially those in the life sciences (e.g. Huneman, 2018; Kostic 2020). These debates are accompanied by the development of a new mechanism that is becoming the major response to the nature of scientific explanation in the life sciences (e.g. Craver, Darden 2013; Craver 2006); and also by the development of a design explanation (e.g. Eck, Mennes 2016) that represents a modern variant of a functional explanation. In this paper, we will methodically: 1. evaluate the plurality of explanatory strategies in contemporary science (chapter 2). 2. describe the mechanical philosophy and mechanistic explanation (Glennan 2016; Craver, Darden 2013, etc.) (chapter 3). 3. explicate the role of mechanisms in code biology (Barbieri 2015, 2002, etc.) and its relation to the new mechanism (chapter 4). 4. fulfill the main goal of the paper - to apply mechanistic explanations in code biology (Barbieri 2019, etc.) and to apply their suitability for this scientific domain (chapter 5).

Design in biology and rational design in vaccinology: A conceptual analysis.

This review discusses the philosophical foundations of what used to be called "the scientific method" and is nowadays often known as the scientific attitude. It used to be believed that scientific theories and methods aimed at the truth especially in the case of physics, chemistry and astronomy because these sciences were able to develop numerous scientific laws that made it possible to understand and predict many physical phenomena. The situation is different in the case of the biological sciences which deal with highly complex living organisms made up of huge numbers of constituents that undergo continuous dynamic processes; this leads to novel emergent properties in organisms that cannot be predicted because they are not present in the constituents before they have interacted with each other. This is one of the reasons why there are no universal scientific laws in biology. Furthermore, all scientific theories can only achieve a restricted level of predictive success because they remain valid only under the limited range of conditions that were used for establishing the theory' in the first place. Many theories that used to be accepted were subsequently shown to be false, demonstrating that scientific theories always remain tentative and can never be proven beyond and doubt. It is ironical that as scientists have finally accepted that approximate truths are perfectly adequate and that absolute truth is an illusion, a new irrational sociological phenomenon called Post-Truth conveyed by social media, the Internet and fake news has developed in the Western world that is convincing millions of people that truth simply does not exist. Misleading information is circulated with the intention to deceive and science denialism is promoted by denying the remarkable achievements of science and technology during the last centuries. Although the concept of intentional design is widely used to describe the methods that biologists use to make discoveries and inventions, it will be argued that the term is not appropriate for explaining the appearance of life on our planet nor for describing the scientific creativity of scientific investigators. The term rational for describing the development of new vaccines is also unjustified. Because the analysis of the COVID-19 pandemic requires contributions from biomedical and psycho-socioeconomic sciences, one scientific method alone would be insufficient for combatting the pandemic.

The "Fourth Musketeer". Bioethics: Within life sciences - or above? A choice to make now.

The article deals with the different organisational units in current bioethics, and their main characteristics concerning life sciences. Three out of four organisational units of current bioethics jeopardise the autonomy of life sciences by establishing rules, regulations and laws, regulating and restricting life sciences. Only the integration of bioethics in life science serves the interests of life sciences.

The FEBS Journal: hidden gems.

Every quarter, The FEBS Journal presents some of its "hidden gems"-original research and review-type articles that provide a significant advance or discuss recent developments in the molecular or cellular life sciences. These articles are of high value to the scientific community, and we like to take the opportunity to promote these contributions from previous issues of the journal, as we feel their scientific content merits a boost in exposure.

MultipleTesting.com: A tool for life science researchers for multiple hypothesis testing correction.

Scientists from nearly all disciplines face the problem of simultaneously evaluating many hypotheses. Conducting multiple comparisons increases the likelihood that a non-negligible proportion of associations will be false positives, clouding real discoveries. Drawing valid conclusions require taking into account the number of performed statistical tests and adjusting the statistical confidence measures. Several strategies exist to overcome the problem of multiple hypothesis testing. We aim to summarize critical statistical concepts and widely used correction approaches while also draw attention to frequently misinterpreted notions of statistical inference. We provide a step-by-step description of each multiple-testing correction method with clear examples and present an easy-to-follow guide for selecting the most suitable correction technique. To facilitate multiple-testing corrections, we developed a fully automated solution not requiring programming skills or the use of a command line. Our registration free online tool is available at www.multipletesting.com and compiles the five most frequently used adjustment tools, including the Bonferroni, the Holm (step-down), the Hochberg (step-up) corrections, allows to calculate False Discovery Rates (FDR) and q-values. The current summary provides a much needed practical synthesis of basic statistical concepts regarding multiple hypothesis testing in a comprehensible language with well-illustrated examples. The web tool will fill the gap for life science researchers by providing a user-friendly substitute for command-line alternatives.

Europe PMC in 2020.

Europe PMC (https://europepmc.org) is a database of research articles, including peer reviewed full text articles and abstracts, and preprints - all freely available for use via website, APIs and bulk download. This article outlines new developments since 2017 where work has focussed on three key areas: (i) Europe PMC has added to its core content to include life science preprint abstracts and a special collection of full text of COVID-19-related preprints. Europe PMC is unique as an aggregator of biomedical preprints alongside peer-reviewed articles, with over 180 000 preprints available to search. (ii) Europe PMC has significantly expanded its links to content related to the publications, such as links to Unpaywall, providing wider access to full text, preprint peer-review platforms, all major curated data resources in the life sciences, and experimental protocols. The redesigned Europe PMC website features the PubMed abstract and corresponding PMC full text merged into one article page; there is more evident and user-friendly navigation within articles and to related content, plus a figure browse feature. (iii) The expanded annotations platform offers ∼1.3 billion text mined biological terms and concepts sourced from 10 providers and over 40 global data resources.

Questionable research practices may have little effect on replicability.

This article examines why many studies fail to replicate statistically significant published results. We address this issue within a general statistical framework that also allows us to include various questionable research practices (QRPs) that are thought to reduce replicability. The analyses indicate that the base rate of true effects is the major factor that determines the replication rate of scientific results. Specifically, for purely statistical reasons, replicability is low in research domains where true effects are rare (e.g., search for effective drugs in pharmacology). This point is under-appreciated in current scientific and media discussions of replicability, which often attribute poor replicability mainly to QRPs.

Forster resonance energy transfer for studying nucleic acids denaturation: A chemical and biological sciences laboratory experiment.

The Förster resonance energy transfer (FRET) melting assay intends to evaluate the unfolding, denaturation process of DNA secondary structures, and its stabilization using compounds known as DNA binders, some of which are highly specific for G-quadruplex DNAs versus duplex DNAs. First, students determined the melting temperature (Tm ) of DNA sequences double labeled with 5'-FAM (fluorescein) and 3'-TAMRA (tetramethylrhodamine) in the absence of DNA binders. Second, they determined the melting temperature of the DNAs in the presence of DNA binders by monitoring fluorescence. After completing this experiment, students understood that this method allows a semiquantitative analysis to test a variety of DNA binders against DNA secondary structures, and it can be used to rapidly identify the most promising drug candidates in the drug development stages at the basic research level.

Practising co-production and interdisciplinarity: Challenges and implications for one health research.

We review the nature of interdisciplinary research in relation to One Health, a perspective on human-animal health which would appear to merit close interdisciplinary cooperation to inform public health policy. We discuss the relationship between biological sciences, epidemiology and the social sciences and note that interdisciplinary work demands attention be given to a range of often neglected epistemological and methodological issues. Epidemiologists may sometimes adopt social science techniques as "bolt-ons"1 to their research without having a complete understanding of how the social sciences work. The paper introduces a range of social science concepts and applies them to the challenges of understanding and practicing participatory and local epidemiology. We consider the problem of co-production of knowledge about One Health and zoonotic diseases in relation to funding structures, working in large international teams and explore some of the often-neglected realities of working across disciplines and cultures. We do this in part by applying the concept of value-chain to the research process.

X-ray computed tomography in life sciences.

Recent developments within micro-computed tomography (µCT) imaging have combined to extend our capacity to image tissue in three (3D) and four (4D) dimensions at micron and sub-micron spatial resolutions, opening the way for virtual histology, live cell imaging, subcellular imaging and correlative microscopy. Pivotal to this has been the development of methods to extend the contrast achievable for soft tissue. Herein, we review the new capabilities within the field of life sciences imaging, and consider how future developments in this field could further benefit the life sciences community.

Integrating the basic sciences in medical curricula: focus on the basic scientists.

Basic science educators are not trained as clinicians, yet are expected to adjust their content to mesh appropriately with its clinical application. While achievable, this is a challenge that requires intentional effort on the part of the basic science educators. A practical solution to facilitate curricular integration is to create experiential opportunities for basic scientists to observe the clinical application of their content and to pair these initiatives with training in effective medical education practices.

Visual Life Sciences Workflow Design Using Distributed and Heterogeneous Resources.

Programming or querying usually presupposes some degree of technical familiarity with the syntax of a language and the peculiarity of the objects it manipulates to produce useful information. The degree of abstractions supported in a language helps lessen the depth of such familiarity needed, and aids in improving access to and usability of these resources. To help biologists concentrate more on their science questions and not on how to compute it, several successful workflow orchestration languages and systems have been proposed. Despite their popularity, significant limitations reduce their usability and limit applicability in novel applications. In this paper, we present a visual language, called VisFlow, for workflow orchestration using heterogeneous and distributed resources. We advance the idea that once resources are minimally described and abstracted, arbitrary workflows can be designed solely using query primitives supported in VisFlow. Its capabilities can be augmented by including computational artifacts in the form of library functions written in R, Python, and Java, or even in SQL and XQuery, making it a truly extensible system. We discuss its salient features and illustrate its capabilities using a substantial set of examples.

Open Source Tools for Biological Image Analysis.

Visiting the Bio Imaging Search Engine (BISE) (Bio, BISE, Engine, http://biii.eu/, Imaging, Search) website at the time of writing this article, almost 1200 open source assets (components, workflows, collections) were found. This overwhelming range of offer difficults the fact of making a reasonable choice, especially to newcomers. In the following chapter, we briefly sketch the advantages of the open source software (OSS) particularly used for image analysis in the field of life sciences. We introduce both the general OSS idea as well as some programs used for image analysis. Even more, we outline the history of ImageJ as it has served as a role model for the development of more recent software packages. We focus on the programs that are, to our knowledge, the most relevant and widely used in the field of light microscopy, as well as the most commonly used within our facility. In addition, we briefly discuss recent efforts and approaches aimed to share and compare algorithms and introduce software and data sharing good practices as a promising strategy to facilitate reproducibility, software understanding, and optimal software choice for a given scientific problem in the future.

Recent advances in the fabrication and application of nanomaterial-based enzymatic microsystems in chemical and biological sciences.

Novel enzymatic microsystems have strong prospects in chemical and biological analysis due to their low consumption of reagents, fast analysis time, easy manipulation and satisfactory portability. Nanomaterials (NMs) provide a favorable platform for integrating enzymes into microsystems with enhanced selectivity and sensitivity. Various NM-enzyme immobilization strategies applied in the fabrication of capillary-based and chip-based enzymatic microsystems are summarized in this manuscript. We focus on highlighting the advantages of employing NM-based enzymatic microsystems for enantioseparation, inhibitor screening, bioreaction and biosensing. Innovative nanocomposites and NM-functionalized monoliths used to construct multienzymatic microsystems are also illustrated. The general development trend identified in this review indicates that the application of NMs has significantly improved enzymatic microsystem performance.