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.

[Reform of experimental course of large-scale life science equipment in universities].

The opening and sharing of large-scale life science equipment in universities is expanding. We must find ways to improve the role of large-scale equipment in cultivating students' practical and innovative abilities, to tap the potential of equipment, then to support scientific research and speed up scientific research output. We established a set of large-scale equipment training and practice teaching system including 15 topics and covering a wide range of technologies. In practice, we constantly innovated personalized courses. According to the differences of students' major and scientific research needs, we classified teaching content and set up parallel classes. Each class had the individualized teaching content and students could select from a menu of courses. In addition, we built up a cloud classroom teaching platform, online and offline teaching method supplemented each other. The teaching system have produced certain effect.

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.

Thiol-Ene Based Polymers as Versatile Materials for Microfluidic Devices for Life Sciences Applications.

While there is a steady growth in the number of microfluidics applications, the search for an optimal material that delivers the diverse characteristics needed for the numerous tasks is still nowhere close to being settled. Often overlooked and still underrepresented, the thiol-ene family of polymer materials has an enormous potential for applications in organs-on-a-chip, droplet productions, microanalytics, and point of care testing. In this review, the main characteristics of the thiol-ene materials are given, and advantages and drawbacks with respect to their potential in microfluidic chip fabrication are critically assessed. Select applications, which exploit the versatility of the thiol-ene polymers, are presented and discussed. It is concluded that, in particular, the rapid prototyping possibility combined with the material's resulting mechanical strength, solvent resistance, and biocompatibility, as well as the inherently easy surface functionalization, are strong factors to make thiol-ene polymers strong contenders for promising future materials for many biological, clinical, and technical lab-on-a-chip applications.

The Internet of Things Is Digitizing and Transforming Science.

Over the past decade, cloud software has transformed numerous industries-from finance to logistics, marketing to manufacturing. The simplified aggregation of data, enabled by cloud computing, empowers individuals to glean insights and make data-driven decisions rapidly. In science, however, such a transformation has yet to emerge. The domain lacks centralized, machine-readable repositories of scientific data; this absence inhibits analytics and expedient decision-making. Recently, the Internet of Things (IoT) has served as a catalyst for digitizing and automating science. IoT enables the centralized collection and analysis of scientific data (e.g., instruments, sensors, and environments). Here, we discuss this new technology trend, its applications in laboratories and promise as a platform for improved efficiency, more innovative capabilities, and machine learning/artificial intelligence.

Achieving Reproducibility and Closed-Loop Automation in Biological Experimentation with an IoT-Enabled Lab of the Future.

A robotic cloud laboratory driven by a state-of-the-art unified laboratory operating system integrates automated hardware, humans, and sensors. This lab of the future system enables researchers to transparently and collaboratively create, optimize, and organize biological experiments to achieve more reproducible results, perform around-the-clock experimentation, and more efficiently navigate the vast parameter space of biology.

BOOTSTRAPPING BIOLOGY.

Sarah Webb explores how DIYbio strategies are helping biologists in traditional labs do better science.

A guide to light-sheet fluorescence microscopy for multiscale imaging.

The impact of light-sheet fluorescence microscopy (LSFM) is visible in fields as diverse as developmental and cell biology, anatomical science, biophysics and neuroscience. Although adoption among biologists has been steady, LSFM has not displaced more traditional imaging methods despite its often-superior performance. One reason for this is that the field has largely conformed to a do-it-yourself ethic, although the challenges of big image data cannot be overstated. With the most powerful implementations of LSFM available to only a few groups worldwide, the scope of this technique is unnecessarily limited. Here we elucidate the key developments and define a simple set of underlying principles governing LSFM. In doing so, we aim to clarify the decisions to be made for those who wish to develop and use bespoke light-sheet systems and to assist in identifying the best approaches to apply this powerful technique to myriad biological questions.

Application of EELS and EFTEM to the life sciences enabled by the contributions of Ondrej Krivanek.

The pioneering contributions of Ondrej Krivanek to the development of electron energy loss spectrometers, energy filters, and detectors for transmission and scanning transmission electron microscopes have provided researchers with indispensible tools across a wide range of disciplines in the physical sciences, ranging from condensed matter physics, to chemistry, mineralogy, materials science, and nanotechnology. In addition, the same instrumentation has extended its reach into the life sciences, and it is this aspect of Ondrej Krivanek's influential contributions that will be surveyed here, together with some personal recollections. Traditionally, electron microscopy has given a purely morphological view of the biological structures that compose cells and tissues. However, the availability of high-performance electron energy loss spectrometers and energy filters offers complementary information about the elemental and chemical composition at the subcellular scale. Such information has proven to be valuable for applications in cell and structural biology, microbiology, histology, pathology, and more generally in the biomedical sciences.

Recent advances in X-ray compatible microfluidics for applications in soft materials and life sciences.

The increasingly narrow and brilliant beams at X-ray facilities reduce the requirements for both sample volume and data acquisition time. This creates new possibilities for the types and number of sample conditions that can be examined but simultaneously increases the demands in terms of sample preparation. Microfluidic-based sample preparation techniques have emerged as elegant alternatives that can be integrated directly into the experimental X-ray setup remedying several shortcomings of more traditional methods. We review the use of microfluidic devices in conjunction with X-ray measurements at synchrotron facilities in the context of 1) mapping large parameter spaces, 2) performing time resolved studies of mixing-induced kinetics, and 3) manipulating/processing samples in ways which are more demanding or not accessible on the macroscale. The review covers the past 15 years and focuses on applications where synchrotron data collection is performed in situ, i.e. directly on the microfluidic platform or on a sample jet from the microfluidic device. Considerations such as the choice of materials and microfluidic designs are addressed. The combination of microfluidic devices and measurements at large scale X-ray facilities is still emerging and far from mature, but it definitely offers an exciting array of new possibilities.

Applications of "Tender" Energy (1-5 keV) X-ray Absorption Spectroscopy in Life Sciences.

The "tender" energy range of 1 to 5 keV, between the energy ranges of most "hard" (>5 keV) and "soft" (<1 keV) synchrotron X-ray facilities, offers some unique opportunities for synchrotron- based X-ray absorption fine structure spectroscopy in life sciences. In particular the K absorption edges of Na through Ca offer opportunities to study local structure, speciation, and chemistry of many important biological compounds, structures and processes. This is an area of largely untapped science, in part due to a scarcity of optimized facilities. Such measurements also entail unique experimental challenges. This brief review describes the technique, its experimental challenges, recent progress in development of microbeam measurement capabilities, and several highlights illustrating applications in life sciences.

Moving Liquids with Sound: The Physics of Acoustic Droplet Ejection for Robust Laboratory Automation in Life Sciences.

Liquid handling instruments for life science applications based on droplet formation with focused acoustic energy or acoustic droplet ejection (ADE) were introduced commercially more than a decade ago. While the idea of "moving liquids with sound" was known in the 20th century, the development of precise methods for acoustic dispensing to aliquot life science materials in the laboratory began in earnest in the 21st century with the adaptation of the controlled "drop on demand" acoustic transfer of droplets from high-density microplates for high-throughput screening (HTS) applications. Robust ADE implementations for life science applications achieve excellent accuracy and precision by using acoustics first to sense the liquid characteristics relevant for its transfer, and then to actuate transfer of the liquid with customized application of sound energy to the given well and well fluid in the microplate. This article provides an overview of the physics behind ADE and its central role in both acoustical and rheological aspects of robust implementation of ADE in the life science laboratory and its broad range of ejectable materials.

Single-walled carbon nanotubes as near-infrared optical biosensors for life sciences and biomedicine.

Single-walled carbon nanotubes that emit photostable near-infrared fluorescence have emerged as near-infrared optical biosensors for life sciences and biomedicine. Since the discovery of their near-infrared fluorescence, researchers have engineered single-walled carbon nanotubes to function as an optical biosensor that selectively modulates its fluorescence upon binding of target molecules. Here we review the recent advances in the single-walled carbon nanotube-based optical sensing technology for life sciences and biomedicine. We discuss the structure and optical properties of single-walled carbon nanotubes, the mechanisms for molecular recognition and signal transduction in single-walled carbon nanotube complexes, and the recent development of various single-walled carbon nanotube-based optical biosensors. We also discuss the opportunities and challenges to translate this emerging technology into biomedical research and clinical use, including the biological safety of single-walled carbon nanotubes. The advances in single-walled carbon nanotube-based near-infrared optical sensing technology open up a new avenue for in vitro and in vivo biosensing with high sensitivity and high spatial resolution, beneficial for many areas of life sciences and biomedicine.

Amino acidic scaffolds bearing unnatural side chains: an old idea generates new and versatile tools for the life sciences.

The unnatural amino acids (UAAs) are members of a class of molecules with relevant impacts in the life sciences. Due to the role of these molecules in the modulation of the chemical and physical properties of biological and inorganic materials, UAAs have attracted increasing interest in recent years. The aim of this review is to highlight (i) the most recent and innovative synthetic routes for the preparation of UAAs, (ii) the recently marketed UAA-based drugs, and (iii) the most promising technological applications involving novel UAA-containing molecular entities.

Teaching about photosynthesis with simple equipment: analysis of light-induced changes in fluorescence and reflectance of plant leaves.

Solar energy absorbed by plants results in either reflection or absorption. The latter results in photosynthesis, fluorescence, or heat. Measurements of fluorescence changes have been used for monitoring processes associated with photosynthesis. A simple method to follow changes in leaf fluorescence and leaf reflectance associated with nonphotochemical quenching and light acclimation of leaves is described. The main equipment needed consists of a green-light emitting laser pointer, a digital camera, and a personal computer equipped with the camera acquisition software and the programs ImageJ and Excel. Otherwise, only commonly available cheap materials are required.