Live Streaming of a Single Cell's Life over a Local pH-Monitoring Nanowire Waveguide.

Spatiotemporal pH monitoring of single living cells across rigid cell and organelle membranes has been challenging, despite its significance in understanding cellular heterogeneity. Here, we developed a mechanically robust yet tolerably thin nanowire waveguide that enables in situ monitoring of pH dynamics at desired cellular compartments via direct optical communication. By chemically labeling fluorescein at one end of a poly(vinylbenzyl azide) nanowire, we continuously monitored pH variations of different compartments inside a living cell, successfully observing organelle-exclusive pH homeostasis and stimuli-selective pH regulations. Importantly, it was demonstrated for the first time that, during the mammalian cell cycle, the nucleus displays pH homeostasis in interphase but a tidal pH curve in the mitotic phase, implying the existence of independent pH-regulating activities by the nuclear envelope. The rapid and accurate local pH-reporting capability of our nanowire waveguide would be highly valuable for investigating cellular behaviors under diverse biological situations in living cells.

Drop impact on hot plates: contact times, lift-off and the lamella rupture.

When a liquid drop impacts on a heated substrate, it can remain deposited, or violently boil in contact, or lift off with or without ever touching the surface. The latter is known as the Leidenfrost effect. The duration and area of the liquid-substrate contact are highly relevant for the heat transfer, as well as other effects such as corrosion. However, most experimental studies rely on side view imaging to determine contact times, and those are often mixed with the time until the drop lifts off from the substrate. Here, we develop and validate a reliable method of contact time determination using high-speed X-ray imaging and total internal reflection imaging. We exemplarily compare contact and lift-off times on flat silicon and sapphire substrates. We show that drops can rebound even without formation of a complete vapor layer, with a wide range of lift-off times. On sapphire, we find a local minimum of lift-off times that is much shorter than expected from capillary rebound in the comparatively low-temperature regime of transition boiling/thermal atomization. We elucidate the underlying mechanism related to spontaneous rupture of the lamella and receding of the contact area.

Progression in the Fountain Pen Approach: From 2D Writing to 3D Free-Form Micro/Nanofabrication.

The fountain pen approach, as a means for transferring materials to substrates, has shown numerous incarnations in recent years for creating 2D micro/nanopatterns and even generating 3D free-form nanostructures using a variety of material "inks". While the idea of filled reservoirs used to deliver material to a substrate via a capillary remains unchanged since antiquity, the advent of precise micromanipulation systems and functional material "inks" allows the extension of this mechanism to more high-tech applications. Herein, the recent growth in meniscus guided fountain pen approaches for benchtop micro/nanofabrication, which has occurred in the last decade, is discussed. Particular attention is given to the theory, equipment, and experimentation encompassing this unique direct writing approach. A detailed exploration of the diverse ink systems and functional device applications borne from this strategy is put forth to reveal its rapid expansion to a broad range of scientific and engineering disciplines. As such, this informative review is provided for researchers considering adoption of this recent advancement of a familiar technology.

Patterning droplets with durotaxis.

Numerous cell types have shown a remarkable ability to detect and move along gradients in stiffness of an underlying substrate--a process known as durotaxis. The mechanisms underlying durotaxis are still unresolved, but generally believed to involve active sensing and locomotion. Here, we show that simple liquid droplets also undergo durotaxis. By modulating substrate stiffness, we obtain fine control of droplet position on soft, flat substrates. Unlike other control mechanisms, droplet durotaxis works without imposing chemical, thermal, electrical, or topographical gradients. We show that droplet durotaxis can be used to create large-scale droplet patterns and is potentially useful for many applications, such as microfluidics, thermal control, and microfabrication.

Alveolar Microdynamics during Tidal Ventilation in Live Animals Imaged by SPring-8 Synchrotron.

It is self-evident that our chests expand and contract during breathing but, surprisingly, exactly how individual alveoli change shape over the respiratory cycle is still a matter of debate. Some argue that all the alveoli expand and contract rhythmically. Others claim that the lung volume change is due to groups of alveoli collapsing and reopening during ventilation. Although this question might seem to be an insignificant detail for healthy individuals, it might be a matter of life and death for patients with compromised lungs. Past analyses were based on static post-mortem preparations primarily due to technological limitations, and therefore, by definition, incapable of providing dynamic information. In contrast, this study provides the first comprehensive dynamic data on how the shape of the alveoli changes, and, further, provides valuable insights into the optimal lung volume for efficient gas exchange. It is concluded that alveolar micro-dynamics is nonlinear; and at medium lung volume, alveoli expand more than the ducts.

Self-pinning by colloids confined at a contact line.

Colloidal particles suspended in a fluid usually inhibit complete wetting of the fluid on a solid surface and cause pinning of the contact line, known as self-pinning. We show differences in spreading and drying behaviors of pure and colloidal droplets using optical and confocal imaging methods. These differences come from spreading inhibition by colloids confined at a contact line. We propose a self-pinning mechanism based on spreading inhibition by colloids. We find a good agreement between the mechanism and the experimental result taken by directly tracking individual colloids near the contact lines of evaporating colloidal droplets.

Spherical alveolar shapes in live mouse lungs.

Understanding how the alveolar mechanics work in live lungs is essential for comprehending how the lung behaves during breathing. Due to the lack of appropriate imaging tools, previous research has suggested that alveolar morphologies are polyhedral rather than spherical based on a 2D examination of alveoli in fixed lungs. Here, we directly observe high-resolution 3D alveoli in live mice lungs utilizing synchrotron x-ray microtomography to show spherical alveolar morphologies from the live lungs. Our measurements from x-ray microtomography show high sphericity, low packing density, big alveolar size, and low osmotic pressure, indicating that spherical alveolar morphologies are natural in living lungs. The alveolar packing fraction is quite low in live lungs, where the spherical alveoli would behave like free bubbles, while the confinement of alveolar clusters in fixed lungs would lead to significant morphological deformations of the alveoli appearing polyhedral. Direct observations of the spherical alveolar shapes will help understand and treat lung disease and ventilation.

Type-Independent 3D Writing and Nano-Patterning of Confined Biopolymers.

Biopolymers are essential building blocks that constitute cells and tissues with well-defined molecular structures and diverse biological functions. Their three-dimensional (3D) complex architectures are used to analyze, control, and mimic various cells and their ensembles. However, the free-form and high-resolution structuring of various biopolymers remain challenging because their structural and rheological control depend critically on their polymeric types at the submicron scale. Here, direct 3D writing of intact biopolymers is demonstrated using a systemic combination of nanoscale confinement, evaporation, and solidification of a biopolymer-containing solution. A femtoliter solution is confined in an ultra-shallow liquid interface between a fine-tuned nanopipette and a chosen substrate surface to achieve directional growth of biopolymer nanowires via solvent-exclusive evaporation and concurrent solution supply. The evaporation-dependent printing is biopolymer type-independent, therefore, the 3D motor-operated precise nanopipette positioning allows in situ printing of nucleic acids, polysaccharides, and proteins with submicron resolution. By controlling concentrations and molecular weights, several different biopolymers are reproducibly patterned with desired size and geometry, and their 3D architectures are biologically active in various solvents with no structural deformation. Notably, protein-based nanowire patterns exhibit pin-point localization of spatiotemporal biofunctions, including target recognition and catalytic peroxidation, indicating their application potential in organ-on-chips and micro-tissue engineering.

Deep 3D reconstruction of synchrotron X-ray computed tomography for intact lungs.

Synchrotron X-rays can be used to obtain highly detailed images of parts of the lung. However, micro-motion artifacts induced by such as cardiac motion impede quantitative visualization of the alveoli in the lungs. This paper proposes a method that applies a neural network for synchrotron X-ray Computed Tomography (CT) data to reconstruct the high-quality 3D structure of alveoli in intact mouse lungs at expiration, without needing ground-truth data. Our approach reconstructs the spatial sequence of CT images by using a deep-image prior with interpolated input latent variables, and in this way significantly enhances the images of alveolar structure compared with the prior art. The approach successfully visualizes 3D alveolar units of intact mouse lungs at expiration and enables us to measure the diameter of the alveoli. We believe that our approach helps to accurately visualize other living organs hampered by micro-motion.

Coalescence preference depends on size inequality.

During bubble or droplet coalescence, there is a puzzling tendency for the coalesced bubble or droplet to be preferentially placed closer to the larger of its two parents. We confirm that this preference is a function of parent size ratio by directly visualizing coalescing air bubbles on an oil-water interface and coalescing water droplets immersed in oil. We find that the final position of the coalesced sphere is controlled by surface energy release and is related to the parent size ratio by a power-law relationship. 

How does an air film evolve into a bubble during drop impact?

When a liquid drop impacts a solid surface, air is generally entrapped underneath. Using ultrafast x-ray phase-contrast imaging, we directly visualized the profile of an entrapped air film and its evolution into a bubble during drop impact. We identified a complicated evolution process that consists of three stages: inertial retraction of the air film, contraction of the top air surface into a bubble, and pinch-off of a daughter droplet inside the bubble. Energy transfer during retraction drives the contraction and pinch-off of a daughter droplet. The wettability of the solid surface affects the detachment of the bubble, suggesting a method for bubble elimination in many drop-impact applications.

Air evolution during drop impact on liquid pool.

We elucidate the evolution of the entrained air in drop impact on a wide range of liquids, using ultrafast X-ray phase-contrast imaging. We elaborate the retraction mechanism of the entrapped air film in terms of liquid viscosity. We found the criterion for deciding if the entrapped air evolves into single or double bubbles, as determined by competition among inertia, capillarity, and viscosity. Low viscosity and low surface tension induce a small daughter droplet encapsulated by a larger air shell bubble, forming an antibubble. We demonstrate a phase diagram for air evolution regarding hydrodynamics.

Theoretical estimation of maximum human lifespan.

The existence of maximum human lifespan remains a puzzle in aging research. Maximum human lifespan is believed to be around 125 years, whereas current demographic trends seem to show no limitation. To reconcile this contrast, the estimation of maximum human lifespan requires an adequate mathematical model. However, sparse data of available old-age mortality pattern make the estimation impossible. Here we suggest an extended Weibull model for the estimation using a proper mathematical method based on survival probability pattern. We find a tendency that survival probability is maximized in modern human survival curves. Based on such tendency, we develop an estimation method for maximum human lifespan and indeed obtain about 126 years from periodic life tables for Swedish female between 1950 and 2005. Despite uncertainty from available mortality data, our approach may offer quantitative biodemographic opportunities linking aging and survival kinetics.

Nondestructive Characterization by Advanced Synchrotron Light Techniques: Spectromicroscopy and Coherent Radiology.

The advanced characteristics of synchrotron light has led in recent years to the development of a series of new experimental techniques to investigate chemical and physical properties on a microscopic scale. Although originally developed for materials science and biomedical research, such techniques find increasing applications in other domains - and could be quite useful for the study and conservation of cultural heritage. Specifically, they can nondestructively provide detailed chemical composition information that can be useful for the identification of specimens, for the discovery of historical links based on the sources of chemical raw materials and on chemical processes, for the analysis of damage, their causes and remedies and for many other issues. Likewise, morphological and structural information on a microscopic scale is useful for the identification, study and preservation of many different cultural and historical specimens. We concentrate here on two classes of techniques: in the first case, photoemission spectromicroscopy. This is the result of the advanced evolution of photoemission techniques like ESCA (Electron Microscopy for Chemical Analysis). By combining high lateral resolution to spectroscopy, photoemission spectromicroscopy can deliver fine chemical information on a microscopic scale in a nondestructive fashion. The second class of techniques exploits the high lateral coherence of modern synchrotron sources, a byproduct of the quest for high brightness or brilliance. We will see that such techniques now push radiology into the submicron scale and the submillisecond time domain. Furthermore, they can be implemented in a tomographic mode, increasing the information and becoming potentially quite useful for the analysis of cultural heritage specimens.

Precise Placement of Microbubble Templates at Single Entity Resolution.

Microbubbles have been used as a soft template to produce hollow structures for diverse applications in chemistry, materials science, and biomedicine. It is a challenge, however, to control their size and position at single-entity level. We report on an on-demand method to produce and place a single microbubble with programmed size and position. The method exploits scanning an electrolyte-filled micropipette to place a hydrogen (H2) bubble, generated by water electrolysis, on the desired position. The bubble growth is self-limited after the bubble size fits to the pipet aperture, yielding well-controlled bubble size. The bubble growth dynamics within the pipet is successfully investigated by a methodology that combines phase-contrast X-ray imaging and electric-current measurement. We show that the microbubbles, accurately controlled in size and position, can be used for the fabrication of various polypyrrole microcontainer arrays. We expect the scanning-pipet strategy could be generalized for manipulating various soft materials at will.

Synchrotron microradiography study on acute lung injury of mouse caused by PM(2.5) aerosols.

In order to investigate FeSO(4), ZnSO(4) (the two of main metal compositions of Shanghai PM(2.5) (particle matter with those aerodynamical diameter <2.5 microm)) effects on acute lung injury, six solutions contained PM(2.5) aerosol particles, FeSO(4), ZnSO(4) and their mixtures were instilled intratracheally into mouse lungs for experiment. By 2 days after instillation, the live mice were checked in vivo by synchrotron refractive index microradiography. In addition after extracted and examined by dissection, the right lobes of lung were fixed by formalin, then imaged by synchrotron microradiography again. Corresponding parts of those lung tissues were embedded in paraffin for histopathologic study. The synchrotron X-ray microradiographs of live mouse lung showed different lung texture changes after instilled with different toxic solutions. Hemorrhage points in lung were observed more from those mice instilled by FeSO(4) contained toxin solutions groups. Bronchial epithelial hyperplasia can be observed in ZnSO(4) contained solution-instilled groups from histopathologic analysis. It was found that the acute lung injury of mice caused by solution of PM(2.5)+FeSO(4)+ZnSO(4) was more serious than other toxin solutions. Results suggested that FeSO(4) mainly induced hemorrhage and ZnSO(4) mainly induced inflammation and bronchiolar epithelial hyperplasia in the early toxicological effects of PM(2.5).

Alcian blue staining method to visualize bonghan threads inside large caliber lymphatic vessels and x-ray microtomography to reveal their microchannels.

BACKGROUND: Novel threadlike structures, so-called Bonghan threads, inside large caliber lymphatic vessels were recently observed by using the Janus Green B staining in the case of rabbits, and by using fluorescent magnetic nanoparticles for rats. They are thought to have channels through which some liquid with abundant hyaluronic acid (HA) flows. In the present work, the authors put forward a hypothesis to test that the Bonghan threads can be visualized in vivo by HA-staining dye, Alcian blue, and that the channels may be revealed by using X-ray microtomography. METHODS AND RESULTS: Alcain blue was injected into a lymph vessel near the caudal vena cava of a rat to make the Bonghan threads visible. Specimens were stained with YoYo-1 and Masson's trichrome. They were also examined by confocal microscopy and X-ray microtomography. A Bonghan thread was well stained by Alcian blue and was about 50 microm thick and 10-20 times thinner than the surrounding lymphatic vessel. It had a broken line shape, with a distribution of rod-shaped nuclei, which is the characteristic of Bonghan threads in general. Whereas lymphatic vessels are surrounded by a collagenous matrix, Bonghan threads do not contain any collagenous component. X-ray microtomography revealed continuous microchannels inside the Bonghan threads. CONCLUSIONS: Bonghan threads contain HA abundantly, harbor continuous microchannels, and have characteristic distribution of the rod-shape nuclei. Thus, they are novel anatomical structures with liquid-carrying microchannels.

Nanowires: Quantitative Probing of Cu(2+) Ions Naturally Present in Single Living Cells (Adv. Mater. 21/2016).

Quantitative probing of the Cu(2+) ions naturally present in single living cells is accomplished by a probe made from a quantum-dot-embedded-nanowire waveguide. After inserting the active nanowire-based waveguide probe into single living cells, J. H. Je and co-workers directly observe photoluminescence (PL) quenching of the embedded quantum dots by the Cu(2+) ions diffused into the probe as described on page 4071. This results in quantitative measurement of intracellular Cu(2+) ions.

Quantitative Probing of Cu(2+) Ions Naturally Present in Single Living Cells.

Quantitative probing of Cu(2+) ions naturally present in single living cells is realized by developing a quantum-dot-embedded nanowire-waveguide probe. The intracellular Cu(2+) ion concentration is quantified by direct monitoring of photoluminescence quenching during the insertion of the nanowire in a living neuron. The measured intracellular Cu(2+) ion concentration is 3.34 ± 1.04 × 10(-6) m (mean ± s.e.m.) in single hippocampal neurons.