Images Of Droplets Research Articles

Reconsideration of the ESIPT off mechanism for fluorescent probe MNC in aqueous solution

Fluorescent probes with excited state intramolecular proton transfer (ESIPT) properties play a significant role in the research of life science and material science. Guo et al. designed 3-hydroxy-2-(6-Methoxynaphthalen-2-yl)-4H-chromen-4-one (MNC) as a control to achieve the dual-color fluorescence imaging of lipid droplets and endoplasmic reticulum (ER). They deemed that the ESIPT process would be turned off in ER with high water content [J. Am. Chem. Soc. 2021, 143, 3169–3179]. However, contrary to the conventional ESIPT off case, the enol* state fluorescence intensity that should have been enhanced was severely quenched in water. Here, combined with ultrafast spectrum, steady-state fluorescence spectrum and potential energy surface, the mechanism of ESIPT process of MNC turned off in water is revised. Furthermore, the formation of aggregated states in water is responsible for the quenching of MNC fluorescence. This work is expected to provide broader ideas for the design of hydrophobic fluorescent probes.

Stability of spent HCl acid-crude oil emulsion

Acid stimulation may cause formation damage due to incompatibility of the injected acid and crude oil, which may form stable emulsion and ultimately acid sludge. In this process, the pH value of the injected live HCl solution increases during reaction with rocks or minerals. The spent acid may, in turn, be incompatible with the reservoir fluid causing formation damage. Due to the prevalent drawbacks of these unwanted emulsions, this study focused on the stability of spent acid-crude oil emulsions affected by pH, ferric ion concentration and acid to mixture volume ratio (AMR) through a series of systematic experiments. Response surface method (RSM) was applied for design of experiments and using crude oil samples with various properties, which helped to get better insight into acid-crude oil interactions. The results showed that crude oil properties, such as TBN, aromaticity, asphaltene to resin ratio and CII, affect the emulsion stability. A crude oil with higher TBN, CII and asphaltene to resin ratio, and lower aromaticity, has more affinity to interact with the spent acid and to form stable emulsion. The pH value of the spent acid was of lower importance than other properties as stated above, while AMR was found to be the most influential factor in such a way that it reversely influenced the emulsion stability. The impact of ferric ion depended on both pH of the spent acid and chemical properties of the crude oil. Based on the microscopic images of the emulsion droplets, it was concluded that the stability of spent acid-crude oil emulsion is closely associated to characteristics of the interfacial film than the droplet size.

A fluorescent and solvatochromic 1,8-naphthalimide probe for detection of lipid droplet trafficking and biogenesis

We present a highly fluorescent, solvatochromic, 1,8-naphthalimide for simultaneous imaging of lipid droplets and endoplasmic reticulum, with distinguishable emission maxima. By delineating organelles based on their emission profiles, lipid droplet biogenesis and intercellular trafficking events are visualised. The probe also stains myelin lipids in ex vivo brain tissue, indicating histochemical applications.

Carbonized Polymer Dot Probe for Two-Photon Fluorescence Imaging of Lipid Droplets in Living Cells and Tissues.

As a dynamic and multifunctional organelle, lipid droplets (LDs) are essential in maintaining lipid balance and transducing biological signals. LD accumulation and catabolism are closely associated with energy metabolism and cell signaling. In order to easily trace LDs in living cells, a novel carbonized polymer dot (CPD)-based fluorescent nanoprobe is reported to serve the needs of LD-targeting imaging. This probe exhibits the advantages of excellent biocompatibility, simple preparation, good lipophilicity, and high compatibility with commercial dyes. Transient absorption spectroscopy was employed to discuss the luminescence mechanism of CPDs, and the results indicate that the excellent fluorescence property and the environment-responsive feature of our CPDs are derived from the intramolecular charge transfer (ICT) characteristics and the D-π-A structure that possibly formed in CPD. This nanoprobe is available for one-photon fluorescence (OPF) and two-photon fluorescence (TPF) imaging and is also practicable for staining LDs in living/fixed cells and lipids in tissue sections. The staining process is completed within several seconds, with no washing step. The intracellular LDs involving the intranuclear LDs (nLDs) can be selectively lit up. This probe is feasible for visualizing dynamic interactions among LDs, which suggests its great potential in revealing the secret of LD metabolism. The in situ TPF spectra were analyzed to determine surrounding microenvironment according to the polarity-responsive feature of our CPDs. This work expands the applications of CPDs in biological imaging, helps design new LD-selective fluorescent probes, and has implications for studying LD-related metabolism and diseases.

Polarity and viscosity-sensitive fluorescence probes for lipid droplet imaging in cancer cells

Fluorescent probes sensitive to microenvironments have constantly been attractive, due to their significant benefits in monitoring changes in the pathophysiological microenvironment and their use in the early detection of associated disorders. In this work, we explored three new molecular rotors that could be potential candidates for detecting lipid droplets (LDs) in cancer cells based on coumarin derivatives connected with electron-donating polycyclic units including, tetraphenylethylene (LD-TPE), pyrene (LD-PYR), and 4,5-diphenylimidazole (LD-DIP). Gratifyingly, LD-PYR showed exceptional polarity and viscosity sensitivity and demonstrated bright fluorescence in highly viscous environments due to the suppression of the twisted intramolecular charge transfer (TICT) phenomenon and intramolecular rotation. Moreover, microenvironment-guided specific LD bioimaging by LD-PYR exhibited a reasonable ability to distinguish between cancer and noncancer cells. These data suggested that LD-PYR has interesting clinical diagnostic and microenvironment-related biological applications.

Optimization of CMT Characteristic Parameters for Swing Arc Additive Manufacturing of AZ91 Magnesium Alloy Based on Process Stability Analysis.

The droplet transfer behavior and stability of the swing arc additive manufacturing process of AZ91 magnesium alloy based on the cold metal transfer (CMT) technique were studied by analyzing the electrical waveforms and high-speed droplet images as well as the forces on the droplet, and the Vilarinho regularity index for short-circuit transfer (IVSC) based on variation coefficients was used to characterize the stability of the swing arc deposition process. The effect of the CMT characteristic parameters on the process stability was investigated; then, the optimization of the CMT characteristic parameters was realized based on the process stability analysis. The results show that the arc shape changed during the swing arc deposition process; thus, a horizontal component of the arc force was generated, which significantly affected the stability of the droplet transition. The burn phase current I_sc_wait presented a linear function relation with IVSC, while the other three characteristic parameters, i.e., boost phase current I_boost, boost phase duration t_I_boost and short-circuiting current I_sc2, all had a quadratic correlation with IVSC. A relation model of the CMT characteristic parameters and IVSC was established based on the rotatable 3D central composite design; then, the optimization of the CMT characteristic parameters was realized using a multiple-response desirability function approach.

Evaporation and Autoignition Characteristics of JP-10 Droplets with Hyperbranched Polyester as Additive

It was found in our previous work that hyperbranched polyester (HPE) can generate radicals and accelerate the chemical reactions of hydrocarbon fuels used as initiators. In this work, the evaporation and autoignition characteristics of JP-10 droplets with or without HPE were investigated using the high-speed backlight imaging technique in detail. The results indicate that the puffing and micro-explosion phenomena of HPE-blended JP-10 droplets can accelerate fuel evaporation and autoignition. When a 0.1% mass concentration of HPE was used, the droplet lifetime was reduced by 16.5% in evaporation at 850 K and 18.0% in autoignition at 900 K. A mechanism of HPE that promotes puffing and micro-explosions was proposed by analyzing droplet images of combustion and SEM images of combustion residues. Overall, this study provides a method for improving the evaporation and autoignition performance of JP-10.

An advanced organic molecular probe for multimodal fluorescence imaging of cellular lipid droplets

Lipid droplets (LDs) are critical cellular organelles associated with various physiological processes in eukaryotic cells. To visualize LDs and research their complex physiological functions, various fluorescence imaging models including the confocal imaging, the lambda scan imaging and the stimulated emission depletion (STED) super-resolution imaging have been successfully employed in recent years. However, since different fluorescence imaging models have different requirements for fluorescent probes, it is very challenging to employ single fluorescent probe for multimodal fluorescence imaging. Herein, an advanced organic fluorescent probe Lipi-DSBOMe has been sophisticatedly developed for purpose. Featuring with the advantages of high staining selectivity towards LDs, polarity sensitive luminescence, high photostability and low STED saturation intensity, this fluorescent probe is well suited for multimodal fluorescence imaging. For instance, employed with this probe for three-dimensional (3D) confocal imaging to quantitatively determine the change of number of LDs under the oleic acid stimulation, lambda scan imaging to precisely measure the polarity of LDs in different types of cells, and STED imaging to measure the size of LDs with the state-of-the-art resolution (42 nm). In overall, employing this single fluorescent probe for multimodal fluorescence imaging has allowed to provide abundant information about LDs, thus effectively promoting the LDs biology studies.

Nanoscale Contact Line Pinning Boosted by Ångström-Scale Surface Heterogeneity.

The pinning effect plays an important role in many fluidic systems but remains poorly understood, especially at the nanoscale. In this study, we measured the contact angles of glycerol nanodroplets on three different substrates using atomic force microscopy. By comparison of the shapes of the three-dimensional images of droplets, we found that a possible origin of the long-discussed deviation of the contact angles of nanodroplets from the macroscopic value is the pinning force induced by ångström-scale surface heterogeneity. It was also revealed that the pinning forces acting on glycerol nanodroplets on a silicon dioxide surface are up to twice as large as those acting on macroscale droplets. On a substrate where the effect of pinning was strong, an unexpected irreversible change from an irregularly shaped droplet to an atomically flat liquid film occurred. This was explained by the transition of the dominant force from liquid/gas interfacial tension to an adsorption force.

Preparation and Emulsifying Properties of Carbon-Based Pickering Emulsifier

Water is increasingly being used as a solvent in place of organic solvent in order to meet the demand for green chemical synthesis. Nevertheless, many of the reaction substrates are organic matter, which have low water solubility, resulting in a low reaction interface and limiting the development of organic-water biphasic systems. A surfactant is typically added to the two-phase system to form an emulsion to increase the contact area between the organic phase and the water. Compared to ordinary emulsion stabilized with the surfactant, Pickering emulsion offers better adhesion resistance, biocompatibility, and environmental friendliness. It possesses unrivaled benefits as an emulsifier and catalyst in a two-phase interfacial catalysis system (PIC). In this study, the amine group (NNDB) was employed to alter the surface of graphene oxide (GO). A stable Pickering emulsion was created by adsorbing GO-NNDB on the toluene–water interface. It was determined that the emulsion system had good stability by analyzing digital photographs and microscope images of droplets at various temperatures, and fluorescence microscopy images of emulsion droplets created by both newly added and recovered emulsifiers. This work provided the groundwork for future applications of Pickering emulsion in interfacial catalysis.

Multiclass Reinforced Active Learning for Droplet Pinch-Off Behaviors Identification in Inkjet Printing

Abstract Inkjet printing (IJP) is one of the promising additive manufacturing techniques that yield many innovations in electronic and biomedical products. In IJP, the products are fabricated by depositing droplets on substrates, and the quality of the products is highly affected by the droplet pinch-off behaviors. Therefore, identifying pinch-off behaviors of droplets is critical. However, annotating the pinch-off behaviors is burdensome since a large amount of images of pinch-off behaviors can be collected. Active learning (AL) is a machine learning technique which extracts human knowledge by iteratively acquiring human annotation and updating the classification model for the pinch-off behaviors identification. Consequently, a good classification performance can be achieved with limited labels. However, during the query process, the most informative instances (i.e., images) are varying and most query strategies in AL cannot handle these dynamics since they are handcrafted. Thus, this paper proposes a multiclass reinforced active learning (MCRAL) framework in which a query strategy is trained by reinforcement learning (RL). We designed a unique intrinsic reward signal to improve the classification model performance. Moreover, how to extract the features from images for pinch-off behavior identification is not trivial. Thus, we used a graph convolutional network for droplet image feature extraction. The results show that MCRAL excels AL and can reduce human efforts in pinch-off behavior identification. We further demonstrated that, by linking the process parameters to the predicted droplet pinch-off behaviors, the droplet pinch-off behavior can be adjusted based on MCRAL.

Structured Illumination Microscopy of Mitochondrial in Mouse Hepatocytes with an Improved Image Reconstruction Algorithm.

In this paper, a structured illumination microscopy (SIM) image reconstruction algorithm combined with notch function (N-SIM) is proposed. This method suppresses the defocus signal in the imaging process by processing the low-frequency signal of the image. The existing super-resolution image reconstruction algorithm produces streak artifacts caused by defocus signal. The experimental results show that the algorithm proposed in our study can well suppress the streak artifacts caused by defocused signals during the imaging process without losing the effective information of the image. The image reconstruction algorithm is used to analyze the mouse hepatocytes, and the image processing tool developed by MATLAB is applied to identify, detect and count the reconstructed images of mitochondria and lipid droplets, respectively. It is found that the mitochondrial activity in oxidative stress induced growth inhibitor 1 (OSGIN1) overexpressed mouse hepatocytes is higher than that in normal cells, and the interaction with lipid droplets is more obvious. This paper provides a reliable subcellular observation platform, which is very meaningful for biomedical work.

Red-emissive Dual-state Fluorogenic Probe for Wash-free Imaging of Lipid Droplets in Living Cells and Fatty Liver Tissues.

Lipid droplet (LD) dysfunction can result in various diseases, such as nonalcoholic fatty liver disease. Imaging agents built on dual-state emission (DSE) molecules that fluoresce in both dilute solutions and the aggregated state are receiving attention as this type of probe could provide bright fluorescence signals at variable concentrations, avoiding false signal readout caused by the concentration fluctuation in living systems. Herein, we identified a red emissive molecule featuring DSE, from three newly synthesized molecules, for specific detection of LDs in live cells. The bioimaging abilities have been well confirmed by optical spectroscopies, theoretical calculations, cell experiments, as well as animal studies. The DSE probe is effective for LD detection at concentrations ranging from 1 μM to 100 μM while retaining high brightness and signal fidelity. This study provides a knowledge base for the future design of DSE-active fluorescent probes for understanding LD-related diseases.

PH-Triggered Charge Reversible Fluorescent Probe for Simultaneous Imaging of Lipid Droplets and Nucleoli in Living Cells.

Cooperation between organelles is essential to maintain the normal functions of cells. Lipid droplets (LDs) and nucleoli, as important organelles, play an important role in the normal activities of cells. However, due to the lack of appropriate tools, in situ observation of the interaction between them has been rarely reported. In this work, taking into full consideration the pH and charge differences between LDs and nucleoli, a pH-triggered charge reversible fluorescent probe (LD-Nu) was constructed based on a cyclization-ring-opening mechanism. The in vitro pH titration experiment and 1H NMR showed that LD-Nu gradually transferred from the charged form to the electroneutral form with the increase of pH, and thus, the conjugate plane was reduced and its fluorescence blue-shifted. Most importantly, the physical contact between LDs and nucleoli was visualized for the first time. Meanwhile, the relationship between LDs and nucleoli was also further investigated, and the results showed that their interaction was more liable to be affected by the abnormality of LDs than those of nucleoli. Moreover, the cell imaging results displayed that the LDs both in the cytoplasm and nucleus were observed using the probe LD-Nu, and interestingly, the LDs in the cytoplasm were more susceptible to external stimuli than those in the nucleus. In a word, the probe LD-Nu can serve as a powerful tool for further exploration of the interaction mechanism between LDs and nucleoli in living cells.

In Vivo Imaging of Lipid Droplets and Oxygen Status in Hepatic Tissues of Nonalcoholic Fatty Liver Model Mice Using a Lipophilic Ir(III) Complex

Nonalcoholic fatty liver disease (NAFLD) is becoming common worldwide. In pathophysiological studies of NAFLD, an in vivo optical probe that enables visualization of lipid droplets (LDs) and imaging of oxygen status in hepatic tissues simultaneously would be very useful. Here, we present the phosphorescent Ir(III) complex BTP ((btp)2Ir(acac) (btp = benzothienylpyridine, acac = acetylacetone)) as the first probe that meets this requirement. BTP was efficiently taken up into cultured 3T3-L1 adipocytes and selectively accumulated into LDs. Quantifying oxygen levels in LDs based on the phosphorescence lifetime of BTP allowed us to track changes in cellular oxygen tension after treatment with metabolic stimulants. Phosphorescence lifetime imaging microscopy combined with intravenously administered BTP in mice enabled specific visualization of LDs in hepatic lobules and simultaneous imaging of the oxygen gradient that decreased from the portal vein (PV) to the central vein (CV). NAFL model mice were created by feeding a high-fat diet (HFD) to mice for 3 or 7 days. The mice fed an HFD showed a marked increase in the amount and size of LDs in hepatocytes compared with those fed a normal diet, leading to abnormal microvascular structures. In addition, HFD-fed mice also exhibited reduced oxygen tension in areas other than the CV. Multicolor imaging with the LD-accumulated oxygen probe BTP and vasculature-staining FITC-lectin suggested that structural distortions of the sinusoidal microvasculature caused by enlarged LDs were associated with partial hypoxia in NAFL.

Conical liquid sheet morphology and 3D droplet distribution of aviation kerosene pressure-swirl spray with digital off-axis holography

In this study, the test performance of high-resolution digital off-axis holography for the non-spherical droplet characteristics and spatial distribution in the primary breakup region of a pressure-swirl nozzle were experimentally investigated, with spatial resolution of 4.5μm and 3.0μm. The field of interest (FOI) was restricted within 9 mm downstream of the nozzle outlet, where the primary and some secondary breakup processes take place. The liquid investigated here is RP-3 kerosene, and the injection pressure and liquid temperature were controlled in the range of 0.5 ∼ 1.0 MPa and 250 ∼ 291 K separately. Clear spray morphology and deformed droplet image were obtained through the holographic reconstruction process. Droplet extraction criteria were proposed to distinguish deformed droplets from ligaments. The overall drop size and ellipticity distribution were quantified and discussed. The results show that the overall drop size distributions near the nozzle are also typical single-peaked curves. The proportions of non-spherical droplet increase as the fuel pressure and temperature increase. The spatial Sauter mean diameter (SMD) distributions of spray at different positions downstream of the nozzle were calculated, with the grid size of 1 × 1 × 1 mm3, showing the doughnut-like annular SMD and droplet number distribution profiles of the hollow conical spray. As the temperature and fuel pressure decrease, SMD distribution becomes uneven and larger. Fine droplets exist in the region where the perforation and primary breakup process occur, and the SMD increases over the axial distance due to the breakup process of ligaments near the nozzle. Results also prove that digital off-axis holography presented here is expected to realize the 3D viewing of the conical liquid sheet and statistical quantification of the droplet where primary and secondary breakup processes mainly occurred.

Long-term in-situ real-time fluorescence imaging of lipid droplets during cell ferroptosis process enabled by an epindolidione-based fluorescent probe

Ferroptosis, as a new form regulation of cell death discovered recently, is triggered by an iron-dependent accumulation of lipid reactive oxygen species and thus closely associated with cellular lipid droplets (LDs). However, directly limited by the un-sufficient photostability and low LDs specificity of common fluorescent probes, visualization of LDs during ferroptosis process particular in a long-term, in-situ, and real-time manner is highly preferred but extremely challenge. To this end, here a new LDs fluorescent probe Lipi-EP featuring with outstanding photostability and high LDs specificity have been rationally developed based on the four-ring-fused epindolidione. Experimentally, the photostability and LDs specificity of this new probe are much higher than the representative LDs probe BODIPY and Nile Red. Consequently, time-lapse multicolor imaging and time-lapse three-dimensional imaging employed this new probe have been successfully conducted to in-situ real-time monitor the LDs change during ferroptosis process up to 6 h. The morphology changes of cellular organelles (LDs, mitochondria and nucleus) and the variations of space distribution of LDs have been impressively visualized. Moreover, the sizes and numbers of LDs have been also determined based on the imaging. This work providing abundant information of LDs would significantly promote the in-depth study of LDs during ferroptosis.

Ferrostatin-1 improves BMSC survival by inhibiting ferroptosis

ObjectiveTo investigate the effect of ferroptosis in BMSCs and explore the protective metabolism of ferrostatin-1 under GSDH treatment. MethodsBMSCs were treated with GSDH to simulate the damaged microenvironment in vivo to establish a cell injury model. Propidium iodide and CCK8 were utilized to detect the ratio of dead cells and cell viability. DCFH-DA and Amplex Red, FerroOrange, and BPDIPY were used to visualize the cellular fluorescent images of ROS, Fe2+, and lipid droplets, respectively. The quantified detection of MDA was conducted by a Lipid Peroxidation MDA Assay Kit. JC-1 staining, Mito-Tracker staining, and TEM were implemented to detect the membrane potential, morphology, and ultrastructure of mitochondria, respectively. The expression levels of ferroptosis-related proteins such as GPX4 and FTH1 were measured by Western blotting. ResultsGSDH treatment induced ferroptosis in BMSCs based on an increased ratio of cell death, Fe2+, ROS, lipid droplets, and MDA in cells plus decreased protein levels of antioxidant systems, such as GPX4, and increased protein levels related to fatty acid synthesis. Compared to the blank group, mitochondria in the GSDH group underwent lower membrane potential, damaged morphology, and shrunken ultrastructure; Ferr-1 rescued the injured BMSCs to a certain extent as the declined ratio of cell death, Fe2+, ROS, lipid droplets, MDA, and the increased level antioxidant protein. AMPK was phosphorylated and activated after Ferr-1 treatment, and its downstream lipid peroxidation and antioxidation proteins changed accordingly. Inhibition of AMPK hindered the curative effect of Ferr-1. ConclusionFerr-1 rescued ferroptosis-induced injury to BMSCs under GSDH conditions, and AMPK might have a relationship with the mitigative effect of Ferr-1.

One Stone, Three Birds: A Smart Single Fluorescent Probe for Simultaneous and Discriminative Imaging of Lysosomes, Lipid Droplets, and Mitochondria.

Complex intracellular life processes are usually completed through the cooperation of multiple organelles. Real-time tracking of the interplays between multiple organelles with a single fluorescent probe (SFP) is very helpful to deepen our understanding of complex biological processes. So far, SFP for simultaneously differentiating and visualizing of more than two different organelles has not been reported. Herein, we report an SFP (named ICM) that can be used for simultaneously differentiating and visualizing three important organelles: mitochondria, lysosomes, and lipid droplets (LDs). The probe can simultaneously light up mitochondria/lysosomes (∼700 nm) and LDs (∼480 nm) at significantly different emission wavelengths with high fidelity, and mitochondria and lysosomes can be effectively distinguished by their different shapes and fluorescence intensities. With this smart probe, real-time and simultaneous tracking of the interplays of these three organelles was successfully achieved for the first time.

Surface deposition characteristics of water-based SiO2 nanofluids on coal

Water-based SiO2 nanofluids, a type of wetting agent for water injection of coal seams, have been well recognized by many scholars due to being environment-friendly and having a stable modification effect. The key of the modification effect of water-based SiO2 nanofluids on coal seam is the deposition behavior of nanofluids on coal surface. Based on this issue, this research conducted real-time monitoring of the morphological evolution characteristics and deposition images of water-based SiO2 nanofluid droplets with different concentrations on coal surface. Moreover, the deposition behavior of water-based SiO2 nanofluids on coal surface was microscopically expounded. The results indicate that the initial contact angle between the nanofluid droplets and coal surface is small, and the spreading degree of water-based SiO2 nanofluids is better than that of deionized water. With the increase of time, the contact angles between water-based SiO2 nanofluid droplets and coal surface tend to linearly decrease. However, the relative length of contact line features flattening at first and then a sharp decline. As the nanoparticles are completely precipitated, water-based SiO2 nanofluid droplets form a deposition pattern with a coffee ring structure on coal surface. In addition, as the particle concentration increases, the relative width of the coffee ring increases stepwise. The particle aggregate on the middle of the deposition pattern gradually connects to coalesce and forms a relatively uniform and flat deposition layer. Based on the research results aforementioned, this research divided the deposition behavior of the water-based SiO2 nanofluid droplets on coal surface into initial stage, the contact angle dominance change, the contact angle and line co-dominance change and complete deposition. Among which, the duration of the angle dominance change stage is longest. With the increase of the particle concentration, the proportion of the angle dominance change stage increases. The research can fill the gap of the research on the deposition behavior of nanofluids on coal surface, which lays a foundation for further research of coal seam water injection using nanofluids.