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At least 91 records · Page 5

Tailored Micromagnet Sorting Gate for Simultaneous Multiple Cell Screening in Portable Magnetophoretic Cell‐On‐Chip Platforms

Abstract Conventional magnetophoresis techniques for manipulating biocarriers and cells predominantly rely on large‐scale electromagnetic systems, which is a major obstacle to the development of portable and miniaturized cell‐on‐chip platforms. Herein, a novel magnetic engineering approach by tailoring a nanoscale notch on a disk micromagnet using two‐step optical and thermal lithography is developed. Versatile manipulations are demonstrated, such as separation and trapping, of carriers and cells by mediating changes in the magnetic domain structure and discontinuous movement of magnetic energy wells around the circumferential edge of the micromagnet caused by a locally fabricated nano‐notch in a low magnetic field system. The motion of the magnetic energy well is regulated by the configuration of the nanoscale notch and the strength and frequency of the magnetic field, accompanying the jump motion of the carriers. The proposed concepts demonstrate that multiple carriers and cells can be manipulated and sorted using optimized nanoscale multi‐notch gates for a portable magnetophoretic system. This highlights the potential for developing cost‐effective point‐of‐care testing and lab‐on‐chip systems for various single‐cell‐level diagnoses and analyses.

36 MATERIALS SCIENCE↗

On Collisions Times of ‘Self-Sorting’ Interacting Particles in One-Dimension with Random Initial Positions and Velocities

We investigate a one-dimensional system of N particles, initially distributed with random positions and velocities, interacting through binary collisions. The collision rule is such that there is a time after which the N particles do not interact and become sorted according to their velocities. When the collisions are elastic, we derive asymptotic distributions for the final collision time of a single particle and the final collision time of the system as the number of particles approaches infinity, under different assumptions for the initial distributions of the particles’ positions and velocities. For comparison, a numerical investigation is carried out to determine how a non-elastic collision rule, which conserves neither momentum nor energy, affects the median collision time of a particle and the median final collision time of the system.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Applying Improved Optical Recognition with Machine Learning on Sorting Cu Impurities in Steel Scrap

Annually, 20–55 million tons of electronic waste (e-waste) is produced worldwide (5% of all municipal solid waste). Although e-waste embodies only 2% of America’s municipal waste, it accounts for a significantly larger proportion of the heavy metals and flame retardants present in the waste stream. Currently, less than 20% of all e-waste is recycled in the United States because the heterogeneity of the feedstock limits the opportunity for reuse in high value products and processing of the waste itself is often too costly to justify handling. To address this concern, the aims of this study are: 1) to identify the major plastic and metal compositions within electronic shredder residue (ESR), 2) to formulate solvents and processing conditions to separate 90% of the plastics targeted from consumer shred ESR, and 3) to develop a process design model to estimate the cost and energy efficiency of the proposed solvent-based processing. In this study, a pre-sorted heterogeneous ESR feedstock (one where aluminum, magnetic components, and hazardous battery materials removed by an e-waste recycling facility) was used, with the major compositions of the ESR characterized. It was found that 25 wt.% of the feedstock was composed of plastics, 6 wt.% rubber, 27 wt.% printed circuit boards, 23% wire, and the remainder metals and capacitors. Within the plastic portion, polystyrene (PS, 40 wt.%), acrylonitrile butadiene styrene (ABS, 25 wt.%), and styrene-acrylonitrile (SAN, 9wt.%) were identified to compose the majority of the screened plastics using Fourier transform infrared spectroscopy (FTIR). Next, selective solvents were screened using Hansen Solubility Parameter Theory (HSP) for dissolving PS and ABS. The pre-screening results show that methylene chloride (dichloromethane, DCM) and tetrahydrofuran (THF) are capable of dissolving the most PS and ABS, while methanol (MeOH) and ethylene glycol (EG) are capable of precipitating the most PS and ABS. These solvents were subsequently used to recover polymers and remove. flame retardants within the ESR feedstock. By optimizing the dissolution time and the solvents used, the highest polymer dissolution yield (99 wt.%%) was achieved using DCM for 48 hr. Both pre-screened anti-solvents (MeOH and EG) showed the highest polymer precipitation yield (71 wt.%). In terms of flame retardant removal rate, EG was found to have a high phosphorus-containing flame retardant removal rate (up to 98%). Characterization shows that the proposed solvent-based processing can preserve a high molecular weight fraction of the polymers and effectively remove flame retardants. Cost analysis indicates that the amount of the solvent/anti-solvent recovered after the reaction would play a critical role in reducing the operating costs. The energy analysis shows that the proposed solvent-based processes can save up to 60% of the embodied energy used to manufacture plastics used in electronics (PS and ABS were used for calculations). The results from this project prove the potential of solvent-based processing to produce secondary materials (plastics and metals) from e-waste for cross-industry reuse.

36 MATERIALS SCIENCE↗

Nonlocal Means Denoising of Self-Gated and k-Space Sorted 4-Dimensional Magnetic Resonance Imaging Using Block-Matching and 3-Dimensional Filtering: Implications for Pancreatic Tumor Registration and Segmentation

Purpose: To denoise self-gated k-space sorted 4-dimensional magnetic resonance imaging (SG-KS-4D-MRI) by applying a nonlocal means denoising filter, block-matching and 3-dimensional filtering (BM3D), to test its impact on the accuracy of 4D image deformable registration and automated tumor segmentation for pancreatic cancer patients. Methods and Materials: Nine patients with pancreatic cancer and abdominal SG-KS-4D-MRI were included in the study. Block-matching and 3D filtering was adapted to search in the axial slices/frames adjacent to the reference image patch in the spatial and temporal domains. The patches with high similarity to the reference patch were used to collectively denoise the 4D-MRI image. The pancreas tumor was manually contoured on the first end-of-exhalation phase for both the raw and the denoised 4D-MRI. B-spline deformable registration was applied to the subsequent phases for contour propagation. The consistency of tumor volume defined by the standard deviation of gross tumor volumes from 10 breathing phases (σ-GTV), tumor motion trajectories in 3 cardinal motion planes, 4D-MRI imaging noise, and image contrast-to-noise ratio were compared between the raw and denoised groups. Results: Block-matching and 3D filtering visually and quantitatively reduced image noise by 52% and improved image contrast-to-noise ratio by 56%, without compromising soft tissue edge definitions. Automatic tumor segmentation is statistically more consistent on the denoised 4D-MRI (σ-GTV = 0.6 cm{sup 3}) than on the raw 4D-MRI (σ-GTV = 0.8 cm{sup 3}). Tumor end-of-exhalation location is also more reproducible on the denoised 4D-MRI than on the raw 4D-MRI in all 3 cardinal motion planes. Conclusions: Block-matching and 3D filtering can significantly reduce random image noise while maintaining structural features in the SG-KS-4D-MRI datasets. In this study of pancreatic tumor segmentation, automatic segmentation of GTV in the registered image sets is shown to be more consistent on the denoised 4D-MRI than on the raw 4D-MRI.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

Testing Pollen Sorted by Flow Cytometry as the Basis for High-Resolution Lacustrine Chronologies

Documenting leads and lags in terrestrial records of past climate change is critical to understanding the behavior of Earth’s natural climate system and making reliable predictions of future climate conditions. However, uncertainties of several hundred years in age models make it difficult to distinguish synchronicity and feedbacks in paleo archives. In lakes this is often due to the lack of terrestrial macrofossils in climate-sensitive locations, such as high alpine or dryland settings. The potential of radiocarbon (14C) dating of pollen has long been recognized, but the difficulty of cleanly separating pollen from other kinds of organic carbon has limited its usefulness. Here we report 14C ages on pollen separated by flow cytometry, from a set of closely spaced samples from Mono Lake, California. The accuracy of the pollen ages is tested using well-dated bracketing tephras, the South Mono and North Mono-Inyo tephras. In spite of the purity of the sorted samples, the pollen dates are older than the bounding tephras by ~400 yr, similar to some other pollen-dating studies. While improvements in sample preparation protocols are planned, understanding the geological processes involved in the production, preservation, and deposition of pollen at each site will be critical to developing robust high-resolution age models.

58 GEOSCIENCES↗

Development of a Fluorescence-Activated Cell Sorting Method Coupled with Whole Genome Amplification To Analyze Minority and Trace Dehalococcoides Genomes in Microbial Communities

Dehalococcoides mccartyi are functionally important bacteria that catalyze the reductive dechlorination of chlorinated ethenes. However, these anaerobic bacteria are fastidious to isolate, making downstream genomic characterization challenging. In order to facilitate genomic analysis, a fluorescence-activated cell sorting (FACS) method was developed in this study to separate D. mccartyi cells from a microbial community, and the DNA of the isolated cells was processed by whole genome amplification (WGA) and hybridized onto a D. mccartyi microarray for comparative genomics against four sequenced strains. First, FACS was successfully applied to a D. mccartyi isolate as positive control, and then microarray results verified that WGA from 10 6 cells or ∼1 ng of genomic DNA yielded high-quality coverage detecting nearly all genes across the genome. As expected, some inter- and intrasample variability in WGA was observed, but these biases were minimized by performing multiple parallel amplifications. Subsequent application of the FACS and WGA protocols to two enrichment cultures containing ∼10% and ∼1% D. mccartyi cells successfully enabled genomic analysis. As proof of concept, this study demonstrates that coupling FACS with WGA and microarrays is a promising tool to expedite genomic characterization of target strains in environmental communities where the relative concentrations are low.

59 BASIC BIOLOGICAL SCIENCES↗

Activity-based cell sorting reveals responses of uncultured archaea and bacteria to substrate amendment

Metagenomic studies have revolutionized our understanding of the metabolic potential of uncultured microorganisms in various ecosystems. However, many of these genomic predictions have yet to be experimentally tested, and the functional expression of genomic potential often remains unaddressed. In order to obtain a more thorough understanding of cell physiology, novel techniques capable of testing microbial metabolism under close to in situ conditions must be developed. Here, we provide a benchmark study to demonstrate that bioorthogonal non-canonical amino acid tagging (BONCAT) in combination with fluorescence-activated cell sorting (FACS) and 16S rRNA gene sequencing can be used to identify anabolically active members of a microbial community incubated in the presence of various growth substrates or under changing physicochemical conditions. We applied this approach to a hot spring sediment microbiome from Yellowstone National Park (Wyoming, USA) and identified several microbes that changed their activity levels in response to substrate addition, including uncultured members of the phyla Thaumarchaeota, Acidobacteria, and Fervidibacteria. Because shifts in activity in response to substrate amendment or headspace changes are indicative of microbial preferences for particular growth conditions, results from this and future BONCAT-FACS studies could inform the development of cultivation media to specifically enrich uncultured microbes. Most importantly, BONCAT-FACS is capable of providing information on the physiology of uncultured organisms at as close to in situ conditions as experimentally possible.

59 BASIC BIOLOGICAL SCIENCES↗

Identification of inulin-responsive bacteria in the gut microbiota via multi-modal activity-based sorting

Prebiotics are defined as non-digestible dietary components that promote the growth of beneficial gut microorganisms. In many cases, however, this capability is not systematically evaluated. Here, we develop a methodology for determining prebiotic-responsive bacteria using the popular dietary supplement inulin. We first identify microbes with a capacity to bind inulin using mesoporous silica nanoparticles functionalized with inulin. 16S rRNA gene amplicon sequencing of sorted cells revealed that the ability to bind inulin was widespread in the microbiota. We further evaluate which taxa are metabolically stimulated by inulin and find that diverse taxa from the phyla Firmicutes and Actinobacteria respond to inulin, and several isolates of these taxa can degrade inulin. Incubation with another prebiotic, xylooligosaccharides (XOS), in contrast, shows a more robust bifidogenic effect. Interestingly, the Coriobacteriia Eggerthella lenta and Gordonibacter urolithinfaciens are indirectly stimulated by the inulin degradation process, expanding our knowledge of inulin-responsive bacteria.

59 BASIC BIOLOGICAL SCIENCES↗

Spatial tuning of acoustofluidic pressure nodes by altering net sonic velocity enables high-throughput, efficient cell sorting

Particle sorting using acoustofluidics has enormous potential but widespread adoption has been limited by complex device designs and low throughput. Here, we report high-throughput separation of particles and T lymphocytes (600 μL min -1 ) by altering the net sonic velocity to reposition acoustic pressure nodes in a simple two-channel device. Finally, the approach is generalizable to other microfluidic platforms for rapid, high-throughput analysis.

59 BASIC BIOLOGICAL SCIENCES↗

High performance transistors via aligned polyfluorene-sorted carbon nanotubes

We evaluate the performance of exceptionally electronic-type sorted, semiconducting, aligned single-walled carbon nanotubes (s-SWCNTs) in field effect transistors (FETs). High on-conductance and high on/off conductance modulation are simultaneously achieved at channel lengths which are both shorter and longer than individual s-SWCNTs. The s-SWCNTs are isolated from heterogeneous mixtures using a polyfluorene-derivative as a selective agent and aligned on substrates via dose-controlled, floating evaporative self-assembly at densities of ∼50 s-SWCNTs μm{sup −1}. At a channel length of 9 μm the s-SWCNTs percolate to span the FET channel, and the on/off ratio and charge transport mobility are 2.2 × 10{sup 7} and 46 cm{sup 2} V{sup −1} s{sup −1}, respectively. At a channel length of 400 nm, a large fraction of the s-SWCNTs directly span the channel, and the on-conductance per width is 61 μS μm{sup −1} and the on/off ratio is 4 × 10{sup 5}. These results are considerably better than previous solution-processed FETs, which have suffered from poor on/off ratio due to spurious metallic nanotubes that bridge the channel. 4071 individual and small bundles of s-SWCNTs are tested in 400 nm channel length FETs, and all show semiconducting behavior, demonstrating the high fidelity of polyfluorenes as selective agents and the promise of assembling s-SWCNTs from solution to create high performance semiconductor electronic devices.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗

Tracking heavy water (D 2 O) incorporation for identifying and sorting active microbial cells

Significance Measuring activity patterns of microbes in their natural environment is essential for understanding ecosystems and the multifaceted interactions of microorganisms with eukaryotes. In this study, we developed a technique that allows fast and nondestructive activity measurements of microbial communities on a single-cell level. Microbial communities were amended with heavy water (D 2 O), a treatment that does not change the available substrate pool. After incubation, physiologically active cells are rapidly identified with Raman microspectroscopy by measuring cellular D incorporation. Using this approach, we characterized the activity patterns of two dominant microbes in mouse cecum samples amended with different carbohydrates and discovered previously unidentified bacteria stimulated by mucin and/or glucosamine by combining Raman microspectroscopy and optical tweezer-based sorting.

59 BASIC BIOLOGICAL SCIENCES↗

Osmotic and phoretic competition explains chemotaxic assembly and sorting

Microscale objects responding to chemical gradients by migrating toward or away from a preferred species is a simple yet constitutive mechanism by which transport occurs in biological organisms. Synthetic chemotaxis provides key physical descriptions of simplified systems that can be used in biological models, or in the creation of advanced responsive material systems. In this article, we provide a quantitative framework for understanding synthetic chemotaxis of microparticles which involves a competition between phoresis and osmosis. We present separate quantitative measurements of phoresis and osmosis acting on individual taxing particles, finding that phoresis follows the long-predicted v ∼ 1 / r 2 scaling while the osmotic contribution depends on the geometry and details of the system, and must be solved on a case-by-case basis. Through this, we are able to develop a more accurate picture of particle transport at the single particle level. Equipped with this approach, we go on to describe how high concentrations of particles in a symmetric chemical gradient grow close-packed hives that reach a steady-state size tunable through light intensity or particle size. Last, we demonstrate that mixed particles experiencing the same chemical gradient will selectively migrate toward or away depending on the nature of the particle surface, thereby locally sorting out a particular species. We anticipate these results will be important in describing both biological and synthetic chemotaxis in phoretic systems and should bring a wealth of studies that take advantage of competing osmotic flows to illicit unexpected dynamic active behavior.

Science & Technology - Other Topics↗

High-resolution lidar observations of sedimentation-induced size sorting of droplets near a laboratory cloud top

Cloud optical properties and precipitation, which are crucial to weather and climate, are strongly influenced by cloud microphysical properties that are still poorly understood. Here, we develop a high-resolution time-correlated single-photon-counting lidar and apply it to observe cloud microphysical properties at one-centimeter range resolution in a convection chamber under well-controlled conditions. Together with concurrent in-situ measurements and theoretical analysis, our lidar observations indicate that although turbulent mixing tends to homogenize the cloud in the bulk region, entrainment and sedimentation cause inhomogeneities in droplet concentrations near the cloud top. Specifically, the topmost region is directly affected by entrainment, and lidar profiles show clear evidence of entrained air and detrained cloud filament. The transition region below exhibits vertical size sorting of cloud droplets caused by sedimentation. Our results suggest that using a single sedimentation velocity for all cloud droplets, as is done in many atmospheric models, overlooks key physics relevant to the microphysical structure near the cloud top. In conclusion, our conceptual model used to describe these measurements can serve as a step toward improving the current modeling of processes in the cloud top region.

54 ENVIRONMENTAL SCIENCES↗

Geometric validation of self-gating k-space-sorted 4D-MRI vs 4D-CT using a respiratory motion phantom

Purpose: MRI is increasingly being used for radiotherapy planning, simulation, and in-treatment-room motion monitoring. To provide more detailed temporal and spatial MR data for these tasks, we have recently developed a novel self-gated (SG) MRI technique with advantage of k-space phase sorting, high isotropic spatial resolution, and high temporal resolution. The current work describes the validation of this 4D-MRI technique using a MRI- and CT-compatible respiratory motion phantom and comparison to 4D-CT. Methods: The 4D-MRI sequence is based on a spoiled gradient echo-based 3D projection reconstruction sequence with self-gating for 4D-MRI at 3 T. Respiratory phase is resolved by using SG k-space lines as the motion surrogate. 4D-MRI images are reconstructed into ten temporal bins with spatial resolution 1.56 × 1.56 × 1.56 mm{sup 3}. A MRI-CT compatible phantom was designed to validate the performance of the 4D-MRI sequence and 4D-CT imaging. A spherical target (diameter 23 mm, volume 6.37 ml) filled with high-concentration gadolinium (Gd) gel is embedded into a plastic box (35 × 40 × 63 mm{sup 3}) and stabilized with low-concentration Gd gel. The phantom, driven by an air pump, is able to produce human-type breathing patterns between 4 and 30 respiratory cycles/min. 4D-CT of the phantom has been acquired in cine mode, and reconstructed into ten phases with slice thickness 1.25 mm. The 4D images sets were imported into a treatment planning software for target contouring. The geometrical accuracy of the 4D MRI and CT images has been quantified using target volume, flattening, and eccentricity. The target motion was measured by tracking the centroids of the spheres in each individual phase. Motion ground-truth was obtained from input signals and real-time video recordings. Results: The dynamic phantom has been operated in four respiratory rate (RR) settings, 6, 10, 15, and 20/min, and was scanned with 4D-MRI and 4D-CT. 4D-CT images have target-stretching, partial-missing, and other motion artifacts in various phases, whereas the 4D-MRI images are visually free of those artifacts. Volume percentage difference for the 6.37 ml target ranged from 5.3% ± 4.3% to 10.3% ± 5.9% for 4D-CT, and 1.47 ± 0.52 to 2.12 ± 1.60 for 4D-MRI. With an increase of respiratory rate, the target volumetric and geometric deviations increase for 4D-CT images while remaining stable for the 4D-MRI images. Target motion amplitude errors at different RRs were measured with a range of 0.66–1.25 mm for 4D-CT and 0.2–0.42 mm for 4D-MRI. The results of Mann–Whitney tests indicated that 4D-MRI significantly outperforms 4D-CT in phase-based target volumetric (p = 0.027) and geometric (p < 0.001) measures. Both modalities achieve equivalent accuracy in measuring motion amplitude (p = 0.828). Conclusions: The k-space self-gated 4D-MRI technique provides a robust method for accurately imaging phase-based target motion and geometry. Compared to 4D-CT, the current 4D-MRI technique demonstrates superior spatiotemporal resolution, and robust resistance to motion artifacts caused by fast target motion and irregular breathing patterns. The technique can be used extensively in abdominal targeting, motion gating, and toward implementing MRI-based adaptive radiotherapy.

60 APPLIED LIFE SCIENCES↗

Sorted-cell proteomics reveals an AT1-associated epithelial cornification phenotype and suggests endothelial redox imbalance in human bronchopulmonary dysplasia

Bronchopulmonary dysplasia (BPD) is a neonatal lung disease characterized by inflammation and scarring leading to long-term tissue damage. Previous whole tissue proteomics identified BPD-specific proteome changes and cell type shifts. Little is known about the proteome-level changes within specific cell populations in disease. Here, we sorted epithelial (EPI) and endothelial (ENDO) cell populations based on their differential surface markers from normal and BPD human lungs. Using a low-input compatible sample preparation method (MicroPOT), proteins were extracted and digested into peptides and subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS) proteome analysis. Of the 4,970 proteins detected, 293 were modulated in abundance or detection in the EPI population and 422 were modulated in ENDO cells. Modulation of proteins associated with actin-cytoskeletal function, such as SCEL, LMO7, and TBA1B was observed in the BPD EPIs. Using confocal imaging and analysis, we validated the presence of aberrant multilayer-like structures comprising SCEL and LMO7, known to be associated with epidermal cornification, in the human BPD lung. This is the first report of the accumulation of cornification-associated proteins in BPD. Their localization in the alveolar parenchyma, primarily associated with alveolar type 1 (AT1) cells, suggests a role in the BPD postinjury response. In the ENDOs, redox balance and mitochondrial function pathways were modulated. Alternative mRNA splicing and cell proliferative functions were elevated in both populations, suggesting potential dysregulation of cell progenitor fate. This study characterized the proteome of epithelial and endothelial cells from the BPD lung for the first time, identifying population-specific changes in BPD pathogenesis.

BPD↗

Aspenplus Model For Msw Sorting Process

Unit operations for the MSW sorting process include shredding, magnets for ferrous separation, screening, air classification, eddy current for non-ferrous separation, and near-infrared detection for plastic recovery. The MIXCINC module within Aspen Plus was selected to handle mixed solid streams containing inert (metals and glass) and non-conventional materials (paper, plastics, biomass, organic waste). The Peng-Robinson equation of state with Boston-Mathias modifications was applied to predict properties for a nonconventional solids mix. Enthalpy and density of the non-conventional materials were defined using non-conventional material properties.

Hu, Hongqiang↗

Synthase-selected sorting approach identifies a beta-lactone synthase in a nudibranch symbiotic bacterium

Background - Nudibranchs comprise a group of > 6000 marine soft-bodied mollusk species known to use secondary metabolites (natural products) for chemical defense. The full diversity of these metabolites and whether symbiotic microbes are responsible for their synthesis remains unexplored. Another issue in searching for undiscovered natural products is that computational analysis of genomes of uncultured microbes can result in detection of novel biosynthetic gene clusters; however, their in vivo functionality is not guaranteed which limits further exploration of their pharmaceutical or industrial potential. To overcome these challenges, we used a fluorescent pantetheine probe, which produces a fluorescent CoA-analog employed in biosynthesis of secondary metabolites, to label and capture bacterial symbionts actively producing these compounds in the mantle of the nudibranch Doriopsilla fulva. Results - We recovered the genome of Candidatus Doriopsillibacter californiensis from the Ca. Tethybacterales order, an uncultured lineage of sponge symbionts not found in nudibranchs previously. It forms part of the core skin microbiome of D. fulva and is nearly absent in its internal organs. We showed that crude extracts of D. fulva contained secondary metabolites that were consistent with the presence of a beta-lactone encoded in Ca. D. californiensis genome. Beta-lactones represent an underexplored group of secondary metabolites with pharmaceutical potential that have not been reported in nudibranchs previously. Conclusions - Altogether, this study shows how probe-based, targeted sorting approaches can capture bacterial symbionts producing secondary metabolites in vivo.

59 BASIC BIOLOGICAL SCIENCES↗