Cell sorting - Automated separation of mammalian cells as a function of intracellular fluorescence
High speed fluorescent cell sorting system for sorting mouse spleen cells from Chinese hamster ovarian tumor cells
Barrett, J.↗
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Order-sorting filter transmittance measured with an array detector
The simultaneous measurement of the spectrally and spatially variant transmittance of a linear variable order-sorting filter in a manner that closely resembles its conditions of actual use is described. The transmittance of a prototype order-sorting filter was measured in the 400- to 880-nm wavelength region by illuminating it with the output beam of a spectrophotometer while the filter was attached to the front of a 30 x 32 pixel silicon array detector. The filter was designed to be used in the output beam of a grating spectrometer to prevent the dispersal of higher diffracted orders onto an array detector. Areas of the filter that were spatially matched to the corresponding detector pixel column had measured peak transmittances of about 90 percent that were uniform to within +/- 1.5 percent along a given column. Transmittances for incident wavelengths shorter than the desired bandpass, corresponding to the order overlap region, were measured in the 0.003 range. Line spread function measurements made with the array detector indicated no significant beam spreading caused by inserting the filter into the beam.
Heaney, James B.↗
Mechanism for Particle Transport and Size Sorting via Low-Frequency Vibrations
There is a need for effective sample handling tools to deliver and sort particles for analytical instruments that are planned for use in future NASA missions. Specifically, a need exists for a compact mechanism that allows transporting and sieving particle sizes of powdered cuttings and soil grains that may be acquired by sampling tools such as a robotic scoop or drill. The required tool needs to be low mass and compact to operate from such platforms as a lander or rover. This technology also would be applicable to sample handling when transporting samples to analyzers and sorting particles by size.
Sherrit, Stewart↗
Shape-induced Gravitational Sorting of Saharan Dust During Transatlantic Voyage: Evidence from CALIOP Lidar Depolarization Measurements
Motivated by the physical picture of shape-dependent air resistance and, consequently, shape-induced differential sedimentation of dust particles, we searched for and found evidence of dust particle asphericity affecting the evolution and distribution of dust-scattered light depolarization ratio (delta). Specifically, we examined a large data set of Cloud-Aerosol Lidar with Orthogonal Polarization (CALIOP) observations of Saharan dust from June to August 2007. Observing along a typical transatlantic dust track, we find that (1) median delta is uniformly distributed between 2 and 5 km altitudes as the elevated dust leaves the west coast of Africa, thereby indicating uniformly random mixing of particle shapes with height; (2) vertical homogeneity of median delta breaks down during the westward transport: between 2 and 5 km delta increases with altitude and this increase becomes more pronounced with westward progress; (3) delta tends to increase at higher altitude (greater than 4 km) and decrease at lower altitude (less than 4 km) during the westward transport. All these features are captured qualitatively by a minimal model (two shapes only), suggesting that shape-induced differential settling and consequent sorting indeed contribute significantly to the observed temporal evolution and vertical stratification of dust properties. By implicating particle shape as a likely cause of gravitational sorting, these results will affect the estimates of radiative transfer through Saharan dust layers.
CALIPSO↗
Sediment Sorting and Rounding in a Basaltic Glacio-Fluvio-Aeolian Environment: Ϸhórisjökull Glacier, Iceland
Sediments and sedimentary rocks preserve a rich history of environment and climate. Identifying these signals requires an understanding of the physical and chemical processes that have affected sedimentary deposits [1]. Such processes include sorting and rounding during transport and chemical alteration through weathering and diagenesis. Although these processes have long been studied in quartz-dominated sedimentary systems [2], a lack of studies of basaltic sedimentary systems limits our interpretations of the environment and climate where mafic source rocks dominate, such as on Mars [3,4]. As part of the SAND-E: Semi-Autonomous Navigation for Detrital Environments project [5], which uses robotic operations to examine physical and chemical changes to sediments in basaltic glacio-fluvialaeolian environments, this research studies changes in sorting and rounding of fluvial-aeolian sediments along a glacier-proximal-to-glacier-distal transect in the outwash-plain of the Ϸόrisjökull glacier in SW Iceland (Fig. 1)
Mason, K. G.↗
A Simulation Study of Bin-and-Sort Policies in a Distributed System for Flights Scheduling
This is a report on the results of a simulation experiment conducted on an automated distributed system for scheduling flights in a shared airspace. The scheduling is carried out by computer systems owned by the private operators of the flights, which interface through a centralized independent server. The scheduling requires determining which of the group of flights at hand (whether owned by the same operator or different ones) are to have the advantage of being scheduled before the others. A chosen method of grouping and prioritizing flights must: have the ability to schedule a flight promptly, prevent scheduling conflicts between pairs of overlapping flight routes, and be computationally feasible. Scheduling flights one by one is the First Come First Served policy. Scheduling too many at a time is infeasible. The type of scheduling policy proposed herein, called bin-and-sort, is an attempt to reach a compromise between these two extremes. Bin-and-sort allows for each operator to choose its criteria for prioritizing its flights, and for the centralized component to use different arbitration criteria. A number of these criteria are used in the simulation presented herein. The results of each criterion choice are reported and analyzed.
flight scheduling↗
A Simulation Study of Bin-and-Sort Policies in a Distributed System for Flights Scheduling
This is a report on the results of a simulation experiment conducted on an automated distributed system for scheduling flights in a shared airspace. The scheduling is carried out by computer systems owned by the private operators of the flights, which interface through a centralized independent server. The scheduling requires determining which of the group of flights at hand (whether owned by the same operator or different ones) are to have the advantage of being scheduled before the others. A chosen method of grouping and prioritizing flights must: have the ability to schedule a flight promptly, prevent scheduling conflicts between pairs of overlapping flight routes, and be computationally feasible. Scheduling flights one by one is the First Come First Served policy. Scheduling too many at a time is infeasible. The type of scheduling policy proposed herein, called bin-and-sort, is an attempt to reach a compromise between these two extremes. Bin-and-sort allows for each operator to choose its criteria for prioritizing its flights, and for the centralized component to use different arbitration criteria. A number of these criteria are used in the simulation presented herein. The results of each criterion choice are reported and analyzed.
flight scheduling↗
Chip-based droplet sorting
A non-contact system for sorting monodisperse water-in-oil emulsion droplets in a microfluidic device based on the droplet's contents and their interaction with an applied electromagnetic field or by identification and sorting.
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
Sorting of plastics
Systems and methods for classifying and sorting of plastic materials utilizing a vision system and one or more sensor systems, which may implement a machine learning system in order to identify or classify each of the materials, which may then be sorted into separate groups based on such an identification or classification.
Kumar, Nalin↗
Symmetry-Breaking and Self-Sorting in Block Copolymer-Based Multicomponent Nanocomposites
Co-assembly of inorganic nanoparticles (NPs) and nanostructured polymer matrix represents an intricate interplay of enthalpic or entropic forces. Particle size largely affects the phase behavior of the nanocomposite. Theoretical studies indicate that new morphologies would emerge when the particles become comparable to the soft matrix's size, but this has rarely been supported experimentally. By designing a multicomponent blend composed of NPs, block copolymer-based supramolecules, and small molecules, a 3-D ordered lattice beyond the native BCP's morphology was recently reported when the particle is larger than the microdomain of BCP. The blend can accommodate various formulation variables. In this work, when the particle size equals the microdomain size, a symmetry-broken phase appears in a narrow range of particle sizes and compositions, which we named the "train track" structure. In this phase, the NPs aligned into a 3-D hexagonal lattice and packed asymmetrically along the c axis, making the projection of the ac and the bc plane resemble train tracks. Computational studies show that the broken symmetry reduces the polymer chain deformation and stabilizes the metastable hexagonally perforated lamellar morphology. Given the mobility of the multicomponent blend, the system shows a self-sorting behavior: segregating into two macroscopic phases with different nanostructures based on only a few nanometers NP size differences. Smaller NPs form "train track" morphology, while larger NPs form a "simple hexagon" structure, where the NPs take a symmetric hexagonal arrangement. Detailed structural evolution and simulation studies confirm the systematic-wide cooperativity across different components, indicating the strong self-regulation of the multicomponent system.
"train track" morphology↗
Extensive Genome-Wide Phylogenetic Discordance Is Due to Incomplete Lineage Sorting and Not Ongoing Introgression in a Rapidly Radiated Bryophyte Genus
The relative importance of introgression for diversification has long been a highly disputed topic in speciation research and remains an open question despite the great attention it has received over the past decade. Gene flow leaves traces in the genome similar to those created by incomplete lineage sorting (ILS), and identification and quantification of gene flow in the presence of ILS is challenging and requires knowledge about the true phylogenetic relationship among the species. We use whole nuclear, plastid, and organellar genomes from 12 species in the rapidly radiated, ecologically diverse, actively hybridizing genus of peatmoss (Sphagnum) to reconstruct the species phylogeny and quantify introgression using a suite of phylogenomic methods. We found extensive phylogenetic discordance among nuclear and organellar phylogenies, as well as across the nuclear genome and the nodes in the species tree, best explained by extensive ILS following the rapid radiation of the genus rather than by postspeciation introgression. Our analyses support the idea of ancient introgression among the ancestral lineages followed by ILS, whereas recent gene flow among the species is highly restricted despite widespread interspecific hybridization known in the group. Our results contribute to phylogenomic understanding of how speciation proceeds in rapidly radiated, actively hybridizing species groups, and demonstrate that employing a combination of diverse phylogenomic methods can facilitate untangling complex phylogenetic patterns created by ILS and introgression.
59 BASIC BIOLOGICAL SCIENCES↗
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↗
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↗
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↗
Laser particle activated cell sorting in microfluidics
A microfluidic device is developed that uses the narrowband optical radiation of laser particle barcodes to sort tagged cells.
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗