Differential Sedimentation Research Articles

The Southern Atlas Front in Tunisia and its foreland basin: Structural style and regional-scale deformation

Surface and subsurface data together with balanced cross-sections are used to emphasize the structural style and regional-scale deformation of the Southern Atlas Front in Tunisia and its foreland basin. This E–trending shaped belt reflects successive tectonic events. Mixed thick- and thin-skinned structural style characterizing this belt is defined by shallow thrusts rooted down into deep E–trending basement thrusts with major decollement level located within Triassic salt. A NW–trending system is also recognized in the front and particularly responsible for the belt's curved shape, which probably acts as transfer areas between the wide-ranging E–trending major structures. The intermediate areas are characterized by salt tectonics probably active during Jurassic and Lower Cretaceous, in response to active synsedimentary normal faults and differential sedimentation loading. The shortening amount varies and decreases from West to East and from North to South: ~7.45% in the West, ~3.3% in west-central part, ~2% in the central area to insignificant (~1.4%) in the east. From north to south, the Northern Chotts Range narrow zone accommodates overall the Cenozoic shortening with ~6.95% and less than 1% in the Chotts region. This variation coincides with the pre-Cenozoic basin's architecture acquired during Mesozoic Tethyan evolution, which generates the main Jurassic E–trending Chotts basin. Inside this basin, distributary sub-basins are formed in relation with NW–trending faults active during the Cretaceous period. The Chotts basin appears, therefore, as Mesozoic rift rollover structure well-preserved in the front of the belt. The compilation of data suggests that the Front of the southeasternmost end of the Atlassic domain is distinguished by the reactivation of inherited fault systems acting during alpine orogeny as major southward thrusts. The foreland shows a structural singularity to be formed by a preserved almost unmodified and well exposed large Mesozoic rollover structure.

Titanium Oxide Microspheres with Tunable Size and Phase Composition.

Due to their unique physical and chemical properties, monodisperse titanium oxide microspheres can be used in dye-sensitized solar cells, as cosmetic pigments, and for other applications. However, the synthesis of microspheres with narrow size distribution, desired phase composition, and porosity is still a challenge. In this work, spherical titania particles with controllable size, crystallinity, and pore size were obtained by Ti(OnBu)4 hydrolysis in ethanol. The influence of NaOH addition on the particles’ size and morphology was investigated for the first time. Particle diameter can be tailored from 300 nm to 1.5 μm by changing water and NaOH concentrations. Particle size was analyzed by the statistical processing of scanning electron microscopy (SEM) images and differential centrifugal sedimentation (DCS) measurements. Optical properties of the microspheres were studied by diffuse reflectance UV-Vis spectroscopy. Thermal and hydrothermal treatment allowed transforming amorphous phase in as-prepared particles into nanocrystalline anatase and/or rutile. Transmission electron microscopy (TEM) study of the lamellae, cut out from spherical particles using focused ion beam (FIB), revealed that as-synthesized microspheres are non-hollow, homogeneous, and crystallize throughout the whole volume of the particle. The spherical particles possess photoprotective properties; the highest sun protection factor (SPF) was observed for amorphous microspheres.

The (relative) insignificance of G revisited to include nanoparticles

AbstractThe collision efficiency functions, α(i,j), of two colliding particles for the three operative transport mechanisms in flocculation (Brownian motion [Br], fluid shear [Sh], and differential sedimentation [DS]) were numerically calculated over a broad size range (including nanoparticles). All computations include the hydrodynamic force and van der Waals (vdW) attraction, while the effects of electrical double‐layer repulsive forces were investigated by their exclusion or inclusion. Flocculation of nanoparticles should be extremely rapid if they are uncharged but will be dramatically reduced when substantially charged. For small neutral particles, α(i,j) values could be greater than those obtained during Sh and DS as vdW attraction outweighs the (negative) hydrodynamic effects. Overall, the results confirm the findings of an earlier article, that Br is the dominant flocculation mechanism when at least one particle is small (<1 μm in diameter), Sh is only dominant when both particles are greater than 1 μm and the ratio of the two particle sizes is less than 10, and DS is dominant in all other cases. Flocculators should be operated with low‐velocity gradient (G) values (in the range of 10–20 s−1), with only sufficient mixing to keep particles in suspension, but proper particle destabilization is essential for effective flocculation.

Click Chemistry on the Surface of Ultrasmall Gold Nanoparticles (2 nm) for Covalent Ligand Attachment Followed by NMR Spectroscopy.

Ultrasmall gold nanoparticles (core diameter 2 nm) were surface-conjugated with azide groups by attaching the azide-functionalized tripeptide lysine(N3)-cysteine-asparagine with ∼117 molecules on each nanoparticle. A covalent surface modification with alkyne-containing molecules was then possible by copper-catalyzed click chemistry. The successful clicking to the nanoparticle surface was demonstrated with 13C-labeled propargyl alcohol. All steps of the nanoparticle surface conjugation were verified by extensive NMR spectroscopy on dispersed nanoparticles. The particle diameter and the dispersion state were assessed by high-resolution transmission electron microscopy (HRTEM), differential centrifugal sedimentation (DCS), and 1H-DOSY NMR spectroscopy. The clicking of fluorescein (FAM-alkyne) gave strongly fluorescing ultrasmall nanoparticles that were traced inside eukaryotic cells. The uptake of these nanoparticles after 24 h by HeLa cells was very efficient and showed that the nanoparticles even penetrated the nuclear membrane to a very high degree (in contrast to dissolved FAM-alkyne alone that did not enter the cell). About 8 fluorescein molecules were clicked to each nanoparticle.

Elastocapillary flow driven lab-on-a-membrane device based on differential wetting and sedimentation effect for blood plasma separation

We report an elastocapillary flow—driven lab on a membrane device based on differential wetting and sedimentation effect for the separation of plasma from whole blood. Interaction between a thin polydimethylsiloxane (PDMS) membrane (thickness ~35 μm) bonded to the edge of a PDMS substrate and a sample blood drop (of volume ~70 μl) gives rise to deformation of the soft membrane due to the capillary forces providing a conduit and consequent elastocapillary flow of blood. The surface of the PDMS membrane is hydrophilic up to a certain length along the flow direction to support the elastocapillary flow and hydrophobic thereafter to impede the flow. In the hydrophobic region, owing to a much smaller sedimentation time scale (~100 s) as compared to the capillary flow time scale (~1000 s), sedimentation of blood cells occurs thus facilitating separation of plasma from the blood cells in the hydrophobic region. The role of differential wetting and sedimentation effects on the blood plasma separation is studied. The effects of membrane width and thickness, length of the hydrophilic region, erythrocyte sedimentation rate (ESR) on the separation of plasma were investigated. Using a membrane of width 3 mm, thickness 35 μm, total length 25 mm and hydrophilic length of 4 mm and 70 μl of whole blood with ESR varying in the range 4 mm h−1 to 40 mm h−1, the volume of plasma was in the range of 7.5 μl to 20 μl, respectively, which corresponds to a plasma recovery of 22%–49%, respectively. Purity of the plasma from the proposed device was compared with that obtained from centrifugation which showed a good match. The device was integrated with a commercially available detection strip to detect the level of glucose present in the plasma from blood samples of healthy and diabetic patients which are in qualitative agreement with that obtained from conventional tests.

Number Concentration of Gold Nanoparticles in Suspension: SAXS and spICPMS as Traceable Methods Compared to Laboratory Methods.

The industrial exploitation of high value nanoparticles is in need of robust measurement methods to increase the control over product manufacturing and to implement quality assurance. InNanoPart, a European metrology project responded to these needs by developing methods for the measurement of particle size, concentration, agglomeration, surface chemistry and shell thickness. This paper illustrates the advancements this project produced for the traceable measurement of nanoparticle number concentration in liquids through small angle X-ray scattering (SAXS) and single particle inductively coupled plasma mass spectrometry (spICPMS). It also details the validation of a range of laboratory methods, including particle tracking analysis (PTA), dynamic light scattering (DLS), differential centrifugal sedimentation (DCS), ultraviolet visible spectroscopy (UV-vis) and electrospray-differential mobility analysis with a condensation particle counter (ES-DMA-CPC). We used a set of spherical gold nanoparticles with nominal diameters between 10 nm and 100 nm and discuss the results from the various techniques along with the associated uncertainty budgets.

Sticky Measurement Problem: Number Concentration of Agglomerated Nanoparticles.

Measuring the number concentration of colloidal nanoparticles (NPs) is critical for assessing reproducibility, enabling compliance with regulation, and performing risk assessments of NP-enabled products. For nanomedicines, their number concentration directly relates to their dose. However, the lack of relevant reference materials and established traceable measurement approaches make the validation of methods for NP number concentration difficult. Furthermore, commercial products often exhibit agglomeration, but guidelines for dealing with nonideal samples are scarce. We have compared the performance of five benchtop measurement methods for the measurement of colloidal number concentration in the presence of different levels of agglomeration. The methods are UV-visible spectroscopy, differential centrifugal sedimentation, dynamic light scattering, particle tracking analysis, and single-particle inductively coupled plasma mass spectrometry. We find that both ensemble and particle-by-particle methods are in close agreement for monodisperse NP samples and three methods are within 20% agreement for agglomerated samples. We discuss the sources of measurement uncertainties, including how particle agglomeration affects measurement results. This work is a first step toward validation and expansion of the toolbox of methods available for the measurement of real-world NP products.

Combining Small-Angle X-ray Scattering and X-ray Powder Diffraction to Investigate Size, Shape and Crystallinity of Silver, Gold and Alloyed Silver-Gold Nanoparticles

The combination of simultaneous measurements of small and wide angle X-ray scattering (SAXS/WAXS–SWAXS) to investigate the overall size, shape, and crystallinity of silver nanoparticles and alloyed silver-gold nanoparticles in the size range of 8 to 80 nm is shown. The obtained results for overall size are in agreement with the particle size obtained by differential centrifugal sedimentation (DCS) and transmission electron microscopy (TEM). In addition to the overall size, SWAXS provided precise information about the crystallographic internal structure of the particles, providing a powerful multi-scale tool for structural characterization of the studied systems.

The Regenerative Potential of Amniotic Fluid Stem Cell Extracellular Vesicles: Lessons Learned by Comparing Different Isolation Techniques

Extracellular vesicles (EVs) derived from amniotic fluid stem cells (AFSCs) mediate anti-apoptotic, pro-angiogenic, and immune-modulatory effects in multiple disease models, such as skeletal muscle atrophy and Alport syndrome. A source of potential variability in EV biological functions is how EV are isolated from parent cells. Currently, a comparative study of different EV isolation strategies using conditioned medium from AFSCs is lacking. Herein, we examined different isolation strategies for AFSC-EVs, using common techniques based on differential sedimentation (ultracentrifugation), solubility (ExoQuick, Total Exosome Isolation Reagent, Exo-PREP), or size-exclusion chromatography (qEV). All techniques isolated AFSC-EVs with typical EV morphology and protein markers. In contrast, AFSC-EV size, protein content, and yield varied depending on the method of isolation. When equal volumes of the different AFSC-EV preparations were used as treatment in a model of lung epithelial injury, we observed a significant variation in how AFSC-EVs were able to protect against cell death. AFSC-EV enhancement of cell survival appeared to be dose dependent, and largely uninfluenced by variation in EV-size distributions, relative EV-purity, or their total protein content. The variation in EV-mediated cell survival obtained with different isolation strategies emphasizes the importance of testing alternative isolation techniques in order to maximize EV regenerative capacity.

Zonal rotor centrifugation revisited: new horizons in sorting nanoparticles.

Density gradient centrifugation is an effective method for the isolation and purification of small particles. Hollow rotors capable of hosting density gradients replace the need for centrifuge tubes and therefore allow separations at large scales. So far, zonal rotors have been used for biological separations ranging from the purification of whole cells down to serum proteins. We demonstrate that the high-resolution separation method opens up exciting perspectives apart from biology, namely in sorting mixtures of synthetic nanoparticles. Loading and unloading, while the rotor is spinning, avoids perturbations during acceleration and deceleration periods, and thus makes a vital contribution to sorting accuracy. Nowadays one can synthesize nanoscale particles in a wide variety of compositions and shapes. A prominent example for this are “colloidal molecules” or, generally speaking, defined assemblies of nanoparticles that can appear in varying aggregation numbers. Fractionation of such multimodal colloids plays an essential role with regard to their organization into hierarchical organized superstructures such as films, mesocrystals and metamaterials. Zonal rotor centrifugation was found to be a scalable method of getting “colloidal molecules” properly sorted. It allows access to pure fractions of particle monomers, dimers, and trimers, just as well as to fractions that are essentially rich in particle tetramers. Separations were evaluated by differential centrifugal sedimentation, which provides high-resolution size distributions of the collected nanoparticle fractions. The performance achieved in relation to resolution, zone widths, sorting efficiencies and recovery rates clearly demonstrate that zonal rotor centrifugation provides an excellent solution to the fractionation of nanoparticle mixtures.

X-ray powder diffraction to analyse bimetallic core-shell nanoparticles (gold and palladium; 7-8 nm).

A comparative X-ray powder diffraction study on poly(N-vinyl pyrrolidone) (PVP)-stabilized palladium and gold nanoparticles and bimetallic Pd–Au nanoparticles (both types of core–shell nanostructures) was performed. The average diameter of Au and Pd nanoparticles was 5 to 6 nm. The two types of core–shell particles had a core diameter of 5 to 6 nm and an overall diameter of 7 to 8 nm, i.e. a shell thickness of 1 to 2 nm. X-ray powder diffraction on a laboratory instrument was able to distinguish between a physical mixture of gold and palladium nanoparticles and bimetallic core–shell nanoparticles. It was also possible to separate the core from the shell in both kinds of bimetallic core–shell nanoparticles due to the different domain size and because it was known which metal was in the core and which was in the shell. The spherical particles were synthesized by reduction with glucose in aqueous media. After purification by multiple centrifugation steps, the particles were characterized with respect to their structural, colloid-chemical, and spectroscopic properties, i.e. particle size, morphology, and internal elemental distribution. Dynamic light scattering (DLS), differential centrifugal sedimentation (DCS), atomic absorption spectroscopy (AAS), ultraviolet-visible spectroscopy (UV-vis), high-angle annular dark field imaging (HAADF), and energy-dispersed X-ray spectroscopy (EDX) were applied for particle characterization.

Solution NMR Spectroscopy with Isotope-Labeled Cysteine (13C and 15N) Reveals the Surface Structure of l-Cysteine-Coated Ultrasmall Gold Nanoparticles (1.8 nm).

Ultrasmall gold nanoparticles with a diameter of 1.8 nm were synthesized by reduction of tetrachloroauric acid with sodium borohydride in the presence of l-cysteine, with natural isotope abundance as well as 13C-labeled and 15N-labeled. The particle diameter was determined by high-resolution transmission electron microscopy and differential centrifugal sedimentation. X-ray photoelectron spectroscopy confirmed the presence of metallic gold with only a few percent of oxidized Au(+I) species. The surface structure and the coordination environment of the cysteine ligands on the ultrasmall gold nanoparticles were studied by a variety of homo- and heteronuclear NMR spectroscopic techniques including 1H-13C-heteronuclear single-quantum coherence and 13C-13C-INADEQUATE. Further information on the binding situation (including the absence of residual or detached l-cysteine in the solution) and on the nanoparticle diameter (indicating the well-dispersed state) was obtained by diffusion-ordered spectroscopy (1H-, 13C-, and 1H-13C-DOSY). Three coordination environments of l-cysteine on the gold surface were identified that were ascribed to different crystallographic sites, supported by geometric considerations of the nanoparticle ultrastructure. The particle size data and the NMR-spectroscopic analysis gave a particle composition of about Au174(cysteine)67.

A high-resolution study of in situ surface-enhanced Raman scattering nanotag behavior in biological systems

The colloidal stability of surface-enhanced Raman scattering (SERS) nanotags (Raman reporter-conjugated plasmonic nanoparticles) significantly affects the accuracy and reproducibility of SERS measurements, particularly in biological systems. Limited understanding of SERS nanotag stability may partly hamper the translation of SERS nanotags from the laboratory to their use in the clinic. In this contribution, we utilized differential centrifugal sedimentation (DCS), a reliable and straightforward technique to comprehensively analyze the colloidal stability of SERS nanotags in biological systems. Compared with other particle characterization techniques, DCS has been shown to have a unique advantage for high-resolution and high-throughput polydisperse particle characterization. DCS data revealed that the universal aggregation prevention practice of coating SERS nanotags with silica or bovine serum albumin layers did not sufficiently stabilize them in common measurement environments (e.g., 1 × PBS). Combined DCS and SERS measurements established a strong correlation between the degrees of nanotag aggregation and signal intensities, further reinforcing the necessity of characterizing SERS nanotag stability for every condition in which they are used. We also found that increasing the protein thickness by the inclusion of extra protein components in the detection environments and antibody functionalization can improve the stability of SERS nanotags. We believe that this study can provide guidelines on appropriate measurement techniques and particle design considerations to assess and improve SERS nanotag stability in complex biological systems.

Comparative biological effects of spherical noble metal nanoparticles (Rh, Pd, Ag, Pt, Au) with 4-8 nm diameter.

For a comparative cytotoxicity study, nanoparticles of the noble metals Rh, Pd, Ag, Pt, and Au (spherical, average diameter 4 to 8 nm) were prepared by reduction in water and colloidally stabilized with poly(N-vinyl pyrrolidone) (PVP). Thus, their shape, size, and surface functionalization were all the same. Size and morphology of the nanoparticles were determined by dynamic light scattering (DLS), analytical disc centrifugation (differential centrifugal sedimentation, DCS), and high-resolution transmission electron microscopy (HRTEM). Cell-biological experiments were performed to determine the effect of particle exposure on the viability of human mesenchymal stem cells (hMSCs). Except for silver, no adverse effect of any of the metal nanoparticles was observed for concentrations up to 50 ppm (50 mg L−1) incubated for 24 h, indicating that noble metal nanoparticles (rhodium, palladium, platinum, gold) that do not release ions are not cytotoxic under these conditions.

Anionic polyacrylamide efficiency in goethite removal from aqueous solutions: goethite suspension destabilization by PAM

The main aim of the study was to investigate the anionic polyacrylamide (PAM) efficiency as a flocculant relative to goethite particles. This mineral is a toxic waste produced during hydrometallurgical processes and, as a result, poses a threat to the environment. In the experiments, the following analytical methods were applied: potentiometric titration, zeta potential and adsorption amount measurements, nitrogen adsorption/desorption isotherms, Fourier transform infrared spectroscopy, differential centrifugal sedimentation, UV/Vis spectrophotometry. The result analysis showed that polyacrylamide adsorbs on the goethite surface; however, its adsorption amount depends on the pH value. Due to the adsorbent–adsorbate electrostatic repulsion, the smallest number of macromolecules adsorbs at pH 9. The PAM adsorption affects the goethite surface charge and zeta potential values. It makes these parameters more negative. What is more regardless of pH value, the polymer addition contributes to clear aggregation of solid particles (suspension destabilization). Thus, PAM can be considered as a potential flocculant improving the goethite removal from aqueous wastes.

Properties of natural rubber reinforced with cellulose nanofibers based on fiber diameter distribution as estimated by differential centrifugal sedimentation

Cellulose nanofibers (CNFs) with different degrees of fibrillation are prepared by the mechanical fibrillation of kraft pulp using wet disk milling, and dispersions of the prepared CNFs were subjected to differential centrifugal sedimentation (DCS) in order to estimate the diameter distributions of the CNFs. The low-fibrillated CNFs (fiber diameter (d): >10 μm) had a weak reinforcing effect on natural rubber (NR), while the medium-fibrillated CNFs (d: 0.1–10 μm) dramatically improve the initial modulus and decrease the elongation at break. The high-fibrillated CNFs (d: <0.1 μm) enhanced the tensile strength even further while maintaining the elongation at break. The reinforcing mechanism of the NR composites reinforced by the CNFs (NR-CNFs) was confirmed by field-emission scanning electron microscopy imaging, dynamic mechanical analysis, and toluene uptake measurements. It was concluded that these characteristic mechanical properties of the NR-CNFs were determined by the morphologies of the CNFs. The branching structure of the medium-fibrillated CNFs affected high improvement of the initial modulus, and the network formed by the high-fibrillated CNFs were involved in enhancement of the tensile strength without compromising viscoelastic properties. Understanding the effect of their diameter distribution can potentially reduce the production cost of CNFs and thus expand their applicability.

Experimental Assessment and Model Validation on How Shape Determines Sedimentation and Diffusion of Colloidal Particles.

Understanding the hydrodynamics of colloids with complex shapes is of equal importance to widespread practical applications and fundamental scientific problems, such as gelation, crystallization, and phase behavior. Building on previous work, we present a comprehensive study of sedimentation, diffusion, intrinsic viscosities, and other shape-dependent quantities of clusters built from spherical nanoparticles. Cluster preparation is accomplished by assembling surface-modified polystyrene particles on evaporating emulsion droplets. This results in supracolloids that exhibit well-defined configurations, which are governed by the number of constituent particles. Sorting into uniform cluster fractions is achieved through centrifugation of the cluster mixture in a density gradient. Sedimentation coefficients are elucidated by differential centrifugal sedimentation. Rotational and translational diffusion of the clusters are investigated by polarized and depolarized dynamic light scattering. The experimental results are compared to data obtained via a bead-shell model suitable for predicting hydrodynamic quantities of particles with arbitrary shapes. The experimental data are in excellent agreement with the predictions from hydrodynamic modeling. The variety of investigated shapes shows the robustness of our approach and provides a complete picture of the hydrodynamic behavior of complex particles.

Protein Corona Formation on Magnetic Nanoparticles Conjugated with Luminescent Europium Complexes

AbstractThe evaluation of the nanoparticles interactions with blood plasma enables a first and important insight on the organization of the adsorbed protein layer. In this context, we studied the formation of protein corona on magneto‐luminescent Fe3O4@calix‐Eu(TTA) nanoparticles in human plasma. The difference in the surface chemistry of F3O4 functionalized with calixarene (+30 mV ζ‐potential) and europium (III) thenoyltrifluoroacetonate (Eu3+‐TTA) complex (+7.4 mV ζ‐potential) affected the colloidal stability and hard corona composition of the nanoparticles, which were monitored by SDS‐PAGE gel electrophoresis, differential centrifugal sedimentation analyses and luminescence spectroscopy. Strikingly, after conjugation with Eu3+ TTA complex, the magnetic nanoparticles show lower adsorption affinity to proteins at higher blood plasma concentration. Moreover, the Eu3+ compound is featured with the narrow emission lines of 5D0→7FJ (J=0–4) transitions, imparting additional characteristics to bifunctional nanoparticles that were used as probes to evaluate the protein corona formation.

The continental extension discrepancy and anomalous subsidence pattern in the western Qiongdongnan Basin, South China Sea

Passive margins are first-order tectonic features on earth, formed by polyphase extension and breakup of continental lithosphere. It is accepted that mildly extended basins in the proximal margin form largely by uniform continental thinning, but the continental extension in deepwater is much debated. Here we choose a line from the western Qiongdongnan Basin in the magma-poor South China Sea margin to further explore the style of continental extension and the corresponding subsidence patterns, where high resolution seismic and heat flow data are available. Structural analysis reveals that the basin architecture is extremely asymmetric and mainly controlled by the SE directed normal faults, soling into a low-angle detachment fault. However, fault timings nicely show that the faults form through polyphase rifting and from inside outwards, and do not follow a simple sequential faulting like the West Iberia margin, where the upper and lower crustal deformation are strongly coupled. Perhaps the fault order here reflects a much weaker and decoupled lower crust. Then we divide the basin into five zones and calculate the average fault derived extension factors. Moreover, the whole crustal extension factors are also estimated after assessing the initial crustal thickness on basis of the backstripped subsidence. And the lithospheric extension factors are numerically modeled using a coupled lithosphere and basin scale model constrained by the heat flow and sedimentation data. Results show that along the profile, the overall lithospheric extension is much less than that of the whole crust, and only at the south end of the profile does lithospheric extension exceeds the whole crust, showing lithospheric depth-dependent thinning. The whole crustal extension is roughly equal to the faults derived extension in the northern part, displaying no extension discrepancy. In the middle part, the whole crustal extension is less than the faults derived extension, denoting inverse extension discrepancy. While the whole crustal extension is in far excess of the faults derived extension in the southern part, showing obvious positive extension discrepancy. We infer that the inverse and positive discrepancy of crustal extension was likely to be caused by landward directed lower crustal flow, driven by the horizontal pressure arising from the combination of the differential extension in the faulted upper crust, in the mantle lithosphere, and the differential sedimentation during the syn-rift period.

Investigating the Relative Contributions of Charge Deposition and Turbulence in Organizing Charge within a Thunderstorm

Abstract Large-eddy-resolving simulations using the Collaborative Model for Multiscale Atmospheric Simulation (COMMAS), which contains microphysical charging and branched-lightning parameterizations, produce much more complex net charge structures than conventionally visualized from previous observations, simulations, and conceptual diagrams. Many processes contribute to the hydrometeor charge budget within a thunderstorm, including advection, hydrometeor differential sedimentation, subgrid turbulent mixing and diffusion, ion drift, microphysical separation, and the attachment of ion charge deposited by the lightning channel. The lightning deposition, sedimentation, and noninductive charging tendencies contribute the most overall charge at relatively large scales, while the advection tendency, from resolved turbulence, provides the most “texture” at small scales to the net charge density near the updraft region of the storm. The scale separation increases for stronger storm simulations. In aggregate, lightning deposition and sedimentation resemble the smoother distribution of the electric potential, while evidence suggests individual flashes could be responding to the fine texture in the net charge. The clear scale separation between the advection and other net charge tendencies suggest the charge advection is most capable of providing net charge texture; however, a clear-cut causality is not obtained from this study.