Biofouling potential of surface-enhanced Raman scattering-based seawater quality sensors by Ulva spp.

This study investigated the biofouling potential of surface-enhanced Raman scattering (SERS)-based sensor materials in the context of marine environments. Uncoated and monolithic commercial gold (Au) silicon nanopillar array SERS substrates, Au-coated carbon black nanoparticle (AuCB NP) substrates, uncoated and Au sputter-coated in-house SERS, and uncoated and Au sputter-coated glass controls were tested for biofouling potential using Ulva spp. as model biofouling organisms. The mean percentages of Ulva spp. zoospores that adhered per mm2 (×103) on the uncoated and coated Au silicon nanopillar array, AuCB NP, uncoated and Au sputter-coated in-house, and uncoated and Au sputter-coated glass substrates were 10.28%, 5.45%, 10.49%, 3.25%, 24.84%, 12.86% and 7.78%, respectively. Results indicated that surface properties such as hydrophobicity, roughness, Au sputter-coating and the presence of micro-refuges on nano- and microstructured substrates were critical to the biofouling formation.

Plasmon-Tuned Particles for the Amplification of Surface-Enhanced Raman Scattering from Analytes.

Inelastic scattering from molecules because of vibrational modes produces unique Raman shifts, allowing these analytes to be detected with high specificity. Because Raman scattering is weak, surface-enhanced Raman scattering (SERS) has been used as a label-free technique for the detection of a variety of analytes at low concentrations. Using simple solution-based colloidal processing techniques, we have fabricated gold-coated carbon-black nanoparticles that show enhanced Raman activity. By varying the fabrication conditions, we create particles of different surface morphologies, allowing control over the peak wavelength for localized surface plasmon resonance (LSPR). By matching the LSPR wavelength to the incident laser wavelength, we get the highest signal from two model analytes, 4-nitrobenzenethiol (4-NBT) and Congo Red (CR). Our straightforward room-temperature-solution-based approach for making tunable SERS-active particles expands the range of incident radiation wavelengths that can be used for the detection of analytes using Raman scattering.

Interaction of Cyanobacteria with Nanometer and Micron Sized Polystyrene Particles in Marine and Fresh Water.

Microplastics and nanoplastics are emerging pollutants, widespread both in marine and in freshwater environments. Cyanobacteria are also ubiquitous in water and play a vital role in natural ecosystems, using photosynthesis to produce oxygen. Using photography, fluorescence microscopy and cryogenic and scanning electron microscopy (cryo-SEM, SEM) we investigated the physicochemical response of one of the most predominant seawater cyanobacteria (Synechococcus elongatus, PCC 7002) and freshwater cyanobacteria (S. elongatus Nageli PCC 7942) when exposed to 10 µm diameter polystyrene (microPS) and 100 nm diameter polystyrene (nanoPS) particles. Marine and freshwater cyanobacteria formed aggregates with the nanoPS, bound together by extracellular polymeric substances (EPS), and these aggregates sedimented. The aggregates were larger, and the sedimentation was more rapid for the marine system. Aggregate morphologies were qualitatively different for the microPS samples, with the bacteria linking up a small number of particles, all held together by EPS. There was no sedimentation in these samples. The cyanobacteria remained alive after exposure to the particles. The particle size- and salt concentration-dependent response of cyanobacteria to these anthropogenic stressors is an important factor to consider for a proper understanding of the fate of the particles as well as the bacteria.

Attachment of Alcanivorax borkumensis to Hexadecane-In-Artificial Sea Water Emulsion Droplets.

Alcanivorax borkumensis (AB) is a marine bacterium that dominates bacterial communities around many oil spills because it enzymatically degrades the oil while using it as a nutrient source. Several dispersants have been used to produce oil-in-water emulsions following a spill. Compared to surface slicks, the additional oil-water surface area produced by emulsification provides greater access to the oil and accelerates its degradation. We deliberately cultured AB cells using hexadecane as the only nutrient source. We then examined the first critical step of the biodegradation process, the attachment of these AB cells to hexadecane-water interfaces, using fluorescence microscopy and cryogenic scanning electron microscopy. The hexadecane-in-artificial sea water (ASW) emulsions were produced by gentle shaking and were stabilized either by AB alone, by Corexit 9500, by Tween 20, or by carbon black particles. When no dispersants were used, AB stabilizes the emulsion, and bacterial cells attach to the hexadecane droplets within the first 3 days. When Corexit 9500 was used as the dispersant, AB did not attach to the hexadecane droplets over 3 days, and many AB cells in the aqueous phase appeared dead. Only limited attachment was observed after 7 days. No AB attachment was observed over 3 days when Tween 20 was used as the dispersant. However, the bacteria used Tween 20 in the ASW as a nutrient. Large amounts of AB attached to carbon black stabilized hexadecane droplets within 3 days. An analysis that accounts for van der Waals and electrostatic interactions is unable to predict all of these observations, indicating that the attachment of AB to the hexadecane is a complex phenomenon that goes beyond simple physiochemical effects. While these experiments do not mimic conditions in the open ocean where the large amount of water dilutes any emulsion stabilizer, they provide important insights on bacteria adhesion to oil, a critical step in the oil degradation process following a marine spill.

Behavior of Marine Bacteria in Clean Environment and Oil Spill Conditions.

Alcanivorax borkumensis is a bacterial community that dominates hydrocarbon-degrading communities around many oil spills. The physicochemical conditions that prompt bacterial binding to oil/water interfaces are not well understood. To provide key insights into this process, A. borkumensis cells were cultured either in a clean environment condition (dissolved organic carbon) or in an oil spill condition (hexadecane as the sole energy source). The ability of these bacteria to bind to the oil/water interface was monitored through interfacial tension measurements, bacterial cell hydrophobicity, and fluorescence microscopy. Our experiments show that A. borkumensis cells cultured in clean environment conditions remain hydrophilic and do not show significant transport or binding to the oil/water interface. In sharp contrast, bacteria cultured in oil spill conditions become partially hydrophobic and their amphiphilicity drives them to oil/water interfaces, where they reduce interfacial tension and form the early stages of a biofilm. We show that it is A. borkumensis cells that attach to the oil/water interface and not a synthesized biosurfactant that is released into solution that reduces interfacial tension. This study provides key insights into the physicochemical properties that allow A. borkumensis to adhere to oil/water interfaces.

Influence of the Oil on the Structure and Electrochemical Performance of Emulsion-Templated Tin/Carbon Anodes for Lithium Ion Batteries.

Tin (Sn) is a useful anode material for lithium ion batteries (LIBs) because of its high theoretical capacity. We fabricated oil-in-water emulsion-templated tin nanoparticle/carbon black (SnNP/CB) anodes with octane, hexadecane, 1-chlorohexadecane, and 1-bromohexadecane as the oil phases. Emulsion creaming, the oil vapor pressure, and the emulsion droplet size distribution all affect drying and thus the morphology of the dried emulsion. This morphology has a direct impact on the electrochemical performance of the anode. SnNP/CB anodes prepared with hexadecane showed very few cracks and had the highest capacities and capacity retention. The combination of low vapor pressure, creaming, which forced the emulsion droplets into a close-packed arrangement on the surface of the continuous water phase, and the small droplets allowed for gentle evaporation of the liquids during drying. This led to lower differential stresses on the sample and reduced cracking. For octane, the vapor pressure was high, the droplet sizes were large for 1-cholorohexadecane, and there was no creaming for 1-bromohexadecane. All of these factors contributed to cracking of the anode surface during drying and reduced the electrochemical performance. Choosing an oil with balanced properties is important for obtaining the best cell performance for emulsion-templated anodes for LIBs.

Near-Infrared Responsive Gold-Layersome Nanoshells.

Anionic liposomes coated with cationic polyelectrolyte poly-l-lysine (PLL), or layersomes, were used as soft, self-assembled templates for synthesizing gold nanoshells that absorb near-infrared radiation. The gold nanoshells were formed using two techniques: (a) direct reduction of tetrachloroauric acid on the layersomes and (b) the reduction of a tetrachloroauric acid/potassium carbonate "growth" solution on nanosized gold seeds bound to the surface of layersomes. The resulting structures were characterized by transmission and scanning electron microscopy and visible-near-infrared spectroscopy. Direct reduction produced discrete gold nanoparticles on the layersomes. The slower reduction from the growth solution on the gold seeds resulted in more complete shells. The absorption spectra of these suspensions were sensitive to the synthesis method. The morphology of the gold shells was tuned for absorption at biologically safe and tissue-penetrating NIR wavelengths, and laser irradiation at 810 nm produced significant heat. These gold-layersome nanoshells have the potential to be used for photothermal therapy, photothermally mediated drug delivery, and biomedical imaging.

Destabilization of Oil-in-Water Emulsions Stabilized by Non-ionic Surfactants: Effect of Particle Hydrophilicity.

We investigate the use of particle hydrophilicity as a tool for emulsion destabilization in Triton-X-100-stabilized hexadecane-in-water emulsions. The hydrophilicity of the particles added to the aqueous phase was found to have a pronounced effect on the stability of the emulsion. Specifically, the addition of hydrophilic fumed silica particles to the aqueous phase resulted in coarsening of the emulsion droplets, with droplet flocculation observed at higher particle concentrations. On the other hand, when partially hydrophobic fumed silica particles were added to the aqueous phase, coarsening of the emulsion droplets was observed at low particle concentrations and phase separation of oil and water was observed at higher particle concentrations. Surface tension and interfacial tension measurements showed significant depletion of the surfactant from the aqueous phase in the presence of the partially hydrophobic particles. The observed changes in the stability of the emulsion and the depletion of the surfactant can be rationalized in terms of changes in the adsorption behavior of the surfactant molecules, from one dominated by hydrogen bonding on hydrophilic particles to one dominated by hydrophobic interactions on partially hydrophobic particles. Our findings also provide, for the first time, an in-depth understanding of antagonistic (destabilizing) effects in mixtures of partially hydrophobic particles and a non-ionic surfactant (Triton X-100) in water.

Patchy Layersomes Formed by Layer-by-Layer Coating of Liposomes with Strong Biopolyelectrolytes.

Layer-by-layer deposition of polyelectrolytes (PEs) onto self-assembled liposomes represents an alternative to PE deposition on solid particles for the formation of hollow nanoscale capsules. This work examines how competition between PE-liposome and inter-PE interactions drives the structure and colloidal stability of layersomes. Unlike solid particles, liposomes respond to adsorbed material through lipid reorganization and changes in size and shape. This responsive nature could yield new types of layered PE structures. We show that sequential deposition of strong biopolyelectrolytes, dextran sulfate-sodium salt (DxS-) and poly-l-arginine (PA+), onto cationic liposomes in water yields the expected charge inversion behavior commonly observed for dispersed particles. However, cryogenic transmission electron microscopy results show that the layersomes formed and their PE coatings were heterogeneous. The PE coatings contained PE complexes (PECs) that were formed when an even number of layers (2 or 4) was deposited. PECs remained attached as patches that were spatially distinguishable. This behavior was confirmed through fluorescence anisotropy measurements of liposome bilayer fluidity, where PA+ counteracted the ordering effects of DxS- on the lipid bilayer through charge neutralization and local PEC desorption. With increased charge screening, DxS- desorbed from the layersomes, whereas the patchy layersomes terminating in PA+ retained their PE coatings and colloidal stability at higher salt concentrations. To our knowledge, this is the first time such patchy layersome structures have been observed.

Interaction of Alcanivorax borkumensis with a Surfactant Decorated Oil-Water Interface.

Alcanivorax borkumensis is a hydrocarbon degrading bacterium linked to oil degradation around oil spill sites. It is known to be a surface bacterium leading to substantial interaction with the oil-water interface. Because of its abundance in oil spill regions, it has great potential to be used actively in oil spill remediation. Dispersants are thought to be important in the creation of oil-in-water emulsions that are meant to aid in the biodegradation process by bacteria. Although it is likely that some sort of dispersant will be used again in the case of another oil spill, to date, no studies have shown the impact of dispersants on the bacteria population. Corexit 9500 was the main dispersant used during the Deepwater Horizon oil spill, but little is known about its effect on the bacteria community. We built an experimental platform to quantitatively measure the transient growth of Alcanivorax borkumensis at the interface of oil and water. To our knowledge, this is the first study of how A. borkumensis interacts with a surfactant decorated oil-water interface. We use COREXIT EC9500A, cetylytrimethylamonium bromide, dioctyl sulfosuccinate sodium salt, l-α-phosphatidylcholine, sodium dodecyl sulfate, and Tween 20 to investigate the impact of dispersants on Alcanivorax borkumensis. We assess the impact of these dispersants on the growth rate, lag time, and maximum concentration of Alcanivorax borkumensis. We show that the charge, structure, and surface activity of these surfactants greatly impact the growth of A. borkumensis. Our results indicated that out of the surfactants tested only Tween 20 assists Acanivorax borkumensis growth. The results of this study will be important in the decision of dispersant use in the future.

Tuning the Wettability of Halloysite Clay Nanotubes by Surface Carbonization for Optimal Emulsion Stabilization.

The carbonization of hydrophilic particle surfaces provides an effective route for tuning particle wettability in the preparation of particle-stabilized emulsions. The wettability of naturally occurring halloysite clay nanotubes (HNT) is successfully tuned by the selective carbonization of the negatively charged external HNT surface. The positively charge chitosan biopolymer binds to the negatively charged external HNT surface by electrostatic attraction and hydrogen bonding, yielding carbonized halloysite nanotubes (CHNT) on pyrolysis in an inert atmosphere. Relative to the native HNT, the oil emulsification ability of the CHNT at intermediate levels of carbonization is significantly enhanced due to the thermodynamically more favorable attachment of the particles at the oil-water interface. Cryogenic scanning electron microscopy (cryo-SEM) imaging reveals that networks of CHNT attach to the oil-water interface with the particles in a side-on orientation. The concepts advanced here can be extended to other inorganic solids and carbon sources for the optimal design of particle-stabilized emulsions.

Radio Frequency-Activated Nanoliposomes for Controlled Combination Drug Delivery.

This work was conducted in order to design, characterize, and evaluate stable liposomes containing the hydrophobic drug raloxifene HCl (RAL) and hydrophilic doxycycline HCl (DOX), two potentially synergistic agents for treating osteoporosis and other bone lesions, in conjunction with a radio frequency-induced, hydrophobic magnetic nanoparticle-dependent triggering mechanism for drug release. Both drugs were successfully incorporated into liposomes by lipid film hydration, although combination drug loading compromised liposome stability. Liposome stability was improved by reducing the drug load and by including Pluronics® (PL) in the formulations. DOX did not appear to interact with the phospholipid membranes comprising the liposomes, and its release was maximized in the presence of radio frequency (RF) heating. In contrast, differential scanning calorimetry (DSC) and phosphorus-31 nuclear magnetic resonance ((31)P-NMR) analysis revealed that RAL developed strong interactions with the phospholipid membranes, most notably with lipid phosphate head groups, resulting in significant changes in membrane thermodynamics. Likewise, RAL release from liposomes was minimal, even in the presence of RF heating. These studies may offer useful insights into the design and optimization of multidrug containing liposomes. The effects of RAL on liposome characteristics and drug release performance underscore the importance of appropriate physical-chemical analysis in order to identify and characterize drug-lipid interactions that may profoundly affect liposome properties and performance early in the formulation development process.

Response of surfactant stabilized oil-in-water emulsions to the addition of particles in an aqueous suspension.

As a model for understanding how surfactant-stabilized emulsions respond to the addition of interacting and noninteracting particles, we investigated the response of dodecane-in-water emulsions stabilized by SDS (anionic), CTAB (cationic), and Triton X-100 (nonionic) surfactants to the addition of an aqueous suspension of negatively charged fumed silica particles. The stability of the emulsion droplets and the concentration of surfactants/particles at the oil-water interfaces are sensitive to surfactant-particle interactions, mixing conditions, and the particle concentration in the bulk. Addition of the particle suspension to the SDS-stabilized emulsions showed no effect on emulsion stability. Coarsening of emulsion droplets is observed when fumed silica particles were added to emulsions stabilized by Triton X-100. Depending on the concentration of silica particles in the suspension, the addition of fumed silica particles to CTAB-stabilized emulsions resulted in droplet coalescence and phase separation of oil and water or formation of particle-coated droplets. Vigorous (vortex) mixing allows the particles to breach the oil-water interfaces and stabilize emulsions. While we have examined a specific particle suspension and a set of three surfactants, these observations can be generalized for other surfactant-particle mixtures.

Phase and steady shear behavior of dilute carbon black suspensions and carbon black stabilized emulsions.

We use para-amino benzoic acid terminated carbon black (CB) as a model particulate material to study the effect of salt-modulated attractive interactions on phase behavior and steady shear stresses in suspensions and particle-stabilized emulsions. Surprisingly, the suspension displayed a yield stress at a CB volume fraction of ϕCB = 0.008. The yield stress scaled with CB concentration with power law behavior; the power law exponent changed abruptly at a critical CB concentration, suggesting a substantial change in network structure. Cryogenic scanning electron microscopy revealed structural differences between the networks found in each scaling regime. Randomly oriented pores with thick CB boundaries were observed in the scaling region above the critical particle concentration, suggesting a strong gel network, and long, oriented pores were found in the scaling region below the critical particle concentration, suggesting a weak network influenced by an induced shear stress. These findings correlate with the existence of gels and transient networks. Transient networks break down under gravitational forces over time periods of 12-24 hours. The yield stresses of CB-gels containing oil emulsion droplets were found to scale with carbon black concentration similar to the CB-gels without oil. These results offer insight into salt-induced attractive colloidal networks and the difference in structure and yield-stress behavior between transient networks and gels. Furthermore, CB offers the ability to stabilize an oil phase in discrete droplets and contain them within a rigid network structure.

Two-dimensional materials as emulsion stabilizers: interfacial thermodynamics and molecular barrier properties.

A new application for two-dimensional (2D) materials is emulsification, where they can serve as ultrathin platelike interfacial stabilizers in two-liquid systems. We present a first detailed thermodynamic analysis of atomically thin 2D materials at organic-aqueous liquid-liquid interfaces and derive expressions for the transfer free energies of emulsion stabilization that account for material geometry, van der Waals transparency or opacity, and variable hydrophobicity. High mass potency is shown to be an intrinsic property of the 2D geometry, which at the atomically thin limit places every atom in contact with both liquid phases, resulting in unit atom efficiency. The thermodynamic model successfully predicts that graphene oxide but not pristine graphene has a favorable hydrophobic-hydrophilic balance for oil-water emulsion stabilization. Multilayer tiling is predicted to occur by the passivation of droplet surface patches left uncovered by packing inefficiencies in the first monolayer, and complete multilayer coverage is confirmed by cryogenic scanning electron microscopy. The molecular barrier function of graphene interfacial films causes a significant suppression of dispersed-phase evaporation rates with potential applications in controlled release. Finally, these emulsions can be used as templates for creating solid graphene foams or graphene microsacks filled with lipophilic cargos. Emerging 2D materials are promising as dispersants or emulsifiers where high mass potency and multifunctional properties are desired.

Release of surfactant cargo from interfacially-active halloysite clay nanotubes for oil spill remediation.

Naturally occurring halloysite clay nanotubes are effective in stabilizing oil-in-water emulsions and can serve as interfacially-active vehicles for delivering oil spill treating agents. Halloysite nanotubes adsorb at the oil-water interface and stabilize oil-in-water emulsions that are stable for months. Cryo-scanning electron microscopy (Cryo-SEM) imaging of the oil-in-water emulsions shows that these nanotubes assemble in a side-on orientation at the oil-water interface and form networks on the interface through end-to-end linkages. For application in the treatment of marine oil spills, halloysite nanotubes were successfully loaded with surfactants and utilized as an interfacially-active vehicle for the delivery of surfactant cargo. The adsorption of surfactant molecules at the interface serves to lower the interfacial tension while the adsorption of particles provides a steric barrier to drop coalescence. Pendant drop tensiometry was used to characterize the dynamic reduction in interfacial tension resulting from the release of dioctyl sulfosuccinate sodium salt (DOSS) from halloysite nanotubes. At appropriate surfactant compositions and loadings in halloysite nanotubes, the crude oil-saline water interfacial tension is effectively lowered to levels appropriate for the dispersion of oil. This work indicates a novel concept of integrating particle stabilization of emulsions together with the release of chemical surfactants from the particles for the development of an alternative, cheaper, and environmentally-benign technology for oil spill remediation.

Homogeneous percolation versus arrested phase separation in attractively-driven nanoemulsion colloidal gels.

We elucidate mechanisms for colloidal gelation of attractive nanoemulsions depending on the volume fraction (ϕ) of the colloid. Combining detailed neutron scattering, cryo-transmission electron microscopy and rheological measurements, we demonstrate that gelation proceeds by either of two distinct pathways. For ϕ sufficiently lower than 0.23, gels exhibit homogeneous fractal microstructure, with a broad gel transition resulting from the formation and subsequent percolation of droplet-droplet clusters. In these cases, the gel point measured by rheology corresponds precisely to arrest of the fractal microstructure, and the nonlinear rheology of the gel is characterized by a single yielding process. By contrast, gelation for ϕ sufficiently higher than 0.23 is characterized by an abrupt transition from dispersed droplets to dense clusters with significant long-range correlations well-described by a model for phase separation. The latter phenomenon manifests itself as micron-scale "pores" within the droplet network, and the nonlinear rheology is characterized by a broad yielding transition. Our studies reinforce the similarity of nanoemulsions to solid particulates, and identify important qualitative differences between the microstructure and viscoelastic properties of colloidal gels formed by homogeneous percolation and those formed by phase separation.

The response of carbon black stabilized oil-in-water emulsions to the addition of surfactant solutions.

We use carboxyl-terminated, negatively charged, carbon black (CB) particles suspended in water to create CB-stabilized octane-in-water emulsions, and examine the consequences of adding aqueous anionic (SOS, SDS), cationic (OTAB, DTAB), and nonionic (Triton X-100) surfactant solutions to these emulsions. Depending upon the amphiphile's interaction with particles, interfacial activity, and bulk concentration, some CB particles get displaced from the octane-water interfaces and are replaced by surfactants. The emulsions remain stable through this exchange. Particles leave the octane-water interfaces by two distinct modes that depend on the nature of particle-surfactant interactions. Both happen over time scales of the order of seconds. For anionic and nonionic surfactants that bind to the CB through hydrophobic interactions, individual particles or small agglomerates stream away steadily from the interface. Cationic surfactants bind strongly to the carboxylate groups, reduce the magnitude of the surface potential, and cause the CB particles to agglomerate into easily visible chunks at the droplet interfaces. These chunks then leave the interfaces at discrete intervals, rather than in a steady stream. For the longer chain cationic surfactant, DTAB, the particle ejection mode reverts back to a steady stream as the concentration is increased beyond a threshold. This change from chunks of particles leaving intermittently to steady streaming is because of the formation of a surfactant bilayer on the particles that reverses the particle surface charge and makes them highly hydrophilic. The charge reversal also suppresses agglomeration. Zeta potentials of CB particles measured after exposure to surfactant solutions support this hypothesis. These results are the first systematic observations of different particle release modes from oil-water interfaces produced by variations in interactions between surfactants and particles. They can be generalized to other particle-surfactant systems and exploited for materials synthesis.

Shear-directed assembly of graphene oxide in aqueous dispersions into ordered arrays.

A wide variety of new carbon-based materials are being developed from graphene oxide (GO) precursor sheets, whose assembly in aqueous phases determines the form, structure, and properties of the resultant carbon. Here we show that graphene oxide forms ordered linear arrays of aggregates when aqueous suspensions are subjected to shear flow in the presence of soluble salts. These linear arrays align along the vorticity direction, normal to the direction of flow. We propose that salt addition screens electrostatic repulsion and allows formation of fractal-like GO sheet aggregates by hydrophobic forces. Fluid shear in a confined gap then guides the assembly of these primary aggregates into optically visible, ordered linear arrays or "superaggregates" whose characteristics are a function of GO concentration, salt valency, salt concentration, and gap confinement. This is the first reported observation of vorticity banding in graphene oxide suspensions and the first reported observation of such banding based on salt-induced interactions. We also demonstrate that simple isometric nanoparticles of carbon or gold do not form such linear superaggregate arrays but can be assembled into such arrays using graphene oxide as a two-dimensional colloidal template.

Shear free and blotless cryo-TEM imaging: a new method for probing early evolution of nanostructures.

Cryogenic transmission electron microscopy (cryo-TEM) is a powerful method to image native state morphologies of nanoscale soft and hard objects suspended in solvents. Sample preparation is a critical step toward producing images at length and time scales of interest. We demonstrate a nearly shear-free sample thinning method which simultaneously allows imaging of evolving nanostructures at subsecond time scales. This device breaks the trade-off between high shear and short time scales typical in current cryo-TEM sample preparation methods. We demonstrate the low-shear feature of the new method by imaging wormlike micelles, showing an interconnected network, in contrast to the traditional sample preparation method which shows aligned micelles at similar time points. The time resolution of this method is demonstrated by imaging morphologies of calcium carbonate (formed through the reaction of calcium chloride with sodium carbonate) at subsecond time scales, capturing its evolution from an amorphous to a crystalline state. The impact of hyperbranched polyglycerol additives on the amorphous to crystalline transition in calcium carbonate at short times is examined. Early images at low shear provide unique fundamental insights into mechanisms of nanostructure evolution, thus offering a new paradigm for research in materials sciences, soft matter, and biological sciences.