Switchable Double Inversion of Chirality in a Helical Polyelectrolyte.

Switchable inversion of chirality between opposite handedness by varying pH is discovered for a histidine pendant polymer, polymethyl (4-vinylbenzoyl) histidinate (PBHis), as demonstrated by the circular dichroism as well as the changes of hydrodynamic radius measured by fluorescence correlation spectroscopy at the single molecular level. The polyelectrolyte is found to take an M-helicity below pH 8.0 and change into P-helicity above pH 8.0. Such helicity further inverses into M-chirality above pH 10.6. All these helical structures with opposite handedness can be switched using pH variations. The mechanism of such a unique phenomenon is attributed to the protonation and deprotonation of the imidazole group and the hydroxide-ion-mediated hydrogen bonding, which determine the mutual orientation between the adjacent side groups under the hydrogen bonds and π-π stacking and therefore the handedness of the helical structure.

Free Space Makes the Polymer "Dead Layer" Alive.

The effect of free space on molecular motion inside the polymer "dead layer" or adsorbed nanolayers on solid surfaces is investigated. Free space is introduced into the nanolayer by choosing a polymer with a relatively big side group, poly n-butyl methacrylate (PnBMA), and polarization-resolved single-molecule fluorescence microscopy is adopted as the method. The rotational motion of the doped fluorescent probes is found to be considerably excited at moderate temperatures, attributed to the free space brought by the side group of the PnBMA. The development of the adsorbed nanolayer by the prolonged annealing of the parent film is carefully monitored, together with the evolution of the molecular motion and the glass transition temperature (Tg). The Tg values of the exposed nanolayers are considerably lower than that of the bulk system, while they become higher than those in the bulk situation when the nanolayer is covered with a polymer top layer. The experimental evidence has demonstrated that the free space made available by the side group and the air-polymer interface has considerably promoted the molecular motion inside the adsorbed nanolayers, even under the situation of overwhelming surface attraction.

Poly(ethylene oxide) Is Positively Charged in Aqueous Solutions.

There have been controversies about the binding of cations to poly(ethylene oxide) (PEO) chains in aqueous solutions. In the current study, single molecular evidence of charging PEO chains by cation binding in aqueous solutions is provided. From the adoption of the photon-counting histogram method, it is discovered that the local pH value at the vicinity of the PEO chain is higher than the bulk solution, showing that the PEO chain is positively charged. Such a situation exists with and without the presence of salt (NaCl) in the solution, presumably due to the binding of cations, such as hydronium and sodium ions. Single molecular electrophoresis experiments using fluorescence correlation spectroscopy demonstrate that the PEO chains are weakly charged with a charging extent of ~5%. In comparison to the salt-free condition, the addition of external salt (NaCl) at moderate concentrations further charges the chain. The charging causes the PEO chains to expand and a further increase in the salt concentration causes the chain to shrink, exhibiting a polyelectrolyte-like behavior, demonstrated by the hydrodynamic radii of a single PEO chain. The effect of ion identity is discovered with alkali cations, with the order of the charging capacity of Li+ < Na+ < Cs+ < K+.

Molecular motion activated by residual stress in a glassy polymer thin film.

The activation, by residual stress, of the fast portion of rotational motion of single fluorescent probe molecules inside a polymer thin film near its glass transition temperature is studied at a single molecular level. Spin-casted poly n-butyl methacrylate thin films without thermal annealing are chosen as the model system and single molecule fluorescence defocused microscopy is adopted as the method. The rotational motion of the probes under residual stress is found to be more activated than that under mere thermal activation, and the kinetic energy exhibits a monotonic increase with the stress strength. A rough linear dependence of rotational kinetic energy at low stress is found, yielding the value of characteristic volume for the residual stress to activate the motion of the probes. The values of the volume are close to the van der Waals volume of the probes, indicating that the activation of the fast dynamics by residual stress is localized. The activation effect is weakened and vanishes at or above the glass transition temperature due to stress relaxation. The effect is also absent at temperatures far below Tg due to the frozen molecular motion with a much higher activation energy.

Effect of Counterion Binding to Swelling of Polyelectrolyte Brushes.

The effect of binding strength of counterions with the polyelectrolyte chain to the swelling of polyelectrolyte brushes is studied, by investigating the swelling of both the polycation and polyanion in response to the variation of the salt concentration under the change of counterion's identity. Two polyelectrolyte brushes grafted on solid substrates are adopted: the cationic poly [2-(methacryloyloxy)ethyltrimethyl ammonium] (PMETA-X, X = F, Cl, Br, and I) and the anionic polystyrene sulfonate (M-PSS, M = Li, Na, K, and Cs). The swelling change with the salt concentration is investigated by ellipsometry, quartz crystal microbalance with dissipation, and dielectric spectroscopy. It is discovered that although the thickness of PMETA-X brushes is larger than that of M-PSS brushes of similar grafting density, the former has much less solvent incorporated than the latter. Such a difference is attributed to the weaker interaction between the PMETA+ chain and its halide counterions than that between the PSS- chain and its alkali counterions, discovered by dielectric spectroscopy. This makes the original charges on the PMETA-X chain less neutralized and therefore have a higher charge density, compared with the M-PSS chain. The results demonstrate that the stronger binding of the counterions to the polyelectrolytes makes the main chains less charged, resulting in the weaker inter-chain electrostatic repulsion and less swelling of the brushes. Investigations into the effect of ion identity show the following order of binding strength: for the cationic PMETA+ chain, F- < Cl- < Br- < I- and for the anionic PSS- chain, Li+ < Na+ < K+ < Cs+.

Clusterin inhibits Aß42 aggregation through a "strawberry model" as detected by FRET-FCS.

Extracellular plaque deposits of ß-amyloid peptide (Aß) are one of the main pathological features of Alzheimer's disease (AD). The aggregation of Aß42 species, especially Aß42 oligomers, is still an active research field in AD pathogenesis. Secretory clusterin protein (sCLU), an extracellular chaperone, plays an important role in AD pathogenesis. Although sCLU interacts directly with Aß42 in vitro and in vivo, the mechanism is not clear. In this paper, His-tagged sCLU (sCLU-His) was cloned, expressed and purified, and we applied florescence resonance energy transfer-fluorescence correlation spectroscopy (FRET-FCS) to investigate the direct interaction of sCLU-His and Aß42 at the single-molecule fluorescence level in vitro. Here, we chose four different fluorescently labeled Aß42 oligomers to form two different groups of aggregation models, easy or difficult to aggregate. The results showed that sCLU-His could form complexes with both aggregation models, and sCLU-His inhibited the aggregation of Aß42/RB  ~ Aß42/Atto647 (easy to aggregate model). The complexes were produced as the Aß42/Label adhered to the sCLU-His, which is similar to a "strawberry model," as strawberry seeds are dotted on the outer surface of strawberries. This work provided additional insight into the interaction mechanism of sCLU and Aß42 .

Polymeric liquid layer densified by surface acoustic wave.

Creating densified and stable liquid is a straightforward strategy for the fabrication of strong and ultra-stable amorphous or glassy materials. The current study has discovered that a liquid polymeric thin film is densified under the application of a high frequency surface acoustic wave (SAW). The experimental evidence is the decrease in film thickness and the increase in refractive index, measured by ellipsometry, of polyisobutylene thin films deposited on the solid substrates, when a high frequency SAW (39.5 MHz) is applied to the system. Further investigations by polarization-resolved single molecule fluorescence microscopy have demonstrated that the rotational motion of fluorescent probes doped inside the liquid film is retarded and the dynamical heterogeneity is reduced. The results demonstrate that the application of SAW of high frequency makes the thin polymeric liquid film densified and more dynamically homogeneous.

Anomalous Diffusion Inside Soft Colloidal Suspensions Investigated by Variable Length Scale Fluorescence Correlation Spectroscopy.

The diffusion of molecules and particles inside the aqueous suspension of soft colloids (polymer microgels) is investigated using variable length scale fluorescence correlation spectroscopy (VLS-FCS). Carbopol 940 is chosen as the model matrix system, and two factors affecting diffusion are investigated: the spatial hindrance and the diffusant-matrix interaction. By studying diffusion of molecules and particles with different sizes inside the suspension, VLS-FCS reveals the restricted motion at a short length scale, that is, in the gaps between the microgels, and normal diffusion at a larger length scale. The information on the gap's length scale is also accessed. On the other hand, by tuning the pH value, the diffusant-matrix electrostatic attraction is adjusted and the results expose a short-time fast diffusion of probe molecules inside the gaps and a long-time restricted diffusion because of trapping inside the microgels. It is proved that VLS-FCS is a powerful method, investigating anomalous diffusion at different length scales and it is a promising approach to investigate diffusion in complex soft matter systems.

Lateral diffusion of single polymer molecules at interfaces between water and oil.

Lateral diffusion of polymer molecules at the interfaces between immiscible oil and water is investigated at the single molecular level. The interfaces between water and alkanes are chosen as the model systems and polyethylene oxide (PEO) is the model polymer. Fluorescence correlation spectroscopy is used to measure the interfacial diffusion of fluorescence-labeled PEO with its molecular weight ranging over more than an order of magnitude. It is discovered that the interfacial diffusion coefficient scales with the molecular weight by the exponent of -0.5. Detailed analysis shows that the PEO chain takes an ideal two-dimensional random coil conformation at these fluidic interfaces and the bigger contribution from water's hydrodynamic friction is discovered.

A negative correlation between water content and protein adsorption on polymer brushes.

The correlation between the water content and protein adsorption on the surface of polymer brushes is investigated quantitatively. Using a number of model systems such as poly N-isopropylacrylamide (PNIPAM), polyethylene glycol (PEG), poly(2-(methacryloyloxy)ethyl phosphorylcholine) (PMPC), poly(2-hydroxyethyl methacrylate) (PHEMA), and poly(2,2,3,4,4,4-hexafluorobutyl)methacrylate (PHFBMA) polymer brushes with various grafting densities, the amount of water hydrodynamically coupled inside the brushes and its correlation with protein adsorption (BSA and Fg proteins as model systems) were determined by quartz crystal microbalance with dissipation (QCM-D) and surface plasma resonance (SPR). The results demonstrate the negative correlation between the water content and protein adsorption - the more water is coupled inside the brushes, the more protein adsorption is suppressed. In particular for PNIPAM brushes with a high enough grafting density and with a water content greater than 250 ng cm-2, the protein adsorption is negligible.

Cell membrane mimetic copolymer coated polydopamine nanoparticles for combined pH-sensitive drug release and near-infrared photothermal therapeutic.

In this study, we modified the well-known photothermal polydopamine nanoparticles with the poly[(2-methacryloyloxy)ethyl phosphorylcholine-b-(2-dimethylamino)ethyl methacrylate] (PMPC-b-PDMAEMA) diblock copolymers, containing both biocompatible cell membrane phosphorylcholine zwitterions segments and pH-responsive dimethylaminoethyl units on the polymer chains, to achieve both the photothermal property and pH-responsive release behavior. The results showed that the obtained nanoparticles had a narrow size distribution with the diameter about 220 nm. Besides, the modified polydopamine nanoparticles showed enhanced colloidal stability, pH-sensitive drug release behavior, excellent biocompatibility and remarkable near-infrared photothermal property. Thus, it is highly anticipated that PMPC-b-PDMAEMA modified polydopamine nanoparticles can serve as a powerful drug delivery system for combined pH-sensitive drug release and near-infrared photothermal therapeutic.

Molecular weight dependence of chain conformation of strong polyelectrolytes.

Using sodium polystyrene sulfonate (NaPSS) and quarternized poly 4-vinylpyridine (QP4VP) as model systems, the chain conformation of polyelectrolytes under finite salt concentrations is investigated at a single molecular level. By fluorescence correlation spectroscopy (FCS), the hydrodynamic radius (R h) of the samples with the molecular weight ranging more than one order of magnitude was measured. The variations of R h as a function of molecular weight reveal the molecular weight dependence: under moderate salt concentrations (such as 10-4 and 0.1M), the shorter chains of both NaPSS and QP4VP take the rod-like conformation, while the longer chains take the coiled conformation (random coil or swelled random coil conformation, respectively). At high enough salt levels, both the charged chains take the coiled conformations. Photon counting histogram (PCH) measurements of the local pH value at the vicinity of the NaPSS chain expose the higher extent of counterion adsorption for longer chains as well as higher salt concentrations, telling that the charge regularization process is the major governing factor.

The effects of fluorescent labels on Aß42 aggregation detected by fluorescence correlation spectroscopy.

Fluorescence-based methods are promising for measuring amyloid beta (Aß) oligomers, given their capacity to analyse a sample at the single-molecule level. As the attachment of fluorescent labels may influence the biochemical properties of the Aß oligomers, the effects of fluorescent labels on Aß oligomers must be evaluated. In this paper, we compared the impacts of five different fluorescent dyes on the aggregation of Aß42 oligomers using fluorescence correlation spectroscopy (FCS). We found that fluorescent labels of BODIPY® FL-C5 (BP), N-hydroxysuccinimide rhodamine B ester (RB) and rhodamine B isothiocyanate (RITC) increased the propensity of labelled Aß42 oligomers to aggregate, whereas 6-(fluorescein-5-carboxamido) hexanoic acid succinimidyl ester (5-SFX) and fluorescein 5(6)-isothiocyanate (5(6)-FITC) decreased the propensity of labelled Aß42 oligomers to aggregate. This difference originated from the different electric charges and hydrophobicity of the fluorescent dyes. These results provide valuable information for establishing different aggregation models for Aß42 oligomers in vitro using FCS.

Response of a Permanently Charged Polyelectrolyte Brush to External Ions: The Aspects of Structure and Dynamics.

Structure and dynamics inside permanently charged polyelectrolyte brushes, sodium polystyrene sulfonate brushes, during their response to the introduction of external ions (NaCl) are investigated by neutron reflectivity and dielectric spectroscopy. Neutron reflectivity measurements show that the segmental density of the inner part of the brushes decreases and that of the outer part increases when the salt level is tuned from the salt-free condition to a moderate level (<10-2 M)-the brushes swell further compared with the salt-free condition. This is attributed to the breakup of the multiplets formed by dipole-dipole pairs, and by this process, the previously constrained chain segments by the multiplets are released. Dielectric spectroscopy discovers a giant dipole by the charge separation of the adsorbed counterions and the PSS- chains, induced by electric field. The dynamics of the induced giant dipole is accelerated with the increase of external salt, as a result of the charge regularization by elevated salt level. At high-enough salt level, the screening effect reduces the electrostatic repulsion between the neighboring chains and makes the brushes shrink.

Charge evolution during the unfolding of a single DNA i-motif.

The effective charge and evolution of single chains of a DNA i-motif during its unfolding process are investigated at the single molecule level. Using fluorescence correlation spectroscopy and photon counting histograms, the single chain dimensions and electrical potential of cytosine-rich human telomeric oligonucleotides are monitored, during their unfolding from the i-motif to the random coil state. It is discovered that the effective charge density of the DNA chain is very sensitive to conformation changes and the results remarkably expose the existence of an intermediate state of the unfolding process. A huge difference in pH value exists in the vicinity of the DNA chain and the bulk solution, depending on the salt concentration, as reflected by a down-shift in the pH value of unfolding. The presence of an external salt in the solution helps to stabilize the i-motif structure at low pH values due to the reduction of the effective charge density. It can also destabilize the folded structure in the pH range of the conformation transition due to the elevation of the local pH value, encouraging the deprotonation of the cytosine groups. These results provide new information for understanding the structure and stability of i-motif DNA, and its biological function, as well as the building blocks for smart nanomaterials.

Light-Switchable Self-Healing Hydrogel Based on Host-Guest Macro-Crosslinking.

A self-healing hydrogel is prepared by crosslinking acrylamide with a host-guest macro-crosslinker assembled from poly(ß-cyclodextrin) nanogel and azobenzeneacrylamide. The photoisomerizable azobenzene moiety can change its binding affinity with ß-cyclodextrin, therefore the crosslinking density and rheology property of the hydrogel can be tuned with light stimulus. The hydrogel can repair its wound autonomously through the dynamic host-guest interaction. In addition, the wounded hydrogel will lose its ability of self-healing when exposed to ultraviolet light, and the self-healing behavior can be recovered upon the irradiation of visible light. The utilizing of host-guest macro-crosslinking approach manifests the as-prepared hydrogel reversible and light-switchable self-healing property, which would broaden the potential applications of self-healing polymers.

Interfacial diffusion of a single cyclic polymer chain.

Lateral diffusion of cyclic polystyrene at the interface between fused silica and dichloromethane is investigated at the single-molecular level. Narrowly distributed cyclic polystyrene (c-PS) of high purity with molecular weights spanning nearly an order of magnitude is prepared via atom transfer radical polymerization (ATRP) and Cu-catalyzed azide/alkyne cycloaddition (CuAAC) click reaction. The diffusion coefficients (D) of c-PS and its linear analogue (l-PS) on the surface are measured by fluorescence correlation spectroscopy (FCS). The diffusion coefficient of c-PS is discovered to have an inverse dependence on molecular weight, D ∼ M-1, in contrast to the case of linear polystyrene, which reproduces a stronger molecular weight dependence, D ∼ M-3/2. The slower interfacial motion of cyclic chains is attributed to their stronger binding to the surface and more importantly, the unique surface diffusion mechanism due to the absence of free chain ends.

Single chains of strong polyelectrolytes in aqueous solutions at extreme dilution: Conformation and counterion distribution.

The molecular conformation of two typical polyelectrolytes, sodium polystyrene sulfonate (NaPSS) and quarternized poly-4-vinylpyridine (QP4VP), was studied in aqueous solutions without salt addition at the single molecular level. By fluorescence correlation spectroscopy, the hydrodynamic radius (Rh) of NaPSS and QP4VP with the molecular weight ranging more than one order of magnitude was measured. The scaling analysis of Rh exhibits scaling exponent of 0.70 and 0.86 for NaPSS and QP4VP in solutions without added salts, respectively, showing the conformation is much more expanded than random coil. Numerical fittings using the model of diffusion of a rod molecule agree with the data well, indicating that the polyelectrolyte chains take the rod-like conformation under the condition without salt addition. Further investigations by determining the electric potential of single PSS- chains using the photon counting histogram technique demonstrate the enhanced counterion adsorption to the charged chain at higher molecular weight.

Examining dynamics in a polymer matrix by single molecule fluorescence probes of different sizes.

Rotational motion of single fluorescence probes with different sizes doped inside films of polyvinylacetate is investigated by defocused single molecule fluorescence microscopy. Discrete vibration modes in the rotational motion of individual probes are clearly exposed in the power spectra of the rotation trajectories, reflecting multiple relaxation dynamics and also the difference in dynamics among separate locations. The power spectra show a strong dependence on the probe size and temperature. By sampling the rotation of a large number of probes, the averaged power spectra show that the activated rotation of the probes falls into the frequency range of the α-relaxation and data analysis shows that activation of the vibration modes with all probes corresponds to the maximum population of rotating probes, and this made the comparison between single molecule data and the ensemble data meaningful (differential scanning calorimetry, as an example). Such an analysis shows a coincidence between the temperature of a significant occurrence of glass transition and the activation of rotational motion of all probes with one specific size, indicating that such a dimension correlates with the size of the cooperative rearrangement region - a volume of approximately 1.0 nm(3).

Huge Differences in the Kinetics of Swelling Enhancement and De-enhancement of Permanently Charged Polyelectrolyte Brushes.

As demonstrated previously (X. Chu et al., Soft Matter 2014, 10, 5568), permanently charged polyelectrolyte brushes can experience an enhancement of swelling by exposure to an external monovalent salt solution in moderate concentrations. Beyond the previous static measurements, the kinetics of the swelling enhancement and de-enhancement were investigated in the current study by using a quartz crystal microbalance with dissipation (QCM-D). By developing an effective approach to quantify the response in QCM-D, a vast difference in swelling enhancement and de-enhancement of a model permanently charged polyelectrolyte brush (sodium polystyrene sulfonate, NaPSS) was discovered. The results indicate new physics of the charged brushes: the difference in the attachment and detachment of counterions to the polyelectrolyte chains.