The single-chain mechanics of two similar thermosensitive polymers, poly(N,N-diethylacrylamide) (PDEAM) and poly(N-isopropylacrylamide) (PNIPAM), have been studied by atomic force microscopy-based single-molecule force spectroscopy (SMFS). In a typical nonpolar organic solvent, octane, both of the polymers show the same inherent elasticity, although they have different substitutional groups. However, the mechanics of the two polymers presents large differences in water. The energies needed for the rearrangement of the bound water during elongation at room temperature are estimated by the SMFS method at the single-chain level, which is ∼1.13 ± 0.10 and ∼5.19 ± 0.10 kJ/mol for PDEAM and PNIPAM, respectively. In addition, PNIPAM shows a temperature-dependent single-chain mechanics when the temperature is increased across the lower critical solution temperature (LCST), while PDEAM does not. These differences observed in aqueous solution originate from the different structures of the two polymers. With a hydrogen bond donor in the amide group, PNIPAM will be more hydrated when T < LCST. When T > LCST, PNIPAM will have larger changes in both conformation and hydration. These findings also suggest that PNIPAM is a good candidate for a thermo-driven single-molecule motor, while PDEAM is not.

Sample preparation is crucial to the studies of polymers on surfaces and interfaces. For studies using single-molecule force spectroscopy (SMFS), sample preparation is the key to obtain high-quality data. In each of the previous SMFS studies, either physisorption or chemisorption was applied in the sample preparation. However, a direct comparison on the same polymer species with both of the two strategies has not been reported yet. With two methods (physisorption, “PS”, or chemisorption, “CS”) and two surfaces (AFM tip or quartz substrate), four types of samples of poly(ethylene glycol) (PEG) can be prepared from the polymer solution. The performance of these samples with the same species of PEG are directly compared by SMFS. It is found that among these samples, two of them, i.e., tip-CS and substrate-PS, are suitable for SMFS. The advantage of substrate-PS is the simple preparation. In contrast, the advantage of tip-CS is the higher rupture force and the lower sample consumption. The former feature will be time saving if a high rupture force is needed in the analysis. The latter feature will be economic when an expensive sample is used. The other two types of samples, i.e., tip-PS and substrate-CS show lower data yield and lower rupture force. In summary, the tip-CS and substrate-PS are recommended for sample preparation in SMFS. The tip-CS is the most promising protocol, if a functionalized polymer sample is available.

A novel environment-friendly system is proposed to fabricate polymer brush, which has the advantages including non-toxic and inexpensive initiator (eosin Y), visible-light exposure (λ = 515 nm), water medium and ambient environment. The experimental results from UV-Vis spectroscopy, AFM-based single molecule force spectroscopy (SMFS) and other measurements indicate that a polymer brush with a living nature is fabricated via free radical polymerization. This polymer brush may find applications in coatings, bio-interfaces and so forth.

Comparative studies of single molecule force spectroscopy and molecular dynamics simulations indicate that natural cellulose is more hydrophobic than amylose at the single-chain level, implying that the hydrophobicities of these polymeric isomers are regulated by only one parameter in the chains, the linkage between the sugar rings.

Dopamine (DA) and its polymer (PDA) have attracted broad interest in recent years. Due to the insolubility of PDA, however, it has been difficult to determine the exact structure and the polymerization mechanism. PDA is usually prepared under alkaline conditions (pH 8.5). In this case, the polymerization process may follow an equilibrium pathway to form indole-like repeating units, which lead to cross-linked structures. However, PDA can be prepared even under acidic conditions (pH 4.0) in the presence of an oxidant, ammonium persulfate (APS), which cannot be interpreted by previous mechanisms. Therefore, there should exist several pathways in parallel on the formation of PDA. Herein, a derivative of DA, 2-(4-methoxy-3-methylphenyl)ethylamine (MOE) that has fewer active sites, is used as a simplified model system to study the polymerization mechanism of DA. Experimental results from UV-Vis spectroscopy, single-molecule force spectroscopy and other measurements indicate that free radical polymerization of MOE occurs under both acidic and alkaline conditions in the presence of a polymerization initiator, APS. According to the mathematical principle of Set Theory and the fact that MOE has fewer active sites than DA, we speculate that free radical polymerization as a possible pathway should exist in parallel along with previously proposed pathways during the formation of PDA. These parallel pathways may be the main reason for the structural complexity of PDA.

The material properties of polylactic acid (PLA) are largely determined by its stereo-regularity (tacticity). To find out the origin at the molecular level, the single-chain mechanics of poly-L-lactic acid (PLLA) and poly-D,L-lactide (PDLLA) were comparatively investigated by single-molecule atomic force microscopy (AFM). At a low concentration, PLLA adopted a random-coil conformation in a good solvent. At a high concentration, however, the PLLA chain can be induced into a helix, which consumed additional energy during unfolding by further stretching. Due to the random arrangement of L- and D-repeating units in the PDLLA chain, PDLLA adopts a random-coil conformation at all concentrations. The difference in single-chain mechanics of PLLA and PDLLA at high concentrations may be the cause of their different macroscopic properties. This is the first report to reveal the stereo-regularity-dependent mechanics of a polymer at the single-molecule level, which may help to bridge the gap between understanding single-molecule and materials properties.

Both of temperature (in water) and composition (in the water/methanol mixed solvent) can induce the coil-to-globule transition of poly(N-isopropylacrylamide) (PNIPAM). The atomic force microscope (AFM) based single molecule force spectroscopy (SMFS) has been exploited to investigate the interactions between the polymer chain and solvent at the single-molecule level. It is found that the single-chain mechanics of PNIPAM show a remarkable dependence on the two external stimuli. A confusing experimental result is that all the force-extension (F-E) curves of unfolding an individual PNIPAM globule present a feature of elastic (monotonically increasing force) stretching but not plateau (constant force) stretching predicted by theory. In this article, we clarify that the presence of the interior solvent molecules in the single-chain globule is the origin of the discrepancy between the F-E curves obtained from theory and experiment. Although both of the external stimuli do tend to lower the solvent quality for PNIPAM, water and the water/methanol mixed solvent will never be the strongly poor solvent for PNIPAM, even at the worst condition.

Polymers with a carbon–carbon (C–C) backbone are an important class of polymers, which can be regarded as the derivatives of polyethylene (PE). To investigate the effect of side chains on the single-chain enthalpic elasticity (SCEE) of polymers with a C–C backbone, several polymers with pendants or side chains of different lengths and shapes have been studied by single-molecule AFM. We find that both length and shape of the side chains count: only the side chains that are both long and bulky (i.e., bulky dendrons of second or higher generation as side chains) affect the SCEE. Thus, only rare polymers have special SCEE. For the vast majority of polymers, the SCEE is identical to that of PE, which means that the SCEE is determined by the nature of the C–C backbone. It is expected that this conclusion can also be popularized to all polymers with various backbones. This study is an important update to the understanding of polymers at the single-chain level.

Most proteins are typically folded into predetermined three-dimensional structures in the aqueous cellular environment. However, proteins can be exposed to a nonpolar environment under certain conditions, such as inside the central cavity of chaperones and unfoldases during protein degradation. It remains unclear how folded proteins behave when moved from an aqueous solvent to a nonpolar one. Here, we employed single-molecule atomic force microscopy and molecular dynamics (MD) simulations to investigate the structural and mechanical variations of a polyprotein, I278, during the change from a polar to a nonpolar environment. We found that the polyprotein was unfolded into an unstructured polypeptide spontaneously when pulled into nonpolar solvents. This finding was corroborated by MD simulations where I27 was dragged from water into a nonpolar solvent, revealing details of the unfolding process at the water/nonpolar solvent interface. These results highlight the importance of water in maintaining folding stability, and provide insights into the response of folded proteins to local hydrophobic environments.

In this paper, we prepare a hydrogel in a green method via the redox reaction between phenol groups and Eosin Y by visible light irradiation. Eosin Y shows significant absorption at 515 nm while the reduced product does not. A series of control experiments show that almost 100% of the consumed Eosin Y reacted with phenol groups to form cross-link points. On the basis of the two results, the chemical cross-link density can be easily determined quantitatively by UV-vis spectroscopy. With this method, specimens from the state of the starting solution to solid gel can be measured in situ and in real time without any pretreatment. The result obtained by this new method is proved by traditional rheological measurements.

Water, the dominant component under the physiological condition, is a complicated solvent which greatly affects the properties of solute molecules. Here, we utilize atomic force microscope-based single-molecule force spectroscopy to study the influence of water on the single-molecule elasticity of an unstructured single-stranded RNA (poly(U)). In nonpolar solvents, RNA presents its inherent elasticity, which is consistent with the theoretical single-chain elasticity calculated by quantum mechanics calculations. In aqueous buffers, however, an additional energy of 1.88 kJ/mol·base is needed for the stretching of the ssRNA chain. This energy is consumed by the bound water rearrangement (Ew) during chain elongation. Further experimental results indicate that the Ew value is uncorrelated to the salt concentrations and stretching velocity. The results obtained in an 8 M guanidine·HCl solution provide more evidence that the bound water molecules around RNA give rise to the observed deviation between aqueous and nonaqueous environments. Compared to synthetic water-soluble polymers, the value of Ew of RNA is much lower. The weak interference of water is supposed to be the precondition for the RNA secondary structure to exist in aqueous solution.

Natural cellulose (NC) is the most abundant biomacromolecule. The fact that each pyranose ring of NC has three hydroxyl groups implies that NC should be highly hydrophilic. The paradoxical water insolubility of NC is usually explained by the strong tendency to form the hydrogen-bonding network by the high content of hydroxyl groups. In this study, we present the first experimental evidence that NC is rather hydrophobic, even if the chains are molecularly dispersed. By single-molecule force spectroscopy, the single-chain mechanics of NC has been studied in various liquid environments. In a common nonpolar solvent, octane, NC shows the elastic force–extension (F–E) curve. We find that this kind of F–E curve can be fitted well by the QM-FJC model, in which the single-chain elasticity obtained from quantum mechanical (QM) calculations is integrated into the freely jointed-chain (FJC) model. However, the result of NC obtained in water is different, which shows a long plateau in the F–E curve. Further study shows that the height of the plateau is temperature dependent. However, the plateau disappears when an 8 M urea solution is used as the liquid environment. AFM images obtained in water show that single NC chains exist in a compact globule conformation on the sample surface. According to the molecular structure, methylcellulose (MC) should be more hydrophobic than NC. However, no plateau can be observed from the MC samples in water. On the basis of all the results above, we can draw a conclusion that both of the hydrophobic effect and the crystallization of NC contribute to the plateau in the F–E curve obtained in water. The experimental observation of the hydrophobic nature of NC at the single-chain level provides new insights into the understanding of NC.

At crystal or larger scales, natural cellulose was generally recognized to be a rigid material. Our single-chain mechanical measurements, however, reveal that the natural cellulose is as flexible as common synthetic polymers at the single-chain level, creating new opportunities in the designing of nano materials.

The unique self-cleaning feature of the lotus-like superhydrophobic (SH) surface attracted worldwide interest in recent years. However, the mechanism of the self-cleaning phenomena remains unclear. Here, we attempt to provide a comprehensive understanding of why self-cleaning of the particles with a broad range of size can be realized on the lotus-like SH surfaces. After measurements and analysis of the force involved at the interface, we conclude that there are four main preconditions for self-cleaning: (1) contact angle (CA) > 90°, (2) low enough sliding angle, (3) low enough adhesion force, and (4) proper particle size. However, as far as the lotus-like SH surface and typical dust are concerned, all the preconditions will be satisfied automatically. We also observe that the particles with a broad range of size (from submicron level to the millimeter level) and density (virtually no limit) can be driven by a water droplet on the lotus-like SH surface. This interesting finding may be helpful for the design of novel engineering system at the micron-millimeter scale in the future.

In this study, we find that visible light can trigger both the loading and the release of N-alkyl substituted 4-picolinium on self-assembled monolayers (SAM). The latter process will result in surface-charge inversion of the SAM, which can be used for controlled release of molecules of interest.

The single-chain mechanics of two similar thermosensitive polymers, poly(N,N-diethylacrylamide) (PDEAM) and poly(N-isopropylacrylamide) (PNIPAM), have been studied by atomic force microscopy-based single-molecule force spectroscopy (SMFS). In a typical nonpolar organic solvent, octane, both of the polymers show the same inherent elasticity, although they have different substitutional groups. However, the mechanics of the two polymers presents large differences in water. The energies needed for the rearrangement of the bound water during elongation at room temperature are estimated by the SMFS method at the single-chain level, which is ∼1.13 ± 0.10 and ∼5.19 ± 0.10 kJ/mol for PDEAM and PNIPAM, respectively. In addition, PNIPAM shows a temperature-dependent single-chain mechanics when the temperature is increased across the lower critical solution temperature (LCST), while PDEAM does not. These differences observed in aqueous solution originate from the different structures of the two polymers. With a hydrogen bond donor in the amide group, PNIPAM will be more hydrated when T < LCST. When T > LCST, PNIPAM will have larger changes in both conformation and hydration. These findings also suggest that PNIPAM is a good candidate for a thermo-driven single-molecule motor, while PDEAM is not.

The variation of the single-chain mechanics of poly(N-isopropyl-acrylamide) (PNIPAM) for the entire range of the methanol molar fraction (χmethanol) (0%–100%) in the water/methanol mixed solvent has been studied by AFM-based single-molecule force spectroscopy at room temperature (RT). In general, the single-chain mechanics of PNIPAM shows a reentrant variation upon the change of χmethanol from 0% to 100%. The possible mechanism for the reentrant variation of the single-chain mechanics is discussed. The current study may cast new light on the design of novel environmental responsive nano-devices in the future, when χmethanol changes from 16% to 0%.

We study the single-chain elasticities of three kinds of neutral polymers with a carbon–carbon (C–C) backbone by atomic force microscopy-based single-molecule force spectroscopy in a nonpolar solvent (octane), aiming at measuring the inherent chain elasticity of this very important class of polymers. The finding that the single-chain elasticities of all three polymers in octane are virtually identical in the entire force region implies that the side chains of the polymers have no detectable effects on the single-chain elasticity. By utilizing the single-chain elasticity from quantum mechanics calculations, the freely rotating chain model can provide the best fitting curve when each C–C bond is set to be the rotating unit. Although there are some exceptions when the side chain is very huge, our work provides a general result for the inherent elasticity of single neutral flexible polymer chains with C–C backbones.

Radical polymerization from a single initiator molecule in a microenvironment is a nearly ideal system in which bimolecular termination, solution concentration, and viscosity changes could be neglected. In this study, we provide two facile methods of preparing polymers via atom-transfer radical polymerization (ATRP) under single-initiator conditions: tether initiators on planar substrates at superlow density through mixed self-assembled monolayers (SAMs) and encapsulated single initiators in microfluidic droplets. The molecular weight (MW) of the resultant polymers characterized by atomic force microscope-based single-molecule force spectroscopy (AFM-based SMFS) showed that the single-chain ATRP had an extraordinarily faster chain propagation rate (2 unit/s) on planar substrates and gave polymers with much higher MWs (105–106 g/mol) than those obtained from traditional ATRP (103–105 g/mol). The former method offered a general platform for single-chain polymer synthesis and investigation, and the latter could be amplified to obtain abundant single-chain polymers with ultrahigh molecular weight (UHMW) for commercial applications.

Poly(N-isopropyl-acrylamide) (PNIPAM) is a paradigm thermally sensitive polymer, which has a lower critical solution temperature (LCST) of ∼32 °C in water. Herein by AFM-based single molecule force spectroscopy (SMFS), we measured the single chain elasticity of PNIPAM across the LCST in water. Below LCST, the force curves obtained at different temperatures have no remarkable difference; while above LCST, an unexpected temperature dependent elasticity is observed, mainly in the middle force regime. We found that 35 °C is a turning point of the variation: from 31 to 35 °C, the middle parts of the force curves drop gradually, whereas from 35 to 40 °C, the middle parts rise gradually. A possible mechanism for the unexpected temperature dependent mechanics is proposed. The single chain contraction against external force upon heating from 35 to 40 °C may cast new light on the design of molecular devices that convert thermal energy to mechanical work.

We prepared robust cross-linked (x-linked) multilayer films under visible light irradiation with the catalysis of a Ru(II) complex. The x-linking is achieved by the coupling reaction between phenol group and primary amine group within the self-assembled multilayer films that were prepared beforehand. Three kinds of polymers, i.e., poly(4-vinylphenol), poly(allylamine) and poly(ethyleneimine), were selected as the model system to illustrate the concept of this strategy. Upon visible light irradiation, the chemical stability of the x-linked films towards solution etching was greatly enhanced. In previous studies, horseradish peroxidase (HRP) is often utilized to catalyze the C–C, C–O and C–N coupling structures, which is useful to prepare polymers, capsules and bulk hydrogels. We also tried to prepare the x-linked films by the catalysis of HRP. The comparison of the two methods suggests that the Ru(II) complex method is more ideal for fabricating x-linked films. In addition, the photo-triggered chemical reaction within the films was confirmed by the solid-state 13C NMR, XPS and FT-IR measurements. Without UV light irradiation or thermal treatment, this strategy brings many advantages. It is anticipated that this approach can be easily extended to the applications of the biological related fields in the future.

Highly stable covalently attached multilayer films were constructed by visible-light irradiation of hydrogen-bonding directed multilayer films of poly(allylamine) and poly(4-vinylphenol).

There is no doubt that water is pivotal to life. Yet, as the emergence of life is still a big challenge in science, the detailed involvement of water in that process is not well recognized. Following the clues provided by recent single-molecule studies on DNA, we attempt to elucidate the possible roles of water in the prebiotic chemical evolution. Water has long been recognized as an important reactant in the Miller-Urey experiment and then as the only solvent of the primitive soup. Besides that, water also played a vital role in the prebiotic chemical evolution: water is the important criterion in the combinatorial library screening for self-assembling macromolecules. With this notion, the uniformity of biochemistry for all terrestrial life may be explained. A possible roadmap from the inorganic world to the origin of life is also discussed.

We utilize the extracted water-soluble wool keratin as the building block for the layer-by-layer (LbL) assembly. By adjusting the solution pH, keratin can be used as polycation or polyanion. Experimental results indicate that multilayer films of keratin together with other synthetic polyelectrolytes have been fabricated successfully. We have found that the average thickness of each bilayer of the keratin containing film is evidently larger than that of usual multilayer films previously reported. This feature is helpful to fabricate relatively thick films in less deposition cycles. The use of keratin in LbL assembly is helpful to prepare a biocompatible surface for tissue engineering.Keratin, which is extracted from wool, has been utilized as a novel building block for layer-by-layer assembly.

We report on the facile preparation of chemically cross-linked microgels in mild conditions by using the reversed microemulsion technique. Sodium alginate has been modified by partially grafting phenol groups to the backbone, on the basis of which microgels have been prepared by the irradiation of visible light in the presence of catalyst Ru(II) complex at room temperature. The irradiation of visible light instead of UV light or gamma rays brings many advantages. The mean diameters of the microgels are 15−40 μm in aqueous solution and 5−15 μm in the dried state. Although the size of the microgel is sensitive to the environment change, it presents excellent size stability in a broad range that covers the physiological condition. The applications of this biocompatible and biodegradable microgel in biology are greatly anticipated.

In this study, we find that visible light can trigger both the loading and the release of N-alkyl substituted 4-picolinium on self-assembled monolayers (SAM). The latter process will result in surface-charge inversion of the SAM, which can be used for controlled release of molecules of interest.

Highly stable covalently attached multilayer films were constructed by visible-light irradiation of hydrogen-bonding directed multilayer films of poly(allylamine) and poly(4-vinylphenol).

There is no doubt that water is pivotal to life. Yet, as the emergence of life is still a big challenge in science, the detailed involvement of water in that process is not well recognized. Following the clues provided by recent single-molecule studies on DNA, we attempt to elucidate the possible roles of water in the prebiotic chemical evolution. Water has long been recognized as an important reactant in the Miller-Urey experiment and then as the only solvent of the primitive soup. Besides that, water also played a vital role in the prebiotic chemical evolution: water is the important criterion in the combinatorial library screening for self-assembling macromolecules. With this notion, the uniformity of biochemistry for all terrestrial life may be explained. A possible roadmap from the inorganic world to the origin of life is also discussed.