In Situ Real-Time Atomic Force Microscopy Observation of the Surface Mobility on Each Domain of a Polystyrene-b-poly(methyl methacrylate) Film at High Temperatures.

The surface chain movements within the microdomains of a polystyrene-b-poly(methyl methacrylate) (PS-b-PMMA) and corresponding homopolymer films were observed via in situ real-time atomic force microscopy (AFM) at high temperatures and analyzed quantitatively using particle image velocimetry (PIV). At low temperatures, mobility within the PS microdomains resembled that within the PS homopolymer film, but movements in the PMMA microdomains were notably accelerated compared to the PMMA homopolymer. Conversely, at high temperatures, mobility within both PS and PMMA microdomains was considerably suppressed compared to their respective homopolymer films, likely owing to the fixed linkage of the block chains at the microdomain interface. This combination of real-time AFM observation and PIV analysis is an effective method for quantitatively evaluating surface chain mobility in real space.

In Situ Real-Time Atomic Force Microscopy Observations of Chain Mobility at Polymer/Water Interfaces of Poly(methyl methacrylate), Poly(2-hydroxyethyl methacrylate), and Poly(2-methoxyethyl methacrylate) Films in Water.

Polymer materials are widely used in water or in contact with an aqueous environment. However, evaluating the chain mobility, a crucial parameter, at a polymer-water interface is challenging. In this study, we, for the first time, observed poly(methyl methacrylate) (PMMA), poly(2-hydroxyethyl methacrylate) (PHEMA), and poly(2-methoxyethyl methacrylate) (PMEMA) film surfaces in water via in situ real-time atomic force microscopy (AFM) in tapping mode and quantified the chain mobility. The average displacement between adjacent images (nm/8.75 min) was evaluated using particle image velocimetry. The displacement of PMMA, which has a high bulk glass-transition temperature (Tg) (108 °C) and exhibits limited water absorption, was low both in air (0.54 nm/8.75 min) and water (0.86), while PHEMA, which has a high bulk Tg (99 °C) and exhibits high water absorption, exhibited low mobility in air (0.40) but two orders of magnitude higher mobility in water (60). PMEMA, which has a low bulk Tg (14 °C) and exhibits limited water absorption, already started to move in air (4.5), and its mobility moderately increased in water (20). These behaviors were reasonable, considering the bulk Tg and water absorption characteristics of the polymers. Further, the chain mobility in water was compared with that of dried samples at high temperatures in air. The mobility of PMMA, PHEMA, and PMEMA in water corresponded to that of the dried samples observed in air below the surface Tg (97 °C) for PMMA, at ∼125 °C for PHEMA, and at ∼35 °C for PMEMA. In situ real-time AFM analysis of polymer materials in water is an effective method for evaluating the chain mobility at the polymer/water interface.

Crystallization of Star-Shaped Poly(l-lactide)s with Arm Chains Aligned in the Same Direction in Two-Dimensional Crystals in a Langmuir Monolayer.

Polylactide (PLA) crystallizes to form extended-chain crystals in a Langmuir monolayer because crystallization is accelerated on the water surface. This is a unique situation where chain packing can be analyzed by simply measuring the lamellar thickness. Herein, star-shaped poly(l-lactide)s (PLLAs) with 2-12 arms were synthesized through the polymerization of l-lactide with various polyols as initiators, and their crystallization behavior in a monolayer was studied via atomic force microscopy. The PLLAs comprising 2-4 arms crystallized with all arms aligned in the same direction and being folded at the central polyol unit. Meanwhile, the PLLAs comprising 6 and 12 arms crystallized with both halves of the arms extended from the center to the opposite directions, most likely due to the steric hindrance of the crowded arms. Considering that the PLLAs crystallized from a once-formed condensed amorphous state during compression, they have a strong tendency to crystallize with the arms aligned in the same direction. The crystallization rate of star-shaped PLAs is known to reduce compared with that of a linear PLA even if the number of arms is as few as 2. This should be closely related to the unique crystallization behavior of the star-shaped PLLAs with the arms aligned in the same direction.

Chain Movements at the Topmost Surface of Poly(methyl methacrylate) and Polystyrene Films Directly Evaluated by In Situ High-Temperature Atomic Force Microscopy.

The surfaces of polymeric materials are thermodynamically unstable, and the glass-transition temperature (Tg) is significantly lower than that in the bulk material. However, the mobility of the chains at the top of the surface has never been directly evaluated. In this study, the movements of the topmost chains of poly(methyl methacrylate) (PMMA) and polystyrene (PS) bulk films were observed in situ at high temperatures with atomic force microscopy in tapping mode. PMMA and PS chains started moving at ∼97 and ∼50 °C, respectively, which were slightly and significantly below the values of their bulk Tg (PMMA, 108 °C; PS, 104 °C), respectively. The activation energies of the apparent diffusion constants of PMMA and PS, derived by particle image velocimetry analysis, were 193 and 151 kJ mol-1, respectively, and reasonable for the glass transition. Movements of isolated PMMA chains deposited on a PMMA film by the Langmuir-Blodgett technique were also observed and confirmed to be essentially the same as those on the PMMA film surface.

Molecular Combing of Various Poly(n-Alkyl Acrylate) Chains on Mica by the Dipping Method.

If polymer chains could be deposited on a substrate as a fully extended chain, a procedure known as "molecular combing," the chain structure could be characterized by atomic force microscopy in more detail than has been possible with the measurements available today. We show here, for the first time, that flexible polymers can be molecularly combed to fully extended chains by the dipping method. We studied the molecular combing of a series of poly(n-alkyl acrylate)s on mica from a chloroform solution by the dipping method and found that poly(n-alkyl acrylate)s with an alkyl group longer than n-octyl can be molecularly combed into straight chains under optimized conditions. With increasing alkyl lengths, the number of chains deposited decreases by four orders of magnitude, and chains become molecularly combed under a wider range of conditions. The length of the molecularly combed chains is ∼80% for poly(n-octyl acrylate) but ∼100% of the all-trans conformation for poly(n-alkyl acrylate)s with an alkyl length longer than n-nonyl.

Gemini Thermotropic Smectic Liquid Crystals for Two-Dimensional Nanostructured Water-Treatment Membranes.

We have developed a two-dimensional (2D) liquid-crystalline (LC) nanostructured water-treatment membrane showing high virus rejection ability (over 99.99997% for bacteriophage Qß) and improved water permeation. Polymerizable gemini amphiphiles have been designed and synthesized. They have H-shaped gemini-type structures of thermotropic smectic liquid crystals composed of cationic imidazolium moieties. One of the gemini amphiphiles shows a smectic A phase with an interdigitated bilayer structure. A cross-linked self-standing 2D nanostructured polymer film has been obtained by in situ photopolymerization of the gemini amphiphile in the smectic phase. The length of linkers in gemini amphiphiles affects the formation of LC phases. The 2D nanostructured membrane also showed selective salt rejection.

In situ AFM Observation of the Movements of Isolated Isotactic Poly(methyl methacrylate) Chains in a Precursor Film of an Oligo(methyl methacrylate) Droplet Spreading on Mica.

Atomic force microscopy (AFM) is a powerful tool to observe polymer chains at the molecular level. In this study, we show that the movements of isolated linear polymer chains in a precursor film of a droplet of an oligomer spreading on a substrate could be visualized in situ at the molecular level by AFM for the first time. The system was an isotactic poly(methyl methacrylate) (it-PMMA) solubilized in an oligo(MMA) matrix (it-PMMA/oligo(MMA) = 1/10,000 w/w) spreading on mica under high humidity. Because of the limited resolution of the AFM instrument, condensed linear polymer chains could not be visualized, but a small amount of it-PMMA chains that were solubilized as isolated chains in the oligo(MMA) matrix could be visualized in the precursor film, the contrast of which came from a large difference in glass transition temperature (Tg) of it-PMMA and oligo(MMA). The it-PMMA chains in the precursor film spread in the radial direction of the droplet with vigorously changing chain conformations. The spreading rate of it-PMMA chains under 72% relative humidity was ∼1/30 of the spreading rate of the oligo(MMA) matrix, which was estimated based on the decrease in the volume of the macroscopic droplet. The spreading of the it-PMMA chains and droplet strongly depended on humidity and was suppressed with the decrease in humidity, most likely because of the increase in friction with the substrate. The difference in the spreading rate of it-PMMA and oligo(MMA) further increased under low humidity. The dynamic molecular information of a precursor film by AFM should help to elucidate the wetting dynamics on a substrate.

Preparation of a Si(111) Atomically Flat Substrate via Wet Etching and Evaluation as an AFM Substrate for Observations of Isolated Chains, Crystals, and Crystallization of Isotactic Poly(methyl methacrylate) at the Molecular Level.

To observe a polymer chain deposited on a substrate by atomic force microscopy (AFM) at the molecular level, the substrate should be atomically flat and stable under laboratory conditions and adsorb polymer chains firmly. Therefore, substrates used under laboratory conditions are practically limited to mica, highly ordered pyrolytic graphite, and atomically stepped sapphire, and polymers observed by AFM at the molecular level are also limited. A silicon wafer is frequently used as a substrate for AFM observation for somewhat macroscopic observations, but the surface of the silicon wafer is too rough to observe polymer chains deposited on it at the molecular level. In this study, we prepared an atomically stepped Si(111) substrate via wet etching in NH4F and evaluated it as an AFM substrate. The Si(111) substrate was stable as an AFM substrate, and isolated poly(methyl methacrylate) (it-PMMA) chains and a crystalline monolayer deposited on the substrate were observed by AFM at the molecular level. An it-PMMA amorphous monolayer deposited on mica crystallized under high humidity, but that on the Si(111) substrate did not because of the difference in the surface nature and the crystal structure of the substrates. The Si(111) substrate was hydrophobic, and the it-PMMA monolayers could be deposited as a multilayer, which could not be formed on hydrophilic mica. The crystallization behavior of an it-PMMA amorphous multilayer and an amorphous/crystalline mixed multilayer on the Si(111) substrate was also evaluated.

Self-Assembly of Linear and Cyclic Polylactide Stereoblock Copolymers with a Parallel and Antiparallel Chain Arrangement Distinguishing Their Directions on a Water Surface.

The self-assembly of molecules into a well-ordered structure is one of the most important processes in fabricating sophisticated materials. Here, we show that polymer chains can be self-assembled, distinguishing their direction (parallel or antiparallel), and could be a new useful scaffold for self-assembly in a controlled direction. The system that was used was a stereocomplex (SC) formation of linear and cyclic polylactide (PLA) stereoblock copolymers with a parallel and antiparallel chain arrangement in a Langmuir monolayer. The linear and cyclic stereoblock copolymers with a parallel arrangement formed a well-ordered lamellar SC in the first and second layers upon compression, but the linear and cyclic stereoblock copolymers with an antiparallel arrangement did not form a first-layer lamella and instead formed only the second-layer lamella. These results were only rationally explained by assuming that the enantiomeric PLA chains selectively assembled in a parallel direction, not in an antiparallel direction, in the SC. A simple polymer chain could be self-assembled, distinguishing the direction without a specific interaction group in it.

Condensed desmin and actin cytoskeletal communication in lipid droplets.

The interplay between intermediate filaments (IFs) and other cytoskeletal components is important for the integrity and motility of cells. The impact of IF assembly on other components and cell morphology is not yet fully understood. Therefore, we examined the effects of combined desmin and actin assembly on cytoskeletal network arrangement in artificial cell-sized droplets. Fluorescently labeled desmin, with or without actin, was enclosed in droplets prepared with 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) using the water-in-oil method. Protein networks were observed using fluorescence microscopy in the presence of 150 mM KCl, 20 mM imidazole-HCl (pH 7.4), 2 mM MgCl2 , and 1 mM adenosine 5'-triphosphate for both desmin and actin assembly. As desmin alone can assemble into filaments within seconds, desmin networks mainly localizing at the inner margins of the droplets were observed within 10 min after assembly initiation. Subsequently, deformations of droplets appeared. Furthermore, a portion of droplets formed desmin-rich protrusions of several micrometers. Notably, actin alone rarely formed protrusions under the same conditions. When 1,2-dioleoyl-sn-glycero-3-phosphocholine was used instead of DOPE, protrusions became less frequent. The combination of desmin and actin increased the number of deformed droplets in which the proteins were considerably colocalized. The assembly process of desmin facilitated colocalization. Atomic force microscopy failed to reveal interactions between the two filament types. These results suggest that the mechanical properties of desmin networks may influence the behavior of actin networks, as well as membrane morphology, possibly reflecting the mechanical function of desmin filaments in muscle cells.

Evaluation of Ring Expansion-Controlled Radical Polymerization System by AFM Observation.

We here present a direct link between the reaction mechanisms for the ring-expansion "vinyl" polymerization system and atomic force microscopy (AFM) observations. The brush-modification clearly discriminates the desired cyclic species with the contour lengths (Lc) of 28-132 nm and molar masses (MAFM) of 60.2-283 kg mol-1 from the other linear ones. The 293 polymer blushes observed in a 1.0 µm × 1.0 µm AFM image are individually characterized, eventually providing clear answers about the mechanisms of this rare polymerization system, which include ring-expansion vinyl polymerizations to generate cyclic polymers, fusions of the generated cycles to form multimers, and their scission to form linear or ring-opened species. The relationship between the molecular chain lengths and the cyclic versus linear morphologies is highlighted.

Molecular Combing of a Flexible Polymer Chain by Simple Spin-Casting.

If polymer chains could be fixed on a substrate as a fully elongated chain, a procedure known as "molecular combing", the chain structure could be analyzed more precisely than has been possible with the characterization techniques available today. Although the molecular combing of a rigid biomolecule, DNA, has been attained for the mapping of genetic information, that of flexible chains has never been achieved as yet. We show here that poly(n-nonyl acrylate) (PNA) can be molecularly combed on mica by a simple spin-casting method, and that the chain lengths were in good agreement with that of the all-trans conformation. One of the key factors for successful molecular combing was found to be the weak adsorption of PNA on mica, indicating that flexible polymers may be molecularly combed by adjusting their affinity to the substrate. The molecular combing of polymer chains may open a new way not only to characterize the chain structures more precisely but also to fabricate new nanomaterials based on polymers.

Morphology control through hierarchical phase separation in Langmuir monolayers of poly(methyl methacrylate)-b-poly(n-butyl acrylate).

Precise morphology control of ultrathin films is one of the important issues in nanotechnology. To this end, we describe that various controlled morphologies of hierarchical phase separation can be attained using poly(methyl methacrylate)-b-poly(n-butyl acrylate) (PMMA-b-PBA) monolayers spread on a water surface. At a low surface pressure, they were miscible, but upon compression, phase separated with a monolayer of the major component block spreading on the water surface, on top of which the minor component block separated out (hierarchical phase separation). The morphology of the minor component block separated out on top of the monolayer varied from a sphere/short string mixture, to long strings, and a mesh-like structure having a regular domain-domain spacing with the increasing content of the minor block, and the hierarchical phase separation was reversible depending on the surface pressure. However, these well-ordered hierarchical phase separations were observed only in the PBA-rich polymers, and a clear morphology was not observed in the PMMA-rich polymers, because of insufficient domain growth after the hierarchical phase separation for the PMMA-rich block copolymers which are more rigid than the PBA-rich polymers. Although the clear hierarchical phase separation was limited in flexible composition, we noted that the hierarchical phase separation provided a unique well-controlled morphology in the Langmuir monolayers.

Crystallization behavior of single isotactic poly(methyl methacrylate) chains visualized by atomic force microscopy.

We have, for the first time, successfully visualized the crystallization behavior of a single isolated polymer chain at the molecular level by atomic force microscopy (AFM). Previously, we found that isotactic poly(methyl methacrylate) (it-PMMA) formed two-dimensional folded chain crystals composed of double-stranded helices upon compression of its Langmuir monolayer on a water surface, and the molecular images of the crystals deposited on mica were clearly visualized by AFM (Kumaki, J.; et al. J. Am. Chem. Soc. 2005, 127, 5788). In the present study, a high-molecular-weight it-PMMA was diluted in a monolayer of an it-PMMA oligomer which cannot crystallize at the experimental temperature due to its low molecular weight. At a low surface pressure, isolated amorphous chains of the high-molecular-weight it-PMMA solubilized in the oligomer monolayer were observed. On compression, the isolated chains converted to crystals composed of a single chain, typically some small crystallites linked by an amorphous chain like a necklace. Detailed AFM observations of the crystals indicated that the crystalline nuclei preferentially formed at the ends of the chains, and the size of the nuclei was almost independent of the molecular weight of it-PMMA over a wide range. At an extremely slow compression, crystallization was promoted, resulting in crystallization of the whole chain. The crystallization behavior of a single isolated chain provides new insights in understanding the polymer crystallization process.

Two-dimensional phase separation of a poly(methyl methacrylate)/poly(L-lactide) mixed Langmuir monolayer via a spinodal decomposition mechanism.

We have found the first evidence that a polymer blend Langmuir monolayer can phase-separate via spinodal decomposition (SD) mechanism. The system was a poly(methyl methacrylate)/poly(L-lactide) mixture. It phase-separated immediately after compression on a water surface and formed a spinodal-like morphology, as observed by atomic force microscopy (AFM). The fast Fourier transform of the AFM images showed a clear spinodal ring with a maximum intensity at a wavenumber of q(m). At the small quench depth at a surface pressure of 2 mN/m, q(m) did not change, but the concentration difference between the domains (ΔΦ) grew with time, corresponding to the early-stage SD. At larger quench depths at 4 and 5 mN/m, q(m) significantly decreased, but ΔΦ was constant with time; this behavior corresponded well to the late-stage SD. Thus, the 2D phase separation in the Langmuir monolayer was basically explained by the SD mechanism well-known in 3D systems. In the late stage SD of the monolayer, q(m) scaled with time much faster than that expected by theories for the 2D state. Phase separation via a SD mechanism is a promising new way to control the lateral morphology of Langmuir monolayers, one of the main issues in nanotechnology that remains difficult to attain even today.

Significant melting point depression of two-dimensional folded-chain crystals of isotactic poly(methyl methacrylate)s observed by high-resolution in situ atomic force microscopy.

The properties of polymer ultrathin films are a subject of intense study from both practical and academic viewpoints. Previously, we found that upon compression, an isotactic poly(methyl methacrylate) (it-PMMA) Langmuir monolayer crystallized to form a two-dimensional (2D) folded-chain crystal, and the molecular image of the crystal with chain folding and tie chains was clearly visualized by atomic force microscopy (AFM). In the present study, the melting behaviors of the it-PMMA 2D crystals were successfully observed in situ by high-temperature AFM at the molecular lever for the first time. The chain-chain distances (~1.2 nm) of the crystals were clearly resolved even at temperatures close to the melting temperatures (Tm) of the 2D crystals. We found that the Tm of the 2D crystals was at most 90 °C lower than the bulk crystals. The Tm depression strongly depended on the molecular weight, while the molecular weight dependence of the bulk Tm was negligible in the molecular weight regime studied. The Tm depression also depended on the substrates, a slightly larger depression being observed on a sapphire substrate compared to that on a mica. The large Tm depressions of the 2D crystals could not be explained by a simple Thomson-Gibbs argument, theoretical developments are necessary to understand the melting of the 2D crystals.

Visualization of two-dimensional single chain conformations solubilized in a miscible polymer blend monolayer by atomic force microscopy.

Polymer Langmuir monolayers spread on a water surface are one of the best models for two-dimensional (2D) polymer and have been extensively studied. However, the most fundamental issue in understanding a 2D film, the polymer chain packing in the film, is still not well-understood, especially from the experimental point of view. Direct observation of the chain packing by microscopy at a molecular level, such as by atomic force microscopy (AFM), might be one of the most promising ways to study this issue; however, because of the limited resolution of the method, the chain packing of polymer cannot be resolved by AFM, except for especially large polymers. Here, we show that a mixed monolayer of vinyl polymers, poly(methyl methacrylate) (PMMA) and poly(n-nonyl acrylate) (PNA), was miscible at a low surface pressure, and if a small amount of PMMA chains was solubilized in a PNA monolayer, the isolated PMMA chains in the PNA monolayer were, for the first time, successfully visualized by AFM with a clear contrast, which originated from a difference of rigidities of the polymers due to their different glass transition temperatures (105 °C(PMMA) and -89 °C(PNA)). The PMMA chains were found to strongly interpenetrate into the PNA monolayer, with a radius of gyration (R(g(PMMA))) that was several times larger than that of the 2D ideal chain (segregated-chain). Furthermore, the radius scaled with the molecular weight of the PMMA (M(PMMA)) as R(g(PMMA)) ∝ M(PMMA)(0.63), which was between the scaling of the 2D ideal chain (segregated chain), R(g) ∝ M(0.5), and the 2D chain in good solvent, R(g) ∝ M(0.75). On the other hand, R(g(PMMA)) was independent of the molecular weight of the PNA matrix over a wide range. These results indicate that the PNA/PMMA monolayer is a strongly miscible system, although the R(g(PMMA)) scaling with M(PMMA) (0.63) is somewhat smaller than that expected for a 2D chain in good solvent systems (0.75). The generation of molecular level information by direct observation of polymer chains in 2D blend films should improve our understanding of polymer 2D films.

Hierarchical amplification of macromolecular helicity of dynamic helical poly(phenylacetylene)s composed of chiral and achiral phenylacetylenes in dilute solution, liquid crystal, and two-dimensional crystal.

Optically active poly(phenylacetylene) copolymers consisting of optically active and achiral phenylacetylenes bearing L-alanine decyl esters (1L) and 2-aminoisobutylic acid decyl esters (Aib) as the pendant groups (poly(1L(m)-co-Aib(n))) with various compositions were synthesized by the copolymerization of the optically active 1L with achiral Aib using a rhodium catalyst, and their chiral amplification of the macromolecular helicity in a dilute solution, a lyotropic liquid crystalline (LC) state, and a two-dimensional (2D) crystal on the substrate was investigated by measuring the circular dichroism of the copolymers, mesoscopic cholesteric twist in the LC state (cholesteric helical pitch), and high-resolution atomic force microscopy (AFM) images of the self-assembled 2D helix-bundles of the copolymer chains. We found that the macromolecular helicity of poly(1L(m)-co-Aib(n))s could be hierarchically amplified in the order of the dilute solution, LC state, and 2D crystal. In sharp contrast, almost no chiral amplification of the macromolecular helicity was observed for the homopolymer mixtures of 1L and Aib in the LC state and 2D crystal on graphite.

Separation of C70 over C60 and selective extraction and resolution of higher fullerenes by syndiotactic helical poly(methyl methacrylate).

A one-handed helical polymer, syndiotactic poly(methyl methacrylate) (st-PMMA), recognizes the size and chirality of higher fullerenes through an induced-fit mechanism and can selectively extract enantiomers of the higher fullerenes, such as C(76), C(80), C(84), C(86), C(88,) C(90), C(92), C(94), and C(96). This discovery will generate a practical and valuable method for selectively extracting the elusive higher fullerenes and their enantiomers and opens the way to developing novel carbon cage materials with optical activities.

Strong compression rate dependence of phase separation and stereocomplexation between isotactic and syndiotactic poly(methyl methacrylate)s in a Langmuir monolayer observed by atomic force microscopy.

The stereocomplex formation between isotactic and syndiotactic poly(methyl methacrylate) (it-PMMA, st-PMMA) in a Langmuir monolayer was studied by surface pressure-area isotherms and atomic force microscopy (AFM). We found that the stereocomplex formation was highly sensitive to the compression rate of the monolayer. At a normal compression rate of 0.5 mm/s provided by the moving barrier, the blend monolayer formed a clear phase separation of the it- and st-PMMA domains at 1 mN/m. Further compression to 15 mN/m resulted in a limited degree of stereocomplexation, mainly at the interface between the two domains. However, at a 1/50 slower compression rate of 0.01 mm/s, the blend did not form a clear phase separation at 1 mN/m and quantitatively formed a stereocomplex at 15 mN/m. This apparent immiscibility observed at the faster compression rate was found to be kinetically induced as a result of the rapid compression of the phase-separated mixture at the dilute state because it-PMMA and st-PMMA form expanded and condensed monolayer, respectively. On the other hand, at the slower compression rate, the blend formed a thermodynamically miscible phase, and as a result, the stereocomplex was quantitatively formed. This apparent phase separation of a mixed monolayer composed of an expanded and a condensed monolayer should be a common phenomenon for similar systems and might have caused misjudgment of the miscibility in such cases. The compression rate dependence should be carefully evaluated in order to determine the precise miscibility of blended monolayers in similar systems.