Cryogenic Single-Molecule Fluorescence Detection of the Mid-Infrared Response of an Intrinsic Pigment in a Light-Harvesting Complex.

We observed the mid-infrared (MIR) response of a single pigment of bacteriochlorophyll a at the B800 binding site of a light-harvesting 2 complex. At a temperature of 1.5 K, a single complex in a spatially isolated spot in a near-infrared (NIR) fluorescence image was selected and was simultaneously irradiated with MIR and NIR light. We found that the temporal behavior of the NIR fluorescence excitation spectrum of individual pigments in a single complex was modulated by the MIR irradiation at 1650 cm-1. The MIR modulation of a single pigment was linearly proportional to the MIR intensity. The MIR linear response was detected in the range from 1580 to 1670 cm-1.

Superfluid helium nanoscope insert with millimeter working range.

A superfluid helium insert was developed for cryogenic microscopy of millimeter-sized specimens. An optical-interferometric position sensor, cryogenic objective mirror, and piezo-driven cryogenic stage were fixed to an insert holder that was immersed in superfluid helium. The single-component design stabilized the three-dimensional position of the sample, with root-mean-square deviations of (x, lateral) 0.33 nm, (y, lateral) 0.29 nm, and (z, axial) 0.25 nm. Because of the millimeter working range of the optical sensor, the working range of the sample under the active stabilization was (x, y) 5 mm and (z) 3 mm in superfluid helium at 1.8 K. The insert was used to obtain the millimeter-sized fluorescence image of cell nuclei at 1.8 K without a sample exchange.

Energy transfer fluctuation observed by single-molecule spectroscopy of red-shifted bacteriochlorophyll in the homodimeric photosynthetic reaction center.

The photosynthetic reaction center of heliobacteria (hRC) is a homodimeric chromoprotein responsible for light harvesting and photoelectric conversion. The fluorescence of the hRC is radiated from a bacteriochlorophyll (Bchl) g having the lowest energy level, called red-Bchl g. The homodimeric architecture of the hRC indicates that it includes two red-Bchls g arranged symmetrically in pairs. Red-Bchl g is a fluorescent probe useful for monitoring the energy transfer network in the RC. Here, we show the fluorescence polarization dependences of two red-Bchls g, individually measured with selective excitation of chlorophyll a serving as the primary electron acceptor. The two red-Bchls g exhibit almost the same polarization dependences. Based on the polarization dependence and structural data of the hRC, we propose a candidate molecule for red-Bchl g. The fluorescence spectra of single hRCs represent the spectral heterogeneity reflecting the local conformational inhomogeneity. A time series of the fluorescence spectra indicates occasional peak shifts between blue- and red-shifted states without significant changes in the fluorescence intensity. The spectral fluctuation is interpreted to be due to the local conformational dynamics around a Bchl g mediating the energy transfer, switching the terminal energy acceptor between two red-Bchls g. In conclusion, while the energy transfer network in the RC can be perturbed by microscopic dynamics, the total energy transfer efficiency, i.e., the light-harvesting function, is rather robust. The functional robustness may be due to multiple energy transfer pathways composed of many antenna pigments in the RC.

Stabilization of Interfacial Polarization and Induction of Polarization Hysteresis in Organic MISIM Devices.

Molecule-based ferroelectrics has attracted much attention because of its advantages, such as flexibility, light weight, and low environmental load. In the present work, we examined an organic metal|insulator|semiconductor|insulator|metal (MISIM) device structure to stabilize the interfacial polarization in the S layer and to induce polarization hysteresis even without bulk ferroelectrics. The MISIM devices with I = parylene C and S = TMB (=3,3',5,5'-tetramethylbenzidine)-TCNQ (=tetracyanoquinodimethane) exhibited hysteresis loops in the polarization-voltage (P-V) curves not only at room temperature but also over a wide temperature range down to 80 K. The presence of polarization hysteresis for MISIM devices was theoretically confirmed by an electrostatic model, which also explained the observed thickness dependence of the I layers on the P-V curves. Polarization hysteresis curves were also obtained in MISIM devices using typical organic semiconductors (ZnPc, C60, and TCNQ) as the S layer, demonstrating the versatility of the interfacial polarization mechanism.

Variable immersion microscopy with a high numerical aperture.

Three-dimensional (3D) optical microscopy with a high numerical aperture (NA) remains challenging for thick biological specimens owing to aberrations arising from interface refractions. We developed a variable immersion lens (VIL) to passively minimize these aberrations. A VIL is a high-NA concentric meniscus lens and was used in combination with an aberration-corrected high-NA reflecting objective (TORA-FUJI mirror). Wave-optics simulation at a wavelength of 488 nm showed that a VIL microscope enables diffraction-limited 1.2-NA imaging in water (refractive index of 1.34) at a depth of 0.3 mm by minimizing aberrations due to refraction of a sample interface. Another aberration due to the refractive index mismatching between a mounting medium, and an object can also be corrected by the VIL system, because various fluids with different refractive indices can be used as mounting media for the VIL. As a result of correcting the two aberrations at the same time, we experimentally demonstrated that a 6 µm diameter fluorescent bead can be imaged to the true dimensions in 3D.

Cryogenic Far-Field Fluorescence Nanoscopy: Evaluation with DNA Origami.

Far-field fluorescence localization nanoscopy of individual fluorophores at a temperature of 1.8 K was demonstrated using DNA origami as a one-nanometer-accurate scaffold. Red and near-infrared fluorophores were modified to the scaffold, and the fluorophores were 11 or 77 nm apart. We performed the localization nanoscopy of these two fluorophores at 1.8 K with a far-field fluorescence microscope. Under the cryogenic conditions, the fluorophores were perfectly immobilized and their photobleaching was drastically suppressed; consequently, the lateral spatial precision (a measure of reproducibility) was increased to 1 nm. However, the lateral spatial accuracy (a measure of trueness) remained tens of nanometers. We observed that the fluorophore centroids were laterally shifted as a function of the axial position. Because the orientation of the transition dipole of the fluorophores was fixed under cryogenic conditions, the anisotropic emission from the single fixed dipole had led to the lateral shift. This systematic error due to the dipole-orientation effect could be corrected by the three-dimensional localization of the individual fluorophores with spatial precisions of (lateral) 1 nm and (axial) 17 nm. In addition, the xy-error arising from the three-dimensional (3D) orientation of the scaffold with the two fluorophores 11 nm apart was estimated to be 0.3 nm. As a result, the individual fluorophores on the DNA origami were localized at the designed position, and the lateral spatial accuracy was quantified to be 4 nm in the standard error.

Cryogenic Single-Molecule Spectroscopy of the Primary Electron Acceptor in the Photosynthetic Reaction Center.

The photosynthetic reaction center (RC) converts light energy into electrochemical energy. The RC of heliobacteria (hRC) is a primitive homodimeric RC containing 58 bacteriochlorophylls and 2 chlorophyll as. The chlorophyll serves as the primary electron acceptor (Chl a-A0) responsible for light harvesting and charge separation. The single-molecule spectroscopy of Chl a-A0 can be used to investigate heterogeneities of the RC photochemical function, though the low fluorescence quantum yield (0.1%) makes it difficult. Here, we show the fluorescence excitation spectroscopy of individual Chl a-A0s in single hRCs at 6 K. The fluorescence quantum yield and absorption cross section of Chl a-A0 increase 2- and 4-fold, respectively, compared to those at room temperature. The two Chl a-A0s in single hRCs are identified as two distinct peaks in the fluorescence excitation spectrum, exhibiting different excitation polarization dependences. The spectral changes caused by photobleaching indicate the energy transfer across subunits in the hRC.

Chemical potentials of electric double layers at metal-electrolyte interfaces: dependence on electrolyte concentration and electrode materials, and application to field-effect transistors.

When a metal is soaked in an electrolyte solution, the metal and solution affect each other through the formation of electric double layers (EDLs) at their interfaces. The EDLs at metal-electrolyte interfaces can realize high-density charge-carrier injections and accumulations, and thus have recently attracted attention for their potential application to energy storage, and electronic and electrochemical devices. In such EDL-based devices, including field-effect transistors (FETs), the potential energy of surface electrons in the metal electrodes (EM) governs the transistor device performance. This is in clear contrast to redox-driven electrochemical devices such as dye-sensitized solar cells and electrochromic devices, whose performance is primarily governed by the potentials of the redox-active species. However, there has been no systematic research to bridge the distance between metal electrons and electrolyte ions. In the present study, we carefully examined the dependence of EM of ITO, Au and Pt electrodes on the concentration of the PEG solutions of LiCl and MgCl2, because it has been well established that the chemical potential of electrolyte solutions is dependent on the solution concentrations. Our results showed that, at the same electrolyte concentration, the values of EM increased in the order of ITO, Au and Pt; moreover, on the same electrode, EM showed linear decreases as a function of the logarithm of the electrolyte concentrations. To understand these behaviors, we developed a theoretical treatment of the EDLs based on the simple Gouy-Chapman model, and obtained the theoretical expressions of EM in terms of the concentration of electrolyte and the work function of the metal electrode (ΦM), which were found to successfully explain the dependences of EM on the electrolyte concentration and the electrode materials. We also examined the EDL-FETs of platinum phthalocyanine (PtPc), with various LiCl-PEG solutions of different concentrations as gate electrolytes. The threshold voltage eVT and EM exhibited a linear relation, which was well explained by the relation between EM and the valence band energy EVB of PtPc. The transfer characteristics at various gate voltage VG were found to be well normalized by a function of eVG + EM.

Nanometer Accuracy in Cryogenic Far-Field Localization Microscopy of Individual Molecules.

We demonstrate the nanometer accuracy of far-field fluorescence localization microscopy at a temperature of 1.8 K using near-infrared and red fluorophores bonded to double-stranded DNA molecules (10.2 nm length). Although each fluorophore was localized with a 1 nm lateral precision by acquiring an image at one axial position within the focal depth of ±0.7 µm, the distance between the two fluorophores on the lateral plane (Dxy) was distributed from 0 to 50 nm. This systematic error was mainly due to detecting with the large focal depth the dipole emission from orientationally fixed fluorophores. Each fluorophore was localized with precisions of ±1 nm (lateral) and simultaneously ±11 nm (axial) by acquiring images every 100 nm in the axial direction from -900 to 900 nm. By correcting the dipole orientation effects, the distribution of Dxy was centered around the DNA length. The average and standard deviation of Dxy were 10 and 5 nm.

Ionic liquid thin layer-induced memory effects in organic field-effect transistors.

We examined the morphologies and structures of pentacene and C60 thin films grown on thin layers of an ionic liquid, N,N-diethyl-N-methyl-N-(2-methoxyethyl)ammonium bis(trifluoromethanesulfonyl)imide (DEME-TFSI), and found that the characteristics of the films depended significantly on the thickness of DEME-TFSI. In addition, we fabricated organic field-effect transistors (OFETs) of pentacene and C60 in which a thin layer of DEME-TFSI was inserted between the organic semiconductor (pentacene or C60) and the gate insulating layer, and measured their performance in situ. We found that 1.5-2 ML (ML: monolayer) DEME-TFSI produced a large hysteresis loop in the transfer characteristics in these OFETs, but 5 ML DEME-TFSI resulted in the formation of normally-on states with far smaller memory effects. The curvatures of the hysteresis loops were caused by the formation of trap states induced by the DEME-TFSI layers. This novel technique provides a simple tool for creating hysteresis behavior and could potentially be applied to transistor memory devices.

Rate-determining process in MISIM photocells for optoelectronic conversion using photo-induced pure polarization current without carrier transfer across interfaces.

Recently, we proposed a [metal|insulator|semiconductor|insulator|metal] (MISIM) photocell, as a novel architecture for high-speed organic photodetectors. The electric polarization in the S layer, induced by modulated light illumination, propagates into the outside circuit as a polarization current through the I layers, without any carrier transfer across the interfaces. In the present work, we examined the MISIM photocells consisting of zinc-phthalocyanine(ZnPc)-C60 bilayers for the S layer and Parylene C for the two I layers, to understand the fundamental aspects of the MISIM photocells, such as current polarity and modulation-frequency dependence. It was found that, in such devices, the current polarity was primarily determined by the polarization in the S layer, which was induced by the donor-acceptor charge-transfer upon illumination. Furthermore, the ON and OFF current, which appeared in the periods of illumination-on and -off, respectively, exhibited significantly different dependence on the modulation frequency. This was well-explained by an imbalance between a quick polarization in the S layer during illumination and its slow relaxation in the dark.

Electron Highways into Nanochannels of Covalent Organic Frameworks for High Electrical Conductivity and Energy Storage.

To enhance the electron transfer within the covalent organic frameworks (COFs), we obtained a nanocomposite of conductive poly(3,4-ethylenedioxythiophene) (PEDOT) and redox-active AQ-COF by performing a facile in situ solid-state polymerization inside the nanochannels of COFs. The PEDOT chains functioned like electron highways within the nanochannels, resulting in a PEDOT@AQ-COF nanocomposite with an excellent electrical conductivity of 1.1 S cm-1 and a remarkably improved performance in faradaic energy storage. The all-organic PEDOT@AQ-COF electrode showed specific capacitance as high as 1663 F g-1 (at 1 A g-1), ultrafast charge/discharge rate performance (998 F g-1 at 500 A g-1), and excellent stability for 10 000 cycles. This research demonstrates a promising strategy for increasing the conductivity of COF-based materials and broadening their applications.

Cryogenic Fluorescence Localization Microscopy of Spectrally Selected Individual FRET Pairs in a Water Matrix.

We prepared a pair with a visible-absorbing donor dye and a near-infrared fluorescing acceptor dye. The donor and the acceptor were covalently linked close enough for Förster resonance energy transfer to occur. Under cryogenic conditions at 1.7 K, we observed the fluorescence excitation spectra of the individual pairs in a water matrix. We tested one rhodamine, two Bodipy, and one carbopyronine derivatives as the donor. Among these donors, Bodipy derivatives show the narrowest spectral width of the individuals with respect to the ensemble width. Thus, Bodipy dyes were favorable as the donor for the spectral selection of individual pairs. At 1.7 K, from the several Bodipy-acceptor pairs in the diffraction-limited volume, an individual pair was selected by the fluorescence excitation spectrum of the donor. The spectrally selected pair was localized using the near-infrared fluorescence of the acceptor.

Multifunctional Dithiadiazolyl Radicals: Fluorescence, Electroluminescence, and Photoconducting Behavior in Pyren-1'-yl-dithiadiazolyl.

The pyren-1'-yl-functionalized dithiadiazolyl (DTDA) radical, C16H9CNSSN (1), is monomeric in solution and exhibits fluorescence in the deep-blue region of the visible spectrum (440 nm) upon excitation at 241 nm. The salt [1][GaCl4] exhibits similar emission, reflecting the largely spectator nature of the radical in the fluorescence process, although the presence of the radical leads to a modest quenching of emission (ΦF = 98% for 1+ and 50% for 1) through enhancement of non-radiative decay processes. Time-dependent density functional theory studies on 1 coupled with the similar emission profiles of both 1+ and 1 are consistent with the initial excitation being of predominantly pyrene π-π* character. Spectroscopic studies indicate stabilization of the excited state in polar media, with the fluorescence lifetime for 1 (τ = 5 ns) indicative of a short-lived excited state. Comparative studies between the energies of the frontier orbitals of pyren-1'-yl nitronyl nitroxide (2, which is not fluorescent) and 1 reveal that the energy mismatch and poor spatial overlap between the DTDA radical SOMO and the pyrene π manifold in 1 efficiently inhibit the non-radiative electron-electron exchange relaxation pathway previously described for 2. Solid-state films of both 1 and [1][GaCl4] exhibit broad emission bands at 509 and 545 nm, respectively. Incorporation of 1 within a host matrix for OLED fabrication revealed electroluminescence, with CIE coordinates of (0.205, 0.280) corresponding to a sky-blue emission. The brightness of the device reached 1934 cd/m2 at an applied voltage of 16 V. The crystal structure of 1 reveals a distorted π-stacked motif with almost regular distances between the pyrene rings but alternating long-short contacts between DTDA radicals. Solid state measurements on a thin film of 1 reveal emission occurs at shorter wavelengths (375 nm) whereas conductivity measurements on a single crystal of 1 show a photoconducting response at longer wavelength excitation (455 nm).

Giant negative magnetoresistance in Ni(quinoline-8-selenoate)2.

The magnetic, structural, conductivity and magnetoresistance properties of [Ni(quinoline-8-selenoate)2] ([Ni(qs)2]) have been studied. Despite the insolubility of the material necessitating its study as a powdered sample, a remarkably high conductivity has been measured. The conductivity is an order of magnitude greater than the thin-film processable thiol analogue previously reported and has been interpreted through the same space-charge limited conduction mechanism with charges injected from the electrodes. The introduction of selenium, results in a material with conductivity approaching metallic due to the enhanced interaction between adjacent molecules. Additionally, under an applied magnetic field, the material displays a negative magnetoresistance effect above 35% at 2 K. The effect can still be observed at 200 K and is interpreted in terms of a double-exchange mechanism.

3D Spin-Liquid State in an Organic Hyperkagome Lattice of Mott Dimers.

We report the first 3D spin liquid state of isotropic organic spins. Structural analysis, and magnetic and heat-capacity measurements were carried out for a chiral organic radical salt, (TBA)_{1.5}[(-)-NDI-Δ] (TBA denotes tetrabutylammonium and NDI denotes naphthalene diimide), in which (-)-NDI-Δ forms a K_{4} structure due to its triangular molecular structure and an intermolecular π-π overlap between the NDI moieties. This lattice was identical to the hyperkagome lattice of S=1/2 Mott dimers, and should exhibit 3D spin frustration. In fact, even though the high-temperature magnetic susceptibility followed the Curie-Weiss law with a negative Weiss constant of θ=-15 K, the low-temperature magnetic measurements revealed no long-range magnetic ordering down to 70 mK, and suggested the presence of a spin liquid state with a large residual paramagnetism χ_{0} of 8.5×10^{-6} emu g^{-1} at the absolute zero temperature. This was supported by the ^{14}N NMR measurements down to 0.38 K. Further, the low-temperature heat capacities c_{p} down to 68 mK clearly indicated the presence of c_{p} for the spin liquid state, which can be fitted to the power law of T^{0.62} in the wide temperature range 0.07-4.5 K.

Three-Dimensional Localization of an Individual Fluorescent Molecule with Angstrom Precision.

Among imaging techniques, fluorescence microscopy is a unique method to noninvasively image individual molecules in whole cells. If the three-dimensional spatial precision is improved to the angstrom level, various molecular arrangements in the cell can be visualized on an individual basis. We have developed a cryogenic reflecting microscope with a numerical aperture of 0.99 and an imaging stability of 0.05 nm in standard deviation at a temperature of 1.8 K. The key optics to realize the cryogenic performances is the reflecting objective developed by our laboratory. With this cryogenic microscope, an individual fluorescent molecule (ATTO647N) at 1.8 K was localized with standard errors of 0.53 nm (x), 0.31 nm (y), and 0.90 nm (z) when 106 fluorescence photons from the molecule were accumulated in 5 min.

Molecular and thin film properties of cobalt half-sandwich compounds for optoelectronic application.

The structure and electronic properties of a novel cobalt half sandwich complex of cyclopentadiene (Cp) and diaminonaphthalene (DAnap) [CpCo(DAnap)] are described and compared to the previously reported diaminobenzene derivative [CpCo(DAbnz)] in view of their potential for (opto)electronic device application. Both complexes show stable redox processes, tunable through the diaminoacene ligand, and show strong absorption in the visible region, with additional transitions stretching into the near infrared (NIR). CpCo(DAnap) crystallises with a particularly large unit cell (9301 Å3), comprising 32 molecules, with a gradual rotation over 8 molecules along the long c-axis. In the solid state the balance of the optical transitions in both complexes is reversed, with a suppression of the visible band and an enhancement of the NIR band, attributed to extensive intermolecular electronic interaction. In the case of CpCo(DAnap), highly crystalline thin films could be formed under physical vapor deposition, which show a photocurrent response stretching into the NIR, and p-type semiconductor behavior in field effect transistors with mobility values of the order 1 × 10-4 cm2 V-1 s-1. The device performance is understood through investigation of the morphology of the grown films.

Transport Characteristics of the Organic Field-Effect Transistors Based on Charge Transfer Complex as Semiconductors.

The n-type organic field-effect transistors are fabricated with using four kinds of charge transfer (CT) complexes with PXX (peri-xanthenoxanthene) as a donor component. The CT complexes with four kinds of acceptors form mixed-stack type one-dimentional columns with different PXX-acceptor overlaps. Comparison of the field-effect properties reveals the correlation between the device performance and intermolecular interaction in the semiconducting CT complexes.