Detection of FRET efficiency in imaging systems by photo-bleaching acceptors

Fluorescence resonance energy transfer (FRET) is widely used to obtain the distance between a donor and an acceptor in biological research. However, the detection of FRET efficiencies with fluorescence microscopy imaging systems remains a great challenge due to the difficulties of transferring gray scales of the images into fluorescence intensities, and the absence of exact quantum yields of donors and acceptors. Herein, we presented a new method to detect the FRET efficiency in imaging systems by analyzing the photo-bleaching-induced changes in fluorescent intensities of quantum dots (QDs, donors) and Cy5 dyes (acceptors). Our method is different from the previous acceptor-photo-bleaching studies in imaging systems by theoretically analyzing the bleaching process, and bringing forward a new parameter which is universal for samples of the same kind. It is convenient for calculating FRET efficiencies. There is hardly any spectral crosstalk between 605QD and Cy5, thus the FRET result is more accurate than that of many other common FRET pairs. The lengths of single-stranded and double-stranded DNA fragments in solution were determined via the analysis of FRET efficiency values. This technique provides a reliable approach to study biomacromolecules in living cells through fluorescent imaging and in situ measurements.     

Talbot holographic illumination nonscanning (THIN) fluorescence microscopy

Optical sectioning techniques offer the ability to acquire three‐dimensional information from various organ tissues by discriminating between the desired in‐focus and out‐of‐focus (background) signals. Alternative techniques to confocal, such as active structured illumination, exist for fast optically sectioned images, but they require individual axial planes to be imaged consecutively. In this article, an imaging technique (THIN), by utilizing active Talbot illumination in 3D and multiplexed holographic Bragg filters for depth discrimination, is demonstrated for imaging in vivo 3D biopsy without mechanical or optical axial scanning.     

Effect of potassium permanganate on morphological,structural and electro-optical properties of graphene oxide thin films

This work investigated the effect of Potassium Permanganate (KMnO₄) on graphene oxide (GO) properties, especially on electrical properties. The GO thin films were deposited on a glass substrate using drop casting technique and were analysed by using various type of spectroscopy (e.g. Scanning Electron Microscopy (SEM), Ultra- Violet Visible (UV–VIS), Fourier Transform Infrared (FTIR), X-Ray Diffraction (XRD), optical band gap, Raman Spectroscopy). Furthermore, the electrical experiments were carried out by using current–voltage (I-V) characteristic. The GO thin film with 4.5 g of KMnO₄ resulted in higher conductivity which is 3.11 × 10^?4 S/cm while GO with 2.5 g and 3.5 g of KMnO₄ achieve 2.47 × 10^?9 S/cm and 1.07 × 10^?7 S/cm, respectively. This further affects the morphological (SEM), optical (band gap, UV–Vis, FTIR, and Raman), and crystalline structural (XRD) properties of the GO thin films. The morphological, elemental, optical, and structural data confirmed that the properties of GO is affected by different amount of KMnO₄ oxidizing agent, which revealed that GO can potentially be implemented for electrical and electronic devices.     

明永乐青花瓷片的显微拉曼光谱分析

利用显微拉曼光谱对明永乐青花瓷釉及釉中的结晶物进行了分析,釉的拉曼谱中Si-O弯曲振动与伸缩振动峰的面积比(Ip=A500/A1000)与陶瓷的烧成温度相关连。同时在釉中还发现有Fe3O4、α-Fe2O3、Co3O4、MnO、Mn2O3、硅酸钙、磷酸钙等结晶物。研究结果表明,显微拉曼光谱是一种简便、可靠的无损检测手段,对古陶瓷的鉴定与分析有重要的应用意义。     

Ultrasensitive Imaging of Cells and Sub-Cellular Entities by Electrochemiluminescence

Here we report on a label-free electrochemiluminescence (ECL) microscopy using exceptionally low concentrations of the [Ru(bpy)₃]²⁺ luminophore. This work addresses the central point of the minimal concentration of the ECL luminophore required to image single entities. We demonstrate the possibility to record ECL images of cells and mitochondria at concentrations down to nM and pM. This is 7 orders of magnitude lower than classically-used concentrations and corresponds to a few hundreds of luminophores diffusing around the biological entities. Yet, it produces remarkably sharp negative optical contrast ECL images, as demonstrated by structural similarity index metric analyses and supported by predictions of the ECL image covering time. Finally, we show that the reported approach is a simple, fast, and highly sensitive method, which opens new avenues for ultrasensitive ECL imaging and ECL reactivity at the single molecule level.     

Synthesis and characterization of dispersions of ZnCrO4 prepared in AOT stabilized water/heptane microemulsion

Dispersions of zinc chromate (ZnCrO₄) were prepared in H₂O/AOT (sodium bis(2-ethylhexyl sulfosuccinate))/n-heptane water-in-oil (W/O) microemulsion medium with various water pool sizes and precursor concentration both without and with sonication. The formation of ZnCrO₄ in the microemulsion was verified by XRD and FTIR measurements. The absorbance of the dispersions formed in different water pool sizes was studied. Their dimension in the microemulsion medium was determined by the dynamic light scattering method. Enthalpy of formation of ZnCrO₄ in W/O microemulsion medium was measured by isothermal titration calorimetry (ITC). The dimension and morphology of the formed ZnCrO₄ colloidal particles examined by transmission electron microscopy (TEM) were strongly dependent on the water pool size, precursor concentration and sonication.     

Synthesis and electro‐optical characterization of new conducting PEDOT/Au‐nanorods nanocomposites

This paper describes a new strategy to obtain PEDOT/Au‐nanorods nanocomposites with different PEDOT: Au ratio. A polymeric ionic liquid (PIL) was used as stabilizer during the chemical synthesis of PEDOT dispersions. PEDOT/Au‐nanorods dispersions in organic media were obtained. Electrochemical characterization of PEDOT/Au‐nanorods nanocomposites revealed that the addition of Au nanorods modified the electroactivity of the conducting films by reducing the oxidation potential from +0.33 to +0.23 V (versus Ag/AgCl). Optical contrast (ΔT%) of the films decreased from 17% for neat PEDOT films to 8% for PEDOT/Au‐nanorods nanocomposites films (3:1 (v/v)) while switching times (from 1 to 4 sec) were similar to neat PEDOT. Conductivity of the films increased from 0.027 S/cm for neat PEDOT to 0.691 S/cm for PEDOT/Au‐nanorods nanocomposites. Nanoscale morphology and contact potential of PEDOT/Au‐nanorods nanocomposites were investigated in detail by Scanning Force Microscopy. Electrical measurements show a clear contact potential difference between the ITO substrate and the PEDOT/Au‐nanorods film. On the film, no contact potential inhomogeneity is observed indicating that the Au‐nanorods are uniformly dispersed in the film. Copyright © 2010 John Wiley & Sons, Ltd.     

Self-quenched Fluorophore Dimers for DNA-PAINT and STED Microscopy

Super-resolution techniques like single-molecule localisation microscopy (SMLM) and stimulated emission depletion (STED) microscopy have been extended by the use of non-covalent, weak affinity-based transient labelling systems. DNA-based hybrid systems are a prominent example among these transient labelling systems, offering excellent opportunities for multi-target fluorescence imaging. However, these techniques suffer from higher background relative to covalently bound fluorophores, originating from unbound fluorophore-labelled single-stranded oligonucleotides. Here, we introduce short-distance self-quenching in fluorophore dimers as an efficient mechanism to reduce background fluorescence signal, while at the same time increasing the photon budget in the bound state by almost 2-fold. We characterise the optical and thermodynamic properties of fluorophore-dimer single-stranded DNA, and show super-resolution imaging applications with STED and SMLM with increased spatial resolution and reduced background.     

Physical properties of dynamic force microscopies in contact and noncontact operation

In the field of Scanning Force Microscopy several dynamical contact and noncontact modes have been introduced increasing the range of detectable surface and interface properties, and allowing to detect material properties such as elasticity and mass density on the nanometer scale. A detailed understanding of tip/surface interactions and the dynamic processes involved is required to understand the origin of a material contrast using these techniques. Here a general method to solve the equation of motion of a vibrating SFM cantilever/tip system in an external force field is presented. Contact modes as well as intermittent contact modes are discussed using a single set of equations describing the cantilever/tip motion, and by varying the size of amplitudes of the vibrating cantilever/tip system. To quantitatively describe the oscillation behavior of the SFM cantilever at large amplitudes the computer simulations are based on the MYD/BHW model providing a realistic contact model with respect to the contact area, the size of the contact forces as well as the transition from repulsive to attractive forces. The results are compared with the experiment and with different approaches based on analytical and numerical models.     

In situ infrared spectroscopy at electrochemical interfaces

An insight into the in situ FTIR spectroscopy method as applied in Electrochemistry is given. The particular aspects inherent to the electrochemical method are described in a concise form. Selected examples cover the results of about the last 8 years, on a variety of systems including carbon monoxide, small organic molecules and double-layer components (hydrogen, anions and water). The experimental data refer mostly to adsorption on well-defined single-crystal surfaces. Analogies and differences with data from the metal/gas interface are discussed.