Design, Syntheses and Applications of Fluorescent Dyes

New methodologies for the efficient syntheses of 4,4-difluoro-4-bora-3a,4adiaza- s-indacenes (BODIPYs) and rosamines were developed. A serendipitous discovery led to a new reaction which afforded BODIPYs in high yields. Systematic studies of the kinetics and mechanisms of the new reaction were performed. A series of BODIPYs were successfully prepared using the new approach. A simple and efficient synthesis of rosamines with cyclic-amine substituents was devised. These new rosamines showed interesting anti-tumor activities. Several types of novel fluorescent compounds were prepared. Highly fluorescent GFP-chromophore analogs were designed and synthesized. The correlation between the optical properties and the structures was investigated. New pyronin dyes with mesoheteroatom substituents were efficiently prepared. The fluorescence properties of these compounds were highly dependent on the nature of the meso-substituents. A set of BODIPY dyes that fluoresce brightly above 600 nm were made. They were then used as acceptors to prepare water-soluble through-bond energy transfer cassettes. All the cassettes had complete energy transfer and high quantum yields in MeOH. A few also had good fluorescence properties in aqueous media and even on proteins. The through-bond energy transfer cassettes were used to monitor protein-protein interactions. In order to test our hypothesis, an artificial protein interaction system was built by utilizing the biotin/(strept)avidin interactions. Thus Atto425-BSA-biotin, streptavidin-cassette1 and avidin-cassette2 were prepared. The interactions between Atto425-BSA-biotin and cassette labeled (strept)avidin were successfully detected in vitro and in living cells by fluorescence techniques

Rosamines Targeting the Cancer Oxidative Phosphorylation Pathway

Reprogramming of energy metabolism is pivotal to cancer, so mitochondria are potential targets for anticancer therapy. A prior study has demonstrated the anti-proliferative activity of a new class of mitochondria-targeting rosamines. This present study describes in vitro cytotoxicity of second-generation rosamine analogs, their mode of action, and their in vivo efficacies in a tumor allografted mouse model. Here, we showed that these compounds exhibited potent cytotoxicity (average IC50<0.5 µM), inhibited Complex II and ATP synthase activities of the mitochondrial oxidative phosphorylation pathway and induced loss of mitochondrial transmembrane potential. A NCI-60 cell lines screen further indicated that rosamine analogs 4 and 5 exhibited potent antiproliferative effects with Log10GI50 = -7 (GI50 = 0.1 µM) and were more effective against a colorectal cancer sub-panel than other cell lines. Preliminary in vivo studies on 4T1 murine breast cancer-bearing female BALB/c mice indicated that treatment with analog 5 in a single dosing of 5 mg/kg or a schedule dosing of 3 mg/kg once every 2 days for 6 times (q2d×6) exhibited only minimal induction of tumor growth delay. Our results suggest that rosamine analogs may be further developed as mitochondrial targeting agents. Without a doubt proper strategies need to be devised to enhance tumor uptake of rosamines, i.e. by integration to carrier molecules for better therapeutic outcome

Adaptive Dynamic Surface Control for a Class of Nonlinear Pure-Feedback Systems with Parameter Drift

In order to solve the problem of unknown parameter drift in the nonlinear pure-feedback system, a novel nonlinear pure-feedback system is proposed in which an unconventional coordinate transformation is introduced and a novel unconventional dynamic surface algorithm is designed to eliminate the problem of “calculation expansion” caused by the use of backstepping in the pure-feedback system. Meanwhile, a sufficiently smooth projection algorithm is introduced to suppress the parameter drift in the nonlinear pure-feedback system. Simulation experiments demonstrate that the designed controller ensures the global and ultimate boundedness of all signals in the closed-loop system and the appropriate designed parameters can make the tracking error arbitrarily small

Piezoelectric ZnO Thin Films for 2DOF MEMS Vibrational Energy Harvesting

Zinc oxide (ZnO) is an environmental-friendly semiconducting, piezoelectric and non-ferroelectric material, and plays an essential role for applications in microelectromechanical systems (MEMS). In this work, a fully integrated two-degree-of-freedom (2DOF) MEMS piezoelectric vibration energy harvester (p-VEH) was designed and fabricated using ZnO thin films for converting kinetic energy into electrical energy. The 2DOF energy harvesting system comprises two subsystems: the primary one for energy conversion and the auxiliary one for frequency adjustment. Piezoelectric ZnO thin film was deposited using a radio-frequency magnetron sputtering method onto the primary subsystem for energy conversion from mechanical vibration to electricity. Dynamic performance of the 2DOF resonant system was analyzed and optimized using a lumped parameter model. Two closely located but separated peaks were achieved by precisely adjusting mass ratio and frequency ratio of the resonant systems. The 2DOF MEMS p-VEH chip was fabricated through a combination of laminated surface micromachining process, double-side alignment and bulk micromachining process. When the fabricated prototype was subjected to an excitation acceleration of 0.5 g, two close resonant peaks at 403.8 and 489.9 Hz with comparable voltages of 10 and 15 mV were obtained, respectively

Investigation of a Thin‐Film Quasi‐Reference Electrode Fabricated by Combined Sputtering‐Evaporation Approach

This paper presents the development of a thin‐film quasi‐reference electrode (tQRE), which was fabricated by sputtering silver (Ag) on a conducting gold layer. The Ag layer was subsequently covered by silver chloride (AgCl) with the aid of e‐beam evaporation. The functionality of the tQREs as reliable reference electrodes was confirmed by observing the potential response in solutions with various chloride ion concentrations. The influence of solution pH on the potential change of the tQREs was evaluated. In the solution with controlled ionic strength, the tQREs were able to provide stable and consistent potential outputs. Experimental investigation of the electrochemical sensors with integrated tQREs demonstrated potential applicability of the tQREs to be incorporated into miniaturized and disposable lab‐on‐a‐chip sensors for point‐of‐care/in‐situ measurements.National Research Foundation (NRF)Singapore-MIT Alliance for Research and Technology (SMART)Accepted versionThis research is supported by the National Research Foundation (NRF), Prime Minister’s Office, Singapore under its Campus for Research Excellence and Techno- logical Enterprise (CREATE) programme. The Center for Environmental Sensing and Modeling (CENSAM) is an interdisciplinary research group (IRG) of the Singapore MIT Alliance for Research and Technology (SMART) centre

High-contrast fluorescence imaging in fixed and living cells using optimized optical switches.

We present the design, synthesis and characterization of new functionalized fluorescent optical switches for rapid, all-visible light-mediated manipulation of fluorescence signals from labelled structures within living cells, and as probes for high-contrast optical lock-in detection (OLID) imaging microscopy. A triazole-substituted BIPS (TzBIPS) is identified from a rational synthetic design strategy that undergoes robust, rapid and reversible, visible light-driven transitions between a colorless spiro- (SP) and a far-red absorbing merocyanine (MC) state within living cells. The excited MC-state of TzBIPS may also decay to the MC-ground state emitting near infra-red fluorescence, which is used as a sensitive and quantitative read-out of the state of the optical switch in living cells. The SP to MC transition for a membrane-targeted TzBIPS probe (C₁₂-TzBIPS) is triggered at 405 nm at an energy level compatible with studies in living cells, while the action spectrum of the reverse transition (MC to SP) has a maximum at 650 nm. The SP to MC transition is complete within the 790 ns pixel dwell time of the confocal microscope, while a single cycle of optical switching between the SP and MC states in a region of interest is complete within 8 ms (125 Hz) within living cells, the fastest rate attained for any optical switch probe in a biological sample. This property can be exploited for real-time correction of background signals in living cells. A reactive form of TzBIPS is linked to secondary antibodies and used, in conjunction with an enhanced scope-based analysis of the modulated MC-fluorescence in immuno-stained cells, for high-contrast immunofluorescence microscopic analysis of the actin cytoskeleton

Hyaluronic Acid Methacrylate Hydrogel-Modified Electrochemical Device for Adsorptive Removal of Lead(II)

This paper presents the development of a compact, three-electrode electrochemical device functionalized by a biocompatible layer of hyaluronic acid methacrylate (HAMA) hydrogel for the adsorptive removal of detrimental lead (Pb(II)) ions in aqueous solutions. An adsorption mechanism pertaining to the observed analytical performance of the device is proposed and further experimentally corroborated. It is demonstrated that both the molecular interactions originating from the HAMA hydrogel and electrochemical accumulation originating from the electrode beneath contribute to the adsorption capability of the device. Infrared spectral analysis reveals that the molecular interaction is mainly induced by the amide functional group of the HAMA hydrogel, which is capable of forming the Pb(II)&ndash;amide complex. In addition, inductively coupled plasma mass spectrometric (ICP-MS) analysis indicates that the electrochemical accumulation is particularly valuable in facilitating the adsorption rate of the device by maintaining a high ion-concentration gradient between the solution and the hydrogel layer. ICP-MS measurements show that 94.08% of Pb(II) ions present in the test solution can be adsorbed by the device within 30 min. The HAMA hydrogel-modified electrochemical devices exhibit reproducible performance in the aspect of Pb(II) removal from tap water, with a relative standard deviation (RSD) of 1.28% (for n = 8). The experimental results suggest that the HAMA hydrogel-modified electrochemical device can potentially be used for the rapid, on-field remediation of Pb(II) contamination