Biexciton-like Auger Blinking in Strongly Confined CsPbBr3 Perovskite Quantum Dots.

Perovskite quantum dots (QDs) with high room-temperature luminescence efficiency have been applied in single-photon sources. While the optical properties of large, weakly confined perovskite nanocrystals have been extensively explored at the single-particle level, few studies have focused on single-perovskite QDs with strong quantum confinement. This is mainly due to their poor surface chemical stability. Here we demonstrate that strongly confined CsPbBr3 perovskite QDs (SCPQDs) embedded in a phenethylammonium bromide matrix exhibit a well-passivated surface and improved photostability under intense photoexcitation. We find that in our SCPQDs, photoluminescence blinking is suppressed at moderate excitation intensities, and increasing the excitation rates leads to weak photoluminescence intensity fluctuations accompanied by an unusual spectral blue-shift. We attribute this to a biexciton-like Auger interaction between excitons and trapped excitons formed by surface lattice elastic distortions. This hypothesis is corroborated by the unique repulsive biexciton interaction observed in the SCPQDs.

Amine Functionalization of Silica Sol-Gel Thin Films via Kinetic Doping: A Novel, Green Approach.

Amine-functionalized thin films are highly desirable technologies for analytical, material, and biochemistry applications. Current functionalization procedures can be costly, environmentally unfriendly, and require many synthetic steps. Here, we present an inexpensive and facile way to functionalize a silica thin film with a 25 000 MW branched polyethylenimine (BPEI), consistent with green chemistry principles. Using UV-vis spectroscopy and scanning electron microscopy, BPEI was determined to be loaded into the film at an approximately 0.5 M concentration, which is a 500× increase from the loading solution used. The films were also tested for copper(II) sequestration to assess their potential for heavy metal sequestration and showed a high loading capacity of 10 ± 6 mmol/g. Films proved to be reusable, using ethylenediaminetetraacetic acid to chelate copper and regenerate the films, with only a 6% reduction in the amount of copper(II) ions sequestered by the third use. The films also proved stable against leaching over the course of 1 week in solution, with less than 1% of the original BPEI lost under various storage conditions (i.e., storage in deionized (DI) water, storage in dilute BPEI solution, storage in DI water after annealing). These films show promise for multiple applications, from heavy metal sequestration to antifouling applications, while being inexpensive, facile, and environmentally friendly to synthesize. To our knowledge, this is the first time that BPEI has been doped into silica thin films.

Multienzyme, Multistep Biosensor Produced through Kinetic Doping.

The recently developed kinetic doping technique has shown promise in loading individual enzymes for use as a biosensor. In this study, the first example of kinetic doping to produce a biosensor loaded with more than one enzyme and using a multistep reaction pathway for detection is presented. Glucose oxidase (GOD) is shown to load both individually and together with horseradish peroxidase (HRP) with the tandem action of the two enzymes proving to be effective at detecting glucose in solution. Using a calculation based on known maximum loadings and experimentally determined activities, the final dual-enzyme thin films of known volume are shown to contain 1.8 ± 0.1 mmol/L of HRP and 0.22 ± 0.01 mmol/L of GOD, which represent 33 and 92% of loading efficiencies that each enzyme is known to be, respectively, capable of in a singularly loaded thin film. With the high loading afforded by the kinetic doping process under benign conditions, the thin films are able to load both enzymes all at once in an amount sufficient to function as an efficient biosensor. The most advantageous aspects of this process are its ease of production, involving only a few steps to produce highly loaded thin films that require no additional processing to function as intended, as well as the protein friendly environment that exists in the sol-gel film at the time of enzyme loading. This removes many typical restrictions on immobilizing protein and opens up a wider range of enzymes amenable to the process that enables the fabrication of more complex multistep biosensors utilizing a large array of proteins in the foreseeable future.

Kinetically Doped Silica Sol-Gel Optical Biosensors: Expanding Potential Through Dip-Coating.

Kinetic doping has previously been shown to be an effective method of doping silica sol-gel thin films with an enzyme to construct biosensors. Until now, kinetic doping has only been applied to films produced through the spin-coating method. In this study, we present the use of dip-coating to produce thin films kinetically doped for biosensor development. In this way, kinetically doped biosensors may benefit from the increased range of substrate material shapes and sizes that may be easily coated through dip-coating but not spin-coating. The biosensors produced through dip-coating continue to show enhanced performance over more conventional enzyme loading methods with horseradish peroxidase and cytochrome C samples, showing an increase of 2400× and 1300× in enzyme concentration over that in their loading solutions, respectively. These correspond to enzyme concentrations of 5.37 and 10.57 mmol/L all while preserving a modest catalytic activity for the detection of hydrogen peroxide by horseradish peroxidase. This leads to a 77% and 88% increase in the total amount of horseradish peroxidase and cytochrome C, respectively, over that from coating the same glass coverslip via spin-coating methods.

Silica Sol-Gel Optical Biosensors: Ultrahigh Enzyme Loading Capacity on Thin Films via Kinetic Doping.

Easy to use and easy to produce biosensors would have a huge range of applications. To reach this goal many see the incorporation of a protein into a sol-gel network as one of the most viable options. The current most prevalent technique of predoping presents inherent limits on the concentration possible for the resulting thin film. In this study we demonstrate a new process utilizing the newly developed kinetic doping method to load silica sol-gel thin films with cytochrome C (CytC) and horseradish peroxidase (HRP). Both enzymes are shown to successfully load and have a concentration increase over their original loading solution by factors of 1300× and 2600×, respectively. Furthermore, each enzyme once loaded retained the ability to act as a catalyst for the detection of hydrogen peroxide. Ultimately the CytC- and HRP-loaded thin films were found to have enzyme concentrations of 11 ± 1 mM and 6.0 ± 0.4 mM, respectively, a considerable step up from any doping method reported in the past.

Energy-dependent motion of TonB in the Gram-negative bacterial inner membrane.

Gram-negative bacteria acquire iron with TonB-dependent uptake systems. The TonB-ExbBD inner membrane complex is hypothesized to transfer energy to outer membrane (OM) iron transporters. Fluorescence microscopic characterization of green fluorescent protein (GFP)-TonB hybrid proteins revealed an unexpected, restricted localization of TonB in the cell envelope. Fluorescence polarization measurements demonstrated motion of TonB in living cells, which likely was rotation. By determining the anisotropy of GFP-TonB in the absence and presence of inhibitors, we saw the dependence of its motion on electrochemical force and on the actions of ExbBD. We observed higher anisotropy for GFP-TonB in energy-depleted cells and lower values in bacteria lacking ExbBD. However, the metabolic inhibitors did not change the anisotropy of GFP-TonB in ΔexbBD cells. These findings demonstrate that TonB undergoes energized motion in the bacterial cell envelope and that ExbBD couples this activity to the electrochemical gradient. The results portray TonB as an energized entity in a regular array underlying the OM bilayer, which promotes metal uptake through OM transporters by a rotational mechanism.

Fluorescence-on response via CB7 binding to viologen-dye pseudorotaxanes.

Fluorescence-on sensors typically rely on disrupting photoinduced electron transfer quenching of the excited state through binding the electron donor. To provide a more general fluorescence-on signaling unit, a quencher-fluorophore dyad has been developed in which quenching by electron transfer to a tethered viologen acceptor can be disrupted through complexation of the viologen by cucurbit[7]uril (CB7). Dyads of benzyl viologen-rhodamine B or a BODIPY fluorophore gave upon CB7 complexation 14- and 30-fold fluorescence enhancement, respectively.

Cucurbit[7]uril disrupts aggregate formation between rhodamine B dyes covalently attached to glass substrates.

Dye aggregation is detrimental to the performance of high optical density dye-doped photonic materials. To overcome this challenge, the ability of cucurbit[7]uril (CB7) as a molecular host to disrupt aggregate formation on glass substrates was examined. Rhodamine B was covalently attached to glass slides by initially coating the surface with azidohexylsiloxane followed by copper-catalyzed "click" triazole formation with rhodamine B propargyl ester. The absorption and emission spectra of rhodamine B coated slides in water indicated diverse heterogeneous properties as surface dye density varied. Fluorescence quenching due to dye aggregation was evident at high surface dye density. Addition of aqueous cucurbit[7]uril (CB7) to the surface-tethered dyes perturbed the spectra to reveal a considerable reduction in heterogeneity, which suggested that the presence of a surface in close proximity does not significantly impair CB7's ability to complex with tethered rhodamine B.

Balance between Coulombic interactions and physical confinement in silica hydrogel encapsulation.

We examined the behavior of various entrapped guest molecules within silica hydrogel and evaluated the effect of Coulombic interactions and physical confinement on molecular mobility. Although rhodamine 6G (R6G) and fluorescein (FL) share similar size and molecular structure, their behavior in silica hydrogel was found to be dramatically different. A good majority of R6G was immobilized with little to no exchangeable molecules, whereas FL displayed a considerable amount of mobility in silica hydrogel. Moreover, silica hydrogel encapsulated R6G failed to gain mobility even under low pH or high ionic strength conditions to minimize Coulombic interactions, implying that encapsulated R6G molecules were inaccessible and likely trapped deep inside the silica matrix of a hydrogel. On the contrary, FL was relatively free to rotate and translate inside a silica hydrogel, implying that FL remained solvated in the solvent phase and was able to maintain its mobility throughout the hydrogel formation process. Fluorescence recovery after photobleaching measurements put the diffusion coefficient of FL in silica hydrogel at ca. 2.1 x 10(-6) cm(2) s(-1), about a factor of 3 slower than that in solution. The substantial difference in mobility between cationic R6G and anionic FL led us to conclude that the effect of Coulombic interactions on mobility is more dominating in hydrogel than in alcogel. Our results also suggest that Coulombic interactions are strong enough to influence the eventual placement of a guest molecule in a silica hydrogel, causing R6G and FL to reside in different microenvironments. This has a profound implication on the use of molecular probes to study silica hydrogel since a slight difference in physical attribute may result in very diverse observations even from identically prepared silica hydrogel samples. As demonstrated, the repulsion between FL and silica renders FL liquid-bound, making FL more suitable for monitoring the change in viscosity and physical confinement during hydrogel formation, whereas other researchers have shown that silica-bound R6G is more suitably used as a reliable probe for monitoring the growth of silica colloids because of its strong attraction toward silica.

Liposomes as protective capsules for active silica sol-gel biocomposite synthesis.

Using liposome to shield an enzyme from hostile chemical environments during the sol-gel formation process has resulted in a novel approach to synthesizing silica sol-gel biocomposite materials. By reporting the encapsulation of horseradish peroxidase and firefly luciferase, we demonstrate that this new protocol can produce silica biocomposites that are more active than trapping the enzymes directly into hydrogels.

Comparative study of guest charge-charge interactions within silica sol-gel.

We investigated the effect of charge-charge interactions on the mobilities of rhodamine 6G (R6G), Nile Red, sulforhodamine B, and Oregon Green 514 (ORG) guest molecules within a silica sol-gel host as the guest charge progressed from positive to neutral to negative. Through classification of the mobility as fixed, tumbling, or intermediate behavior, we were able to distinguish differences in surface attraction as the guest charge was varied. On the basis of our results, an attractive charge (as tested by cationic R6G) does not contribute significantly to mobility within dry films. However, an increase in the cationic influence is observed in water-equilibrated environments. A comparison of ORG in dry and water- and phosphate-buffer-equilibrated films indicates that charge repulsion does significantly increase dye rotational mobility (to a maximum of 24 +/- 3% tumbling molecules). However, in view of the percentage of tumbling molecules found, charge-charge interactions do not appear to be the dominant force controlling guest mobility.

Coulombic interactions on the deposition and rotational mobility distributions of dyes in polyelectrolyte multilayer thin films.

We employed negatively charged fluorescein (FL), positively charged rhodamine 6G (R6G), and neutral Nile Red (NR) as molecular probes to investigate the influence of Coulombic interaction on their deposition into and rotational mobility inside polyelectrolyte multilayer (PEM) films. The entrapment efficiency of the dyes reveals that while Coulombic repulsion has little effect on dye deposition, Coulombic attraction can dramatically enhance the loading efficiency of dyes into a PEM film. By monitoring the emission polarization of single dye molecules in polyethylenimine (PEI) films, the percentages of mobile R6G, NR, and FL were determined to be 87 +/- 4%, 76 +/- 5%, and 68 +/- 3%, respectively. These mobility distributions suggest that cationic R6G enjoys the highest degree of rotational freedom, whereas anionic FL shows the least mobility because of Coulombic attraction toward cationic PEI. Regardless of charges, this high percentage of mobile molecules is in stark contrast to the 5-40% probe mobility reported from spun-cast polymer films, indicating that our PEI films contain more free volume and display richer polymer dynamics. These observations demonstrate the potential of using isolated fluorescent probes to interrogate the internal structure of a PEM film at a microscopic level.

Probing the dynamic guest-host interactions in sol-gel films using single molecule spectroscopy.

Organic dyes usually exhibit enhanced photostability when trapped inside sol-gel silicates. The enhanced photostability is attributed to the reduction of intramolecular motions that facilitate photodegradation. We report the simultaneous detection of mobility and photostability of sol-gel encapsulated didodecyl-3,3,3',3'-tetramethylindocarbocyanine (DiI) using single molecule spectroscopy. Fluorescence from DiI was resolved into parallel and perpendicular polarization components and separately detected. On the basis of the calculated fluorescence polarization, single DiI molecules were classified into "tumbling" and "fixed". Out of 212 molecules investigated, 52% were found to be fixed. For the first time, the mobility of a guest molecule in sol-gel silicate can be directly correlated with its own photostability. Both tumbling and fixed molecules have shown to exhibit nonuniform photostability, indicative of the very heterogeneous guest-host interactions within each subgroup. The survival lifetimes for the majority of the tumbling and fixed molecules were found to be 4.3 and 13.1 s, respectively, demonstrating unequivocally that fixed molecules exhibit a higher photostability than tumbling molecules. These results are in accordance with a recent study on rhodamine B encapsulated in dried sol-gel silicates.