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New highly sensitive and selective catalytic DNA biosensors for metal ions.

While remarkable progress has been made in developing sensors for metal ions such as Ca(II) and Zn(II), designing and synthesizing sensitive and selective metal ion sensors remains a significant challenge. Perhaps the biggest challenge is the design and synthesis of a sensor capable of specific and strong metal binding. Since our knowledge about the construction of metal-binding sites in general is limited, searching for sensors in a combinatorial way is of significant value. Therefore, we have been able to use a combinatorial method called in vitro selection to obtain catalytic DNA that can bind a metal ion of choice strongly and specifically. The metal ion selectivity of the catalytic DNA was further improved using a 'negative selection' strategy where catalytic DNA that are selective for competing metal ions are discarded in the in vitro selection processes. By labeling the resulting catalytic DNA with a fluorophore/quencher pair, we have made a new class of metal ion fluorescent sensors that are the first examples of catalytic DNA biosensors for metal ions. The sensors combine the high selectivity of catalytic DNA with the high sensitivity of fluorescent detection, and can be applied to the quantitative detection of metal ions over a wide concentration range and with high selectivity. The use of DNA sensors in detection and quantification of lead ions in environmental samples such as water from Lake Michigan has been demonstrated. DNA is stable, cost-effective, environmentally benign, and easily adaptable to optical fiber and microarray technology for device manufacture. Thus, the DNA sensors explained here hold great promise for on-site and real-time monitoring of metal ions in the fields of environmental monitoring, developmental biology, clinical toxicology, wastewater treatment, and industrial process monitoring.

Biosensing Techniques↗

Shah and sine convolution Fourier transform detection for microchannel electrophoresis with a charge coupled device.

This paper describes an improved format for Shah convolution Fourier transform (SCOFT) detection that utilizes the spatial resolution of a charge-coupled device (CCD) rather than a fixed optical mask to perform a Shah or sine convolution over a fluorescence signal. The laser-induced fluorescence from a 9-mm section of microfabricated channel is collected with a CCD at 28 Hz. Each image frame is multiplied by a convolution function to modulate the collected signal through space. Each frame is then summed to generate an intensity-versus-time data set for Fourier analysis. The fluorescence signal oscillates at a frequency dependent upon both the convolution function multiplied across each data frame and the velocity of fluorescent microspheres or a plug of fluorescent dye flowing through the channel. This SCOFT technique affords more flexibility over formats that employ a physical mask and provides data that can be optimized for signal-to-noise (S/N) or resolution information. A 1,000-fold improvement in S/N is demonstrated for a plug of fluorescein dye. Detection of fluorescent beads exhibited frequency signals that were dependent upon the bead size distribution, the electric field, and the electrophoresis buffer concentration. Data are presented demonstrating the quantitation of fluorescent microspheres.

Journal Article↗

DNA detection on plastic: surface activation protocol to convert polycarbonate substrates to biochip platforms.

A mild and efficient surface activation protocol to convert polycarbonate (PC) substrates, e.g., plastic bases of compact disks, to biochip platforms for DNA probe immobilization and target detection is described. The preparation procedure (activation, patterning, and coupling) is simple and effective; the on-chip hybridization is sensitive and selective. Particularly, UV/ozone treatment of PC sheets produces a hydrophilic surface with a high density of reactive carboxylic acid groups [(4.8 +/- 0.2) x 10-10 mol/cm2] in less than 10 min at ambient conditions, and no significant aging or physical damage to the substrate is observed. Covalent immobilization of DNA probes via both passive (reagent-less photopatterning and coupling in bulk solution phase) and flow-through (creation of microarrays with microfluidic channel plates) procedures has been demonstrated. Subsequent hybridization shows uniform and strong fluorescent signals for complementary target DNA and allows clear discrimination between fully complementary targets and strands with a single base-pair mismatch. The surface chemistry described herein will facilitate the development of disposable plastic biochips (not limited to DNA microarrays) and the fabrication of biomedical devices that are readable with conventional optical drives.

DNA↗

Microelectrochemical logic circuits.

We report microelectrochemical devices that mimic some functions of solid-state circuit components such as diodes and transistors. The operation of this new family of devices is enabled by a network of microfluidic electrochemical cells that communicate with one-another via conductive solutions and, in some cases, bipolar electrodes. The output of the devices can be either electrochemical or optical. The approach reported here is a first step toward the development of large-scale integrated electrochemical systems having parallel processing capabilities.

Journal Article↗

Mediator-template assembly of nanoparticles.

The ability to construct size- and shape-controllable architectures using nanoparticles as building blocks is essential for the exploration of nanoparticle-structured properties. This paper reports findings of an investigation of a mediator-template strategy for the size-controllable assembly of nanoparticles. This strategy explores multidentate thioether ligands as molecular mediators and tetraalkylammonium-capped gold nanoparticles (5 nm) as templates toward the preparation of size-controllable and monodispersed spherical assemblies ( approximately 20-300-nm diameters). The combination of the mediation force of the multidentate thioether and the hydrophobic force of the tetraalkylammonium template establishes the interparticle linkage and stability. The morphological properties of the spherical assemblies have been characterized using TEM, AFM, and SAXS techniques. The finding of the soft-hard nature of the nanoparticle assemblies and their interactions with contacting substrates could form the basis of a new strategy for manipulating nanoscale linkages between nanoparticle assemblies, soldering nanoelectronics, and constructing nanosensor devices. The intriguing light scattering and optical absorption properties in response to assembly, disassembly, sizing, and interparticle spacing parameters have been characterized by dynamic light scattering and spectrophotometric measurements. The discovery of the controlled disassembly into individual nanoparticles and the size regulation by a third capping component could form the basis for applications in controlled drug delivery. The fundamental basis for the mediator-template strategy as a versatile assembly technique is further discussed in terms of experimental and theoretical correlations of the morphological and optical properties.

Journal Article↗

Alkyl chain barriers for kinetic optimization in dye-sensitized solar cells.

The optimization of interfacial charge transfer is crucial to the design of dye-sensitized solar cells. In this paper we address the dynamics of the charge separation and recombination in liquid-electrolyte and solid-state cells employing a series of amphiphilic ruthenium dyes with varying hydrocarbon chain lengths, acting as an insulating barrier for electron-hole recombination. Dynamics of electron injection, monitored by time-resolved emission spectroscopy, and of charge recombination and regeneration, monitored by transient optical absorption spectroscopy, are correlated with device performance. We find that increasing dye alkyl chain length results in slower charge recombination dynamics to both the dye cation and the redox electrolyte or solid-state hole conductor (spiro-OMeTAD). These slower recombination dynamics are however paralleled by reduced rates for both electron injection into the TiO2 electrode and dye regeneration by the I-/I3- redox couple or spiro-OMeTAD. Kinetic competition between electron recombination with dye cations and dye ground state regeneration by the iodide electrolyte is found to be a key factor for liquid electrolyte cells, with optimum device performance being obtained when the dye regeneration is just fast enough to compete with electron-hole recombination. These results are discussed in terms of the minimization of kinetic redundancy in solid-state and liquid-electrolyte dye-sensitized photovoltaic devices.

Journal Article↗

Duplication of photoinduced azo polymer surface-relief gratings through a soft lithographic approach.

In this work, a soft lithographic approach has been developed to duplicate photoinduced surface-relief-gratings (SRGs) of azo polymer films to generate the surface pattern replicas composed of different materials on various substrates. For this purpose, thin films of an epoxy-based azo polymer (BP-AZ-CA) were prepared by spin-coating, and SRGs with different structures were inscribed by exposing the films to interference patterns of Ar(+) laser beams at modest intensity (150 mW/cm(2)). Using the azo polymer films as masters, stamps of poly(dimethylsiloxane) (PDMS) were prepared by replica molding. The PDMS stamps were then used to transfer the solutions of poly(3-hexylthiophene) (P3HT), multiwalled carbon nanotube (MWNT), and BP-AZ-CA to different substrates by contact printing. Through this process, surface pattern replicas made of the functional materials were obtained. The pattern formation and quality depended on the factors such as the solution concentration, contacting time in the printing process, and printing pressure. Under the proper conditions, the printed patterns showed the same grating periods as the masters and the same relief depths as the stamps (replicas of the masters). This approach, showing some attractive characteristics such as the easiness of master preparation and the versatility of soft fabrication processes, can be applied to the fabrications of optical functional surfaces, sensors, and photonic devices.

Journal Article↗

Electroluminescence from single monolayers of nanocrystals in molecular organic devices.

The integration of organic and inorganic materials at the nanometre scale into hybrid optoelectronic structures enables active devices that combine the diversity of organic materials with the high-performance electronic and optical properties of inorganic nanocrystals. The optimization of such hybrid devices ultimately depends upon the precise positioning of the functionally distinct materials. Previous studies have already emphasized that this is a challenge, owing to the lack of well-developed nanometre-scale fabrication techniques. Here we demonstrate a hybrid light-emitting diode (LED) that combines the ease of processability of organic materials with the narrow-band, efficient luminescence of colloidal quantum dots (QDs). To isolate the luminescence processes from charge conduction, we fabricate a quantum-dot LED (QD-LED) that contains only a single monolayer of QDs, sandwiched between two organic thin films. This is achieved by a method that uses material phase segregation between the QD aliphatic capping groups and the aromatic organic materials. In our devices, where QDs function exclusively as lumophores, we observe a 25-fold improvement in luminescence efficiency (1.6 cd A(-1) at 2,000 cd m(-2)) over the best previous QD-LED results. The reproducibility and precision of our phase-segregation approach suggests that this technique could be widely applicable to the fabrication of other hybrid organic/inorganic devices.

Journal Article↗

Direct observation of defect-mediated cluster nucleation.

Ion implantation is widely used to introduce electrically or optically active dopant atoms into semiconductor devices. At high concentrations, the dopants can cluster and ultimately form deactivating precipitates, but deliberate nanocrystal formation offers an approach to self-assembled device fabrication. However, there is very little understanding of the early stages of how these precipitates nucleate and grow, in no small part because it requires imaging an inhomogenous distribution of defects and dopant atoms buried inside the host material. Here we demonstrate this, and address the long-standing question of whether the cluster nucleation is defect-mediated or spontaneous. Atomic-resolution illustrations are given for the chemically dissimilar cases of erbium and germanium implanted into silicon carbide. Whereas interstitial loops act as nucleation sites in both cases, the evolution of nanocrystals is strikingly different: Erbium is found to gather in lines, planes and finally three-dimensional precipitates, whereas germanium favours compact, three-dimensional structures.

Carbon Compounds, Inorganic↗

Ferromagnetism above room temperature in bulk and transparent thin films of Mn-doped ZnO.

The search for ferromagnetism above room temperature in dilute magnetic semiconductors has been intense in recent years. We report the first observations of ferromagnetism above room temperature for dilute (<4 at.%) Mn-doped ZnO. The Mn is found to carry an average magnetic moment of 0.16 mu(B) per ion. Our ab initio calculations find a valance state of Mn(2+) and that the magnetic moments are ordered ferromagnetically, consistent with the experimental findings. We have obtained room-temperature ferromagnetic ordering in bulk pellets, in transparent films 2-3 microm thick, and in the powder form of the same material. The unique feature of our sample preparation was the low-temperature processing. When standard high-temperature (T > 700 degrees C) methods were used, samples were found to exhibit clustering and were not ferromagnetic at room temperature. This capability to fabricate ferromagnetic Mn-doped ZnO semiconductors promises new spintronic devices as well as magneto-optic components.

Journal Article↗

Difficult tracheal intubation induced by maxillary distraction devices in craniosynostosis syndromes.

BACKGROUND: Difficult intubation occurred during anaesthesia for removal of maxillary distraction devices in five of seven children with syndromal craniosynostoses (four with Apert, two with Pfeiffer and one with Crouzon syndrome). METHODS: Intubation was assessed in terms of laryngeal view and an established intubation difficulty score and had been straightforward before device insertion. Difficulty was induced by trismus due to device insertion and by increased maxillary prominence. This was compounded by preexisting mandibular hypoplasia. Cephalometric analysis, with each child acting as their own control, demonstrated anterior displacement of the maxilla and increased maxillary vertical height, as well as increased protuberance of the maxillary incisors. RESULTS: All five difficult tracheal intubations were associated with preoperative Mallampati scores of 3 or 4 and the nine straightforward intubations with scores of 1 or 2. Maximal interincisor distance was less than the lower 95% confidence limit for age in all five children who were difficult to intubate at the time of device removal. No child had a failed intubation, but all had significantly increased intubation difficulty. CONCLUSIONS: In view of the risks of trauma, hypoxia and aspiration associated with difficult direct laryngoscopy, we recommend elective fibreoptic intubation at anaesthesia for removal of maxillary distraction osteogenesis devices in these children.

Adolescent↗

Electronic endoscopy of the gastrointestinal tract. Initial experience with a new type of endoscope that has no fiberoptic bundle for imaging.

We report on 31 endoscopic investigations using a new type of electronic endoscope which has no fiberoptic bundle for imaging. The device carries on its tip a light-sensitive CCD chip which functions as a TV camera. The signals received are transformed by a processor and displayed on a TV monitor. The upper digestive tract was inspected in 23 patients with the electronic endoscope and in the same session, a fiberoptic-endoscopic esophagogastroduodenoscopy was carried out on 16 of them. The information obtained using the 2 systems differed neither quantitatively nor qualitatively. Because of its external diameter, it was not possible to introduce the electronic endoscope past esophageal stenoses in 2 patients. Preliminary experience with 2 types of colonoscopes in 8 patients strongly indicates that these devices are equivalent, in respect of optical quality, to the fiberoptic endoscopes. The quality and color reproduction of the TV image of the upper and lower digestive tract are excellent. Electronic endoscopy offers great potential for consultation, training and documentation. The patients tolerated the procedure very well.

Adult↗

Structural basis of human erythrocyte glucose transporter function in proteoliposome vesicles: circular dichroism measurements.

The secondary structural compositions of the human erythrocyte glucose transporter in proteoliposome vesicles were assessed on the basis of circular dichroism (CD) spectra measured in the absence and in the presence of D-glucose or an inhibitor, cytochalasin B. We designed and used a scattered-light-collecting device, which corrects CD spectra for optical artifacts originating from light scattering. Relative contents of eight types of secondary structure were estimated by using basis spectra generated by the eigenvector method based on CD spectra of 15 proteins of known structure. Results indicate that the glucose transporter is composed of approximately 82% alpha-helices, 10% beta-turns, and 8% other random structure, with no beta-strands. In the presence of an excess of D-glucose, the alpha-helical content is reduced by more than 10% and there is a significant increase in the random structure content. Cytochalasin B does not appear to affect the secondary structural composition of the transporter to any significant degree.

Circular Dichroism↗

Contraction of neuronal branching volume: an anatomic correlate of Pavlovian conditioning.

Associative memory of the mollusc Hermissenda crassicornis, previously correlated with changes of specific K+ currents, protein phosphorylation, and increased synthesis of mRNA and specific proteins, is here shown to be accompanied by macroscopic alteration in the structure of a single identified neuron, the medial type B photoreceptor cell. Four to five days after training, terminal arborizations of B cells iontophoretically injected with Ni2+ ions and then treated with rubeanic acid were measured with charge-coupled device (CCD)-digitized pseudocolor images of optical sections under "blind" conditions. Boundary volumes enclosing medial-type B-cell arborizations from classically conditioned animals were unequivocally reduced compared with volumes for naive animals or those trained with unpaired stimuli. Branch volume magnitude was correlated with input resistance of the medial type B-cell soma. Such associative learning-induced structural changes may share function with "synapse elimination" described in developmental contexts.

Animals↗

A comparison of x-ray detectors for mouse CT imaging.

There is significant interest in using computed tomography (CT) for in vivo imaging applications in mouse models of disease. Most commercially available mouse x-ray CT scanners utilize a charge-coupled device (CCD) detector coupled via fibre optic taper to a phosphor screen. However, there has been little research to determine if this is the optimum detector for the specific task of in vivo mouse imaging. To investigate this issue, we have evaluated four detectors, including an amorphous selenium (a-Se) detector, an amorphous silicon (a-Si) detector with a gadolinium oxysulphide (GOS) screen, a CCD with a 3:1 fibre taper and a GOS screen, and a CCD with a 2:1 fibre taper and both GOS and thallium-doped caesium iodide (CsI:Tl) screens. The detectors were evaluated by measuring the modulation transfer function (MTF), noise power spectrum (NPS), detective quantum efficiency (DQE), stability over multiple exposures, and noise in reconstructed CT images. The a-Se detector had the best MTF and the highest DQE (0.6 at 0 lp mm(-1)) but had the worst stability (45% reduction after 2000 exposure frames). The a-Si detector and the CCD with the 3:1 fibre, both of which used the GOS screen, had very similar performance with a DQE of approximately 0.30 at 0 lp mm(-1). For the CCD with the 2:1 fibre, the CsI:Tl screen resulted in a nearly two-fold improvement in DQE over the GOS screen (0.4 versus 0.24 at 0 lp mm(-1)). The CCDs both had the best stability, with less than a 1% change in pixel values over multiple exposures. The pixel values of the a-Si detector increased 5% over multiple exposures due to the effects of image lag. Despite the higher DQE of the a-Se detector, the reconstructed CT images acquired with the a-Si detector had lower noise levels, likely due to the blurring effects from the phosphor screen.

Algorithms↗

A prospective analysis of diagnostic laparoscopy in trauma.

OBJECTIVE: This study was performed to assess current and potential future application for laparoscopy (DL) in the diagnosis of penetrating and blunt injuries. Efficacy, safety, and cost analyses were performed. SUMMARY BACKGROUND DATA: Diagnostic peritoneal lavage (DPL) and computed tomography (CT) have been the mainstays in recent years for diagnosis of equivocal nontherapeutic laparotomy, whereas CT is not helpful for the vast majority of penetrating wounds. DL may be a useful adjunct to fill in these gaps. METHODS: Hemodynamically stable patients with equivocal evidence of intraabdominal injury were prospectively entered into the protocol. DL was performed under general anesthesia; patients with wounds penetrating the peritoneum or blunt injury with significant organ injury underwent laparotomy. RESULTS: Over 19 months, 182 patients (55% stab, 36% GSW, 9% blunt) were studied. No peritoneal penetration was found at DL in 55% of penetrating wounds with 66% of the remainder having therapeutic laparotomy, 17% nontherapeutic laparotomy, and 17% negative laparotomy. Therapeutic laparotomy was performed in 53% of blunt injuries after DL. Tension pneumothorax occurred in one patient and one had an iatrogenic small bowel injury. Charges for DL were $3,325 per patient compared with $3,320 for a similar group undergoing negative laparotomy before this protocol. CONCLUSIONS: DL is a safe modality for trauma. With current technology, DL is most efficacious for evaluation of equivocal penetrating wounds. Significant cost savings would be gained by performance under local anesthesia. Development of miniaturized optics, bowel clamps, retractors, and stapling devices will reduce overall costs and permit some therapeutic applications for laparoscopy in trauma management.

Abdominal Injuries↗

Visualization, tracking, and navigation of instruments for magnetic resonance imaging-guided endovascular procedures.

Interventional procedures using magnetic resonance imaging (MRI) guidance have increased in interest during the last few years. Central to the success and safety of MRI-guided procedures is the accurate visualization of the interventional instruments relative to the surrounding anatomy. A variety of methodologies for visualizing and automatically tracking instruments, including needles, radiofrequency and laser ablation devices, endoscopes, catheters, and guidewires have been developed and introduced to help the interventionalist to safely guide the device toward the target region. This article describes and compares characteristics of the four most commonly used localization and tracking systems used for MRI-guided interventional procedures: those based on the susceptibility artifact of the device, those that intentionally create field inhomogeneity along the device, those that rely on an optical tracking system, and active tracking systems using micro receive coils.

Animals↗

A whole-body registration-free navigation system for image-guided surgery and interventional radiology.

RATIONALE AND OBJECTIVES: To develop and test an image-guided navigation system in which the base of reference is taken from the imaging modality, here, a helical CT scanner. METHODS: An optical digitizer together with a calibration device is used to measure the transformation matrix between the digitizer reference system and a CT reference system. During intervention, it tracks radiological and surgical tools with tool references. A specific software visually integrates the current tool position with the corresponding image information. In vitro accuracy tests were performed. RESULTS: With helical CT, freehand positioning accuracy was 1.9 +/- 1.1 mm (mean +/- SD) in vitro (n = 718). CONCLUSIONS: The navigation system developed by the authors appears to be feasible for radiological interventions as well as for minimally invasive surgery. It is not limited to a certain procedure, can be used in every region of the body, and is functional after imaging. Intraprocedural scans can be integrated immediately.

Equipment Design↗