Current trends in COVID-19 diagnosis and its new variants in physiological fluids: Surface antigens, antibodies, nucleic acids, and RNA sequencing.

Rapid, highly sensitive, and accurate virus circulation monitoring techniques are critical to limit the spread of the virus and reduce the social and economic burden. Therefore, point-of-use diagnostic devices have played a critical role in addressing the outbreak of COVID-19 (SARS-CoV-2) viruses. This review provides a comprehensive overview of the current techniques developed for the detection of SARS-CoV-2 in various body fluids (e.g., blood, urine, feces, saliva, tears, and semen) and considers the mutations (i.e., Alpha, Beta, Gamma, Delta, Omicron). We classify and comprehensively discuss the detection methods depending on the biomarker measured (i.e., surface antigen, antibody, and nucleic acid) and the measurement techniques such as lateral flow immunoassay (LFIA), enzyme-linked immunosorbent assay (ELISA), reverse transcriptase-polymerase chain reaction (RT-PCR), reverse transcription loop-mediated isothermal amplification (RT-LAMP), microarray analysis, clustered regularly interspaced short palindromic repeats (CRISPR) and biosensors. Finally, we addressed the challenges of rapidly identifying emerging variants, detecting the virus in the early stages of infection, the detection sensitivity, selectivity, and specificity, and commented on how these challenges can be overcome in the future.

Ferrocene-Containing DNA Monolayers: Influence of Electrostatics on the Electron Transfer Dynamics.

A 153-mer target DNA was amplified using ethynyl ferrocene dATP and a tailed forward primer resulting in a duplex with a single-stranded DNA tail for hybridization to a surface-tethered probe. A thiolated probe containing the sequence complementary to the tail as well as a 15 polythimine vertical spacer with a (CH2)6 spacer was immobilized on the surface of a gold electrode and hybridized to the ferrocene-modified complementary strand. Potential step chronoamperometry and cyclic voltammetry were used to probe the potential of zero charge, PZC, and the rate of heterogeneous electron transfer between the electrode and the immobilized ferrocene moieties. Chronoamperometry gives three, well-resolved exponential current-time decays corresponding to ferrocene centers located within 13 Å (4 bases) along the duplex. Significantly, the apparent standard heterogeneous electron transfer rate constant, kappo, observed depends on the initial potential, i.e., the rate of electron transfer at zero driving force is not the same for oxidation and reduction of the ferrocene labels. Moreover, the presence of ions, such as Sr2+, that strongly ion pair with the negatively charged DNA backbone modulates the electron transfer rate significantly. Specifically, kappo = 246 ± 23.5 and 14 ± 1.2 s-1 for reduction and oxidation, respectively, where the Sr2+ concentration is 10 mM, but the corresponding values in 1 M Sr2+ are 8 ± 0.8 and 150 ± 12 s-1. While other factors may be involved, these results are consistent with a model in which a low Sr2+ concentration and an initial potential that is negative of the PZC lead to electrostatic repulsion of the negatively charged DNA backbone and the negatively charged electrode. This leads to the DNA adopting an extended configuration (concertina open), resulting in a slow rate of heterogeneous electron transfer. In contrast, for ferrocene reduction, the initial potential is positive of PZC and the negatively charged DNA is electrostatically attracted to the electrode (concertina closed), giving a shorter electron transfer distance and a higher rate of heterogeneous electron transfer. When the Sr2+ concentration is high, the charge on the DNA backbone is compensated by the electrolyte and the charge on the electrode dominates the electron transfer dynamics and the opposite potential dependence is observed. These results open up the possibility of electromechanical switching using DNA superstructures.

Composition of bacterial and archaeal communities in the rumen of dromedary camel using cDNA-amplicon sequencing.

The camel is known to survive in harsh environmental conditions, due to its higher digestive efficiency of high-fiber diets compared with other ruminants. However, limited data are available on the microbial community in the rumen of a camel. In this study, the Illumina sequencing of V4 region of 16S rRNA genes based on RNA isolation was employed to get insight into the bacterial and archaeal communities associated with liquid and solid rumen fractions in eight camels under different feeding systems. Camels in group C1 were fed Egyptian clover hay plus concentrates mixture and camels of group C2 were fed fresh Egyptian clover. The results showed that liquid fraction has higher operational taxonomic units (OTUs) than solid fraction, and camel group C1 showed a higher microbial diversity than C2. The UniFrac analysis indicated that the microbial communities in camel groups are distinct. Moreover, phylum Firmicutes and Bacteroidetes dominated the bacterial community and Candidatus Methanomethylophilus dominated the archaeal community with a significant difference in the relative abundance between camel groups. Dominant bacterial genera were Prevotella, Fibrobacteres, Ruminococcus, and Butyrivibrio. There were many negative and positive correlations between and within bacterial and archaeal genera. The composition of microbial community in the rumen of a camel is similar to other ruminants with differences in the abundance.

Electrochemical detection of viruses and antibodies: A mini review.

Near patient detection of viral infection represents a powerful approach for the control of emerging threats to global health. Moreover, the ability to identify individuals who have contracted the disease and developed antibodies that confer immunity is central to a return to normal daily activities. This review presents some of the recent advances in electrochemical sensors for the detection of viruses and their associated antibody profiles. Given the speed, portability, sensitivity and selectivity achieved using electrochemical detection, these sensor systems hold the promise of transformative change in clinical practice.

Electrochemical Properties of Screen-Printed Carbon Nano-Onion Electrodes.

The properties of carbon nano-onions (CNOs) make them attractive electrode materials/additives for the development of low-cost, simple to use and highly sensitive Screen Printed Electrodes (SPEs). Here, we report the development of the first CNO-based ink for the fabrication of low-cost and disposable electrodes, leading to high-performance sensors. Achieving a true dispersion of CNOs is intrinsically challenging and a key aspect of the ink formulation. The screen-printing ink formulation is achieved by carefully selecting and optimising the conductive materials (graphite (GRT) and CNOs), the polymer binder, the organic solvent and the plasticiser. Our CNO/GRT-based screen-printed electrodes consist of an interconnected network of conducting carbon particles with a uniform distribution. Electrochemical studies show a heterogeneous electron transfer rate constant of 1.3 ± 0.7 × 10-3 cm·s-1 and a higher current density than the ferrocene/ferrocenium coupled to a commercial graphite SPEs. In addition, the CNO/GRT SPE can detect dopamine in the concentration range of 10.0-99.9 µM with a limit of detection of 0.92 µM (N = 3). They exhibit a higher analytical sensitivity than the commercial graphite-based SPE, with a 4-fold improvement observed. These results open up the possibility of using high-performing CNO-based SPEs for electrochemical applications including sensors, battery electrodes and electrocatalysis.

Community structure and fibrolytic activities of anaerobic rumen fungi in dromedary camels.

Anaerobic fungi colonize the rumen and degrade cellulose and hemicellulose, which enable them to be key players in the lignocellulose fermentation. Consequently, an expansion of knowledge about rumen fungi could increase animal productivity, utilization of lignified forages like alfalfa hay, and enhance fibrolytic enzymes production. Here, we used an Internal Transcribed Spacer 1 (ITS1) clone library to investigate the anaerobic rumen fungi in camel and to investigate their ability to produce cellulase and xylanase in vitro. Rumen fluid was collected from camels fed Egyptian clover (n = 14), and wheat straw (n = 7) and fecal samples were collected from camels fed wheat straw and concentrates (n = 5), or natural grazing plants (n = 10). Neocallimastix and Cyllamyces were the most abundant anaerobic fungi in all camel groups. An anaerobic rumen fungi media containing alfalfa hay as a carbon source was inoculated by rumen and fecal samples to assess the ability of anaerobic rumen fungi in camel gut to produce cellulase and xylanase. The anaerobic gut fungi in the camel is diverse and has cellulolytic and xylanolytic activities, fungal culture from rumen samples of camel fed wheat straw (R2) exhibited highest cellulase production. In addition, many of the sequences in the current study have no equivalent cultured representative, indicating a novel diversity within the camel gut.

Discovery and characterization of family 39 glycoside hydrolases from rumen anaerobic fungi with polyspecific activity on rare arabinosyl substrates.

Enzyme activities that improve digestion of recalcitrant plant cell wall polysaccharides may offer solutions for sustainable industries. To this end, anaerobic fungi in the rumen have been identified as a promising source of novel carbohydrate active enzymes (CAZymes) that modify plant cell wall polysaccharides and other complex glycans. Many CAZymes share insufficient sequence identity to characterized proteins from other microbial ecosystems to infer their function; thus presenting challenges to their identification. In this study, four rumen fungal genes (nf2152, nf2215, nf2523, and pr2455) were identified that encode family 39 glycoside hydrolases (GH39s), and have conserved structural features with GH51s. Two recombinant proteins, NF2152 and NF2523, were characterized using a variety of biochemical and structural techniques, and were determined to have distinct catalytic activities. NF2152 releases a single product, ß1,2-arabinobiose (Ara2) from sugar beet arabinan (SBA), and ß1,2-Ara2 and α-1,2-galactoarabinose (Gal-Ara) from rye arabinoxylan (RAX). NF2523 exclusively releases α-1,2-Gal-Ara from RAX, which represents the first description of a galacto-(α-1,2)-arabinosidase. Both ß-1,2-Ara2 and α-1,2-Gal-Ara are disaccharides not previously described within SBA and RAX. In this regard, the enzymes studied here may represent valuable new biocatalytic tools for investigating the structures of rare arabinosyl-containing glycans, and potentially for facilitating their modification in industrial applications.

Fibrinogen Motif Discriminates Platelet and Cell Capture in Peptide-Modified Gold Micropore Arrays.

Human blood platelets and SK-N-AS neuroblastoma cancer-cell capture at spontaneously adsorbed monolayers of fibrinogen-binding motifs, GRGDS (generic integrin adhesion), HHLGGAKQAGDV (exclusive to platelet integrin αIIbß3), or octanethiol (adhesion inhibitor) at planar gold and ordered 1.6 µm diameter spherical cap gold cavity arrays were compared. In all cases, arginine/glycine/aspartic acid (RGD) promoted capture, whereas alkanethiol monolayers inhibited adhesion. Conversely only platelets adhered to alanine/glycine/aspartic acid (AGD)-modified surfaces, indicating that the AGD motif is recognized preferentially by the platelet-specific integrin, αIIbß3. Microstructuring of the surface effectively eliminated nonspecific platelet/cell adsorption and dramatically enhanced capture compared to RGD/AGD-modified planar surfaces. In all cases, adhesion was reversible. Platelets and cells underwent morphological change on capture, the extent of which depended on the topography of the underlying substrate. This work demonstrates that both the nature of the modified interface and its underlying topography influence the capture of cancer cells and platelets. These insights may be useful in developing cell-based cancer diagnostics as well as in identifying strategies for the disruption of platelet cloaks around circulating tumor cells.

Wireless Electrochemiluminescence at Nafion-Carbon Microparticle Composite Films.

Thin films of a composite of nafion and carbon microparticles have been deposited on nonconducting substrates and their conductivity as well as their ability to generate electrochemiluminescence investigated. The films exhibit very low conductivity (<6 × 103 S m-1) for low particle loadings, but once the percolation threshold is reached (volume percentage of 71 ± 8% carbon particles), the conductivity increases dramatically and a maximum conductivity of 2.0 ± 0.1 × 107 S m-1 is achieved. The electrochemical properties of the composites, including heterogeneous electron transfer rates, were probed using cyclic voltammetry. Significantly, bipolar, or wireless, electrochemiluminescence can be generated with films that contain >65% (by volume) carbon particles using [Ru(bpy)3]2+ as the luminophore and tripropylamine as the coreactant, at an electric field of 14 V cm-1. Under these conditions, the complete film is sufficiently conducting to become polarized in the external electric field and the electrochemiluminescence intensity correlates strongly with the film conductivity. These results demonstrate the usefulness of particle arrays for the wireless generation of electrochemiluminescence at relatively low electric field strengths.

Evaluating Metabolite-Related DNA Oxidation and Adduct Damage from Aryl Amines Using a Microfluidic ECL Array.

Damage to DNA from the metabolites of drugs and pollutants constitutes a major human toxicity pathway known as genotoxicity. Metabolites can react with metal ions and NADPH to oxidize DNA or participate in SN2 reactions to form covalently linked adducts with DNA bases. Guanines are the main DNA oxidation sites, and 8-oxo-7,8-dihydro-2-deoxyguanosine (8-oxodG) is the initial product. Here we describe a novel electrochemiluminescent (ECL) microwell array that produces metabolites from test compounds and measures relative rates of DNA oxidation and DNA adduct damage. In this new array, films of DNA, metabolic enzymes, and an ECL metallopolymer or complex assembled in microwells on a pyrolytic graphite wafer are housed in dual microfluidic chambers. As reactant solution passes over the wells, metabolites form and can react with DNA in the films to form DNA adducts. These adducts are detected by ECL from a RuPVP polymer that uses DNA as a coreactant. Aryl amines also combine with Cu2+ and NADPH to form reactive oxygen species (ROS) that oxidize DNA. The resulting 8-oxodG was detected selectively by ECL-generating bis(2,2'-bipyridine)-(4-(1,10-phenanthrolin-6-yl)-benzoic acid)Os(II). DNA/enzyme films on magnetic beads were oxidized similarly, and 8-oxodG determined by LC/MS/MS enabled array standardization. The array limit of detection for oxidation was 720 8-oxodG per 106 nucleobases. For a series of aryl amines, metabolite-generated DNA oxidation and adduct formation turnover rates from the array correlated very well with rodent 1/TD50 and Comet assay results.

Hemispherical platinum : silver core : shell nanoparticles for miRNA detection.

Defects within a self-assembled monolayer (SAM) of dodecanethiol on gold have been used as nucleation sites for the electrodeposition of mushroom shaped platinum nanoparticles (PtNPs). The top surfaces of these PtNPs were then decorated with a layer of silver creating a hemispherical - platinum : silver core : shell nanoparticle (Pt-AgNP). Thiolated probe strand miRNA was then immobilised onto the upper silver surface. These regioselectively modified particles were desorbed by applying a current jump to yield nanoparticles capable of hybridising to a complementary miRNA target with electrocatalysis occurring on the non-functionalized lower surface. A second electrode was functionalized with single stranded capture miRNA that has a sequence that is complementary to an miRNA, miR-132, associated with the childhood cancer, Neuroblastoma but leaves a section of the target available to bind the nucleic acid sequence on the core : shell Pt-AgNPs. Following hybridization of the target and capture strands the surface was exposed to the miRNA labelled electrocatalytic Pt-AgNPs. The concentration of the target was then determined by monitoring the current associated with the reduction of hydrogen peroxide in a solution of H2SO4. Calibration plots of the log[miRNA] vs. faradaic current were linear from 1 aM to 1 µM and aM concentrations could be detected without the need for chemical amplification of the target, e.g., using PCR or NASBA. The regioselectively modified particles were also immobilised within the interior of gold microcavity arrays via miRNA hybridisation and their Raman properties investigated.

Detection and Live-Cell Imaging of a Micro-RNA Associated with the Cancer Neuroblastoma.

The ability of a molecular beacon to detect miR-132, a microRNA associated with the childhood cancer neuroblastoma, is reported in solution and within live cells. The stem-loop structure comprises a sequence complementary to miR-132, modified with a 6-FAM dye and dabcyl quencher on either end. In the absence of the target, self-binding occurs bringing the luminophore and quencher into close proximity, significantly decreasing the emission intensity. In the presence of miR-132, the signal is greatly enhanced, with a linear increase in intensity for mole ratios of beacon-to-target between 0.25 and 2.00. The structure differentiates between target and mismatched nucleic acid sequences, e.g., in the presence of a single-base mismatch, no increase in emission intensity beyond the background is observed. The stem-loop can be introduced into neuroblastoma cancer cells by electroporation, allowing miR-132 to be imaged within live cells. miR-132 appears to be localized within the nucleus of the cells, where its concentration is of the order of 1 µM. Significantly, transfection of the cells with a miR-132 mimic causes the emission intensity to more than double, demonstrating the sensitivity of the approach to changes in miR-132 concentration in live cells. This behavior opens up significant theranostic applications, such as the possibility of rapidly identifying retinoic acid resistant patients as well as providing a means to monitor therapeutic efficacy.

Aqueous-filled polymer microcavity arrays: versatile & stable lipid bilayer platforms offering high lateral mobility to incorporated membrane proteins.

A key prerequisite in an ideal supported lipid bilayer based cell membrane model is that the mobility of both the lipid matrix and its components are unhindered by the underlying support. This is not trivial and with the exception of liposomes, many of even the most advanced approaches, although accomplishing lipid mobility, fail to achieve complete mobility of incorporated membrane proteins. This is addressed in a novel platform comprising lipid bilayers assembled over buffer-filled, arrays of spherical cap microcavities formed from microsphere template polydimethoxysilane. Prior to bilayer assembly the PDMS is rendered hydrophilic by plasma treatment and the lipid bilayer prepared using Langmuir Blodgett assembly followed by liposome/proteoliposome fusion. Fluorescence Lifetime Correlation Spectroscopy confirmed the pore suspended lipid bilayer exhibits diffusion coefficients comparable to free-standing vesicles in solution. The bilayer modified arrays are highly reproducible and stable over days. As the bilayers are suspended over deep aqueous reservoirs, reconstituted membrane proteins experience an aqueous interface at both membrane interfaces and attain full lateral mobility. Their utility as membrane protein platforms was exemplified in two case studies with proteins of different dimensions in their extracellular and cytoplasmic domains reconstituted into DOPC lipid bilayers; Glycophorin A, and Integrin αIIbß3. In both cases, the proteins exhibited 100% mobility with high lateral diffusion coefficients.

Peptide-bridged dinuclear Ru(II) complex for mitochondrial targeted monitoring of dynamic changes to oxygen concentration and ROS generation in live mammalian cells.

A novel mitochondrial localizing ruthenium(II) peptide conjugate capable of monitoring dynamic changes in local O2 concentrations within living cells is presented. The complex is comprised of luminescent dinuclear ruthenium(II) polypyridyl complex bridged across a single mitochondrial penetrating peptide, FrFKFrFK-CONH2 (r = D-arginine). The membrane permeability and selective uptake of the peptide conjugate at the mitochondria of mammalian cells was demonstrated using confocal microscopy. Dye co-localization studies confirmed very precise localization and preconcentration of the probe at the mitochondria. This precision permitted collection of luminescent lifetime images of the probe, without the need for co-localizing dye and permitted semiquantitative determination of oxygen concentration at the mitochondria using calibration curves collected at 37 °C for the peptide conjugate in PBS buffer. Using Antimycin A the ability of the probe to respond dynamically to changing O2 concentrations within live HeLa cells was demonstrated. Furthermore, based on lifetime data it was evident that the probe also responds to elevated reactive oxygen species (ROS) levels within the mitochondria, where the greater quenching capacity of these species led to luminescent lifetimes of the probe at longer Antimycin A incubation times which lay outside of the O2 concentration range. Although both the dinuclear complex and a mononuclear analogue conjugated to an octaarginine peptide sequence exhibited some cytotoxicity over 24 h, cells were tolerant of the probes over periods of 4 to 6 h which facilitated imaging. These metal-peptide conjugated probes offer a valuable opportunity for following dynamic changes to mitochondrial function which should be of use across domains in which the metabolic activity of live cells are of interest from molecular biology and drug discovery.

RGD labeled Ru(II) polypyridyl conjugates for platelet integrin αIIbß3 recognition and as reporters of integrin conformation.

The ability of two novel ruthenium(II) polypyridyl-Arg-Gly-Asp (RGD) peptide conjugates to act as molecular probes for reporting on the presence and conformation of integrin αIIbß3 in solution and in live cells was described. The compounds are [Ru(bpy)2PIC-RGD](2+), bpy-RGD, and [Ru(dpp)2PIC-RGD](2+), dpp-RGD, where dpp is 4,7-diphenyl-1,10-phenanthroline, bpy is 2,2'-bipyridine, and PIC is 2-(4-carboxyphenyl)imidazo[4,5-f][1,10]phenanthroline. Bpy-RGD is hydrophilic, whereas dpp-RGD is comparatively hydrophobic. Both probes exhibited good affinity and high specificity for purified αIIbß3 in solution. Binding of either complex to the resting integrin resulted in an approximately 8-fold increase of emission intensity from the metal center with dissociation constants (Kd) in the micromolar range for each complex. The Kd for each conjugate/αIIbß3 assembly were compared following treatment of the integrin with the activating agents, Mn(2+) and dithiothreitol (DTT), which are commonly used to induce active-like conformational changes in the integrin. For bpy-RGD/αIIbß3 Kd showed relatively little variation with integrin activation, presenting the following trend: denatured αIIbß3 > resting αIIbß3 = pretreated DTT = pretreated Mn(2+). Kd for dpp-RGD/ αIIbß3 showed greater variation with integrin activation and the following trend was followed: denatured αIIbß3 > resting αIIbß3 > pretreated Mn(2+) = pretreated DTT. Time resolved luminescence anisotropy was carried out to obtain the rotational correlation time of bpy-RGD and dpp-RGD bound to resting or nominally activated integrin. The rotational correlation times of bpy-RGD and dpp-RGD, too fast to measure unbound, decreased to 1.50 ± 0.03 µs and 2.58 ± 0.04 µs, respectively, when the complexes were bound to resting integrin. Addition of Mn(2+) to bpy-RGD/αIIbß3 or dpp-RGD/αIIbß3 reduced the rotational correlation time of the ruthenium center to 1.29 ± 0.03 µs and to 1.72 ± 0.03 µs, respectively. Following treatment, the rotational correlation time decreased to 1.04 ± 0.01 µs and 1.29 ± 0.03 µs for bpy-RGD/αIIbß3, and dpp-RGD/αIIbß3, respectively. The large relative changes in rotational correlation times observed for Mn(2+) or DTT activated integrin indicates significant change in protein conformation compared with the resting integrin. The results also indicated that the metal complex itself affects the final conformational and/or aggregation status of the protein obtained. Furthermore, the extent of conformational change was influenced by whether the probe was bound to the integrin before or after activator treatment. Finally, in vitro studies indicated that both probes selectively bind to CHO cells expressing the resting form of αIIbß3. In each case the probe colocalized with αIIb specific SZ22 antibody. Overall, this work indicates that bpy-RGD and dpp-RGD may be useful peptide-probes for rapid assessment of integrin structural status and localization in solution and cells.

Electron transfer to covalently immobilized Keggin polyoxotungstates on gold.

Spontaneously adsorbed monolayers have been formed on gold electrodes using a Keggin polyoxotungstate with covalently attached alkanethiol linkers of two different lengths. Films of both polyoxotungstates show two well-defined reduction processes associated with the polyoxotungstate centers where the ionic liquid, [BMIM][BF4], acts as supporting electrolyte. The surface coverages are both less than that expected for a close-packed monolayer. For the short and long linkers, the voltammetric response can be described in terms of the Butler-Volmer response involving a surface confined species using standard heterogeneous electron transfer rate constants of 170 and 140 s(-1) for the first reduction and 150 and 100 s(-1) for the second reduction processes, respectively. The rate of electron transfer to a solution phase redox probe, ferrocyanide, is significantly more sensitive to the length of the linker than the rate of electron transfer to the tungstate centers. This behavior probably arises due to potential-induced changes in the film structure.

Electrodeposited gold-copper core-shell nanowires for high sensitivity DNA detection.

Gold-Copper core shell nanowires have been electrodeposited and their electrochemical and Raman properties probed. First, hollow copper nanotubes, 3.2 ± 0.1 µm long, with a uniform diameter of 70 ± 22 nm, were electrodeposited within the pores of a track etched polycarbonate membrane filter. Second, gold was then electrodeposited within these copper cylinders to yield the gold-copper core-shell nanowires. Nanowires, functionalised with probe strand DNA, that is complementary to that of the pathogen Staph. Aureus, only on their ends, can be immobilised onto an electrode surface in a DNA sandwich assay. Significantly, the charge associated with the selective oxidation of the copper shell depends linearly on the target DNA concentration from 1 nM to 100 µM.

Label-free impedance detection of cancer cells.

Ovarian cancer cells, SKOV3, have been immobilized onto platinum microelectrodes using anti-EPCAM capture antibodies and detected with high sensitivity using electrochemical impedance. The change in impedance following cell capture is strongly dependent on the supporting electrolyte concentration. By controlling the concentration of Dulbecco's phosphate buffered saline (DPBS) electrolyte, the double layer thickness can be manipulated so that the interfacial electric field interacts with the bound cells, rather than simply decaying across the antibody capture layer. Significantly, the impedance changes markedly upon cell capture over the frequency range from 3 Hz to 90 kHz. For example, using an alternating-current (ac) amplitude of 25 mV, a frequency of 81.3 kHz, and an open circuit potential (OCP) as the direct-current (dc) voltage, a detection limit of 4 captured cells was achieved. Assuming an average cell radius of 5 µm, the linear dynamic range is from 4 captured cells to 650 ± 2 captured cells, which is approximately equivalent to fractional coverages from 0.1% to 29%. An equivalent circuit that models the impedance response of the cell capture is discussed.

Insights into electrochemiluminescent enhancement through electrode surface modification.

The electrochemiluminescent (ECL) properties of a luminescent metal centre, [Ru(bpy)(3)](2+), can be significantly modulated through its electronic interaction with neighbouring centres and the polymer backbone used to confine it on an electrode surface. From the perspective of ECL based sensing devices, an increase in the ECL efficiency of a metallopolymer film can result in enhanced sensor sensitivity and selectivity. This work probes the ECL properties of both conjugated, [Ru(bpy)(2)(PPyBBIM)(10)](2+), and non-conjugated, [Ru(bpy)(2)(PVP)(10)](2+), ruthenium based metallopolymer films based on a well documented reaction with sodium oxalate, where bpy is 2,2'-bipyridyl, PPYBBIM is poly[2-(2-pyridyl)-bibenzimidazole] and PVP is poly(4-vinylpyridine). Through a combination of ground state electrochemical studies and ECL measurements, the ECL efficiency for each film is determined. This study reveals that despite a dramatic influence in charge transfer rates between metal centres, as observed for the conducting polymer, mediated through the conducting polymer backbone, a corresponding increase in ECL efficiency is not always observed. The degree of communication between the adjacent excited state metal centres are an important consideration for ECL enhancement however self quenching, luminophore distribution and film porosity must also be considered.

Detection of sub-femtomolar DNA based on double potential electrodeposition of electrocatalytic platinum nanoparticles.

Suspensions of electrocatalytic platinum nanoparticles with radii as small as 78.9 ± 3.5 nm that are functionalised with DNA only in one region have been created using templated electrodeposition. The integrity of the bound DNA following nanoparticle desorption from the electrode is demonstrated by detecting attomolar concentrations of DNA without the need for molecular, e.g., PCR or NASBA, amplification. Double potential step approaches coupled with interface engineering via nucleation sites allows PtNPs to be created with controlled particle size and density in a facile and reproducible manner.