Analysis of the photodecomposition products of Ru(bpy)3(2+) in various buffers and upon zeolite encapsulation

The photodecomposition products of Ru(bpy)3(2+) in water, in aqueous buffered solutions and encapsulated in zeolite-Y have been analyzed by chromatography and UV-visible spectroscopy. The chromatographic method is found to be capable of separating species with the same charge but slightly different ligands as well as geometrical isomers. In all the systems investigated, photodecomposition proceeded via photoaquation resulting in the formation of cis- and trans-Ru(bpy)2(OH2)2(2+). In the case of acetate and phthalate buffers, a third species, Ru(bpy)2(L)(OH2)+, where L is the buffer anion, was found to be the dominant product. For a given pH, the extent of decomposition was found to be dependent on both the buffer anion, following the trend, phosphate < acetate < phthalate and buffer concentration. The presence of the electron-transfer quenching agent, N,N'-dimethyl-4,4'-bipyridinium ion in the medium led to a decrease of the photodecomposition and closely followed the quenching efficiency as measured by intensity and lifetime quenching studies. Encapsulation of Ru(bpy)3(2+) in the supercages of zeolite-Y did not lead to a substantial decrease in photodecomposition as compared to an aqueous solution, suggesting that the expected enhanced stability of Ru(bpy)3(2+*) due to the destabilization of 3dd orbitals and the cage effect was being negated by the close proximity and intrazeolite packing of H2O molecules around the Ru center.     

Determination of Dansyl Amino Acids and Oxalate by HPLC with Electrogenerated Chemiluminescence Detection Using Tris(2,2'-bipyridyl)ruthenium(II) in the Mobile Phase

A new electrogenerated chemiluminescence detection method is investigated for use in detection in reversed-phase and reversed-phase ion-pair HPLC with Ru(bpy)(3)(2+) in the mobile phase. In this method, different concentrations of Ru(bpy)(3)(2+) are dissolved in the mobile phase and the HPLC column flushed with the mobile phase for 1 h until the column is saturated with Ru(bpy)(3)(2+). The separated analytes along with Ru(bpy)(3)(2+) pass through an optical-electrochemical flow cell which has a dual platinum electrode held at a potential of 1250 mV vs a Ag/AgCl reference electrode. On the surface of the electrode, Ru(bpy)(3)(2+) is oxidized to Ru(bpy)(3)(3+) which reacts with the analytes to emit light. The retention times, retention orders, detection limits, and linearity in working curves are compared to those obtained with the conventional postcolumn Ru(bpy)(3)(2+) addition method. The retention times for dansyl amino acids with Ru(bpy)(3)(2+) in the mobile phase are longer than those obtained with the postcolumn addition approach. This may be caused by π-to-π interactions between the aromatic groups of the dansyl derivatives and the bipyridyl groups of Ru(bpy)(3)(2+) in the Ru(bpy)(3)(2+)-saturated reversed-phase column. Similarly, oxalate is separated from urine and blood plasma samples by reversed-phase ion-pair HPLC. Plasma samples are obtained using ultrafiltration to remove proteins from whole blood. Retention times for oxalate with the two detection techniques are identical, and detection limits for these techniques are compared.     

Electrogenerated chemiluminescence from R(bpy)3(2+) ion-exchanged in carbon nanotube/perfluorosulfonated ionomer composite films

The electrochemistry and electrogenerated chemiluminescence (ECL) of ruthenium(II) tris(bipyridine) (Ru(bpy)(3)(2+)) ion-exchanged in carbon nanotube (CNT)/Nafion composite films were investigated with tripropylamine (TPA) as a coreactant at a glassy carbon (GC) electrode. The major goal of this work was to investigate and develop new materials and immobilization approaches for the fabrication of ECL-based sensors with improved sensitivity, reactivity, and long-term stability. Ru(bpy)(3)(2+) could be strongly incorporated into Nafion film, but the rate of charge transfer was relative slow and its stability was also problematic. The interfusion of CNT in Nafion resulted in a high peak current of Ru(bpy)(3)(2+) and high ECL intensity. The results indicated that the composite film had more open structures and a larger surface area allowing faster diffusion of Ru(bpy)(3)(2+) and that the CNT could adsorb Ru(bpy)(3)(2+) and also acted as conducting pathways to connect Ru(bpy)(3)(2+) sites to the electrode. In the present work, the sensitivity of the ECL system at the CNT/Nafion film-modified electrodes was more than 2 orders of magnitude higher than that observed at a silica/Nafion composite film-modified electrode and 3 orders of magnitude higher than that at pure Nafion films. The CNT/Nafion composite film-modified GC electrodes also exhibited long-term stability.     

Synthesis and properties of the diruthenium(II) complexes with diimine-linked polypyridine bridges

The diruthenium complexes, [(bpy)2Ru(II)-(bpy-DPDA)-Ru(II)(bpy)2][PF6]4 3, (bpy: 2,2'-bipyridiyl; bpy-DPDA: Bis(2,2'-dipyridylketenylidene)-N,N-1,4-phenylenediamine}, and [(bpy)2Ru(II)-(Dbpy-DPDA)-Ru(II)(bpy)2][PF6]4 4 {Dbpy-DPDA: Bis(2,2'-dipyridyl ketenylidene)-N,N-1,1'-(4,4'-diphenylene)diamine}, were prepared by the reaction of (bpy)2Ru(II)Cl2 with diimine-linked polypyridine bridges. These complexes were characterized by NMR, IR, UV/VIS, PL and cyclic voltammetry. In the 13C-NMR spectra of 3 and 4, the carbon peaks of the Schiff Bases (>C=N-) were shifted to lower fields, and emissions were observed at 689 and 693 nm with quantum yields of 0.004 and 0.006, respectively. The electrochemistry of complexes 3 and 4 showed four-reversible waves (one oxidation wave and three reduction waves). The electrochemically measured band gaps for complexes 3 and 4 were 2.01 and 2.10 eV, respectively.     

Electrochemiluminescence of Tris(2,2'-bipyridine)ruthenium in Water at Carbon Microelectrodes

Electrochemiluminescence (ECL) of Ru(bpy)(3)(2+) in water only, without any added electrolyte or reducing agents, has been obtained at carbon interdigitated microelectrode arrays (C-IDAs) of 2 μm width and spacing. In a generation/collection biasing mode, ECL can be clearly seen with the naked eye in normal room lighting at concentrations greater than 1 mM. Using a conventional photomultiplier tube (PMT), a detection limit of 10(-)(7) M Ru(bpy)(3)(2+) has been achieved for an electrode area of 0.25 mm(2). In comparison, the ECL intensity produced at Pt-IDA of the same geometry, under identical experimental conditions, was more than 300 times less. The ECL obtained at C-IDAs is attributed to the annihilation reaction of the reduced and oxidized forms of the Ru(bpy)(3)(2+) made possible due to the small electrode spacing.     

Enhanced electrogenerated chemiluminescence in the presence of fluorinated alcohols

The electrochemistry, UV-vis absorption, photoluminescence (PL), and coreactant electrogenerated chemiluminescence (ECL) of Ru(bpy)3(2+) (where bpy=2,2'-bipyridine) have been obtained in a series of hydroxylic solvents. The solvents included fluorinated and nonfluorinated alcohols and alcohol/water mixtures. Tri-n-propylamine was used as the oxidative-reductive ECL coreactant. Blue shifts of up to 30 nm in PL emission wavelength maximums are observed compared to a Ru(bpy)3(2+)/H2O standard due to interactions of the polar excited state (i.e., *Ru(bpy)3(2+)) with the solvent media. For example, Ru(bpy)3(2+) in water has an emission maximum of 599 nm while in the more polar hexafluoropropanol and trifluoroethanol it is 562 and 571 nm, respectively. ECL spectra are similar to PL spectra, indicating the same excited state is formed in both experiments. The difference between the electrochemically reversible oxidation (Ru(bpy)3(2+/3+)) and first reduction (Ru(bpy)2(2+/1+)) correlates well with the energy gap observed in the luminescence experiments. Although the ECL is linear in all solvents with [Ru(bpy)3(2+)] ranging from 100 to 0.1 nm, little correlation between the polarity of the solvent and the ECL efficiency (phiecl=number of photons per redox event) was observed. However, dramatic increases in phiecl ranging from 6- to 270-fold were seen in mixed alcohol/water solutions.     

The effects of nonionic surfactants on the tris(2,2'-bipyridyl)ruthenium(II)--tripropylamine electrochemiluminescence system

The electrochemistry and electrogenerated chemiluminescence (ECL) of Ru(bpy)3(2+) (bpy = 2,2'-bipyridyl) were studied in the presence of the nonionic surfactants Triton X-100, Thesit, and Nonidet P40. The anodic oxidation of Ru(bpy)3(2+) produces ECL in the presence of tri-n-propylamine in both aqueous and surfactant solutions. Increases in both ECL efficiency (> or =8-fold) and duration of the ECL signal were observed in surfactant media. A shift to lower energies of the Ru(bpy)3(2+) ECL emission by approximately 8 nm was also observed. The one-electron oxidation of Ru(bpy)3(2+) to Ru(bpy)3(3t) occurs at + 1.03 V vs Ag/AgCl in aqueous buffered (0.2 M potassium phosphate) solution as found by square wave voltammetry. This potential did not shift in surfactant systems, indicating that the redshifts in ECL emission are due to stabilization of ligand pi* orbitals in the metal-to-ligand charge-transfer excited state. These results are consistent with hydrophobic interactions between Ru(bpy)3(2+) and the nonionic surfactants.     

Scanning electrochemical microscopy. 56. Probing outside and inside single giant liposomes containing Ru(bpy)32+

Giant liposomes containing Ru(bpy)(3)(2+) (bpy = 2,2'-bipyridine) were prepared as model systems for biomembranes and cells and studied by scanning electrochemical microscopy (SECM). Conical carbon fiber tips of submicrometer size were used to approach, image, and puncture individual liposomes immobilized on glass substrates. SECM images of the liposomes were obtained, and the leakage of Ru(bpy)(3)(2+) through the lipid membrane was probed. The tip was also pushed into liposomes and characteristic breakthrough transients, corresponding to liposomes with different compartmental configurations, were obtained. Voltammograms were obtained with the tip inside a single liposome after breaking through the membrane, and the influx of mediator and efflux of encapsulant after puncture could be observed.     

Surfactant chain length effects on the light emission of tris(2,2'-bipyridyl)ruthenium(II)/ tripropylamine electrogenerated chemiluminescence

The effects of nonionic surfactant chain length on the properties of tris(2,2'-bipyridyl)ruthenium(II) (Ru(bpy)3(2+) where bpy = 2,2'-bipyridine) electrochemiluminescence (ECL) have been investigated. The electrochemistry, photophysics, and ECL of Ru(bpy)3(2+) in the presence of a series of nonionic surfactants are reported (Triton X-100, 114, 165, 405, 305, and 705-70). These surfactants differ in the number of poly(ethylene oxide) units incorporated into the surfactant molecule. The anodic oxidation of Ru(bpy)3(2+) produces ECL in the presence of tri-n-propylamine (TPrA) in aqueous surfactant solution. Increases in ECL efficiency (> or = 5-fold) and TPrA oxidation current (> or = 2-fold) have been observed in surfactant media. Slight decreases in ECL intensity are observed as the chain length of the nonionic surfactant increases. The data supports adsorption of surfactant on the electrode surface, thus facilitating TPrA and Ru(bpy)3(2+) oxidation and leading to higher ECL efficiencies.     

Chromatographic detection of nitroaromatic and nitramine compounds by electrochemical reduction combined with photoluminescence following electron transfer

The oxidizing agent tris(bipyridyl)ruthenium(III), or Ru-(bpy)(3)3+, is used as a postcolumn reagent for the detection of nitroaromatic and nitramine explosive compounds. After separation, the explosives are reduced electrochemically to oxidizable products such as hydroxlamines and nitrosamines, and these products react readily with Ru-(bpy)(3)3+ and Ru(bpy)(3)2+. The photoluminescence from the latter is used for detection. A porous carbon electrode was used for on-line analyte reduction following chromatography. Another porous carbon electrode was used to generate the nonluminescent Ru(bpy)(3)3+ from Ru(bpy)(3)3+ on-line at high efficiency. The two streams were combined, and the Ru(bpy)(3)2+ produced by oxidation of the reduced analytes was detected by laser illumination and light detection. Reductive hydrodynamic voltammograms of nitrobenzene, 2,4,6-trinitrotoluene, and hexahydro-1,3,5-trinitro-1,3,5-triazine indicated that a potential of - 1500 mV vs Ag/AgCl was sufficient to achieve a maximum signal from the reduced analytes. HPLC with a water/acetonitrile gradient on a C-18 reversed-phase column was then used to determine these three compounds plus the four additional examples, 1,3,5,7-tetrazocine, 2,4-dinitrotoluene; 2,6-dinitrotoluene, and 4-nitrotoluene. For both hydrodynamic voltammetry and HPLC detection, the photoluminescence following electron-transfer signal was calibrated using the one-electron standards ferrocene and ferrocenecarboxylic acid. Detection limits were in the low-nanomolar range for 20-microL injections of nonpreconcentrated nitro compounds.     

Multilabeling biomolecules at a single site. 1. Synthesis and characterization of a dendritic label for electrochemiluminescence assays

Ultrasensitive bioanalytical assays are of great value for early detection of human diseases and pathogens. The sensitivities of immunoassays and DNA probing can be enhanced by multilabeling the biorecognition partner used for affinity-based assays. However, the bioreactivity of biomolecules is affected by a high degree of multilabeling at multiple functional sites. It is proposed that dendritic scaffoldings be used to link multiple signal-generating units to a single site with potentially minimum impact on the bioaffinity. A prototype label, a zeroth-generation dendron, bearing three [Ru(bpy)(3)](2+) units for electrochemiluminescence (ECL) assays was synthesized and characterized preliminarily by spectroscopic, electrochemical, and ECL methods. No evidence of interaction between the neighboring [Ru(bpy)(3)](2+) units in the label molecule was found from these characterizations. Both the photoluminescence and ECL of the prototype label have features very similar to those of mononuclear [Ru(bpy)(3)](2+) compounds. Labeling a model protein with a triad of [Ru(bpy)(3)](2+) at one NH(2) position was demonstrated. The results reported here provide support to applying the proposed multilabeling strategy to affinity-based bioanalytical assays.     

Method for effective immobilization of Ru(bpy)(3)2+ on an electrode surface for solid-state electrochemiluminescene detection

A novel method for effective immobilization of Ru(bpy)3(2+) on an electrode surface is developed. The whole process involves two steps: the electrostatic interactions between citrate-capped gold nanoparticles (AuNPs) and Ru(bpy)3Cl2 in aqueous medium were used to fabricate Ru(bpy)(3)2+-AuNP aggregates (Ru-AuNPs) first, and then the Au-S interactions between as-formed Ru-AuNPs and sulfhydryl groups were used to effectively immobilize the Ru-AuNPs on a sulfhydryl-derivated indium tin oxide (ITO) electrode surface. As-prepared ITO electrode shows excellent stability, and the ECL active species Ru(bpy)3(2+) contained therein exhibit excellent ECL behaviors.     

Electrochemiluminescent detection of metal cations using a ruthenium(II) bipyridyl complex containing a crown ether moiety

The effects of metal ions on the electrochemiluminescence (ECL) properties of (bpy)2Ru(AZA-bpy) (bpy = 2,2'-bipyridine; AZA-bpy = 4-(N-aza-18-crown-6-methyl-2,2'-bipyridine) have been investigated. The electrochemistry, photophysics and ECL of Ru(bpy)3(2+) in the presence of Pb2+, Hg2+, Cu2+, and K+ are reported. The anodic oxidation of Ru(bpy)3(2+) produces ECL in the presence of tri-n-propylamine (TPrA) in 50:50 (v/v) CH3CN:H2O solution. Increases in ECL efficiency (photons generated per redox event) up to 20-fold that depend on both the concentration and nature of the metal ion have been observed, making this an interesting system for electrochemiluminescence metal ion sensing.     

Spectroelectrochemical sensing based on multimode selectivity simultaneously achievable in a single device. 16. Sensing by fluorescence

A fluorescence spectroelectrochemical sensor capable of detecting very low concentrations of metal complexes is described. The sensor is based on a novel spectroelectrochemical sensor that incorporates multiple internal reflection spectroscopy at an optically transparent electrode (OTE) coated with a selective film to enhance detection limits by preconcentrating the analyte at the OTE surface. Nafion was used as the selective cation exchange film for detecting Ru(bpy)(3)(2+), the model analyte, which fluoresces at 605 nm when excited with a 441.6-nm HeCd laser. The unoptimized linear dynamic range of the sensor for Ru(bpy)(3)(2+) is between 1 x 10(-)(11) and 1 x 10(-)(7) M with a calculated 2 x 10(-)(13) M detection limit. The sensor employs extremely thin films ( approximately 12 nm) without significantly sacrificing its sensitivity. The sensor response is demonstrated with varying film thicknesses. A state-of-the-art flow cell design allows variable cell volumes as low as approximately 4 microL. Fluorescence of the sample can be controlled by electromodulation between 0.7 and 1.3 V. Sensor operation is not reversible for the chosen model film (Nafion) and sample (Ru(bpy)(3)(2+)) but it can be regenerated with ethanol for multiple uses.     

Electrochemiluminescent metallopolymer-nanoparticle composites: nanoparticle size effects

Metallopolymer-gold nanocomposites have been synthesized in which the metal complex-Au nanoparticle (NP) mole ratio is systematically varied by mixing solutions of 4-(dimethylamino) pyridine protected gold nanoparticles and a [Ru(bpy)(2)PVP(10)](2+) metallopolymer; bpy is 2,2'-bipyridyl and PVP is poly-(4-vinylpyridine). The impact of changing the gold nanoparticle diameter ranging from 4.0 ± 0.5 to 12.5 ± 1 nm has been investigated. The photo induced emission of the metallopolymer undergoes static quenching by the metal nanoparticles irrespective of their size. When the volume ratio of Au NP-Ru is 1, the quenching efficiency increases from 38% to 93% on going from 4.0 ± 0.5 to 12.5 ± 1 nm diameter nanoparticles while the radius of the quenching sphere remains unaffected at 75 ± 5 ?. The conductivity of thin films is initially unaffected by nanoparticle incorporation until a percolation threshold is reached at a mole ratio of 4.95 × 10(-2) after which the conductivity increases before reaching a maximum. For thin films of the nanocomposites on electrodes, the electrochemiluminescence intensity of the nanocomposite initially increases as nanoparticles are added before decreasing for the highest loadings. The electrochemiluminescence intensity increases with increasing nanoparticle diameter. The electrochemiluminescence (ECL) emission intensity of the nanocomposite formed using 12.5 nm particles at mole ratios between 5 × 10(-3) and 10 × 10(-3) is approximately 7-fold higher than that found for the parent metallopolymer. The application of these materials for low cost ECL-based point of care devices is discussed.     

Effect of surface immobilization on the electrochemiluminescence of ruthenium-containing metallopolymers

The effect of surface confinement on the electrochemiluminescence (ECL) properties of metallopolymer [Ru(bpy)2(PVP)10]2+, where bpy is 2,2'-bipyridyl and PVP is poly(4-vinylpyridine), is reported. Immobilizing a luminescent material on an electrode surface can substantially modulate its photophysical properties. Significantly, our study revealed that the overall efficiency of the ECL reaction for the metallopolymer film is almost four times higher, at 0.15%, than the highest value obtained for [Ru(bpy)2(PVP)10]2+ dissolved in solution, (phi(ECL) = 0.04%). Electrochemistry has been used to create well-defined concentrations of the quencher Ru3+ within the film. Analysis of both the steady-state luminescence and lifetimes of the film reveals that static quenching by electron transfer between the photoexcited Ru2+ and the Ru3+ centers is the dominant quenching mechanism. The bimolecular rate of electron transfer is (2.5 +/- 0.4) x 10(6) M(-1) s(-1). The implications of these findings for ECL-based sensors, in terms of optimum luminophore loading, is considered.     

A single calibration graph for the direct determination of ascorbic and dehydroascorbic acids by electrogenerated luminescence based on Ru(bpy)(3)2+ in aqueous solution

Ascorbic (H2A) and dehydroascorbic (DA) acids were for the first time directly determined in a single chromatographic run by means of the tris(2,2'-bipyridine)ruthenium(II) (Ru(bpy)(3)2+) based electrogenerated chemiluminescence (ECL) detection. For the first time, it was demonstrated that DA, a nonelectroactive compound, is ECL active and is responsible for the ECL behavior of H2A. This fact, together with the lack of a DA standard, suggested the use of a calibration graph obtained for H2A, for determining both analytes. The proven ECL activity of DA, together with literature data relative to the standard redox potentials of the different species coming from H2A, led to a reconsideration of the proposed ECL reaction mechanism for H2A. The role of the OH- ion in the reaction mechanism of the two analytes appeared to be crucial. H2A and DA could be separated by a suitable C18-reversed-phase HPLC column using an aqueous 30 mM H3PO4 solution as the mobile phase. The optimal ECL response was achieved by polarizing the working electrode at 1.150 Vvs SCE (standard calomel electrode) (oxidation diffusion limiting potential for both H2A and Ru(bpy)(3)2+). The Ru(bpy)(3)2+ solution, at pH 10 for carbonate buffer, was mixed to the eluent solution in a postcolumn system, obtaining, still at pH 10, the final 0.25 mM Ru(bpy)(3)2+ concentration. The detection limit found for the two analytes was 1 x 10(-7) M. The method was successfully applied to the determination of the analytes in a commercially available orange fruit juice.     

Spectroelectrochemical sensing based on multimode selectivity simultaneously achievable in a single device. 2. Demonstration of selectivity in the presence of direct interferences

Three modes of selectivity based on charge-selective partitioning, electrolysis potential, and spectral absorption wavelength were demonstrated simultaneously in a new type of spectroelectrochemical sensor. Operation and performance of the three modes of selectivity for detection of analytes in the presence of direct interferences were investigated using binary mixture systems. These binary mixtures consisted of Fe(CN)(6)(3-) and Ru(bpy)(3)(2+) and of Fe(CN)(6)(4-) and Ru(CN)(6)(4)(-) in aqueous solutions. Results on the Fe(CN)(6)(3-)/Ru(bpy)(3)(2+) binary mixture showed that an anion-exchange coating consisting of PDMDAAC-SiO(2) [where PDMDAAC is poly(dimethyldiallylammonium chloride)] and a cation-exchange coating consisting of Nafion-SiO(2) can trap and preconcentrate analytes with charge selection. At the same time, such coatings exclude interferences carrying the same type of charge as that of the exchange sites in the sensor coating. Using the Fe(CN)(6)(4-)/Ru(CN)(6)(4-) binary mixture, the Fe(CN)(6)(4-) component can be selectively detected by restricting the modulation potential cycled to a range specific to the redox-active Fe(CN)(6)(4-) component and simultaneously monitoring the optical response at the overlapping wavelength of 420 nm. It was also shown that, when the wavelength for optical monitoring was chosen as 500 nm, which is specific to the Ru(CN)(6)(4-) component, interference from the Fe(CN)(6)(4-) component for spectroelectrochemical detection of Ru(CN)(6)(4-) was significantly suppressed, even though the cyclic modulation potential encompassed the redox range for the Fe(CN)(6)(4-) component.     

Nafion-tris(2-2'-bipyridyl)ruthenium(II) Ultrathin Langmuir-Schaefer films: redox catalysis and electrochemiluminescent properties

A simple procedure to incorporate tris(2-2'-bipyridyl)ruthenium(II), [Ru(bpy)3]2+, into Nafion Langmuir-Schaefer (LS) films is described. Nafion LS films (tens of nanometers thick) were formed on quartz glass and indium tin oxide (ITO) directly from Nafion-[Ru(bpy)3]2+ Langmuir films assembled at the water-air interface. This procedure allowed the direct incorporation of [Ru(bpy)3]2+ into Nafion films without the need for subsequent loading. UV-vis spectroscopy confirmed the successful incorporation of [Ru(bpy)3]2+ within the LS films and showed that the amount of [Ru(bpy)3]2+ immobilized in this way scaled with film thickness. Voltammetric studies on ITO-modified electrodes confirmed the successful incorporation of [Ru(bpy)3]2+ and demonstrated that [Ru(bpy)3]2+ was retained within the ultrathin films over a long time scale. These electrodes were tested for the electrocatalytic reduction of tripropylamine. Significant catalysis was observed due to the rapid turnover of [Ru(bpy)3]2+/3+ between the electrode surface and outer boundary of the film, as a direct consequence of the ultrathin film dimensions. Concomitant electrochemiluminescence (ECL) was demonstrated highlighting the potential of this material for sensing applications.     

Microchip device with 64-site electrode array for multiplexed immunoassay of cell surface antigens based on electrochemiluminescence resonance energy transfer

This paper describes a novel on-chip microarray platform based on an electrochemiluminescence resonance energy transfer (ECL-RET) strategy for rapid assay of cancer cell surface biomarkers. This platform consists of 64 antigen-decorated CdS nanorod spots with the diameter of 1.0 cm uniformly distributed on 16 indium tin oxide (ITO) strips, which is coated with a multichannel decorated polydimethylsiloxane (PDMS) slice to realize multiplexed determination of antigens. To shorten the immune reaction time in the microchannels and simplify the device, magnetic stirring and four-channel universal serial bus (USB) ports for plug-and-play were used. When Ru(bpy)(3)(2+) labeled antibodies were selectively captured by the corresponding antigens on the CdS nanorod spot array, ECL-RET from the CdS nanorod (donor) by cathodic emission in the presence of K(2)S(2)O(8) to Ru(bpy)(3)(2+) (acceptor) occurred. With signal amplification of Ru(bpy)(3)(2+) and competitive immunoassay, carcinoembryonic antigen (CEA), α-fetoprotein (AFP), and prostate specific antigen (PSA) as models were detected on this microfluidic device via recording the increased ECL-RET signals on electrode surfaces. Furthermore, this multiplexed competitive immunoassay was successfully used for detecting cancer cell surface antigens via the specific antibody-cell interactions and cell counting via cell surface receptors and antigens on the CdS nanorod surface. This platform provides a rapid and simple but sensitive approach with microliter-level sample volume and holds great promise for multiplexed detection of antigens and antigen-specific cells.