RESUMO
Flexible transparent electrodes for touch panels, solar cells, and wearable electronics are in great demand in recent years, and the silver nanowire (AgNW) flexible transparent electrode (FTE) is among the top candidates due to its excellent light transmittance and flexibility and the highest conductivity of silver among all metals. However, the conductivity of an AgNWs network has long been limited by the large contact resistance. Here we show a room-temperature solution process to tackle the challenge by nanojoining AgNWs with two-dimensional graphene oxide (GO). The conductivity of the AgNWs network is improved 18 times due to the enhanced junctions between AgNWs by the coated GOs, and the AgNW-GO FTE exhibits a low sheet resistance of 23 Ohm sq-1and 88% light transmittance. It is stable under high temperature and current and their flexibility is intact after 1000 cycles of bending. Measurements of a bifunctional electrochromic device shows the high performance of the AgNW-GO FTE as a FTE.
RESUMO
The potassium ion (K+) plays significant roles in many biological processes. To date, great efforts have been devoted to the development of K+ sensors for colorimetric, fluorescent, and photoacoustic detection of K+ separately. However, the development of molecular K+ probes for colorimetric detection of urinary K+, monitoring K+ fluxes in living cells by fluorescence imaging, and photoacoustic imaging of K+ dynamics in deep tissues still remains an open challenge. Herein, we report the first molecular K+ probe (NK2) for colorimetric, fluorescent, and photoacoustic detection of K+. NK2 is composed of 2-dicyanomethylene-3-cyano-4,5,5-trimethyl-2,5-dihydrofuran (TCF) as the chromophore and phenylazacrown-6-lariat ether (ACLE) as the K+ recognition unit. Predominate features of NK2 include a short synthetic procedure, high K+ selectivity, large detection range (5-200 mM), and triple-channel detection manner. NK2 shows good response to K+ with obvious color changes, fluorescence enhancements (about threefold), and photoacoustic intensity changes. The existence of other metal ions (including Na+, Mg2+, Ca2+, Fe2+) and pH changes (6.5-9.0) have no obvious influence on K+ sensing of NK2. Portable test strips stained by NK2 can be used to qualitatively detect urinary K+ by color changes for self-diagnosis of diseases induced by high levels of K+. NK2 can be utilized to monitor K+ fluxes in living cells by fluorescent imaging. We also find its excellent performance in photoacoustic imaging of different K+ concentrations in the mouse ear. NK2 is the first molecular K+ probe for colorimetric, fluorescent, and photoacoustic detection of K+ in urine, in living cells, and in the mouse ear. The development of NK2 will broaden K+ probes' design and extend their applications to different fields. Graphical abstract.
Assuntos
Colorimetria/métodos , Sondas Moleculares/química , Técnicas Fotoacústicas/métodos , Potássio/análise , Espectrometria de Fluorescência/métodos , Animais , Células HeLa , Humanos , CamundongosRESUMO
Electrically switchable photonic crystals are demonstrated based on TiO2 inverse opals infiltrated with liquid crystals. Macroporous anatase TiO2 inverse opals are fabricated from polystyrene opal templates through a sandwich vacuum backfilled method and followed by calcination. Upon liquid crystal infiltration, the optical properties of the hybrid organic/inorganic structure are characterized by reflectance measurements of the Bragg peak, the position of which can be switched using an external electric field. The physical mechanism underlying this switchable behavior is the reorientation of the liquid crystal molecules inside the spherical voids by the applied electric field, resulting in a significant change of the refractive index contrast between the liquid crystal and the TiO2 inverse opal. With advantageous features of cost-effective fabrication, easy integration, and electric control, such TiO2 inverse opals infiltrated with liquid crystals could play an important role in future development of active photonic devices.
RESUMO
New amphiphilic star or multi-arm block copolymers with different structures were synthesized for enabling the use of hydrophobic oxygen probe of platinum (II)-tetrakis (pentafluorophenyl) porphyrin (PtTFPP) for bioanalysis. The amphiphilic star polymers were prepared through the Atom Transfer Radical Polymerization (ATRP) method by using hydrophilic 4-arm polyethylene glycol (4-arm-PEG) as an initiator. Among the five block copolymers, P1 series (P1a, P1b, and P1c) and P3 possess fluorine-containing moieties to improve the oxygen sensitivity with its excellent capacity to dissolve and carry oxygen. A polymer P2 without fluorine units was also synthesized for comparison. The structure-property relationship was investigated. Under nitrogen atmosphere, high quantum efficiency of PtTFPP in fluorine-containing micelles could reach to 22% and long lifetime could reach to 76 µs. One kind of representative PtTFPP-containing micelles was used to detect the respiration of Escherichia coli (E. coli) JM109 and macrophage cell J774A.1 by a high throughput plate reader. In vivo hypoxic imaging of tumor-bearing mice was also achieved successfully. This study demonstrated that using well-designed fluoropolymers to load PtTFPP could achieve high oxygen sensing properties, and long lifetime, showing the great capability for further in vivo sensing and imaging.
Assuntos
Flúor/química , Interações Hidrofóbicas e Hidrofílicas , Hipóxia/diagnóstico , Oxigênio/metabolismo , Platina/química , Porfirinas/química , Teoria Quântica , Tensoativos/química , Animais , Linhagem Celular , Linhagem Celular Tumoral , Respiração Celular , Sobrevivência Celular , Difusão Dinâmica da Luz , Escherichia coli/citologia , Humanos , Imageamento Tridimensional , Macrófagos/citologia , Camundongos Endogâmicos BALB C , Camundongos Nus , Micelas , Polímeros/síntese química , Polímeros/química , Espectroscopia de Prótons por Ressonância MagnéticaRESUMO
The inflammasome is a caspase-1-activating complex that is implicated in a growing number of acute and chronic pathologies. Interest has increased in identifying small molecular inhibitors of inflammasome signaling because of its role in clinically relevant diseases. It was recently reported that the protein tyrosine kinase, Syk, regulates pathogen-induced inflammasome signaling by phosphorylating a molecular switch on the adapter protein ASC. However, several aspects of the role of Syk in inflammasome signaling and the effects of its inhibition remain unclear. The aim of the present study is to explore in detail the effects of the oxindole Syk inhibitor OXSI-2 on various aspects of nigericin-induced inflammasome signaling. Our results indicate that OXSI-2 inhibits inflammasome assembly, caspase-1 activation, IL-1ß processing and release, mitochondrial ROS generation, and pyroptotic cell death. Using a novel live cell potassium sensor we show that Syk inhibition with OXSI-2 has no effect on potassium efflux kinetics and that blockade of potassium efflux with extracellular potassium alters Syk phosphorylation. The effects of OXSI-2 identified in this study provide context for the role of Syk in inflammasome signaling and demonstrate its importance in oxidative signaling upstream of inflammasome activation and downstream of ion flux.
Assuntos
Indóis/administração & dosagem , Inflamassomos/metabolismo , Potássio/metabolismo , Piroptose/fisiologia , Transdução de Sinais/fisiologia , Sulfonamidas/administração & dosagem , Animais , Linhagem Celular , Relação Dose-Resposta a Droga , Interações Medicamentosas , Peptídeos e Proteínas de Sinalização Intracelular , Macrófagos/efeitos dos fármacos , Macrófagos/patologia , Macrófagos/fisiologia , Taxa de Depuração Metabólica/efeitos dos fármacos , Camundongos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Nigericina/administração & dosagem , Oxindóis , Proteínas Tirosina Quinases , Piroptose/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Quinase SykRESUMO
An ideal fluorescent dye for staining cell organelles should have multiple properties including specificity, stability, biocompatibility, and a large Stokes shift. Tunable photophysical properties enable 1,8-naphthalimide to serve as an excellent fluorophore in biomedical applications. Many naphthalimide derivatives have been developed into drugs, sensors, and other dyes. In this study, a series of 1,8-naphthalimide derivatives targeting live cell mitochondria were synthesized. Among these probes, Mt-4 was characterized as the best one, with highly specific mitochondrial localization, low cytotoxicity, and a large Stokes shift. More importantly, Mt-4 stood out as a potential mitochondrial dye for living-cell experiments involving induced mitochondrial stress arising from the treatments because Mt-4 shows enhanced fluorescence in mitochondrial stress situations.
Assuntos
Corantes Fluorescentes/química , Mitocôndrias/metabolismo , Naftalimidas/química , Sobrevivência Celular , Corantes Fluorescentes/síntese química , Células HeLa , Humanos , Estrutura MolecularRESUMO
Regulation of intracellular potassium (K(+) ) concentration plays a key role in metabolic processes. So far, only a few intracellular K(+) sensors have been developed. The highly selective fluorescent K(+) sensor KS6 for monitoring K(+) ion dynamics in mitochondria was produced by coupling triphenylphosphonium, borondipyrromethene (BODIPY), and triazacryptand (TAC). KS6 shows a good response to K(+) in the range 30-500â mM, a large dynamic range (Fmax /F0 ≈130), high brightness (Ïf =14.4 % at 150â mM of K(+) ), and insensitivity to both pH in the range 5.5-9.0 and other metal ions under physiological conditions. Colocalization tests of KS6 with MitoTracker Green confirmed its predominant localization in the mitochondria of HeLa and U87MG cells. K(+) efflux/influx in the mitochondria was observed upon stimulation with ionophores, nigericin, or ionomycin. KS6 is thus a highly selective semiquantitative K(+) sensor suitable for the study of mitochondrial potassium flux in live cells.
Assuntos
Compostos de Boro/química , Corantes Fluorescentes/química , Mitocôndrias/metabolismo , Potássio/análise , Cátions Monovalentes/análise , Cátions Monovalentes/metabolismo , Linhagem Celular , Células HeLa , Humanos , Microscopia de Fluorescência/métodos , Imagem Óptica/métodos , Potássio/metabolismo , Canais de Potássio/metabolismoRESUMO
A fluorescent colorimetric pH sensor was developed by a polymerization of a monomeric fluorescein based green emitter (SM1) with a monomeric 2-dicyanomethylene-3-cyano-4,5,5-trimethyl-2,5-dihydrofuran derived red emitter (SM2) in poly(2-hydroxyethyl methacrylate)-co-polyacrylamide (PHEMA-co-PAM) matrices. Polymerized SM1 (PSM1) in the polymer matrices showed bright emissions at basic conditions and weak emissions at acidic conditions. Polymerized SM2 (PSM2) in the polymer matrices exhibited a vastly different response when compared to PSM1. The emissions of PSM2 are stronger under acidic conditions than those under basic conditions. When SM1 and SM2 were polymerized in the same polymer matrix, a dual emission sensor acting as a ratiometric pH sensor (PSM1,2) was successfully developed. Because the PSM1 and PSM2 exhibited different pH responses and separated emission windows, the changes in the emission colors were clearly observed in their dual color sensor of PSM1,2, which changed emission colors dramatically from green at pH 7 to red at pH 4, which was detected visually and/or by using a color camera under an excitation of 488 nm. In addition to the development of the dual color ratiometric pH sensor, we also studied the effects of different matrix compositions, crosslinkers, and charges on the reporting capabilities of the sensors (sensitivity and pKa).
RESUMO
We report a novel method for wafer level, high throughput optical chemical sensor patterning, with precise control of the sensor volume and capability of producing arbitrary microscale patterns. Monomeric oxygen (O(2)) and pH optical probes were polymerized with 2-hydroxyethyl methacrylate (HEMA) and acrylamide (AM) to form spin-coatable and further crosslinkable polymers. A micro-patterning method based on micro-fabrication techniques (photolithography, wet chemical process and reactive ion etch) was developed to miniaturize the sensor film onto glass substrates in arbitrary sizes and shapes. The sensitivity of fabricated micro-patterns was characterized under various oxygen concentrations and pH values. The process for spatially integration of two sensors (Oxygen and pH) on the same substrate surface was also developed, and preliminary fabrication and characterization results were presented. To the best of our knowledge, it is the first time that poly (2-hydroxylethyl methacrylate)-co-poly (acrylamide) (PHEMA-co-PAM)-based sensors had been patterned and integrated at the wafer level with micron scale precision control using microfabrication techniques. The developed methods can provide a feasible way to miniaturize and integrate the optical chemical sensor system and can be applied to any lab-on-a-chip system, especially the biological micro-systems requiring optical sensing of single or multiple analytes.
RESUMO
In this paper, we present our results from process development and characterization of optical oxygen sensors that are patterned by traditional UV lithography. An oxygen sensitive luminescent probe, platinum octaethylporphyrin (PtOEP), was encapsulated in commercially purchased photoresist (AZ5214) to form uniform thin sensor films on fused silica substrates. Plasticizer ethoxylated trimethylolpropane triacrylate (SR454) was added to the dye-photoresist sensor mixtures to improve the oxygen sensitivity. The optimum sensor mixture composition that can be patterned with maximum sensitivity was identified. The microfabrication process conditions, cell adherence and oxygen sensitivity results from patterned structures were characterized in detail. Down to 3 µm features have been fabricated on fused silica substrates using the developed techniques. The result implies the developed methods can provide a feasible way to miniaturize the optical sensor system for single cell analysis with precise control of sensor volume and response.
RESUMO
Scaffolded DNA origami, a method to create self-assembled nanostructures with spatially addressable features, has recently been used to develop water-soluble molecular chips for label-free RNA detection, platforms for deterministic protein positioning, and single molecule reaction observatories. These applications highlight the possibility of exploiting the unique properties and biocompatibility of DNA nanostructures in live, cellular systems. Herein, we assembled several DNA origami nanostructures of differing shape, size and probes, and investigated their interaction with lysate obtained from various normal and cancerous cell lines. We separated and analyzed the origami-lysate mixtures using agarose gel electrophoresis and recovered the DNA structures for functional assay and subsequent microscopic examination. Our results demonstrate that DNA origami nanostructures are stable in cell lysate and can be easily separated from lysate mixtures, in contrast to natural, single- and double-stranded DNA. Atomic force microscope (AFM) and transmission electron microscope (TEM) images show that the DNA origami structures are fully intact after separation from cell lysates and hybridize to their targets, verifying the superior structural integrity and functionality of self-assembled DNA origami nanostructures relative to conventional oligonucleotides. The stability and functionality of DNA origami structures in cell lysate validate their use for biological applications, for example, as programmable molecular rafts or disease detection platforms.
Assuntos
Sistema Livre de Células/química , DNA/química , DNA/ultraestrutura , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Humanos , Teste de Materiais , Tamanho da PartículaRESUMO
The concentration of potassium ion is an important indicator for human health, and its abnormality is often accompanied by various diseases. However, most tools currently used to study potassium ion transport are low throughput. Herein, we reported a new K+ fluorescent nanoprobe CP1-KS with high selectivity and sensitivity to K+ (fluorescence enhanced factor was up to 9.91 at 20 mM K+). The polymeric fluorescent probe CP1-KS was composed of the small-molecular K+ indicator KS and amphiphilic copolymer CP1. This sensor can be easily and uniformly dispersed in cell culture medium and is suitable for high throughput analysis. To assess the utility of the probe CP1-KS in biological field, this probe was employed as an extracellular fluorescent probe to monitor the efflux of K+ from cells (E coli, B. Subtilis 168, Hela and MCF-7 cells) under various stimulation including lysozyme, nigericin, digitonin, and ATP. Results demonstrated that CP1-KS is an effective analysis tool for extracellular K+ concentration. We believe that the nanoprobe has great potential in antibacterial drug screening, K+ ionophore function, K+ channel activity, cell membrane permeability analysis or other K+ related field in the future.
Assuntos
Corantes Fluorescentes , Potássio , Bioensaio , Escherichia coli/metabolismo , Humanos , Ionóforos , Íons , Potássio/análiseRESUMO
We describe the synthesis, properties, and application of a new fluorescent potassium chemosensor, KS2, for K(+) sensing and imaging in live cells. By virtue of a strong electron-withdrawing group, 2-dicyanomethylene-3-cyano-4,5,5-trimethyl-2,5-dihydrofuran (TCF), with a triazacryptand ligand, the new sensor can respond to K(+) up to 1.6 M. This is the first highly selective intracellular sensor suitable for sensing K(+) over a broad and high concentration range. Confocal fluorescence microscopy has established the utility of KS2 for live-cell K(+) detection. The application of KS2 combined with other sensors will be of great benefit for investigating cellular metabolism, detecting and diagnosing diseases including cancer, and monitoring responses to therapy.
Assuntos
Corantes Fluorescentes/química , Corantes Fluorescentes/síntese química , Potássio/análise , Linhagem Celular Tumoral , Células Cultivadas , Humanos , Microscopia Confocal , Modelos Moleculares , Estrutura Molecular , Potássio/químicaRESUMO
A random copolymer, poly(NIPAAm-co-PtPorphyrin), consisting of N-isopropylacrylamide (NIPAAm) and platinum (II) porphyrin units, behaves as an optical dual sensor for oxygen and temperature. The dual sensor is designed by incorporating an oxygen-sensitive platinum (II) porphyrin (M1) into a temperature-sensitive polymer (PNIPAAm). The polymer exhibited low critical solution temperature (LCST) property at 31.5 °C. This LCST affected the polymer's aggregation status, which in turn affected the nanostructures, fluorescence intensities, and responses to dissolved oxygen. This enables the polymer to functionalize as a dual temperature and dissolved oxygen sensor. Oxygen response of the platinum (II) porphyrin probes in the polymer followed a two-site Stern-Volmer model, indicating the nonuniform distribution of the probes. The copolymer was used to preliminarily monitor the oxygen consumption of Escherichia coli (E. coli) bacteria. The results indicate a potential application of the polymer in biological fields.
RESUMO
Electrochromic devices can modulate their light absorption under a small driving voltage, but the requirement for external electrical supplies causes response-lag. To address this problem, self-powered electrochromic devices have been studied recently. However, insensitivity to the surrounding light and unsatisfactory stability of electrochromic devices have hindered their critical applications. Herein, novel perovskite solar cell-powered all-in-one gel electrochromic devices have been assembled and studied in order to achieve automatic light adjustment. Two alkynyl-containing viologen derivatives are synthesized as electrochromic materials, the devices with very high stability (up to 70000 cycles) serves as the energy storage and smart window, while the perovskite solar cell with power-conversion-efficiency up to 18.3% serves as the light detector and power harvester. The combined devices can automatically switch between bleached and colored state to adjust light absorption with variable surrounding light intensity in real-time swiftly, which establish significant potentials for applications as modern all-day intelligent windows.
RESUMO
Highly selective fluorescent K+ sensors are of great importance for monitoring K+ fluctuations in various biological processes. In particular, highly efficient ratiometric K+ sensors that can emit in dual wavelengths and facilitate the quantitative determination of K+ are highly anticipated. Herein, we present the first polymer-based ratiometric fluorescent K+ indicator (PK1) for quantitatively detecting K+ in aqueous solutions and high-throughput monitoring K+ fluctuations in living cells. PK1 was synthesized by conjugating a small molecular K+ probe and a red emission reference dye to a hydrophilic polymer skeleton. The newly synthesized PK1 can form highly stable nanoparticles in aqueous solutions and work in 100% water without the aid of any organic solvents or surfactants. PK1 is sensitive to K+ with a fluorescence enhancement of sevenfold after interactions with K+ at 1000 mM and inert to other metal ions, physiological pH, or dye concentration vibrations. More importantly, the fluorescence intensity ratio at 572 and 638 nm is linearly correlated with log [K+] in the range of 2-500 mM (R2 = 0.998), which will facilitate the quantitative detection of K+. Practical application of PK1 in detecting different K+-rich samples demonstrates its great potential in quantitative detection of K+. PK1 can be quickly internalized by live cells and shows no obvious cytotoxicity. We also demonstrate that PK1 could be used for monitoring K+ fluctuations under different stimulations by using a confocal microscope and especially a microplate reader, which is high throughput and time saving. The rational design of PK1 will broaden the design concept of ratiometric fluorescent K+ sensors and facilitate the quantitative detection of K+.
Assuntos
Materiais Biocompatíveis/química , Corantes Fluorescentes/química , Polímeros/química , Potássio/análise , Materiais Biocompatíveis/síntese química , Linhagem Celular , Corantes Fluorescentes/síntese química , Humanos , Teste de Materiais , Estrutura Molecular , Polímeros/síntese químicaRESUMO
A potassium ionoxygen (K+-O2) dual fluorescent sensing film was developed. The film contains three probes, which are K+ probe (KS), O2 probe (OS), and reference probe (RP) in a polymer film composed of poly(ethylene glycol) methyl ether methacrylate (PEGMA), poly(ethylene glycol) dimethacrylate (PEGDMA) and methacrylic acid (MAA). The RP showed blue emission, the KS exhibited green emission, and the OS showed red emission. The emission peaks of three probes do not interfere with each other, which enable the sensing film to be used for ratiometrically and quantitatively detecting the concentrations of K+ and dissolved oxygen (DO). The sensing films showed high sensitivity and selectivity to potassium ions over other metal ions and also good sensitivity for DO from deoxygenated to oxygenated conditions. The sensing film was demonstrated to be capable of analyzing K+ and DO concentrations with experimental errors smaller than ±8.5% in aqueous solutions, showing the potential applications of the sensing films.
RESUMO
The first NIR fluorescent mitochondria-targeting K+ sensor, denoted as TAC-Rh, was developed. The produced sensor consists of a rhodamine analog as the fluorophore and triazacryptand (TAC) as the K+ recognition unit. Compared to the K+ sensors reported previously, TAC-Rh exhibits two unique optical properties: the largest Stokes shifts (120 nm) and the longest emission peak wavelength (720 nm). With the assistance of this novel sensor, real-time changes of K+ concentrations in mitochondria during apoptosis were monitored for the first time. Moreover, it was also the first time that the relationship between mitochondrial K+ flux and apoptosis was investigated in real time using fluorescence imaging.
Assuntos
Apoptose/fisiologia , Corantes Fluorescentes/química , Mitocôndrias/metabolismo , Potássio/análise , Compostos Azabicíclicos/química , Linhagem Celular Tumoral , Humanos , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Potássio/metabolismo , Rodaminas/químicaRESUMO
A two-photon absorbing (2PA) red emitter group was chemically conjugated onto amphiphilic poly(ethylene glycol)-block-poly(ε-caprolactone) (PEG-b-PCL) copolymers, and further grafted with cyclo (Arg-Gly-Asp) (cRGD) peptide to form micelle 1. Micelle 1 with cRGD targeting groups were used for targeted bioimaging. For comparison, micelle 2 without the cRGD targeting groups were also prepared and investigated. The micelles were characterized using dynamic light scattering (DLS), showing average diameters of around 77 nm. The cRGD targeting group is known to bind specifically with α v ß 3 integrin in cancer cells. In this study, α v ß 3 integrin overexpressed human glioblastoma U87MG cell line and α v ß 3 integrin deficient human cervical cancer HeLa cell line were chosen. Results showed that the cRGD targeting group enhanced the cellular uptake efficiency of the micelles significantly in α v ß 3 integrin rich U87MG cells. Higher temperature (37 °C versus 4 °C) and calcium ions (with 3M calcium chloride in the cell culture medium versus no addition of calcium ions) enhanced the cellular uptake efficiency, suggesting that the uptake of the micelles is through the endocytosis pathway in cells. A 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) viability assay was used to evaluate the cytotoxicity of the micelles and no significant cytotoxicity was observed. The BTD-containing two-photon absorbing emitter in the micelles showed a two-photon absorbing cross-section of 236 GM (1GM = 1 × 10-50 cm4 s photon-1 molecule-1) at 820 nm, which is among the highest values reported for red 2PA emitters. Because of the two-photon absorbing characteristics, micelle 1 was successfully used for two-photon fluorescence imaging targeted to U87MG cells under a two-photon fluorescence microscope. This study is the first report regarding the targeted imaging of a specific cancer cell line (herein, U87MG) using the BTD-conjugated-fluorophore-containing block copolymers.
RESUMO
Carcinoembryonic antigen (CEA) is an oncofoetal cell-surface glycoprotein that serves as an important tumor marker for colorectal and some other carcinomas. In this work, a CEA immunoassay using a surface plasmon resonance (SPR) biosensor has been developed. SPR could provide label-free, real-time detection with high sensitivity, though its ability to detect CEA in human serum was highly dependent on the analytical conditions employed. We investigated the influences of various analytical conditions including immobilization methods for anti-CEA antibody and composition of sensor surface on the selective and sensitive detection of CEA. The results show that anti-CEA antibody immobilized via Protein A or Protein G caused a large increase in the resonance signal upon injection of human serum due to the interactions with IgGs in serum, while direct covalent immobilization of anti-CEA antibody could substantially reduce it. An optimized protocol based on further kinetic analysis and the use of 2nd and 3rd antibodies for the sandwich assay allowed detecting spiked CEA in human serum as low as 25 ng/mL. Furthermore, a self-assembled monolayer of mixed ethylene-glycol terminated alkanethiols on gold was found to have a comparable ability in detecting CEA as CM5 with thick dextran matrix and C1 with short flat layer on gold.