RESUMO
Alkaline phosphatase is an important biomarker for medical diagnosis. An enzymatic fluorescence supramolecular hydrogel with AIE properties was developed and used for sensing alkaline phosphatase in vitro and in living cells. In the presence of ALP, K(TPE)EFYp was partially converted to the hydrogelator K(TPE)EFY and self-assembled into nanofibers to form Hydrogel. With the sol-gel transition and the AIE effect, the fluorescence emission was turned on. The linear concentration range of ALP activity in vitro quantified by this method was determined as 0-3 U/L with aLODat 0.02 U/L. In addition, cell imaging and serum experiment showed that K(TPE)EFYp could also be used to detect ALP activity in living cells and biological samples.
Assuntos
Fosfatase Alcalina , Hidrogéis , Espectrometria de Fluorescência , Fosfatase Alcalina/metabolismo , Fosfatase Alcalina/análise , Humanos , Hidrogéis/química , Corantes Fluorescentes/químicaRESUMO
Circulating tumor cells in human peripheral blood play an important role in cancer metastasis. In addition to the size-based and antibody-based capture and separation of cancer cells, their electrical characterization is important for rare cell detection, which can prove fatal in point-of-care testing. Herein, an organic electrochemical transistor (OECT) biosensor made of solution-gated carboxyl graphene mixed with PEDOT:PSS for the detection of cancer cells in situ is reported. Carboxyl graphene was used in this work to modulate cancer cell morphology, which differs significantly from normal blood cells, to achieve rare cancer cell detection. When the concentration of carboxyl graphene mixed in PEDOT:PSS was increased from 0 to 5 mg mL-1, the cancer cell surface area increased from 218 µm2 to 530 µm2, respectively. A change in cell morphology was also detected by the OECT. Negative charges in the cancer cells induced a positive shift in gate voltage, which was approximately 40 mV for spherical-shaped cells. When the cell surface area increased, transfer curves of transistor revealed a negative shift in gate voltage. Therefore, the sensor can be used for in situ detection of cancer cell morphology during the cell capture process, which can be used to identify whether the captured cells are deformable.
Assuntos
Técnicas Biossensoriais , Grafite , Células Neoplásicas Circulantes , Humanos , Grafite/química , Técnicas Eletroquímicas , Membrana CelularRESUMO
Organic electrochemical transistor (OECT) was applied in chemical and biological sensing. In this work, we developed a simple and repeatable method to fabricate OECT array, which had been successfully used to detect cancer cells. PEDPT:PSS conductive film between source and drain electrodes were patterned through photolithography, which can achieve uniform devices with same electrical characterization. When MCF-7 cancer cells are captured on the PEDOT:PSS surface via specifical antibody, the transfer characteristic of OECT shifts to higher gate electrode voltage due to the electrostatic interaction between cancer cells and device. The effective gate voltage shift can reach about 63 mV when the concentration of cancer cells increased to 5000. The shift of effective gate voltage is related to the cancer cell morphology, which is increased in the first 1 h and decreased when the capture time was larger than 1 h. The device of OECT array can increase the sample flux and make the detection result more accurate. It is expected that OECT array will have promising practical applications in single cancer cell detection in the future.
RESUMO
A new type of carbon dot (CD)-functionalized solution-gated graphene transistor (SGGT) sensor was designed and fabricated for the highly sensitive and highly selective detection of glutathione (GSH). The CDs were synthesized via a one-step hydrothermal method using DL-thioctic acid and triethylenetetramine (TETA) as sources of S, N, and C. The CDs have abundant amino and carboxyl groups and were used to modify the surface of the gate electrode of SGGT as probes for detecting GSH. Remarkably, the CDs-SGGT sensor exhibited excellent selectivity and ultrahigh sensitivity to GSH, with an ultralow limit of detection (LOD) of up to 10-19 M. To the best of our knowledge, the sensor outperforms previously reported systems. Moreover, the CDs-SGGT sensor shows rapid detection and good stability. More importantly, the detection of GSH in artificial serum samples was successfully demonstrated.
Assuntos
Grafite , Pontos Quânticos , Carbono , Limite de Detecção , GlutationaRESUMO
Hydrophobic conjugated polymers have poor ionic transport property, so hydrophilic side chains are often grafted for their application as organic electrochemical transistors (OECTs). However, this modification lowers their charge transport ability. Here, an ionic gel interfacial layer is applied to improve the ionic transport while retaining the charge transport ability of the polymers. By using the ionic gels comprising gel matrix and ionic liquids as the interfacial layers, the hydrophobic polymer achieves the OECT feature with high transconductance, low threshold voltage, high current on/off ratio, short switching time, and high operational stability. The working mechanism is also revealed. Moreover, the OECT performance can be tuned by varying the types and ratios of ionic gels. With the proposed ionic gel strategy, OECTs can be effectively realized with hydrophobic conjugated polymers.
RESUMO
Immunosorbent assay is one of the most popular immunological screening techniques which has been widely used for the clinical diagnosis of alpha-fetoprotein (AFP). While traditional immunosorbent assay (ELISA) suffers from low detection sensitivity due to its low intensity of colorimetric signal. To improve the sensitivity of AFP detection, we developed a new and sensitive immunocolorimetric biosensor by combining Ps-Pt nanozyme with terminal deoxynucleotidyl transferase (TdT)-mediated polymerization reaction. The determination of AFP was achieved by measuring the visual color intensity produced by the catalytic oxidation reaction of the 3,3',5,5'-tetramethylbenzidine (TMB) solution with Ps-Pt and horseradish peroxidase (HRP). Owing to the synergistic catalysis of Ps-Pt and horseradish peroxidase HRP enriched in polymerized amplification products, this biosensor exhibited a significant color change within 25 s in the presence of 10-500 pg/mL AFP. This proposed method allowed for the specific detection of AFP with a detection limit of 4.30 pg/mL and even 10 pg/mL target protein could be distinguished clearly by visual observation. Furthermore, this biosensor could be applied to analysis of AFP in the complex sample and could be easily extended to the detection of other proteins.
Assuntos
Técnicas Biossensoriais , alfa-Fetoproteínas , alfa-Fetoproteínas/análise , Colorimetria/métodos , Imunoadsorventes , Peroxidase do Rábano Silvestre/metabolismo , Técnicas Biossensoriais/métodos , Peróxido de Hidrogênio , Limite de DetecçãoRESUMO
A 3D network capture substrate based on poly(lactic-co-glycolic acid) (PLGA) nanofibers was studied and successfully used for high-efficiency cancer cell capture. The arc-shaped glass micropillars were prepared by chemical wet etching and soft lithography. PLGA nanofibers were coupled with micropillars by electrospinning. Given the size effect of the microcolumn and PLGA nanofibers, a three-dimensional of micro-nanometer spatial network was prepared to form a network cell trapping substrate. After the modification of a specific anti-EpCAM antibody, MCF-7 cancer cells were captured successfully with a capture efficiency of 91%. Compared with the substrate composed of 2D nanofibers or nanoparticles, the developed 3D structure based on microcolumns and nanofibers had a greater contact probability between cells and the capture substrate, leading to a high capture efficiency. Cell capture based on this method can provide technical support for rare cells in peripheral blood detection, such as circulating tumor cells and circulating fetal nucleated red cells.
RESUMO
Contact electrification between water and a solid surface is crucial for physicochemical processes at water-solid interfaces. However, the nature of the involved processes remains poorly understood, especially in the initial stage of the interface formation. Here we report that H2O2 is spontaneously produced from the hydroxyl groups on the solid surface when contact occurred. The density of hydroxyl groups affects the H2O2 yield. The participation of hydroxyl groups in H2O2 generation is confirmed by mass spectrometric detection of 18O in the product of the reaction between 4-carboxyphenylboronic acid and 18O-labeled H2O2 resulting from 18O2 plasma treatment of the surface. We propose a model for H2O2 generation based on recombination of the hydroxyl radicals produced from the surface hydroxyl groups in the water-solid contact process. Our observations show that the spontaneous generation of H2O2 is universal on the surfaces of soil and atmospheric fine particles in a humid environment.
Assuntos
Eletricidade , Peróxido de Hidrogênio , Radical Hidroxila , Água , Atmosfera/química , Umidade , Peróxido de Hidrogênio/síntese química , Peróxido de Hidrogênio/química , Radical Hidroxila/química , Espectrometria de Massas , Isótopos de Oxigênio/análise , Isótopos de Oxigênio/química , Tamanho da Partícula , Solo/química , Água/químicaRESUMO
The purpose of this experiment was to study the effects of fenugreek seed extract (FSE) on the growth performance, intestinal morphology, intestinal immunity and cecal micro-organisms in yellow-feathered broilers. A total of 240 one-day-old male yellow-feathered broilers were selected and randomly assigned to four treatments with 6 replicates per group and ten broilers per replicate. Started from the third day, birds were fed with basal diet (CON group) or basal diet supplemented with 30 mg/kg Zinc bacitracin (ZB group), or basal diet supplemented with 50 (D-FSE group) or 100 (H-FSE group) mg/kg FSE, respectively. The experiment lasted for 56 d. The results showed that dietary FSE supplementation improved average daily weight gain (ADG) and ratio of feed to weight gain (F: G) (P < 0.01), increased intestinal villus height (VH), villus height to crypt depth ratio (V/C) (P < 0.05), serum concentrations of IL-10, and the contents of secretory immunoglobulin A (sIgA) (P < 0.05), as well as decreased the activity of iNOS (P < 0.05). The high-throughput sequencing results showed that dietary FSE supplementation increased the alpha diversity of cecal microbes, and Firmicutes, Bacteroidetes, Verrucomicrobia and Proteobacteria taken up 95% of all phyla detected, FSE significantly reduced Campylobacter, Synergistes, and Lachnoclostridium abundance (P ≤ 0.05). There were significant difference in more than 30 KEGG pathways between FSE added group and control group or ZB group. FSE supplementation, in other words, maintained gut microbiota homeostasis while improving broiler growth performance. As a result, FSE has the potential to replace prophylactic antibiotic use in poultry production system.
Assuntos
Trigonella , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Galinhas , Dieta/veterinária , Suplementos Nutricionais , Masculino , Extratos Vegetais/farmacologia , Aumento de PesoRESUMO
The heterogeneity of cancer has become a major obstacle to treatment, and the development of an efficient, fast, and accurate drug delivery system is even more urgent. In this work, we designed a device that integrated multiple functions of cell capture, in situ manipulation, and non-destructive release on a single device. With an applied electric field, an intelligent device based on MnO2 nanomaterials was used to realize efficient and rapid capture of cancer cells in both patients' blood and artificial blood samples. This device could capture cancer cells with high efficiency (up to about 93%) and strong specificity in blood samples, the capture time was nearly 50 min faster than that of natural sedimentation, and reduce the effects on cells caused by long-time in vitro culture. In addition, Mn3+ on the surface of the MnO2 substrate was reduced to Mn2+ by an electrochemical method, partial dissolution occurred, and then the captured cells were non-destructively released with rapid speed (about 8 s) and high efficiency (about 94 ± 2%). For in situ regulation, upon applying a pulse electric field, the captured cells were perforated nondestructively, and extracellular molecules could be delivered to the captured cells with well-performed dose and temporal controls. As a proof-of-concept application, we proved that the device could capture circulating tumor cells in peripheral blood faster and achieve in situ drug delivery. Finally, it can also quickly release circulating tumour cells for subsequent analysis, highlighting its accuracy, due to which it is widely used in medical treatment, basic tumor research and drug development.
Assuntos
Nanoestruturas , Células Neoplásicas Circulantes , Linhagem Celular Tumoral , Separação Celular/métodos , Humanos , Compostos de Manganês , Células Neoplásicas Circulantes/metabolismo , ÓxidosRESUMO
MicroRNAs play important roles in disease diagnosis and therapy. However, current methods for microRNA detection suffer from low sensitivity and cannot directly detect short microRNAs. Herein, we have developed a highly sensitive and selective fluorescent method for direct microRNA detection by combining the duplex-specific nuclease-assisted recycling amplification and the nicking enzyme-powered three-dimensional DNA walker. Target microRNA initiates duplex-specific nuclease-assisted recycling amplification, releasing numerous bipedal walking strands. The released bipedal walking strands hybridize with carboxyfluorescein-labeled track DNA and form nicking recognition site. Driven by the hydrolysis of the nicking enzyme, the bipedal walking strand autonomously moves along the track strand, releasing a large number of carboxyfluorescein-labeled DNA fragments and generating obvious fluorescence signals. This dual-signal amplification method can directly detect microRNA 21 as low as 130 fM and has good selectivity. The proposed method is not only simple for nucleic acid design, but also can be used as a universal method for the highly sensitive detection of all RNAs.
Assuntos
MicroRNAs , Técnicas de Amplificação de Ácido Nucleico , DNA , Endonucleases , MicroRNAs/análise , MicroRNAs/biossíntese , MicroRNAs/química , Técnicas de Amplificação de Ácido Nucleico/métodosRESUMO
Planar-type perovskite solar cells (p-PSCs) based on SnO2 have garnered further attention due to their simple and low-temperature fabrication. Improving the critical properties of the electron transport layer (ETL) is an effective way to enhance the performance of p-PSC devices. Here, a brand-new method is developed to relieve the contact recombination caused by the rough fluorine-doped tin oxide (FTO) surface and further boosts the electrical concentration of the ETL. A SnO2-ethylene diamine tetraacetic acid (EDTA) acylamide compound (SEAC) with hydrogen bond-induced adjustable cluster size is reported for the first time. The rational choice of the SEAC cluster size is the key for obtaining the smooth interfacial morphology of the ETL on the rugged FTO substrate. In addition, the energy band gap decreases with the increasing cluster size, and consequently, results in improved electrical conductivity of the SEAC. The upshifted Fermi energy level leads to higher electron concentration, which is an important physical quantity of the ETL. The PSC devices based on the optimized SEAC achieve an improved power conversion efficiency of 21.29% with negligible J-V hysteresis due to significantly enhanced electron transport and reduced contact charge recombination at the ETL/perovskite interface. In general, this paper comes up with a unique strategy to improve the quality of the SnO2-based ETL.
RESUMO
Cell-based high-throughput screening is a key step in the current disease-based research, drug development, and precision medicine. However, it is challenging to establish a rapid culture and screening platform for rare cells (patient-derived) due to the obvious differences between the traditional 2D cell model and the tumor microenvironment, as well as the lack of a low-consumption screening platform for low numbers of cells. Here, we developed an acoustic drop-assisted superhydrophilic-superhydrophobic microarray platform for the rapid culture and screening of a few cells. By employing hydrophilic and hydrophobic microarrays, we can automatically distribute the cell suspension into uniform droplets, and these cells can spontaneously form compact 3D cell spheroids within 36 h (similar to the microenvironment of tumors in vivo). By using the acoustic droplet ejection device, we can accurately inject a drug solution with a volume of â¼pL to â¼nL into the droplet, and the whole process can be completed within 20 ms (one print). By using three different cell lines (Caco-2, MCF-7, and HeLa) to optimize the platform, the culture and screening of five patients' colon cancer were subsequently realized. Using three conventional chemotherapeutics (5-fluorouracil, cetuximab, and panitumumab) of various concentrations, the best treatment was screened out and compared with the actual treatment effect of the patients, and the results were extremely similar. As a proof-of-concept application, we have proved that our platform can quickly cultivate patient samples and effectively screen the best treatment methods, highlighting its wide application in precision medicine, basic tumor research, and drug development.
Assuntos
Antineoplásicos/farmacologia , Neoplasias do Colo/patologia , Ensaios de Seleção de Medicamentos Antitumorais , Ensaios de Triagem em Larga Escala , Análise em Microsséries , Acústica , Idoso , Células CACO-2 , Sobrevivência Celular/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais/instrumentação , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Feminino , Ensaios de Triagem em Larga Escala/instrumentação , Ensaios de Triagem em Larga Escala/métodos , Humanos , Interações Hidrofóbicas e Hidrofílicas , Análise em Microsséries/instrumentação , Análise em Microsséries/métodos , Esferoides Celulares , Células Tumorais CultivadasRESUMO
Solar energy-powered adsorption-based atmospheric water harvesting (ABAWH) is an emerging technology for freshwater production, especially in water-scarce regions that are remote and landlocked. Numerous water adsorbents have been used in ABAWH devices to convert molecule to liquid water. However, it is still challenging to harvest water from the air in cold winter, owing to the water adsorption of sorbents decreasing significantly at low temperature. Herein, we designed and fabricated an ABAWH device by integrating composited ionic liquids (CILs) with carbon nanotubes (CNTs) photothermal materials on the surface of cotton rod fibers. CILs extract water from the air. CNTs enable light-to-heat conversion and drive the solar evaporation process. Importantly, the cotton rods offer a backbone porous structure to maintain its internal temperature at 20 °C under solar irradiation, and thus promote the water adsorption performance of CILs at low environmental temperature. Freshwater is successfully harvested under environment temperature of 6 °C, 30% RH and solar irradiation intensity of 0.6 kW m-2. The water yield can achieve 1.49 kg per m2 per day in an outdoor environment. We believe that the ABAWH device offers a promising approach to effectively harvest water from the air at low temperature and humidity conditions.
RESUMO
Highly sensitive detection of DNA is of great importance for the detection of genetic damage and errors for the diagnosis of many diseases. Traditional highly sensitive organic electrochemical transistor (OECT)-based methods mainly rely on good conductivity materials, which may be limited by complex synthesis and modification steps. In this work, DNA biosensor based on OECT and hybridization chain reaction (HCR) signal amplification was demonstrated for the first time. Au nanoparticles were electrochemically deposited on the Au gate electrode to increase the surface area. Then, the HCR products, long negatively charged double-stranded DNA, were connected to the target by hybridization, which can increase the effective gate voltage offset of OECT. This sensor exhibited high sensitivity and even 0.1 pM target DNA could be directly detected with a significant voltage shift. In addition, it could discriminate target DNA from the mismatched DNA with good selectivity. This proposed method based on HCR in DNA detection exhibited an efficient amplification performance on OECT, which provided new opportunities for highly sensitive and selective detection of DNA.
RESUMO
Li metal is considered a highly desirable anode for next-generation high-energy-density rechargeable lithium batteries. However, irregular Li dendrite formation and infinite relative volume changes prevent the commercial adoption of Li-metal anodes. Here, electrophoretic deposition of black phosphorus (BP) on commercial Cu foam (BP@Cu foam) is reported to regulate Li nucleation for the first time. First-principles calculations reveal that the unique two-dimensional (2D) structure of BP is beneficial to Li intercalation and propagation. Compared with the random Li nucleation and growth on bare Cu foam, Li ions are preferably confined into the BP layers, which induces uniform Li nucleation at the early stage of the Li deposition and guides the following lateral Li growth on BP@Cu foam. In addition, the three-dimensional (3D) porous and conductive framework of Cu foams further mitigate the volume change and dissipate the current density. Attributing to these merits, the BP@Cu foam exhibits significantly enhanced Coulombic efficiency and cycling stability compared with bare Cu foam. In the full-cell configuration paired with a Li4Ti5O12 or LiFePO4 cathode, the BP@Cu foam also boosts the battery performances. This work provides new insights into the development of BP and other elaborate 2D materials for achieving dendrite-free Li-metal anodes.
RESUMO
A new dual-recognition fluorescent biosensor for circulating tumor DNA (ctDNA) detection has been developed, which combines the clamping function of peptide nucleic acid (PNA) and terminal protection of small-molecule-linked DNA (TPSMLD). Taking the tumor-specific E542K mutation and methylation of the PIK3CA gene as the target ctDNA, a low detection limit of 0.3161 pM ctDNA is achieved with good selectivity. This study not only offers a sensitive, selective and accurate ctDNA detection method, but can also be used to detect the target in complex biological samples.
Assuntos
Técnicas Biossensoriais , DNA Tumoral Circulante , Ácidos Nucleicos Peptídicos , DNA Tumoral Circulante/genética , Constrição , DNA/genética , MutaçãoRESUMO
This paper presents a total phosphorus online real-time monitoring system integrated with on-chip digestion based on the merits of optofluidic technology. The integrated optofluidic device contains a hollow optical fiber employed for pretreatment and digestion of phosphorus solution samples, a polydimethylsiloxane (PDMS)-based micromixer with convergent-divergent walls designed to enable sufficient mixing and chromogenic reaction, and a couple of optical fiber collimators attached with a Z-shaped flow cell for optical detection. Details of system design and fabrication are introduced in this paper. In the experiment, on-chip digestion of four typical phosphates in aqueous solution including organophosphorus and inorganic phosphorus is investigated under different reaction conditions, such as digestion temperature, concentration of oxidant and pH value, and the optimal reaction parameters are explored under different conditions. Meanwhile, we demonstrate the online real-time monitoring function of the optofluidic device, and the digestion mechanisms of four different phosphates are analyzed and discussed. Compared with the national standard method, we find that the measurement accuracy and sensitivity are acceptable when the concentration of total phosphorus is between 0.005-0.9 mg/L (by weight of P) in aqueous solution, which covers the range defined in the national standard. The traditional digestion time of several hours is greatly reduced to less than 10 s, and the content of total phosphorus can be obtained in a few minutes. The integrated optofluidic device can significantly shorten the test time and reduce the sample amount, and also provides a versatile platform for the real-time detection and analysis of many biochemical samples.
RESUMO
Non-adherent cells play key roles in various biological processes. Studies on this type of cell, especially at single-cell resolution, help reveal molecular mechanisms underlying many biological and pathological processes. The emerging microfluidics technology has developed effective methods for analyzing cells. However, it remains challenging to treat and monitor single live non-adherent cells in an in situ, long-term, and real-time manner. Herein, a microfluidic platform was set up to generate and anchor cell-laden water-in-oil-in-water (W/O/W) double emulsions (DEs) to investigate these cells. Within the device, W/O/W DEs encapsulating non-adherent cells were generated through two adjacent flow-focusing structures and subsequently anchored in an array of microchambers. These droplets maintained the W/O/W structure and the anchorage status in the continuous perfusion fluid for at least one week. The mass transfer of different molecules with suitable molecular weights and partition coefficients between the interior and exterior of W/O/W DEs could be regulated by perfusion fluid flow rates. These features endow this platform with potential to continuously supply encapsulated non-adherent cells with nutrients or small-molecule stimuli/drugs through fluid perfusion. Meanwhile, the confinement of cells in the anchored DEs favored long-term monitoring of cellular dynamic behaviors and responses. As a proof of concept, fluorescein diacetate (FDA) was employed to visualize the cellular uptake and biochemical metabolism of TF-1 human erythroleukemia cells. We believe that this W/O/W DE anchorage and perfusion platform would benefit single-cell-level studies as well as small-molecule drug discovery requiring live non-adherent cells.
Assuntos
Dispositivos Lab-On-A-Chip , Óleos/química , Análise de Célula Única/instrumentação , Água/química , Adesão Celular , Linhagem Celular Tumoral , Emulsões , Humanos , Fenômenos Mecânicos , Propriedades de SuperfícieRESUMO
Detection of circulating tumor cells (CTCs) in peripheral blood is of paramount significance for cancer diagnosis, progress evaluation, and individualized therapy. However, the rareness and heterogeneity of CTCs introduces significant challenges in the capture of cancer cells as well as downstream genetic analysis. In this work, a microwell-assisted multiaptamer immunomagnetic platform (MMAIP) is proposed for highly efficient capture of CTCs with minimum influence of heterogeneity. Assisted by a microwell chip, the purity of CTCs is greatly improved, thus meeting the requirement of downstream gene analysis. This is, as far as is known, the first aptamer based platform enabling mutation analysis of the captured CTCs from cancer patients, which will contribute to the practical application of aptamers in clinics.