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To enhance the precision and reliability of early disease detection, especially in malignancies, an exhaustive investigation of multi-target biomarkers is essential. In this study, an advanced integrated electrochemical biosensor array that demonstrates exceptional performance was constructed. This biosensor was developed through a controllable porous-size mechanism and in-situ modification of carbon nanotubes (CNTs) to quantify multiplex biomarkers-specifically, C-reaction protein (CRP), carbohydrate antigen 125 (CA125), and carcinoembryonic antigen (CEA)-in human serum plasma. The fabrication process involved creating a highly ordered three-dimensional inverse-opal structure with the CNTs (pCNTs) modifier through microdroplet-based microfluidics, confined spatial self-assembly of nanoparticles, and chemical wet-etching. This innovative approach allowed for direct in-situ modification of nanomaterial onto the surface of electrode array, eliminating secondary transfer and providing exceptional control over structure and stability. The outstanding electrochemical performance was achieved through the synergistic effect of the pCNTs nanomaterial, aptamer, and horseradish peroxidase-labeled (HRP-) antibody. Additionally, the integrated biosensor array platform comprised multiple individually addressable electrode units (n = 11), enabling simultaneous multi-parallel/target testing, thereby ensuring accuracy and high throughput. Crucially, this integrated biosensor array accurately quantified multiplex biomarkers in human serum, yielding results comparable to commercial methods. This integrated technology holds promise for point-of-care testing (POCT) in early disease diagnosis and biological analysis.
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Biomarcadores Tumorais , Técnicas Biossensoriais , Antígeno Ca-125 , Antígeno Carcinoembrionário , Técnicas Eletroquímicas , Nanotubos de Carbono , Neoplasias , Nanotubos de Carbono/química , Humanos , Técnicas Biossensoriais/métodos , Biomarcadores Tumorais/sangue , Neoplasias/sangue , Neoplasias/diagnóstico , Antígeno Carcinoembrionário/sangue , Antígeno Ca-125/sangue , Técnicas Eletroquímicas/métodos , Limite de Detecção , Desenho de Equipamento , Aptâmeros de Nucleotídeos/químicaRESUMO
Reflective displays have stimulated considerable interest because of their friendly readability and low energy consumption. Herein, we develop a reflective display technique via an electro-microfluidic assembly of particles (eMAP) strategy whereby colored particles assemble into annular and planar structures inside a dyed water droplet to create "open" and "closed" states of a display pixel. Water-in-oil droplets are compressed within microwells to form a pixel array. The particles dispersed in droplets are driven by deformation-strengthened dielectrophoretic force to achieve fast and reversible motion and assemble into multiple structures. This eMAP based device can display designed information in three primary colors with ≥170° viewing angle, ~0.14 s switching time, and bistability with an optimized material system. This proposed technique demonstrates the basis of a high-performance and energy-saving reflective display, and the display speed and color quality could be further improved by structure and material optimization; exhibiting a potential reflective display technology.
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Single-cell analysis has important implications for understanding the specificity of cells. To analyze the specificity of rare cells in complex blood and biopsy samples, selective lysis of target single cells is pivotal but difficult. Microfluidics, particularly droplet microfluidics, has emerged as a promising tool for single-cell analysis. In this paper, we present a smart droplet microfluidic system that allows for single-cell selective lysis and real-time sorting, aided by the techniques of microinjection and image recognition. A custom program evolved from Python is proposed for recognizing target droplets and single cells, which also coordinates the operation of various parts in a whole microfluidic system. We have systematically investigated the effects of voltage and injection pressure applied to the oil-water interface on droplet microinjection. An efficient and selective droplet injection scheme with image feedback has been demonstrated, with an efficiency increased dramatically from 2.5% to about 100%. Furthermore, we have proven that the cell lysis solution can be selectively injected into target single-cell droplets. Then these droplets are shifted into the sorting area, with an efficiency for single K562 cells reaching up to 73%. The system function is finally explored by introducing complex cell samples, namely, K562 cells and HUVECs, with a success rate of 75.2% in treating K562 cells as targets. This system enables automated single-cell selective lysis without the need for manual handling and sheds new light on the cooperation with other detection techniques for a broad range of single-cell analysis.
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Técnicas Analíticas Microfluídicas , Microfluídica , Humanos , Microfluídica/métodos , Microinjeções , Hidrolases , Análise de Célula Única/métodos , Células K562 , Técnicas Analíticas Microfluídicas/métodosRESUMO
Colloidal assembly is a key strategy in nature and artificial device. Hereby, an electromicrofluidic assembly platform (eMAP) is proposed and validated to achieve 3D colloidal assembly and manipulation within water droplets. The water-in-oil emulsion droplets autoposition in the eMAP driven by dielectrophoresis, where the (di)electrowetting effect induces droplet deformation, facilitating quadratic growth of the electric field in water droplet to achieve "far-field" dielectrophoretic colloidal assembly. Reconfigurable 3D colloidal configurations are observed and dynamically programmed via applied electric fields, colloidal properties, and droplet size. Binary and ternary colloidal assemblies in one droplet allow designable chemical and physical anisotropies for functional materials and devices. Integration of eMAP in high throughput enables mass production of functional microcapsules, and programmable optoelectronic units for display devices. This eMAP is a valuable reference for expanding fundamental and practical exploration of colloidal systems.
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Microfluídica , Água , Emulsões/química , Microfluídica/métodos , Água/química , Eletroumectação , EletricidadeRESUMO
Electrowetting display (EWD) is a new type of paper-like reflective display based on colored oil, which has gradually become one of the most potential electronic papers with low power consumption, fast response, and full color. However, oil backflow can occur in EWDs, which makes it difficult to maintain a stable aperture ratio. In order to improve the stability of the aperture ratio of EWDs, a new driving waveform was proposed based on analyzing the phenomenon of oil backflow. The driving waveform was composed of a shrinking stage and a driving stage. Firstly, a threshold voltage of oil splitting was calculated by analyzing the luminance curve of EWDs, which were driven by different direct current (DC) voltages. Then, an exponential function waveform, which increased from the threshold voltage, was applied to suppress oil splitting. Finally, a periodic signal combined with a reset signal with a DC signal was applied during the driving stage to maintain a stable aperture ratio display. Experimental results showed that the charge trapping effect could be effectively prevented by the proposed driving waveform. Compared with an exponential function waveform, the average luminance value was increased by 28.29%, and the grayscale stability was increased by 13.76%. Compared to a linear function waveform, the aperture ratio was increased by 10.44% and the response time was reduced by 20.27%.
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Detection of circulating tumor cells (CTCs) has provided a noninvasive and efficient approach for early diagnosis, treatment, and prognosis of cancer. However, efficient capture of CTCs in the clinical environment is very challenging because of the extremely rare and heterogeneous expression of CTCs. Herein, we fabricated a multimarker microfluidic chip for the enrichment of heterogeneous CTCs from peripheral blood samples of breast cancer patients. The multimarker aptamer cocktail DNA nanostructures (TP-multimarker) were modified on a deterministic lateral displacement (DLD)-patterned microfluidic chip to enhance the capture efficiency through the size selection effect of DLD arrays and the synergistic effect of multivalent aptamers. As compared to a monovalent aptamer-modified chip, the multimarker chip exhibits enhanced capture efficiency toward both high and low epithelial cell adhesion molecule expression cell lines, and the DNA nanostructure-functionalized chip enables the accurate capture of different phenotypes of CTCs. In addition, the DNA nanoscaffold makes nucleases more accessible to the aptamers to release cells with molecular integrity and outstanding cell viability.
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Células Neoplásicas Circulantes , Linhagem Celular Tumoral , Separação Celular , DNA/química , Humanos , Microfluídica , Células Neoplásicas Circulantes/metabolismoRESUMO
A microfluidic magnetic analyte delivery (µMAD) technique was developed to realize sample preparation and ultrasensitive biomarker detection. A simply designed microfluidic device was employed to carry out this technique, including a poly(dimethylsiloxane)-glass hybrid microchip having four straight rectangular channels and a permanent magnet. In the µMAD process, functionalized magnetic beads (MBs) were used to recognize and isolate analytes from a complex sample matrix, deliver analytes into tiny microchannels, and preconcentrate analytes in the magnetic trapping/detection region for in situ fluorescence detection. In the feasibility study and sensitivity optimization, horseradish peroxidase-labeled MBs were used, and critical parameters for the signal amplification performance of µMAD were carefully evaluated. At optimized conditions, a sensitivity improvement of at least 2 orders of magnitude was achieved. As a proof of concept, µMAD was combined with the enzyme-linked immunosorbent assay (ELISA), while carcinoembryonic antigen (CEA), prostate-specific antigen (PSA), and interleukin 6 (IL-6) were selected as model biomarkers. The limits of detection (LODs) of µMAD-ELISA were as low as 0.29 pg/mL for CEA, 0.047 pg/mL for PSA, and 0.021 pg/mL for IL-6, which corresponded to an over 200-fold reduction compared to their commercial ELISA results. Meanwhile, µMAD-ELISA revealed high selectivity and reproducibility. In clinical sample analysis, good accuracy was acquired for human serum analysis relative to commercial ELISA kits, and satisfied recoveries of 85.1-102% with RSDs of 1.7-9.8% for IL-6 and 84.7-113% with RSDs of 3.2-8.3% for interferon-γ were obtained. This ultrasensitive and easy operation technique provides a valuable approach for trace-level biomarker detection for practical applications.
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Biomarcadores/análise , Microfluídica , Antígeno Carcinoembrionário/análise , Ensaio de Imunoadsorção Enzimática , Humanos , Interleucina-6/análise , Limite de Detecção , Masculino , Antígeno Prostático Específico/análise , Reprodutibilidade dos TestesRESUMO
Prefocusing of cell mixtures through sheath flow is a common technique used for continuous and high-efficiency dielectrophoretic (DEP) cell separation. However, it usually limits the separation flow velocity and requires a complex multichannel fluid control system that hinders the integration of a DEP separator with other microfluidic functionalities for comprehensive biomedical applications. Here, we propose and develop a high-efficiency, sheathless particle/cell separation method without prefocusing based on flow-field-assisted DEP by combining the effects of AC electric field (E-field) and flow field (F-field). A hollow lemon-shaped electrode array is designed to generate a long-range E-field gradient in the microchannel, which can effectively induce lateral displacements of particles/cells in a continuous flow. A series of arc-shaped protrusion structures is designed along the microchannel to form a F-field, which can effectively guide the particles/cells toward the targeted E-field region without prefocusing. By tuning the E-field, two distinct modes can be realized and switched in one single device, including the sheathless separation (ShLS) and the adjustable particle mixing ratio (AMR) modes. In the ShLS mode, we have achieved the continuous separation of breast cancer cells from erythrocytes with a recovery rate of 95.5% and the separation of polystyrene particles from yeast cells with a purity of 97.1% at flow velocities over 2.59 mm/s in a 2 cm channel under optimized conditions. The AMR mode provides a strategy for controlling the mixing ratio of different particles/cells as a well-defined pretreatment method for biomedical research studies. The proposed microchip is easy to use and displays high versatility for biological and medical applications.
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Técnicas Analíticas Microfluídicas , Separação Celular , Eletrodos , Eletroforese , Microfluídica , Fenômenos FísicosRESUMO
BACKGROUND: The treatment of tendon injuries remains a challenging problem in clinical due to their slow and insufficient natural healing process. Scaffold-based tissue engineering provides a promising strategy to facilitate tendon healing and regeneration. However, many tissue engineering scaffolds have failed due to their poor and unstable mechanical properties. To address this, we fabricated nanofibrous polycaprolactone/methacrylated poly(trimethylene carbonate) (PCL/PTMC-MA) composite scaffolds via electrospinning. MATERIALS AND METHODS: PTMC-MA was characterized by nuclear magnetic resonance. Fiber morphology of composite scaffolds was evaluated using scanning electron microscopy. The monotonic tensile test was performed for determining the mechanical properties of composite scaffolds. Cell viability and collagen deposition were assessed via PrestoBlue assay and enzyme-linked immunosorbent assay, respectively. RESULTS: These PCL/PTMC-MA composite scaffolds had an increase in mechanical properties as PTMC-MA content increase. After photo-crosslinking, they showed further enhanced mechanical properties including creep resistance, which was superior to pure PCL scaffolds. It is worth noting that photo-crosslinked PCL/PTMC-MA (1:3) composite scaffolds had a Young's modulus of 31.13 ± 1.30 MPa and Max stress at break of 23.80 ± 3.44 MPa that were comparable with the mechanical properties of native tendon (Young's modulus 20-1200 MPa, max stress at break 5-100 MPa). In addition, biological experiments demonstrated that PCL/PTMC-MA composite scaffolds were biocompatible for cell adhesion, proliferation, and differentiation.
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Células-Tronco Mesenquimais/citologia , Nanofibras/química , Fotoquímica/métodos , Alicerces Teciduais/química , Animais , Adesão Celular , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Colágeno/metabolismo , Dioxanos/química , Teste de Materiais , Camundongos Endogâmicos C57BL , Poliésteres/química , Polímeros/química , Regeneração , Tendões/fisiologiaRESUMO
Janus particles (JNPs) with controlled anisotropies are regarded as promising materials for sophisticated building blocks and assembly. Herein a straightforward method was proposed for the synthesis of uniformly distributed JNPs with controllable anisotropies, showing two compartmental bulbs with different surface wettability. The synthetic strategy is based on the phase separation-induced styrene liquid protrusion on seed poly(styrene-co-acrylic acid) (CPSAA) nanoparticles via controlled swelling, with the formed polystyrene (PS) and CPSAA compartments corresponding to the amount of monomers. The size (lateral length) ratio of formed PS and CPSAA bulbs, DPS/DCPSAA, defined as "Janusity", has been precisely tuned in the range of 0-0.91 by controlling the mass ratio of two monomers. Obtained JNPs with tunable amphiphilicity are utilized as colloid surfactants to prepare Pickering-emulsions of both water-in-oil (W/O) and oil-in-water (O/W) with proper Janusity. The stability of achieved W/O and O/W Pickering-emulsions is dependent on the adhesion energy of a JNP at the water-oil interfaces. Prepared JNPs have also being utilized to prepare and stabilize monodisperse droplets in microfluidic devices, demonstrating their high potential for fundamental research and practical applications.
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Fiber gas sensing techniques have been applied for a wide range of industrial applications. In this paper, the basic fiber gas sensing principles and the development of different fibers have been introduced. In various specialty fibers, hollow-core photonic crystal fibers (HC-PCFs) can overcome the fundamental limits of solid fibers and have attracted intense interest recently. Here, we focus on the review of HC-PCF gas sensing, including the light-guiding mechanisms of HC-PCFs, various sensing configurations, microfabrication approaches, and recent research advances including the mid-infrared gas sensors via hollow core anti-resonant fibers. This review gives a detailed and deep understanding of HC-PCF gas sensors and will promote more practical applications of HC-PCFs in the near future.
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Optofluidic manipulation of droplets is critical in droplet-based microfluidic systems for chemistry, biology, and medicine. Here, we reported a thermocapillary microvortices-based manipulation platform for controlling oil-in-water droplets through integrating a photothermal waveguide into a microfluidic chip. The sizes and shapes of the droplets can be controlled by adjusting optical power or positions of the water-oil interface. Here, teardrop-shaped droplets, which can encapsulate and accumulate mesoscopic matters easily, were generated when the water-oil interface and the channel boundaries approached the photothermal waveguide center simultaneously. The results showed that the thermocapillary microvortices have good controllability of droplet positions, droplet volumes, and encapsulated-particle distribution and thus it will be a powerful droplet manipulation strategy for microreactors and microcapsules.
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Photothermal effects in SiO2@Au core-shell nanoparticles have demonstrated great potential in various applications for drug delivery, thermo-photovoltaics and photothermal cancer therapy, etc. However, the photothermal conversion of SiO2@Au nanoparticles partially covered by disconnected gold clusters has rarely been investigated systematically. Here, we control the surface morphology of gold clusters on the photothermal conversion performance of SiO2@Au core-shell nanoparticles by means of chemically adjusting the synthesis parameters, including amounts of gold salts, pH value and reducing agent. The macroscopic variations of the photothermal heating of different nanoparticle dispersions are significantly influenced by the nanoscale differences of gold cluster morphologies on the silica core. The temperature rise can be enhanced by the strong near-field coupling and collective heating among gold clusters with a relatively uniform distribution on the silica core. A numerical model of the simplified photothermal system is formulated to interpret the physical mechanism of the experimental observation, and shows a similar trend of temperature rise implying a reasonably good agreement with experimental data. Our work opens new possibilities for manipulating the light-to-heat conversion performance of SiO2@Au core-shell nanoparticles and potential applications of heat delivery with spatial resolution on the nanoscale.
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We established an efficient method for single-cell miRNA analysis by droplet microfluidics, which has high sensitivity of single molecule detection and high throughput. Single-cell analysis of multiple miRNAs in various cells shows that miRNA expression is closely related to cancer type. CTC analysis shows that the method is applicable for rare cell analysis.
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Sequenciamento de Nucleotídeos em Larga Escala/métodos , MicroRNAs/genética , Neoplasias/genética , Análise de Célula Única , Linhagem Celular Tumoral , Fluorescência , Humanos , Limite de DetecçãoRESUMO
Single-cell phenotypic profiling of circulating tumor cells (CTCs) in the blood of cancer patients can reveal vital tumor biology information. Even though various approaches have been provided to enrich and detect CTCs, it remains challenging for consecutive CTC sorting, enumeration, and single-cell characterizations. Here, we report an integrated microfluidic device (IMD) for single-cell phenotypic profiling of CTCs that enables automated CTCs sorting from whole blood following continuous single-cell phenotypic analysis while satisfying the requirements of both high purity (92 ± 3%) of cell sorting and high-throughput processing capacity (5 mL whole blood/3 h). Using this new technique we test the phenotypes of individual CTCs collected from xenograft tumor-bearing mice and colorectal (CRC) patients at different tumor stages. We obtained a correlation between CTC characterization and clinical tumor stage and treatment response. The developed IMD offers a high-throughput, convenient, and rapid strategy to study individual CTCs toward minimally invasive cancer therapy prediction and disease monitoring and has the potential to be translated to clinic for liquid biopsy.
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Antígenos CD/sangue , Biomarcadores Tumorais/sangue , Caderinas/sangue , Neoplasias Colorretais/sangue , Molécula de Adesão da Célula Epitelial/sangue , Dispositivos Lab-On-A-Chip , Células Neoplásicas Circulantes/metabolismo , Análise de Célula Única/métodos , Animais , Antígenos CD/genética , Biomarcadores Tumorais/genética , Caderinas/genética , Contagem de Células , Linhagem Celular Tumoral , Separação Celular/instrumentação , Separação Celular/métodos , Neoplasias Colorretais/diagnóstico , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Molécula de Adesão da Célula Epitelial/genética , Feminino , Fluoresceína-5-Isotiocianato/química , Corantes Fluorescentes/química , Glicoconjugados/química , Xenoenxertos , Humanos , Biópsia Líquida , Camundongos , Camundongos Endogâmicos BALB C , Estadiamento de Neoplasias , Células Neoplásicas Circulantes/patologiaRESUMO
An integrated immunodetection platform employing a simple, reusable, centrifugal microchannel array chip and a smartphone as detection unit was developed. The applicability of the platform to the detection of HIV p24 antigen was demonstrated. The microchip was made of polycarbonate and contained 4â¯×â¯8 zigzag microchannels. After the monoclonal antibody of HIV p24 was adsorbed onto the channel surfaces, HIV p24 was introduced into the microchannel to react with the antibody. A biotin linked polyclonal antibody was then brought in to react with HIV p24, and SP80 (containing streptavidin and horseradish peroxidase) was introduced to react with the biotin. Finally, a solution containing 3,3',5,5'-tetramethylbenzidine and other reagents was passed through the above channels, horseradish peroxidase catalyzed the oxidation of tetramethylbenzidine (to 3,3',5,5'- tetramethylbenzidine diamine) forming a dark color. The color intensity, indicating HIV p24 antigen quantity, was then photographed via a smartphone, and the color of each microchannel was processed via a computer to determine the HIV p24 antigen concentration. Under the optimized conditions, limits of detection (LODs) of 0.17â¯ng/ml and 0.11â¯ng/ml were obtained for p24 antigen in a buffer solution and human serum, respectively. Channel washing/rinsing was implemented via a centrifugal force. An economic portable centrifugal device that could accommodate up to 4 microchips was assembled, and multi-step solution loading and rinsing involved in this sandwich immunoassay were performed conveniently. The microchip could be reused after a simple regeneration process. The low-cost polycarbonate microchip and centrifugal device together with the simple but efficient operation make the method a promising tool for HIV screening in resource limited areas.
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Proteína do Núcleo p24 do HIV/análise , Dispositivos Lab-On-A-Chip , Smartphone , Animais , Anticorpos Monoclonais Murinos/imunologia , Armoracia/enzimologia , Centrifugação , Ensaio de Imunoadsorção Enzimática/métodos , Desenho de Equipamento , Reutilização de Equipamento , HIV/química , Proteína do Núcleo p24 do HIV/imunologia , Peroxidase do Rábano Silvestre/química , Humanos , Limite de Detecção , Camundongos , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Estudo de Prova de Conceito , CoelhosRESUMO
In advanced biomedicine and microfluidics, there is a strong desire to sort and manipulate various cells and bacteria based on miniaturized microfluidic chips. Here, by integrating fiber tweezers into a T-type microfluidic channel, we report an optofluidic chip to selectively trap Escherichia coli in human blood solution based on different sizes and shapes. Furthermore, we simulate the trapping and pushing regions of other cells and bacteria, including rod-shaped bacteria, sphere-shaped bacteria, and cancer cells based on finite-difference analysis. With the advantages of controllability, low optical power, and compact construction, the strategy may be possibly applied in the fields of optical separation, cell transportation, and water quality analysis.
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Separação Celular/instrumentação , Miniaturização/instrumentação , Fibras Ópticas , Pinças Ópticas , Animais , Desenho de Equipamento , Eritrócitos/microbiologia , Escherichia coli/citologia , HumanosRESUMO
As a common physicochemical phenomenon, protonation can cause molecules, atoms or ions with lone-pair electrons to become charged, and can further cause some changes in their physical and chemical properties. Our study first focused on the molecular protonation process and accompanying transitions of the oil/water interface properties in an electric field. The relationship between the protonation degree increment and applied voltage was proposed as a guide for controlling the protonation via applying an electric field. Besides the protonation degree, the water solubility of the oily target molecule obviously increased at 30 V for 600 s along with electric field-driven protonation. At the same time, the electrical conductivity and the underwater interface wettability of oil phase transitioned. These property transitions are anticipated to guide the further improvement and updating of promising protonation functions.
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Cancer has become one of the primary causes of death worldwide. Current cancer-therapy schemes are progressing relatively slowly in terms of reducing mortality, prolonging survival, time and enhancing cure rate, owing to the enormous obstacles of cancer pathophysiology. Therefore, specific diagnosis and therapy for malignant tumors are becoming more and more crucial and urgent, especially for early cancer diagnosis and cancer-targeted therapy. Derived theranostics that combine several functions into one "package" could further overcome undesirable differences in biodistribution and selectivity between distinct imaging and therapeutic agents. In this article, we discuss a chief clinical diagnosis tool - MRI - focusing on recent progress in magnetic agents or systems in multifunctional polymer nanoassemblies for combing cancer theranostics. We describe abundant polymeric MRI-contrast agents integrated with chemotherapy, gene therapy, thermotherapy, and radiotherapy, as well as other developing directions.
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Imageamento por Ressonância Magnética/métodos , Magnetismo , Nanopartículas/química , Neoplasias/diagnóstico por imagem , Polímeros/química , Nanomedicina Teranóstica/métodos , Terapia Genética , Humanos , Nanopartículas/ultraestrutura , Neoplasias/tratamento farmacológicoRESUMO
Oil motion control is the key for the optical performance of electro-fluidic displays (EFD). In this paper, we introduced an extra pinning structure (EPS) into the EFD pixel to control the oil motion inside for the first time. The pinning structure canbe fabricated together with the pixel wall by a one-step lithography process. The effect of the relative location of the EPS in pixels on the oil motion was studied by a series of optoelectronic measurements. EPS showed good control of oil rupture position. The properly located EPS effectively guided the oil contraction direction, significantly accelerated switching on process, and suppressed oil overflow, without declining in aperture ratio. An asymmetrically designed EPS off the diagonal is recommended. This study provides a novel and facile way for oil motion control within an EFD pixel in both direction and timescale.