RESUMO
Many advanced microfluidic Lab-on-disc (LOD) devices require an on-board power supply for powering active components. LODs with an on-board electrical power supply are called electrified-LODs (eLODs) and are the subject of the present review. This survey comprises two main parts. First, we discuss the different means of delivering electrical energy to a spinning disc including slip-ring, wireless power transmission, and on-board power supply. In the second part, we focus on utilizing electrical power on eLODs for three electrokinetic microfluidic processes: electrophoresis, electroosmotic flow, and dielectrophoresis. Electrokinetic phenomena enable propulsion, separation, and manipulation of different fluids and various types of microparticles/cells. We summarize the theoretical and experimental results for all three electrokinetic phenomena enacted on centrifugal platforms. While extensive numerical modeling and experimental research are available for electrokinetics on stationary platforms, there is a noticeable lack of development in this area when executed on rotating platforms. The review concludes by comparing the strengths and weaknesses of different electrokinetic techniques implemented on centrifugal platforms, and additionally, the most promising applications of electrokinetic-assisted eLOD devices are singled out.
Assuntos
Técnicas Biossensoriais , Dispositivos Lab-On-A-Chip , Técnicas Analíticas Microfluídicas/métodos , Eletro-Osmose , Eletroforese , Técnicas Analíticas Microfluídicas/normas , Técnicas Analíticas Microfluídicas/tendênciasRESUMO
BACKGROUND: Immunosuppressant therapeutic drug monitoring (TDM) usually requires outpatient travel to hospitals or phlebotomy sites for venous blood collection; however Mitra® Microsampling Device (MSD) sampling could allow self-collection and shipping of samples to a laboratory for analysis. This study examined the feasibility of using volumetric microsampling by MSD for TDM of tacrolimus (TaC) and cyclosporin A (CsA) in transplant patients, along with their feedback on the process. METHODS: MSD was used to collect TaC and CsA from venous (VB) or capillary (CB) blood. The MSDs were rehydrated, extracted, and analyzed using on-line solid phase extraction coupled to tandem mass spectrometry (SPE-MS/MS). We report an abbreviated method validation of the MSD including: accuracy, precision, linearity, carry-over, and stability using residual venous whole blood (VB) samples. Subsequent clinical validation compared serially collected MSD + CB against VB (200 µL) from transplant patients. RESULTS: Accuracy comparing VB vs. MSD+VB showed high clinical concordance (TaC = 89% and CsA = 98%). Inter- and intra-precision was ≤11.5 %CV for TaC and CsA. Samples were stable for up to 7 days at room temperature with an average difference of <10%. Clinical validation with MSD+CB correlated well with VB for CsA (slope = 0.95, r2 = 0.88, n = 47) and TaC (slope = 0.98, r2 = 0.82, n = 49). CB vs. VB gave concordance of 94% for CsA and 79% for TaC. A satisfaction survey showed 82% of patients preferred having the capillary collection option. CONCLUSION: Transplant patients favored having the ability to collect capillary samples at home for TaC/CsA monitoring. Our results demonstrate good concordance between MSD+CB and VB for TaC and CsA TDM, but additional studies are warranted.
Assuntos
Ciclosporina/farmacocinética , Monitoramento de Medicamentos/métodos , Técnicas Analíticas Microfluídicas , Tacrolimo/farmacocinética , Idoso , Monitoramento de Medicamentos/instrumentação , Monitoramento de Medicamentos/normas , Feminino , Humanos , Masculino , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Técnicas Analíticas Microfluídicas/normas , Pessoa de Meia-Idade , Satisfação do Paciente , Reprodutibilidade dos Testes , Espectrometria de Massas em TandemRESUMO
We present a versatile tool for the generation of monodisperse water-in-fluorinated-oil droplets in standard reaction tubes by centrifugal step emulsification. The microfluidic cartridge is designed as an insert into a standard 2 mL reaction tube and can be processed in standard laboratory centrifuges. It allows for droplet generation and subsequent transfer for any downstream analysis or further use, does not need any specialized device, and manufacturing is simple because it consists of two parts only: A structured substrate and a sealing foil. The design of the structured substrate is compatible to injection molding to allow manufacturing at large scale. Droplets are generated in fluorinated oil and collected in the reaction tube for subsequent analysis. For sample sizes up to 100 µL with a viscosity range of 1 mPa·s-4 mPa·s, we demonstrate stable droplet generation and transfer of more than 6 × 105 monodisperse droplets (droplet diameter 66 µm ± 3 µm, CV ≤ 4%) in less than 10 min. With two application examples, a digital droplet polymerase chain reaction (ddPCR) and digital droplet loop mediated isothermal amplification (ddLAMP), we demonstrate the compatibility of the droplet production for two main amplification techniques. Both applications show a high degree of linearity (ddPCR: R2 ≥ 0.994; ddLAMP: R2 ≥ 0.998), which demonstrates that the cartridge and the droplet generation method do not compromise assay performance.
Assuntos
Centrifugação , Emulsões , Gotículas Lipídicas , Técnicas Analíticas Microfluídicas , Bioensaio/instrumentação , Bioensaio/métodos , Bioensaio/normas , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Técnicas Analíticas Microfluídicas/normas , Microfluídica/instrumentação , Microfluídica/métodos , Microfluídica/normas , Reação em Cadeia da Polimerase/métodos , Padrões de Referência , Viscosidade , Fluxo de TrabalhoRESUMO
BACKGROUND: Great deformability allows red blood cells (RBCs) to flow through narrow capillaries in tissues. A number of microfluidic devices with capillary-like microchannels have been developed to monitor storage-related impairment of RBC deformability during blood banking operations. This proof-of-concept study describes a new method to standardize and improve reproducibility of the RBC deformability measurements using one of these devices. STUDY DESIGN AND METHODS: The rate of RBC flow through the microfluidic capillary network of the microvascular analyzer (MVA) device made of polydimethylsiloxane was measured to assess RBC deformability. A suspension of microbeads in a solution of glycerol in phosphate-buffered saline was developed to be used as an internal flow rate reference alongside RBC samples in the same device. RBC deformability and other in vitro quality markers were assessed weekly in six leukoreduced RBC concentrates (RCCs) dispersed in saline-adenine-glucose-mannitol additive solution and stored over 42 days at 4°C. RESULTS: The use of flow reference reduced device-to-device measurement variability from 10% to 2%. Repeated-measure analysis using the generalized estimating equation (GEE) method showed a significant monotonic decrease in relative RBC flow rate with storage from Week 0. By the end of storage, relative RBC flow rate decreased by 22 ± 6% on average. CONCLUSIONS: The suspension of microbeads was successfully used as a flow reference to increase reproducibility of RBC deformability measurements using the MVA. Deformability results suggest an early and late aging phase for stored RCCs, with significant decreases between successive weeks suggesting a highly sensitive measurement method.
Assuntos
Deformação Eritrocítica/fisiologia , Eritrócitos/citologia , Eritrócitos/fisiologia , Dispositivos Lab-On-A-Chip/normas , Técnicas Analíticas Microfluídicas , Bancos de Sangue/normas , Velocidade do Fluxo Sanguíneo/fisiologia , Preservação de Sangue/efeitos adversos , Preservação de Sangue/métodos , Preservação de Sangue/normas , Criopreservação , Contagem de Eritrócitos/instrumentação , Contagem de Eritrócitos/métodos , Contagem de Eritrócitos/normas , Citometria de Fluxo/instrumentação , Citometria de Fluxo/métodos , Citometria de Fluxo/normas , Hemólise , Humanos , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Técnicas Analíticas Microfluídicas/normas , Estudo de Prova de Conceito , Reprodutibilidade dos Testes , Fatores de Tempo , Armazenamento de Sangue/métodosRESUMO
Upregulated expression of P-selectin on activated endothelium and platelets significantly contributes to the initiation and progression of vaso-occlusive crises (VOC), a major cause of morbidity in sickle cell disease (SCD). Crizanlizumab (ADAKVEO®), a humanized monoclonal antibody against P-selectin, primarily inhibits the interaction between leukocytes and P-selectin, and has been shown to decrease the frequency of VOCs in clinical trials. However, the lack of reliable in vitro assays that objectively measure leukocyte adhesion to P-selectin remains a critical barrier to evaluating and improving the therapeutic treatment in SCD. Here, we present a standardized microfluidic BioChip whole blood adhesion assay to assess leukocyte adhesion to P-selectin under physiologic flow conditions. Our results demonstrated heterogeneous adhesion by leukocytes to immobilized P-selectin, and dose-dependent inhibition of this adhesion following pre-exposure to Crizanlizumab. Importantly, treatment with Crizanlizumab following adhesion to P-selectin promoted detachment of rolling, but not of firmly adherent leukocytes. Taken together, our results suggest that the microfluidic BioChip system is a promising in vitro assay with which to screen patients, monitor treatment response, and guide current and emerging anti-adhesive therapies in SCD.
Assuntos
Anemia Falciforme/tratamento farmacológico , Anticorpos Monoclonais Humanizados/farmacologia , Adesão Celular/efeitos dos fármacos , Leucócitos/efeitos dos fármacos , Selectina-P/antagonistas & inibidores , Adulto , Idoso , Feminino , Humanos , Dispositivos Lab-On-A-Chip/normas , Leucócitos/citologia , Masculino , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Técnicas Analíticas Microfluídicas/normas , Pessoa de Meia-Idade , Adulto JovemRESUMO
Understanding the contribution of relevant factors to the analytical variability of the micro-flow imaging (MFI) technique is of prime importance because of the significance of the subvisible particulate data in biopharmaceutical product development. The current study was performed to determine the contribution of several key variables to the variability of the subvisible particle counts (e.g., day-to-day, vial-to-vial, sample-to-sample, and measurement-to-measurement variabilities) using a nested statistical analysis. The variability was measured in the <10 µm, ≥10 µm, ≥25 µm, and ≥50 µm size ranges along with the total particle count and the maximum and the mean particle size. The contribution of the vial to the variability of the subvisible particle counts was found to be greater than those of the other factors evaluated in the current study. The analytical method variability in terms of percent relative standard deviation with respect to the particle count in the <10 µm, ≥10 µm, and ≥25 µm size ranges was found to be 16%, 40%, and 44%, respectively. A thorough understanding of the contribution of key factors to the analytical variability revealed how the corresponding contribution can be minimized, that is, by increasing the number of vials, samples, and measurements. The results of the current study may be leveraged for the optimization of the analytical method or for minimization of the analytical variability with the MFI technique.
Assuntos
Anticorpos Monoclonais/análise , Interpretação Estatística de Dados , Técnicas Analíticas Microfluídicas/métodos , Tamanho da Partícula , Liofilização/métodos , Liofilização/normas , Técnicas Analíticas Microfluídicas/normas , Agregados ProteicosRESUMO
The field of microfluidics-based three-dimensional (3D) cell culture system is rapidly progressing from academic proof-of-concept studies to valid solutions to real-world problems. Polydimethylsiloxane (PDMS)-based platform has been widely adopted as in vitro platforms for mimicking tumor microenvironment. However, PDMS has not been welcomed as a standardized commercial application for preclinical screening due to inherent material limitations that make it difficult to scale-up production. Here, we present an injection-molded plastic array 3D spheroid culture platform (Sphero-IMPACT). The platform is made of polystyrene (PS) in a standardized 96-well plate format with a user-friendly interface. This interface describes a simpler design that incorporates a tapered hole in the center of the rail to pattern a large spheroid with 3D extracellular matrix and various cell types. This hole is designed to accommodate standard pipette tip for automated system. The platform that mediate open microfluidics allows implement spontaneous fluid patterning with high repeatability from the end user. To demonstrate versatile use of the platform, we developed 3D perfusable blood vessel network and tumor spheroid assays. In addition, we established a tumor spheroid induced angiogenesis model that can be applicable for drug screening. Sphero-IMPACT has the potential to provide a robust and reproducible in vitro assay related to vascularized cancer research. This easy-to-use, ready-to-use platform can be translated into an enhanced preclinical model that faithfully reflects the complex tumor microenvironment.
Assuntos
Técnicas de Cultura de Células/normas , Glioblastoma/patologia , Técnicas Analíticas Microfluídicas/normas , Neovascularização Patológica/patologia , Esferoides Celulares/patologia , Técnicas de Cultura de Células/economia , Técnicas de Cultura de Células/instrumentação , Células Cultivadas , Células Endoteliais da Veia Umbilical Humana/citologia , Humanos , Técnicas Analíticas Microfluídicas/economia , Técnicas Analíticas Microfluídicas/instrumentação , Padrões de ReferênciaRESUMO
VereBeef™ Detection Kit, incorporating both multiplex PCR and microarray technologies on a lab-on-chip platform, is intended for qualitative detection and differentiation of Escherichia coli O157:H7, E. coli O26, E. coli O45, E. coli O103, E. coli O111, E. coli O121, E. coli O145, Shiga toxin-producing E. coli (STEC) virulence factors (stx1A, stx2A, eae), and Salmonella species in one test using raw beef trim samples. This product underwent extensive evaluations, including inclusivity-exclusivity, method comparison, robustness, lot-to-lot variability, and stability studies. The inclusivity/exclusivity study demonstrated that VereBeef Detection Kit specifically detects and identifies target analytes without occurrence of false-positive and false-negative detection. In the method comparison study, the performance of the VereBeef Detection Kit was compared with U.S. Department of Agriculture Food Safety and Inspection Service Microbiology Laboratory Guidebook's methods for target organism detection in raw beef trim using E. coli O157:H7 single inoculation and Salmonella and non-O157 STEC dual inoculation. Data demonstrated equivalence in both methods. The robustness study showed that changes in the test parameters do not impact assay performance. Collectively, VereBeef Detection Kit is able to detect target pathogens in raw beef trim with a minimum enrichment time of 8 h for E. coli O157:H7 detection and 10 h for Salmonella and non-O157 STEC detection.
Assuntos
Microbiologia de Alimentos , Carne/microbiologia , Técnicas Analíticas Microfluídicas/normas , Reação em Cadeia da Polimerase Multiplex/normas , Animais , Bovinos , Escherichia coli/classificação , Escherichia coli/isolamento & purificação , Salmonella/classificação , Salmonella/isolamento & purificaçãoRESUMO
The low specificity of the prostate-specific antigen (PSA) for early detection of prostate cancer (PCa) is a major issue worldwide. The aim of this study to examine whether the serum PCa-associated α2,3-linked sialyl N-glycan-carrying PSA (S2,3PSA) ratio measured by automated micro-total immunoassay systems (µTAS system) can be applied as a diagnostic marker of PCa. The µTAS system can utilize affinity-based separation involving noncovalent interaction between the immunocomplex of S2,3PSA and Maackia amurensis lectin to simultaneously determine concentrations of free PSA and S2,3PSA. To validate quantitative performance, both recombinant S2,3PSA and benign-associated α2,6-linked sialyl N-glycan-carrying PSA (S2,6PSA) purified from culture supernatant of PSA cDNA transiently-transfected Chinese hamster ovary (CHO)-K1 cells were used as standard protein. Between 2007 and 2016, fifty patients with biopsy-proven PCa were pair-matched for age and PSA levels, with the same number of benign prostatic hyperplasia (BPH) patients used to validate the diagnostic performance of serum S2,3PSA ratio. A recombinant S2,3PSA- and S2,6PSA-spiked sample was clearly discriminated by µTAS system. Limit of detection of S2,3PSA was 0.05 ng/mL and coefficient variation was less than 3.1%. The area under the curve (AUC) for detection of PCa for the S2,3PSA ratio (%S2,3PSA) with cutoff value 43.85% (AUC; 0.8340) was much superior to total PSA (AUC; 0.5062) using validation sample set. Although the present results are preliminary, the newly developed µTAS platform for measuring %S2,3PSA can achieve the required assay performance specifications for use in the practical and clinical setting and may improve the accuracy of PCa diagnosis. Additional validation studies are warranted.
Assuntos
Biomarcadores Tumorais , Polissacarídeos/química , Antígeno Prostático Específico/sangue , Antígeno Prostático Específico/química , Neoplasias da Próstata/sangue , Neoplasias da Próstata/diagnóstico , Idoso , Idoso de 80 Anos ou mais , Animais , Células CHO , Cricetulus , Detecção Precoce de Câncer/métodos , Detecção Precoce de Câncer/normas , Glicosilação , Humanos , Imunoensaio , Lectinas , Masculino , Técnicas Analíticas Microfluídicas/métodos , Técnicas Analíticas Microfluídicas/normas , Pessoa de Meia-Idade , Proteínas Recombinantes , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
A microfluidic platform or "microfluidic mapper" is demonstrated, which in a single experiment performs 36 parallel biochemical reactions with 36 different combinations of two reagents in stepwise concentration gradients. The volume used in each individual reaction was 36 nl. With the microfluidic mapper, we obtained a 3D enzyme reaction plot of horseradish peroxidase (HRP) with Amplex Red (AR) and hydrogen peroxide (H2O2), for concentration ranges of 11.7 µM to 100.0 µM and 11.1 µM to 66.7 µM for AR and H2O2, respectively. This system and methodology could be used as a fast analytical tool to evaluate various chemical and biochemical reactions especially where two or more reagents interact with each other. The generation of dual concentration gradients in the present format has many advantages such as parallelization of reactions in a nanoliter-scale volume and the real-time monitoring of processes leading to quick concentration gradients. The microfluidic mapper could be applied to various problems in analytical chemistry such as revealing of binding kinetics, and optimization of reaction kinetics.
Assuntos
Peroxidase do Rábano Silvestre/metabolismo , Peróxido de Hidrogênio/metabolismo , Dispositivos Lab-On-A-Chip , Técnicas Analíticas Microfluídicas , Calibragem , Desenho de Equipamento , Peroxidase do Rábano Silvestre/química , Imageamento Tridimensional , Cinética , Dispositivos Lab-On-A-Chip/normas , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Técnicas Analíticas Microfluídicas/normas , Microscopia de Fluorescência , Imagem com Lapso de TempoAssuntos
Coagulação Sanguínea , Vasos Sanguíneos/metabolismo , Dispositivos Lab-On-A-Chip/normas , Técnicas Analíticas Microfluídicas/normas , Modelos Anatômicos , Modelos Cardiovasculares , Trombose/sangue , Animais , Vasos Sanguíneos/patologia , Vasos Sanguíneos/fisiopatologia , Desenho de Equipamento , Humanos , Cinética , Técnicas Analíticas Microfluídicas/instrumentação , Fluxo Sanguíneo Regional , Reprodutibilidade dos Testes , Estresse Mecânico , Trombose/patologia , Trombose/fisiopatologiaRESUMO
A digital microfluidic biochip (DMFB) is an emerging technology that enables miniaturized analysis systems for point-of-care clinical diagnostics, DNA sequencing, and environmental monitoring. A DMFB reduces the rate of sample and reagent consumption, and automates the analysis of assays. In this paper, we provide the first assessment of the security vulnerabilities of DMFBs. We identify result-manipulation attacks on a DMFB that maliciously alter the assay outcomes. Two practical result-manipulation attacks are shown on a DMFB platform performing enzymatic glucose assay on serum. In the first attack, the attacker adjusts the concentration of the glucose sample and thereby modifies the final result. In the second attack, the attacker tampers with the calibration curve of the assay operation. We then identify denial-of-service attacks, where the attacker can disrupt the assay operation by tampering either with the droplet-routing algorithm or with the actuation sequence. We demonstrate these attacks using a digital microfluidic synthesis simulator. The results show that the attacks are easy to implement and hard to detect. Therefore, this work highlights the need for effective protections against malicious modifications in DMFBs.
Assuntos
Segurança Computacional/normas , Cibernética/normas , Técnicas Analíticas Microfluídicas/normas , Glicemia/análise , Humanos , Modelos TeóricosRESUMO
Echogenic particles, such as microbubbles and volatile liquid micro/nano droplets, have shown considerable potential in a variety of clinical diagnostic and therapeutic applications. The accurate prediction of their response to ultrasound excitation is however extremely challenging, and this has hindered the optimisation of techniques such as quantitative ultrasound imaging and targeted drug delivery. Existing characterisation techniques, such as ultra-high speed microscopy provide important insights, but suffer from a number of limitations; most significantly difficulty in obtaining large data sets suitable for statistical analysis and the need to physically constrain the particles, thereby altering their dynamics. Here a microfluidic system is presented that overcomes these challenges to enable the measurement of single echogenic particle response to ultrasound excitation. A co-axial flow focusing device is used to direct a continuous stream of unconstrained particles through the combined focal region of an ultrasound transducer and a laser. Both the optical and acoustic scatter from individual particles are then simultaneously recorded. Calibration of the device and example results for different types of echogenic particle are presented, demonstrating a high throughput of up to 20 particles per second and the ability to resolve changes in particle radius down to 0.1 µm with an uncertainty of less than 3%.
Assuntos
Microbolhas , Técnicas Analíticas Microfluídicas/métodos , Calibragem , Fluorocarbonos/química , Gases/química , Hidrodinâmica , Lasers , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/normas , Tamanho da Partícula , Razão Sinal-Ruído , SonicaçãoRESUMO
The objective of the study has been the development of a new sensing platform, called Real-time Electrochemical Profiling (REP) that relies on real-time electrochemical immunoassay detection. The proposed REP platform consists of new electrode arrays that are easy to fabricate, has a small imprint allowing microfluidic system integration, enables multiplexed amperometric measurements and performs well in terms of electrochemical immunoassay detection as shown through the deoxynivalenol detection assays. The deoxynivalenol detection has been conducted according to an optimised REP assay protocol using deoxynivalenol standards at varying concentrations and a standard curve was obtained (y=-20.33ln(x)+124.06; R(2)=0.97) with a limit of detection of 6.25 ng/ml. As both ELISA and REP detection methods use horse radish peroxidase as the label and 3.3',5.5'-Tetramethylbenzidine as the substrate, the performance of the REP platform as an ELISA reader has also been investigated and a perfect correlation between the deoxynivalenol concentration and the current response was obtained (y=-14.56ln(x)+101.02; R(2)=0.99). The calibration curves of both assays have been compared to conventional ELISA tests for confirmation. After assay optimisation using toxin spiked buffer, the deoxynivalenol detection assay has also been performed to detect toxins in wheat grain.
Assuntos
Técnicas Biossensoriais/métodos , Contaminação de Alimentos/análise , Técnicas Analíticas Microfluídicas/métodos , Micotoxinas/análise , Triticum/química , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/normas , Sistemas Computacionais , Técnicas Eletroquímicas , Ensaio de Imunoadsorção Enzimática , Humanos , Imunoensaio/instrumentação , Imunoensaio/métodos , Imunoensaio/normas , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/normas , Micotoxinas/normas , Padrões de Referência , Tricotecenos/análise , Tricotecenos/normasRESUMO
Cavity optomechanics experiments that parametrically couple the phonon modes and photon modes have been investigated in various optical systems including microresonators. However, because of the increased acoustic radiative losses during direct liquid immersion of optomechanical devices, almost all published optomechanical experiments have been performed in solid phase. This paper discusses a recently introduced hollow microfluidic optomechanical resonator. Detailed methodology is provided to fabricate these ultra-high-Q microfluidic resonators, perform optomechanical testing, and measure radiation pressure-driven breathing mode and SBS-driven whispering gallery mode parametric vibrations. By confining liquids inside the capillary resonator, high mechanical- and optical- quality factors are simultaneously maintained.
Assuntos
Técnicas Analíticas Microfluídicas/instrumentação , Oscilometria/instrumentação , Desenho de Equipamento , Técnicas Analíticas Microfluídicas/normas , Óptica e Fotônica/instrumentação , Óptica e Fotônica/métodos , Óptica e Fotônica/normas , Oscilometria/métodos , Oscilometria/normasRESUMO
An extremely simple, self-standing microfluidic cell culture system is reported. The whole system is confined in a 35 mm culture dish, and requires only a standard CO2 incubator. The culture medium is perfused by gravity. We successfully cultured NIH3T3-derived cells up to 10 days with a viability of â¼90%.
Assuntos
Técnicas de Cultura de Células/instrumentação , Técnicas Analíticas Microfluídicas/instrumentação , Animais , Técnicas de Cultura de Células/normas , Proliferação de Células , Meios de Cultura , Camundongos , Técnicas Analíticas Microfluídicas/normas , Células NIH 3T3 , Perfusão/métodosRESUMO
Acoustofluidic devices for manipulating microparticles in fluids are appealing for biological sample processing due to their gentle and high-speed capability of sorting cell-scale objects. Such devices are generally limited to moving particles toward locations at integer fractions of the fluid channel width (1/2, 1/4, 1/6, etc.). In this work, we introduce a unique approach to acoustophoretic device design that overcomes this constraint, allowing us to design the particle focusing location anywhere within the microchannel. This is achieved by fabricating a second fluid channel in parallel with the sample channel, separated from it by a thin silicon wall. The fluids in both channels participate to create the ultrasound resonance, while only one channel processes the sample, thus de-coupling the fluidic and acoustic boundaries. The wall placement and the relative widths of the adjacent channels define the particle focusing location. We investigate the operating characteristics of a range of these devices to determine the configurations that enable effective particle focusing and separation. The results show that a sufficiently thin wall negligibly affects focusing efficiency and location compared to a single channel without a wall, validating the success of this design approach without compromising separation performance. Using these principles to design and fabricate an optimized device configuration, we demonstrate high-efficiency focusing of microspheres, as well as separation of cell-free viruses from mammalian cells. These "transparent wall" acoustic devices are capable of over 90% extraction efficiency with 10 µm microspheres at 450 µL min(-1), and of separating cells (98% purity), from viral particles (70% purity) at 100 µL min(-1).
Assuntos
Acústica , Vírus da Dengue/isolamento & purificação , Técnicas Analíticas Microfluídicas/métodos , Tamanho da Partícula , Animais , Bioengenharia/métodos , Chlorocebus aethiops , Técnicas Analíticas Microfluídicas/normas , Microesferas , Células VeroAssuntos
Microbiologia do Ar , Bactérias/isolamento & purificação , Fungos/isolamento & purificação , Técnicas Analíticas Microfluídicas/estatística & dados numéricos , Vírus/isolamento & purificação , Humanos , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/normas , Microfluídica , Técnicas de Amplificação de Ácido NucleicoRESUMO
Mesoporous silica nanoparticles are promising drug delivery agents; however, their interaction with various in vivo biological components is still under investigation. In this work, the impact of sub-50 nm diameter mesoporous silica nanoparticles on platelet function is investigated using a microfluidic platform to model blood vessel characteristics. Platelet adhesion and aggregation in the presence of mesoporous silica nanoparticles is investigated, controlling whether or not platelets are activated ahead of nanoparticle exposure. The results indicate that nanoparticles slightly compromise platelet adhesion to endothelial cells at low nanoparticle doses, but that high nanoparticle doses significantly increase the number of platelet adhesion events, leading to higher probability for uncontrolled platelet actions (e.g. clot formation in vivo). High nanoparticle doses also induced platelet aggregation. While platelet activation and aggregation occurred, in no case did nanoparticle exposure result in significant loss of platelet viability; as such, this work clearly demonstrates that aspects besides viability, such as cellular adhesion and interaction with other cell types, have to be considered in the context of nanotoxicology. This simple and highly adaptable analytical platform will be useful for further nanotoxicity studies involving other nanoparticle and cell types.
Assuntos
Adesão Celular/fisiologia , Técnicas Analíticas Microfluídicas/normas , Nanopartículas/toxicidade , Agregação Plaquetária/fisiologia , Dióxido de Silício/toxicidade , Adesão Celular/efeitos dos fármacos , Células Endoteliais/química , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Humanos , Técnicas Analíticas Microfluídicas/métodos , Agregação Plaquetária/efeitos dos fármacos , Porosidade/efeitos dos fármacosRESUMO
The human signal-molecule-profiling database (HSMPD) is designed as a prospective medical database for translational bioinformatics (TBI). To explore the feasibility of low-cost database construction, we studied the roadmap of HSMPD. A HSMPD-oriented tool, called "signal-molecule-profiling (SMP) chip" was developed for data acquisition, which can be employed in the routine blood tests in hospitals; the results will be stored in the HSMPD system automatically. HSMPD system can provide data services for the TBI community, which generates a stable income to support the data acquisition. The small-scale experimental test was performed in the hospital to verify SMP chips and the demo HSMPD software. One hundred and eighty nine complete SMP records were collected, and the demo HSMPD system was also evaluated in the survey study on patients and doctors. The function of SMP chip was verified, whereas the demo HSMPD software needed to be improved. The survey study showed that patients would only accept free tests of SMP chips when they originally needed blood examinations. The study indicated that the construction of HSMPD relies on the self-motivated cooperation of the TBI community and the traditional healthcare system. The proposed roadmap potentially provides an executable solution to build the HSMPD without high costs.