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1.
Lab Chip ; 10(23): 3284-90, 2010 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-20938545

RESUMO

This paper reports a novel microfluidic-chip based platform using "phase-transfer magnetophoresis" enabling continuous biomolecule processing. As an example we demonstrate for the first time continuous DNA extraction from cell lysate on a microfluidic chip. After mixing bacterial Escherichia coli culture with superparamagnetic bead suspension, lysis and binding buffers, DNA is released from cells and captured by the beads. These DNA carrying beads are continuously transported across the interfaces between co-flowing laminar streams of sample mixture, washing and elution buffer. Bead actuation is achieved by applying a time-varying magnetic field generated by a rotating permanent magnet. Flagella-like chains of magnetic beads are formed and transported along the microfluidic channels by an interplay of fluid drag and periodic magnetic entrapment. The turnover time for DNA extraction was approximately 2 minutes with a sample flow rate of 0.75 µl s(-1) and an eluate flow rate of 0.35 µl s(-1). DNA recovery was 147% (on average) compared to bead based batch-wise extraction in reference tubes within a dilution series experiment over 7 orders of magnitude. The novel platform is suggested for automation of various magnetic bead based applications that require continuous sample processing, e.g. continuous DNA extraction for flow-through PCR, capture and analysis of cells and continuous immunoassays. Potential applications are seen in the field of biological safety monitoring, bioprocess control, environmental monitoring, or epidemiological studies such as monitoring the load of antibiotic resistant bacteria in waste water from hospitals.


Assuntos
Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Eletroforese/instrumentação , Dispositivos Lab-On-A-Chip , Procedimentos Analíticos em Microchip/métodos , Automação , DNA/química , Eletroforese/métodos , Desenho de Equipamento , Escherichia coli/metabolismo , Humanos , Imunoensaio/métodos , Magnetismo , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo
2.
Nucleic Acids Res ; 36(5): 1443-9, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18203749

RESUMO

The interaction of cationic surfactants with single dsDNA molecules has been studied using force-measuring optical tweezers. For hydrophobic chains of length 12 and greater, pulling experiments show characteristic features (e.g. hysteresis between the pulling and relaxation curves, force-plateau along the force curves), typical of a condensed phase (compaction of a long DNA into a micron-sized particle). Depending on the length of the hydrophobic chain of the surfactant, we observe different mechanical behaviours of the complex (DNA-surfactants), which provide evidence for different binding modes. Taken together, our measurements suggest that short-chain surfactants, which do not induce any condensation, could lie down on the DNA surface and directly interact with the DNA grooves through hydrophobic-hydrophobic interactions. In contrast, long-chain surfactants could have their aliphatic tails pointing away from the DNA surface, which could promote inter-molecular interactions between hydrophobic chains and subsequently favour DNA condensation.


Assuntos
DNA/química , Tensoativos/química , Cátions/química , Cetrimônio , Compostos de Cetrimônio/química , Modelos Químicos , Pinças Ópticas , Compostos de Amônio Quaternário/química
3.
Anal Chim Acta ; 929: 1-22, 2016 07 27.
Artigo em Inglês | MEDLINE | ID: mdl-27251944

RESUMO

The last decade has witnessed tremendous advances in employing microfluidic solutions enabling Continuous Processing and Monitoring of Biological Samples (CPMBS), which is an essential requirement for the control of bio-processes. The microfluidic systems are superior to the traditional inline sensors due to their ability to implement complex analytical procedures, such as multi-step sample preparation, and enabling the online measurement of parameters. This manuscript provides a backgound review of microfluidic approaches employing laminar flow, hydrodynamic separation, acoustophoresis, electrophoresis, dielectrophoresis, magnetophoresis and segmented flow for the continuous processing and monitoring of biological samples. The principles, advantages and limitations of each microfluidic approach are described along with its potential applications. The challenges in the field and the future directions are also provided.


Assuntos
Técnicas de Química Analítica/métodos , Técnicas Analíticas Microfluídicas/métodos , Animais , Técnicas de Química Analítica/instrumentação , Desenho de Equipamento , Humanos , Hidrodinâmica , Técnicas Analíticas Microfluídicas/instrumentação
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