We demonstrate a highly efficient method for gene delivery into clinically relevant human cell types, such as induced pluripotent stem cells (iPSCs) and fibroblasts, reducing the protocol time by one full day. To preserve cell physiology during gene transfer, we designed a microfluidic strategy, which facilitates significant gene delivery in a short transfection time (<1 min) for several human cell types. This fast, optimized and generally applicable cell transfection method can be used for rapid screening of different delivery systems and has significant potential for high-throughput cell therapy applications.
Polyaniline nanoparticles were synthesized by simple electrochemical polymerization of aniline in systems (a) aniline-sodium dodecyl sulfate (NaDS)-dodecylbenzene sulfonic acid (DBSA)-H(2)O and (b) aniline-NaDS-DBSA-cetyltrimethyl ammonium bromide (CTAB)-H(2)O. Different morphologies including compact and fractal/dendrimer were observed at different experimental conditions. Fractal dimension was calculated by the box counting method. Growth kinetics during electropolymerization of aniline in both of the systems was studied by measuring the weight of polymer aggregates as a function of time. Growth rate was found to be reduced in system (b) due to coordination of CTAB with the growing polyaniline chain. The weight of polymer aggregates was found to depend on field intensity and attains a maximum value at a critical field intensity 4.0 V/cm. Beyond this critical field intensity, the growth rate was decreased due to loss of conjugation and degradation of the polymer backbone. Electropolymerized aggregates were characterized by transmission electron microscopy (TEM), powder X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), and electrical conductivity measurements. Nanosized polyaniline was formed, with particle diameters in the range of 10-200 nm, as evident by TEM studies and supported by XRD studies. FT-IR spectroscopy established the formation of hyperbranched polyaniline chains. During electropolymerization, oscillations in potential were monitored as a function of time at different experimental conditions. A suitable mechanism for fractal growth of polyaniline was also proposed.
Long-read genomic applications, such as genome mapping in nanochannels, require long DNA that is free of small-DNA impurities. We have developed a chip-based system based on entropic trapping that can simultaneously concentrate and purify a long DNA sample under the alternating application of an applied pressure (for sample injection) and an electric field (for filtration and concentration). In contrast, short DNA tends to pass through the filter owing to its comparatively weak entropic penalty for entering the nanoslit. The single-stage prototype developed here, which operates in a continuous pulsatile manner, achieves selectivities of up to 3.5 for λ-phage DNA (48.5 kilobase pairs) compared to a 2 kilobase pair standard based on experimental data for the fraction filtered using pure samples of each species. The device is fabricated in fused silica using standard clean-room methods, making it compatible for integration with long-read genomics technologies.
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