Yougen Li

Dendrimer-like DNA-based fluorescence nanobarcodes.

A major challenge in clinical diagnostics and environmental analysis is the difficulty in rapid and sensitive detection of multiple target molecules simultaneously (i.e., multiplexed detections). Our group has designed and synthesized a dendrimer-like DNA (DL-DNA) that is multivalent and anisotropic; using this unique DNA structure, we have developed a fluorescence-tagged nanobarcode system for multiplex detection. This nanobarcode system allows the rapid and sensitive detection of multiple pathogens simultaneously using the ratios of two different fluorescent dyes, green and red, with which different DL-DNAs are labeled. The key step of our nanobarcode model lies in the monodisperse preparation of DL-DNA. Two methods, solution phase and solid phase, are presented here. With slight modifications, this platform technology can also be extended to the multiplexed detection of RNA and proteins. This protocol can be completed in 2–5 d.

A new helper phospholipid for gene delivery.

Enhanced gene transfection activity is observed when using a new helper lipid with DOTAP, compared to DOPE.

Single-molecule mobility and spectral measurements in submicrometer fluidic channels

Electrophoretic mobility differences of biological molecules are frequently exploited to physically separate and subsequently identify the components of a mixture. We present a method to rapidly identify single molecules by measuring both their mobility and fluorescence emission under continuous flow without separation. Submicrometer fluidic channels were used to detect individual nucleic-acid-engineered fluorescent labels driven electrokinetically in free solution. Two separate focal volumes along the length of the fluidic channel collected spectral, spatial, and temporal information from the passage of fluorescent labels through the channel. One focal volume was defined by a focused 488-nm-wavelength laser and the other by a focused 568-nm laser. The subfemtoliter focal volumes resulted in signal-to-noise ratios sufficient for single-fluorophore detection, and the two excitation wavelengths enabled detection of multicolor fluorescent labels and discrimination of single-color detection events. Each fluor...

A dendrimer-like DNA-based vector for DNA delivery: a viral and nonviral hybrid approach.

DNA can be used as a generic delivery vector in addition to its genetic role as a antigen expression vector. This is inspired in part by the fact that DNA molecules are true polymers. Surprisingly, DNA molecules have not been used as a delivery vector material. This is probably due to the fact that almost all DNA have only two shapes: linear or circular. This chapter details our efforts in fabricating highly branched dendrimer-like DNA (DL-DNA) that may serve as a multivalent DNA delivery vector. Just like chemical dendrimers, DL-DNA is multi-valent and monodisperse. However, unlike traditional chemical dendrimers, DL-DNA is much larger (~100 nm, generation 4) and can be designed to be nonsymmetric as well. Most importantly, DL-DNA possesses two unique properties: anisotropicity and biodegradability, making multiple, specific conjugations of viral peptides possible. Our method suggests that viral-pep-tide conjugated DL-DNA vectors can deliver genes into cells without any other transfection reagents. This viral-nonviral hybrid system can be further tailored to specific cells by conjugating specific ligands. We believe that such a DL-DNA-based, viral, and nonviral hybrid assembly will provide a new platform for drug delivery in general and gene delivery in particular.

Improvement of the low-frequency sensitivity of MgO-based magnetic tunnel junctions by annealing

Magnetic tunnel junctions can serve as ultrasensitive low-frequency magnetic sensors, however, their low-frequency performance is limited by low-frequency noise, i.e., 1/f noise. In this paper, we investigate the 1/f noise in MgO magnetic tunnel junctions (MTJs) with a tunneling magnetoresistance (TMR) of 160%, and examine the influence of annealing and MTJ size. The results show that the annealing process can not only dramatically improve the TMR, but can also strongly decrease the MTJ noise. The effect is discussed in terms of the structure of MgO barriers and tunneling probabilities. Increasing the MTJ area to 6400 μm2 yields a voltage spectral density as low as 11 nV/Hz1/2 at 1000 Hz. The possible reasons for the area dependence are discussed.

Multiplexed molecular detection using encoded microparticles and nanoparticles

Signal-encoded microparticles and nanoparticles have been used to label many reactions simultaneously for target identification in assays, and thus are an indispensable part of multiplex technologies. With the increasing demand for multiplexed molecular detection, encoded particles have evolved from pattern encoding to signal-intensity encoding, and also from signal-molecule encapsulation to signal-molecule tagging. The fabrication and utilization of such nano- and microparticles should advance multiplexed analysis. This short review focuses on how these encoded particles work and briefly touches on their applications in multiplexed molecular detection.

Design and Validation of High-Efficiency Chopper for Magnetoresistive Sensors

Low-frequency noise, i.e., 1/f noise severely limits the low-frequency performance of magnetoresistive sensors for applications as ultrasensitive magnetic sensors. The combination of a flux concentrator and chopping system has been proposed to mitigate the effect of 1/f noise and improve the field sensitivity. This paper reports a high-efficiency chopping system for magnetoresistive sensors. A simple reluctance analysis allows us to estimate the chopping efficiency and optimize the design, and finite-element simulations confirm the operation. Experimental results based on the optimized design using an anisotropic magnetoresistance sensor validate the concept. The sensitivity of the sensor with the chopper at OFF and ON positions is determined to be 0.009 and 0.067%/Oe, respectively, which yields a high sensitivity chopping efficiency, namely 72%.

Non-viral Charge Reversal Vectors for pDNA Delivery

A synthetic vector that transform from a cationic to an anionic lipid intracellularly is described. This charge-reversal lipid was synthesized and characterized, including the supramolecular complex it forms with DNA. Enhanced gene transfection was observed using this synthetic vector compared to current cationic lipids