Molecular biology on a microfluidic chip

We have developed microfluidic chips for automating molecular biology processes such as gene ligation and gene transformation from nanolitre sample volumes with parallel architecture. Unlike conventional tube methods with cumbersome pipetting procedures, all processes, including metering of samples, ligation and transformation, were carried out in the microfluidic chips through pneumatic control of the nanofluid. The microfluidic devices presented here offer an illustration of some of the basic physics that arises when trying to miniaturize and automate biological techniques

Electrical microfluidic pressure gauge for elastomer microelectromechanical systems

We report on an electrical microfluidic pressure gauge. A polydimethylsiloxane microvalve closes at a characteristic applied pressure determined by the material's properties and the valve's dimensions. Hence, when the same pressure is applied to all valves of a heterogeneous valve array, some valves close while others remain open. The state of the array is combined with knowledge of the respective characteristic closing pressures of the individual valves to yield an estimate of the applied pressure. The state of each valve is obtained by electrical measurements, since the electrical resistance of the respective underlying fluid-filled channel increases by at least two orders of magnitude as the valve closes and its insulating elastomer material interrupts the electrical circuit. The overall system functions as a pressure gauge with electrical readout. This device would be a critical component in active pressure-regulation loops in future integrated microfluidic systems

Experimentally validated quantitative linear model for the device physics of elastomeric microfluidic valves

A systematic experimental study and theoretical modeling of the device physics of polydimethylsiloxane “pushdown” microfluidic valves are presented. The phase space is charted by 1587 dimension combinations and encompasses 45–295 µm lateral dimensions, 16–39 µm membrane thickness, and 1–28 psi closing pressure. Three linear models are developed and tested against the empirical data, and then combined into a fourth-power-polynomial superposition. The experimentally validated final model offers a useful quantitative prediction for a valve's properties as a function of its dimensions. Typical valves (80–150 µm width) are shown to behave like thin springs

Parallel Picoliter RT-PCR Assays Using Microfluidics

The development of microfluidic tools for high-throughput nucleic acid analysis has become a burgeoning area of research in the post-genome era. Here, we have developed a microfluidic chip to perform 72 parallel 450-pL RT-PCRs. We took advantage of Taqman hydrolysis probe chemistry to detect RNA templates as low as 34 copies. The device and method presented here may enable highly parallel single cell gene expression analysis

Scaling properties of a low-actuation pressure microfluidic valve

Using basic physical arguments, we present a design and method for the fabrication of microfluidic valves using multilayer soft lithography. These on-off valves have extremely low actuation pressures and can be used to fabricate active functions, such as pumps and mixers in integrated microfluidic chips. We characterized the performance of the valves by measuring both the actuation pressure and flow resistance over a wide range of design parameters, and compared them to both finite element simulations and alternative valve geometries

Microfluidic Single-Cell mRNA Isolation and Analysis

Single-cell gene expression analysis holds great promise for studying diverse biological systems, but methodology to process these precious samples in a reproducible, quantitative, and parallel fashion remains challenging. Here, we utilize microfluidics to isolate picogram and subpicogram mRNA templates, as well as to synthesize cDNA from these templates. We demonstrate single-cell mRNA isolation and cDNA synthesis, provide quantitative calibrations for each step in the process, and measure gene expression in individual cells. The techniques presented here form the foundation for highly parallel single-cell gene expression studies

Parallel Picoliter RT-PCR Assays Using Microfluidics

The development of microfluidic tools for high-throughput nucleic acid analysis has become a burgeoning area of research in the post-genome era. Here, we have developed a microfluidic chip to perform 72 parallel 450-pL RT-PCRs. We took advantage of Taqman hydrolysis probe chemistry to detect RNA templates as low as 34 copies. The device and method presented here may enable highly parallel single cell gene expression analysis

A nanoliter-scale nucleic acid processor with parallel architecture

The purification of nucleic acids from microbial and mammalian cells is a crucial step in many biological and medical applications1. We have developed microfluidic chips for automated nucleic acid purification from small numbers of bacterial or mammalian cells. All processes, such as cell isolation, cell lysis, DNA or mRNA purification, and recovery, were carried out on a single microfluidic chip in nanoliter volumes without any pre- or postsample treatment. Measurable amounts of mRNA were extracted in an automated fashion from as little as a single mammalian cell and recovered from the chip. These microfluidic chips are capable of processing different samples in parallel, thereby illustrating how highly parallel microfluidic architectures can be constructed to perform integrated batch-processing functionalities for biological and medical applications

Some comments on "The Mathematical Universe"

I discuss some problems related to extreme mathematical realism, focusing on a recently proposed "shut-up-and-calculate" approach to physics (arXiv:0704.0646, arXiv:0709.4024). I offer arguments for a moderate alternative, the essence of which lies in the acceptance that mathematics is (at least in part) a human construction, and discuss concrete consequences of this--at first sight purely philosophical--difference in point of view.Comment: 11 page