Continuous particle separation through deterministic lateral displacement.

We report on a microfluidic particle-separation device that makes use of the asymmetric bifurcation of laminar flow around obstacles. A particle chooses its path deterministically on the basis of its size. All particles of a given size follow equivalent migration paths, leading to high resolution. The microspheres of 0.8, 0.9, and 1.0 micrometers that were used to characterize the device were sorted in 40 seconds with a resolution of approximately 10 nanometers, which was better than the time and resolution of conventional flow techniques. Bacterial artificial chromosomes could be separated in 10 minutes with a resolution of approximately 12%.

Tilted brownian ratchet for DNA analysis.

In this paper, we report a factor of 3 improvement in the resolution and a factor of 10 improvement in the speed of fractionation of approximately 100-kb DNA molecules in Brownian ratchet arrays. In our device, the electrophoretic flow is tilted at a small angle relative to the array axis. Tilting accelerates the fractionation speed because a higher fraction of the diffusing molecules is "ratcheted" at each step in the array. Molecules of lengths 48.5 and 164 kb can be separated in approximately 70 min with a resolution of approximately 3.8, using a 12-mm-long array. The Brownian ratchet arrays are not limited to DNA separation, but can, in principle, be used for any particle in this size range.

Role of molecular size in ratchet fractionation.

We show the importance of finite particle size in microfluidic asymmetric continuous-flow diffusion arrays, specifically the critical nature of the particle size with respect to the barrier gaps. We show that particles much smaller than the barrier gap follow individual field lines through narrow gaps and are poorly fractionated. In contrast, particles comparable to the gap size lose memory of their incoming field line and can be fractionated with high resolution. We demonstrate this effect using a new technological approach to create very straight and narrow injection bands in such arrays, and completely resolve bands of DNA of lengths 48,500 and 16,7000 base pairs.

A DNA prism for high-speed continuous fractionation of large DNA molecules.

The analysis and fractionation of large DNA molecules plays a key role in many genome projects. The standard method, pulsed-field gel electrophoresis (PFGE), is slow, with running times ranging from 10 hours to more than 200 hours. In this report, we describe a thumbnail-sized device that sorts large DNA fragments (61-209 kilobases (kb)) in 15 seconds, with a resolution of approximately 13%. An array of micron-scale posts serves as the sieving matrix, and integrated microfluidic channels spatially shape the electric fields over the matrix. Asymmetric pulsed fields are applied for continuous-flow operation, which sorts DNA molecules in different directions according to their molecular masses, much as a prism deflects light of different wavelengths at different angles. We demonstrate the robustness of the device by using it to separate large DNA inserts prepared from bacterial artificial chromosomes, a widely used DNA source for most genomics projects.