Microfluidic methods for barcoding nucleic acid target molecules to be analyzed, e.g., via nucleic acid sequencing techniques, are provided. Also provided are microfluidic, droplet-based methods of preparing nucleic acid barcodes for use in various barcoding applications. The methods described herein facilitate high-throughput sequencing of nucleic acid target molecules as well as single cell and single virus genomic, transcriptomic, and/or proteomic analysis/profiling. Systems and devices for practicing the subject methods are also provided.

Various aspects of the present invention relate to the control and manipulation of fluidic species, for example, in microfluidic systems. In one aspect, the invention relates to systems and methods for making droplets of fluid surrounded by a liquid, using, for example, electric fields, mechanical alterations, the addition of an intervening fluid, etc. In some cases, the droplets may each have a substantially uniform number of entities therein. For example, 95% or more of the droplets may each contain the same number of entities of a particular species. In another aspect, the invention relates to systems and methods for dividing a fluidic droplet into two droplets, for example, through charge and/or dipole interactions with an electric field. The invention also relates to systems and methods for fusing droplets according to another aspect of the invention, for example, through charge and/or dipole interactions. In some cases, the fusion of the droplets may initiate or determine a reaction. In a related aspect of the invention, systems and methods for allowing fluid mixing within droplets to occur are also provided. In still another aspect, the invention relates to systems and methods for sorting droplets, e.g., by causing droplets to move to certain regions within a fluidic system. Examples include using electrical interactions (e.g., charges, dipoles, etc.) or mechanical systems (e.g., fluid displacement) to sort the droplets. In some cases, the fluidic droplets can be sorted at relatively high rates, e.g., at about 10 droplets per second or more. Another aspect of the invention provides the ability to determine droplets, or a component thereof, for example, using fluorescence and/or other optical techniques (e.g., microscopy), or electric sensing techniques such as dielectric sensing.

A multimodal micromixer obstacle for intensification of mixing and performing the reaction in a continuous manner is disclosed herein. The micromixer 100 comprises of plurality of inlets, an outlet and a plurality of channels. The end channelsof the channels, have pluralityof converging sections having width, to depth ratio ranging 1:1 to 20:1. The intermediate channels have at least, one obstacle having non-circular shape. Each converging section is incomplete ellipse, prolate or oblate shaped having, angle of curvature in the range of 90 to 270. Axes of the inlets are coplanar and perpendicular to the channels. All the components of the micromixer are coplanar.

Disclosed herein are fluidic mixers having bifurcated fluidic flow through toroidal mixing elements. The mixers operate, at least partially, by Dean vortexing. Accordingly, the mixers are referred to as Dean Vortex Bifurcating Mixers (DVBM). The DVBM utilize Dean vortexing and asymmetric bifurcation of the fluidic channels that form the mixers to achieve the goal of optimized microfluidic mixing. The disclosed DVBM mixers can be incorporated into any fluidic (e.g., microfluidic) device known to those of skill in the art where mixing two or more fluids is desired. The disclosed mixers can be combined with any fluidic elements known to those of skill in the art, including syringes, pumps, inlets, outlets, non-DVBM mixers, heaters, assays, detectors, and the like.

The invention describes a method for the synthesis of compounds comprising the steps of: (a) compartmentalizing two or more sets of primary compounds into microcapsules; such that a proportion of the microcapsules contains two or more compounds; and (b) forming secondary compounds in the microcapsules by chemical reactions between primary compounds from different sets; wherein one or both of steps (a) and (b) is performed under microfluidic control; preferably electronic microfluidic control The invention further allows for the identification of compounds which bind to a target component of a biochemical system or modulate the activity of the target, and which is co-compartmentalized into the microcapsules.

The invention describes a method for isolating one or more genetic elements encoding a gene product having a desired activity, comprising the steps of: (a) compartmentalising genetic elements into microcapsules; and (b) sorting the genetic elements which express the gene product having the desired activity; wherein at least one step is under microfluidic control. The invention enables the in vitro evolution of nucleic acids and proteins by repeated mutagenesis and iterative applications of the method of the invention.

Ancillary embodiments and modifications to a homogenizer unit (PPH), and methods of use directed to purification of biogas or other raw methane streams. The apparatus includes a homogenizer body, one or more stream inlets (for the raw methane), one or more chilled water inlets, a mixing zone where the water stream is commingled with the raw methane stream, and a venturi immediately downstream from the mixing zone such that the commingled streams are pulled into the venturi resulting in homogenization. The PPH components are insulated to maintain the chilled water of the various streams at a cooled, below ambient temperature, increasing dissolution of the contaminant gases into the chilled water, and producing a purified methane stream including little or no H.sub.2S and CO.sub.2.

The invention describes a method for isolating one or more genetic elements encoding a gene product having a desired activity, comprising the steps of: (a) compartmentalising genetic elements into microcapsules; and (b) sorting the genetic elements which express the gene product having the desired activity; wherein at least one step is under microfluidic control. The invention enables the in vitro evolution of nucleic acids and proteins by repeated mutagenesis and iterative applications of the method of the invention.

The present invention relates to a microfluidic separation device, a separation method using the same and a kit for separating circulating rare cells from blood using the same, and more particularly, to a microfluidic-based separation technology for fixing target particles of a sample, which have a specific affinity for magnetic nanoparticles, to a device by use of a magnetic material, and for isolating the sample from which the target particles have been removed. The present invention may be effectively applied to remove leukocytes from a blood sample in order to isolate circulating rare cells (CRCs), particularly circulating tumor cells (CTCs).

A static mixer device comprising a housing having a proximal end, a distal end, and an opening extending between the proximal and distal ends. In certain embodiments, a plurality of metal frits is positioned within the opening of the housing, each of the metal frits extending across a cross-sectional dimension of the opening and having interconnected porosity. In other embodiments, one or more mixer elements fabricated using laser additive manufacturing technology and having novel configurations are positioned within the opening of the housing. In yet other embodiments, the housing comprises multiple openings having different diameters from each other, with each opening either extending through the housing with a constant diameter or with one or more of the openings having a varying diameter.