The invention generally relates to methods for quantifying an amount of enzyme molecules. Systems and methods of the invention are provided for measuring an amount of target by forming a plurality of fluid partitions, a subset of which include the target, performing an enzyme-catalyzed reaction in the subset, and detecting the number of partitions in the subset. The amount of target can be determined based on the detected number.

The invention generally relates to methods for quantifying an amount of enzyme molecules. Systems and methods of the invention are provided for measuring an amount of target by forming a plurality of fluid partitions, a subset of which include the target, performing an enzyme-catalyzed reaction in the subset, and detecting the number of partitions in the subset. The amount of target can be determined based on the detected number.

Methods and devices are provided for manipulating droplets on a support using surface tension properties, moving the droplets along a predetermined path and merging two droplets together enabling a number of chemical reactions. Disclosed are methods for controlling the droplets volumes. Disclosed are methods and devices for synthesizing at least one oligonucleotide having a predefined sequence. Disclosed are methods and devices for synthesizing and/or assembling at least one polynucleotide product having a predefined sequence from a plurality of different oligonucleotides having a predefined sequence. In exemplary embodiments, the methods involve synthesis and/or amplification of different oligonucleotides immobilized on a solid support, release of synthesized/amplified oligonucleotides in solution to form droplets, recognition and removal of error-containing oligonucleotides, moving or combining two droplets to allow hybridization and/or ligation between two different oligonucleotides, and further chain extension reaction following hybridization and/or ligation to hierarchically generate desired length of polynucleotide products.

Methods for the automated template-free synthesis of user-defined sequence controlled biopolymers using microfluidic devices are described. The methods facilitate simultaneous synthesis of up to thousands of uniquely addressed biopolymers from the controlled movement and combination of regents as fluid droplets using microfluidic and EWOD-based systems. In some forms, biopolymers including nucleic acids, peptides, carbohydrates, and lipids are synthesized from step-wise assembly of building blocks based on a user-defined sequence of droplet movements. In some forms, the methods synthesize uniquely addressed nucleic acids of up to 1,000 nucleotides in length. Methods for adding, removing and changing barcodes on biopolymers are also provided. Biopolymers synthesized according to the methods, and libraries and databases thereof are also described. Modified biopolymers, including chemically modified nucleotides and biopolymers conjugated to other molecules are described.

The present invention generally relates to droplet libraries and to systems and methods for the formation of libraries of droplets. The present invention also relates to methods utilizing these droplet libraries in various biological, chemical, or diagnostic assays.

The invention generally relates to methods for quantifying an amount of enzyme molecules. Systems and methods of the invention are provided for measuring an amount of target by forming a plurality of fluid partitions, a subset of which include the target, performing an enzyme-catalyzed reaction in the subset, and detecting the number of partitions in the subset. The amount of target can be determined based on the detected number.

Microarray platforms and methods of fabricating said microarrays without traditional high aspect ratio barriers used to define individual array elements are described herein. Self-assembled nanoshells were stabilized with a polymerized scaffold to enhance the stability in physiological conditions and serve as an optical transducer upon molecular recognition events. Soft photolithography combined with surface chemistry was developed for covalent immobilization of nanoshells onto the pre-patterned arrayed microspots for rapid multiplexed detection of membrane-binding analytes. This robust fabrication methodology is amenable for general lipid structures, and thus facilitates the integration of stable membrane architectures into diagnostic and prognostic platforms. In particular, the microarray platform may be used in diverse applications ranging from the detection of pathogens, such bacterial toxin in biological matrices, to cellular membrane studies.

The invention provides a means of generation of high numbers of droplets containing content while still diluting the interfering molecules to allow generating large arrays of isolated droplets without increased need to sort more droplets while allowing a higher content. The invention generates a larger group of droplets containing a library of compounds, then removes the empty droplets and retain the contents of full droplets by size exclusion filtration.

Embodiments disclosed herein are directed to microfluidic devices that allow for scalable on-chip screening of combinatorial libraries and methods of use thereof. Droplets comprising individual molecular species to be screened are loaded onto the microfluidic device. The droplets are labeled by methods known in the art, including but not limited to barcoding, such that the molecular species in each droplet can be uniquely identified. The device randomly sorts the droplets into individual microwells of an array of microwells designed to hold a certain number of individual droplets in order to derive combinations of the various molecular species. The paired droplets are then merged in parallel to form merged droplets in each microwell, thereby avoiding issues associated with single stream merging. Each microwell is then scanned, e.g., using microscopy, such as high content imaging microscopy, to detect the optical labels, thereby identifying the combination of molecular species in each microwell.

A novel miniaturized and highly automated method for the controlled printing of large arrays of nano- to femtoliter droplets is presented by actively transporting mother droplets over hydrophilic-in-hydrophobic micropatches. The proposed technology consists of single plate or double-plate devices where mother droplets can be actuated and hydrophilic-in-hydrophobic micropatches on one or both plates of the device where nano- to femtoliter droplets are printed. Due to the selective wettability of the more wettable hydrophilic micropatches in a hydrophobic matrix, large nano- to femtoliter droplet arrays are created when mother droplets are transported over these arrays. The parent droplets can be moved by different droplet actuation principles, for example, by using the principle of electrowetting-on-dielectric droplet actuation. We propose another method that uses two plates that are placed on top of each other while being separated by a spacer. One plate is dedicated to confirming and guiding of parent droplets by using hydrophilic patches in a hydrophobic matrix, while the other plate contains hydrophilic-in-hydrophobic arrays dedicated to the printing of nano- to femtoliter droplets. When the plate dedicated to parent droplet guiding is rotated over the plate dedicated to printing of nano- to femtoliter droplets, nano- to femtoliter droplets are dispensed inside the hydrophilic-in-hydrophobic array due to their selective wettability. All these proposed methods allow the parent droplets to be moved over the hydrophilic-in-hydrophobic arrays many times, providing unique advantages for performing bio-assays or miniaturized materials synthesis in nano- to femtoliter sized droplets. Upon the controlled evaporation of the dispensed droplets of solution, large arrays of the printed material can be generated on an automated way in seconds of time on a very flexible way. The method disclosed herein provides a distinct nano- to femtoliter droplet printing technique for a wide variety of applications such as protein- or cell-based bio-assays or printing of crystalline structures, suspensions of nanoparticles or components for microelectronics.