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 sunset. The amount of target can be determined based on the detected number.

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.

A miniaturized, automated method for controlled printing of large arrays of nano- to femtoliter droplets by actively transporting mother droplets over hydrophilic-in-hydrophobic (“HIH”) micropatches. The technology uses single or double-plate devices where mother droplets can be actuated and HIH micropatches on one or both plates of the device where the droplets are printed. Due to the selective wettability of the hydrophilic micropatches in a hydrophobic matrix, large nano- to femtoliter droplet arrays are created when mother droplets are transported over the arrays. The parent droplets are moved by various droplet actuation principles. Also, a method using two plates placed one top another while being separated by a spacer. One plate is dedicated to confirming and guiding parent droplets by using hydrophilic patches in a hydrophobic matrix, while the other plate contains HIH arrays for printing of the droplets. When the parent droplet guidance plate is rotated over the plate dedicated to printing of nano- to femtoliter droplets, the droplets are dispensed inside the HIH array utilizing their selective wettability. The methods allow the parent droplets to move over the HIH arrays many times, providing advantages for performing bio-assays or miniaturized materials synthesis in nano- to femtoliter sized droplets. With controlled evaporation of the dispensed droplets of solution, large arrays of printed material can be generated in seconds. The methods provide a nano- to femtoliter droplet printing technique for a wide variety of applications, e.g., protein- or cell-based bio-assays or printing of crystalline structures, suspensions of nanoparticles or microelectronic components.

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.

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.

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.

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 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.

A miniaturized, automated method for controlled printing of large arrays of nano- to femtoliter droplets by actively transporting mother droplets over hydrophilic-in-hydrophobic (HIH) micropatches. The technology uses single or double-plate devices where mother droplets can be actuated and HIH micropatches on one or both plates of the device where the droplets are printed. Due to the selective wettability of the hydrophilic micropatches in a hydrophobic matrix, large nano- to femtoliter droplet arrays are created when mother droplets are transported over the arrays. The parent droplets are moved by various droplet actuation principles. Also, a method using two plates placed one top another while being separated by a spacer. One plate is dedicated to confirming and guiding parent droplets by using hydrophilic patches in a hydrophobic matrix, while the other plate contains HIH arrays for printing of the droplets. When the parent droplet guidance plate is rotated over the plate dedicated to printing of nano- to femtoliter droplets, the droplets are dispensed inside the HIH array utilizing their selective wettability. The methods allow the parent droplets to move over the HIH arrays many times, providing advantages for performing bio-assays or miniaturized materials synthesis in nano- to femtoliter sized droplets. With controlled evaporation of the dispensed droplets of solution, large arrays of printed material can be generated in seconds. The methods provide a nano- to femtoliter droplet printing technique for a wide variety of applications, e.g., protein- or cell-based bio-assays or printing of crystalline structures, suspensions of nanoparticles or microelectronic components.