Abstract: Systems, methods, and apparatuses are provided for self-contained nucleic acid preparation, amplification, and analysis.

Abstract: Systems, methods, and apparatuses are provided for self-contained nucleic acid preparation, amplification, and analysis.

Abstract: Systems, methods, and apparatuses are provided for self-contained nucleic acid preparation, amplification, and analysis.

Abstract: A method of quantifying nucleic acids in a sample includes generating a plurality of droplets in oil within a microfluidic device, wherein at least some of the droplets comprise a nucleic acid, amplification reagents, and a fluorescent probe or dyes contained therein. The droplets are delivered to a collection chamber to form an array of droplets. The droplets are subject to thermal cycling within the collection chamber a plurality of times to perform nucleic acid amplification within the droplets. The array of droplets is imaged during the plurality of thermal cycles as well as at a thermal cycle endpoint. An initial concentration of nucleic acid in the sample is calculated based on at least one of: a ratio of aqueous phase droplets exhibiting fluorescence within the array at the thermal cycle endpoint or a cycle threshold (Ct) of one or more aqueous phase droplets within the array.

Abstract: A method of quantifying nucleic acids in a sample includes generating a plurality of droplets in oil within a microfluidic device, wherein at least some of the droplets comprise a nucleic acid, amplification reagents, and a fluorescent probe or dyes contained therein. The droplets are delivered to a collection chamber to form an array of droplets. The droplets are subject to thermal cycling within the collection chamber a plurality of times to perform nucleic acid amplification within the droplets. The array of droplets is imaged during the plurality of thermal cycles as well as at a thermal cycle endpoint. An initial concentration of nucleic acid in the sample is calculated based on at least one of: a ratio of aqueous phase droplets exhibiting fluorescence within the array at the thermal cycle endpoint or a cycle threshold (Ct) of one or more aqueous phase droplets within the array.

Abstract: An apparatus for passive sorting of microdroplets including a main flow channel, a flow stream of microdroplets in the main flow channel wherein the microdroplets have substantially the same diameter and wherein the flow stream of microdroplets includes first microdroplets having a first degree of stiffness and second microdroplets having a second degree of stiffness wherein the second degree of stiffness is different than the first degree of stiffness. A second flow channel is connected to the main flow channel for the second microdroplets having a second degree of stiffness. A separator separates the second microdroplets having a second degree of stiffness from the first microdroplets and directs the second microdroplets having a second degree of stiffness into the second flow channel.

Abstract: A multilayer well device includes a first substrate comprising an array of wells having a first pattern disposed therein and a second substrate comprising an array of wells having a second pattern, complementary to the first pattern disposed therein, wherein the second substrate is secured adjacent to a face of the first substrate. A common channel is interposed between the array of wells of the respective first and second substrates and is coupled to an inlet and an outlet.

Abstract: An apparatus for passive sorting of microdroplets including a main flow channel, a flow stream of microdroplets in the main flow channel wherein the microdroplets have substantially the same diameter and wherein the flow stream of microdroplets includes first microdroplets having a first degree of stiffness and second microdroplets having a second degree of stiffness wherein the second degree of stiffness is different than the first degree of stiffness. A second flow channel is connected to the main flow channel for the second microdroplets having a second degree of stiffness. A separator separates the second microdroplets having a second degree of stiffness from the first microdroplets and directs the second microdroplets having a second degree of stiffness into the second flow channel.

Abstract: An apparatus for passive sorting of microdroplets including a main flow channel, a flow stream of microdroplets in the main flow channel wherein the microdroplets have substantially the same diameter and wherein the flow stream of microdroplets includes first microdroplets having a first degree of stiffness and second microdroplets having a second degree of stiffness wherein the second degree of stiffness is different than the first degree of stiffness. A second flow channel is connected to the main flow channel for the second microdroplets having a second degree of stiffness. A separator separates the second microdroplets having a second degree of stiffness from the first microdroplets and directs the second microdroplets having a second degree of stiffness into the second flow channel.

Abstract: Embodiments of the claimed subject matter are directed to the ability to further multiplex in the digital regime using combinatorial color, temporal, and intensity encoding of probe sequences for a greater number of total signal readouts. A digital PCR solution is provide which enables the unique ability to identify a greater number of fluorescent probe sequences by using multiple color, temporal, and intensity combinations to encode each unique probe sequence. Furthermore, less expensive real-time PCR amplification indicators such as PicoGreen can be used to achieve multiplexed digital PCR based on temporal cues, intensity cues, or intensity and temporal cues combined, thus distinguishing primer pairs at greater degrees with significant cost reductions. These can also be used to enhance controls and normalize results for greater accuracy.

Abstract: A multilayer well device includes a first substrate comprising an array of wells having a first pattern disposed therein and a second substrate comprising an array of wells having a second pattern, complementary to the first pattern disposed therein, wherein the second substrate is secured adjacent to a face of the first substrate. A common channel is interposed between the array of wells of the respective first and second substrates and is coupled to an inlet and an outlet.

Abstract: A method and device is disclosed for increasing droplet and micro-well reactor densities per unit area for microfluidic platforms. The device and method use controlled Height to Droplet Diameter Ratios (HDR) of the collection region which can produce different crystalline packing formations. HDR ratios above unity and less than about 2.65 are used to create a variety of three-dimensional packing schemes with increased density over conventional single layer hexagonal packing.

Abstract: An apparatus for passive sorting of microdroplets including a main flow channel, a flow stream of microdroplets in the main flow channel wherein the microdroplets have substantially the same diameter and wherein the flow stream of microdroplets includes first microdroplets having a first degree of stiffness and second microdroplets having a second degree of stiffness wherein the second degree of stiffness is different than the first degree of stiffness. A second flow channel is connected to the main flow channel for the second microdroplets having a second degree of stiffness. A separator separates the second microdroplets having a second degree of stiffness from the first microdroplets and directs the second microdroplets having a second degree of stiffness into the second flow channel.