Abstract: In some aspects, the present disclose provides methods for amplifying and quantifying nucleic acids. Methods for amplifying and quantifying nucleic acids comprise isolating a sample comprising nucleic acid molecules into a plurality of microchambers, performing a polymerase chain reaction on the plurality of microchambers, and analyzing the results of the polymerase chain reaction. In some aspects, the present disclosure provides devices consistent with the methods herein.

Abstract: The present disclose provides methods and systems for amplifying and quantifying nucleic acids and for detecting the presence or absence of a target in a sample. The methods and systems provided herein may utilize a device comprising a plurality of partitions separated from an external environment by a gas-permeable barrier. Certain methods disclosed herein involve subjecting nucleic acid molecules in the plurality of partitions to conditions sufficient to conduct nucleic acid amplification reactions. The nucleic acid molecules may be subjected to controlled heating in the plurality of partitions to generate data indicative of a melting point(s) of the nucleic acid molecules.

Abstract: Various embodiments may provide a method of illuminating a sample plane. The method may include providing an illumination subsystem, the illumination subsystem including an optical source and at least one lens, having an optic axis at an incident angle greater than 0° and less than 90° to a normal of the sample plane. The method may also include rotating the illumination subsystem about a pivot point between the optical source and the sample plane along the optic axis so that an adjusted illumination distribution generated by the illumination subsystem at the sample plane has greater symmetry compared to a reference illumination distribution generated by the illumination subsystem at the sample plane without the rotation about the pivot point.

Abstract: In some aspects, the present disclose provides methods for amplifying and quantifying nucleic acids. Methods for amplifying and quantifying nucleic acids comprise isolating a sample comprising nucleic acid molecules into a plurality of microchambers, performing a polymerase chain reaction on the plurality of microchambers, and analyzing the results of the polymerase chain reaction. In some aspects, the present disclosure provides devices consistent with the methods herein.

Abstract: Various embodiments may provide a method of illuminating a sample plane. The method may include providing an illumination subsystem, the illumination subsystem including an optical source and at least one lens, having an optic axis at an incident angle greater than 0° and less than 90° to a normal of the sample plane. The method may also include rotating the illumination subsystem about a pivot point between the optical source and the sample plane along the optic axis so that an adjusted illumination distribution generated by the illumination subsystem at the sample plane has greater symmetry compared to a reference illumination distribution generated by the illumination subsystem at the sample plane without the rotation about the pivot point.

Abstract: The present disclose provides methods and systems for amplifying and quantifying nucleic acids and for detecting the presence or absence of a target in a sample. The methods and systems provided herein may utilize a device comprising a plurality of partitions separated from an external environment by a gas-permeable barrier. Certain methods disclosed herein involve subjecting nucleic acid molecules in the plurality of partitions to conditions sufficient to conduct nucleic acid amplification reactions. The nucleic acid molecules may be subjected to controlled heating in the plurality of partitions to generate data indicative of a melting point(s) of the nucleic acid molecules.

Abstract: In some aspects, the present disclose provides methods for amplifying and quantifying nucleic acids. Methods for amplifying and quantifying nucleic acids comprise isolating a sample comprising nucleic acid molecules into a plurality of microchambers, performing a polymerase chain reaction on the plurality of microchambers, and analyzing the results of the polymerase chain reaction. In some aspects, the present disclosure provides devices consistent with the methods herein.

Abstract: A microfluidic device can have a plurality of microchambers connected to a microchannel via siphon apertures. The microfluidic device may be formed from a thermoplastic and capped by a thermoplastic thin film. The microfluidic device may be used for digital polymerase chain reactions by forcing reagent into the microchambers at low pressure via an inlet, forcing any gas trapped in the microfluidic device to outgas through the thin film by applying high pressure to the microfluidic device via inlets and outlets of the microfluidic device, and applying air at low pressure to the inlets in order to digitize the chambers such that the reagent in each chamber is isolated from the reagent in other chambers by an air gap. After digitization, the device may be used to run a digital polymerase chain reaction process.