University of Pennsylvania

Peter Wilding

Larry J Kricka

Jay N Zemel

Disclosed are devices for amplifying a preselected polynucleotide in a sample by conducting a polynucleotide amplification reaction. The devices are provided with a substrate microfabricated to include a polynucleotide amplification reaction, chamber, having at least one cross-sectional dimension of about 0.1 to 1000 &#x3bc;m. The device also includes at least one port in fluid communication with the reaction chamber, for introducing a sample to the chamber, for venting the chamber when necessary, and, optionally, for removing products or waste material from the device. The reaction chamber may be provided with reagents required for amplification of a preselected polynucleotide. The device also may include means for thermally regulating the contents of the reaction chamber, to amplify a preselected polynucleotide. Preferably, the reaction chamber is fabricated with a high surface to volume ratio, to facilitate thermal regulation.

Mesoscale polynucleotide amplification devices

Disclosed are devices for detecting the presence of a preselected analyte in a fluid sample. The devices comprise a substrate microfabricated to define a sample inlet port, and a mesoscale flow system that includes a sample flow channel extending from the inlet port. The mesoscale flow system further includes an analyte detection region in fluid communication with the flow channel comprised of a binding moiety for specifically binding the analyte. The detection region is constructed with a mesoscale dimension sufficiently small to enhance binding of the binding moiety and the analyte. The binding moiety may be immobilized in the detection region. The mesoscale detection systems of the invention may be used in a wide range of applications, including the detection of cells or macromolecules, or for monitoring reactions or cell culture growth.

Mesoscale detection structures

Disclosed are devices for amplifying a preselected polynucleotide in a sample by conducting a polynucleotide polymerization reaction. The devices comprise a substrate microfabricated to define a sample inlet port and a mesoscale flow system, which extends from the inlet port. The mesoscale flow system includes a polynucleotide polymerization reaction chamber in fluid communication with the inlet port which is provided with reagents required for polymerization and amplification of a preselected polynucleotide. In one embodiment the devices may be utilized to implement a polymerase chain reaction (PCR) in the reaction chamber (PCR chamber). The PCR chamber is provided with the sample polynucleotide, polymerase, nucleoside triphosphates, primers and other reagents required for the polymerase chain reaction, and the device is provided with a device for thermally controlling the temperature of the contents of the reaction chamber at a temperature controlled to dehybridize double stranded polynucleotide, to anneal the primers, and to polymerize and amplify the polynucleotide.

Mesoscale polynucleotide amplification analysis

Device and method for the detection of an analyte utilizing mesoscale flow systems

Disclosed are devices for amplifying a preselected polynucleotide in a sample by conducting a polynucleotide polymerization reaction. The devices comprise a substrate microfabricated to define a sample inlet port and a mesoscale flow system, which extends from the inlet port. The mesoscale flow system includes a polynucleotide polymerization reaction chamber in fluid communication with the inlet port which is provided with reagents required for polymerization and amplification of a preselected polynucleotide. In one embodiment the devices may be utilized to implement a polymerase chain reaction (PCR) in the reaction chamber (PCR chamber). The PCR chamber is provided with the sample polynucleotide, polymerase, nucleoside triphosphates, primers and other reagents required for the polymerase chain reaction, and the device is provided with means for thermally controlling the temperature of the contents of the reaction chamber at a temperature controlled to dehybridize double stranded polynucleotide, to anneal the primers, and to polymerize and amplify the polynucleotide.

Mesoscale polynucleotide amplification device and method

Disclosed are devices for amplifying a preselected polynucleotide in a sample by conducting a polynucleotide amplification reaction. The devices are provided with a substrate microfabricated to include a polynucleotide amplification reaction chamber, having at least one cross-sectional dimension of about 0.1 to 1000 &mgr;m. The device also includes at least one port in fluid communication with the reaction chamber, for introducing a sample to the chamber, for venting the chamber when necessary, and, optionally, for removing products or waste material from the device. The reaction chamber may be provided with reagents required for amplification of a preselected polynucleotide. The device also may include means for thermally regulating the contents of the reaction chamber, to amplify a preselected polynucleotide. Preferably, the reaction chamber is fabricated with a high surface to volume ratio, to facilitate thermal regulation.

A mesoscale sample preparation device capable of providing microvolume test samples, separated into a cell-enriched fraction and a fraction of reduced cell content, for performing various analyses, such as binding assays, determinations involving polynucleotide amplification and the like. Analytical systems including such devices are also disclosed.

Mesoscale sample preparation device and systems for determination and processing of analytes

Mesoscale sample preparation device and systems for determination and

Apparatus for the detection of an analyte utilizing mesoscale flow

Devices and methods are provided to facilitate the rapid, accurate analysis of a sample having cells characterized by their motility. The devices feature a solid substrate microfabricated to define a flow system including one or more ports or chambers, connected by elongate channels of various shapes having a mescoscale cross-sectional dimension on the order of 0.1 .mu.m to 1000 .mu.m. In one embodiment, the devices are fitted with various flow-regulating features to facilitate unimpeded movement of the motile cells of interest along the flow channels. In another embodiment, devices are provided for conducting replicate motile cell assays, or for conducting a series of different assays using a single test sample. In another embodiment, preparative devices are provided for separating and collecting selected motile cell types of interest. In another embodiment, a device designed for performing an in vitro fertilization is provided in a portable incubator, which maintains the in vitro fertilization under optimum conditions. The devices of the invention may be used in a wide range of applications, and are particularly applicable for analyses and preparation of sperm, as well as in vitro fertilization. The devices and methods of the invention are also suitable for various analyses of a wide variety of motile cells, such as motile microorganisms and chemotactic cells, and can be used in clinical tests as well as laboratory and field tests.

Growing community of inventors

Paoli, PA, United States of America

Peter Wilding