Nanogen, Inc.

Genoptix, Inc.

Celula, Inc.

Gamida for Life B.v.

William Frank Butler

Eugene Tu

Mark Wang

Paul David Swanson

Donald E Ackley

Michael James Heller

James P O'Connell

Kristie L Lykstad

Carl Frederick Edman

Ronald George Sosnowski

Michael I Nerenberg

Haichuan Zhang

Philippe Marchand

Norbert D Hagen

Rong Yang

Scott O Graham

Timothy L LeClair

Mirianas Chachisvilis

Daniel E Raymond

Elizabeth L Mather

Robert Dees

Joon Mo Yang

Yi Zhang

Lei Wu

Robert H Cormack

Keunho Ahn

Robert David Juncosa

John Francis

Benjamin Lai

Mark M Wang

Apparatus and Methods are provided for a microfabricated fluorescence activated cell sorter based on a switch for rapid, active control of cell routing through a microfluidic channel network. This sorter enables low-stress, highly efficient sorting of populations of small numbers of cells (i.e., 1000-100,000 cells). The invention includes packaging of the microfluidic channel network in a self-contained plastic cartridge that enables microfluidic channel network to macro-scale instrument interconnect, in a sterile, disposable format. Optical and/or fluidic switching forces are used alone or in combination to effect switching.

Cell sorting system and methods

Apparatus and Methods are provided for a microfabricated fluorescence activated cell sorter based on an optical switch for rapid, active control of cell routing through a microfluidic channel network. This sorter enables low-stress, highly efficient sorting of populations of small numbers of cells (i.e., 1000-100,000 cells). The invention includes packaging of the microfluidic channel network in a self-contained plastic cartridge that enables microfluidic channel network to macro-scale instrument interconnect, in a sterile, disposable format.

Methods and apparatus for sorting cells using an optical switch in a microfluidic channel network

A method for electronically stabilizing hybridization of nucleic acids bound at a test site of a microelectronic device is described. First and second negatively charged nucleic acids are provided, the second nucleic acid being bound to the test site. A zwitterionic buffer having a conductance of less than 100 mS/cm is applied to the microelectronic device. A current is applied to the test site to positively bias the test site, such that the first negatively charged nucleic acid is transported to the positively biased test site having the bound the second negatively charged nucleic acid. At the test site, the first and second negatively charged nucleic acids hybridize. The zwitterionic buffer acquires a net positive charge under influence of the current, such that the positively charged zwitterionic buffer stabilizes the hybridization by reducing the repulsion between the first and second negatively charged nucleic acids.

Self-addressable self-assembling microelectronic integrated systems, component devices, mechanisms, methods, and procedures for molecular biological analysis and diagnostics

A method for sorting a particle of interest from a plurality of particles includes the steps of determining an absorption maxima of the particle of interest, providing a light source for generating a beam of coherent light at a wavelength correlating to the absorption maxima, providing a plurality of particles on a support surface, and imparting relative motion between the beam of coherent light and the plurality of particles so as to cause differential movement between the particle of interest and the plurality of particles. The particle of interest is then collected.

Method for sorting particles

An electronic device for performing active biological operations includes an optically confining region including a support substrate having a via and a chip disposed in a facing arrangement with the support substrate, the chip including an array of electrodes disposed thereon. An optically accessible top member is disposed in a facing arrangement with the support substrate opposite the chip. The device further includes a source of illumination and an edge illumination layer having an input adapted to receive illumination from the source, and a terminal edge that outputs the illumination, the edge illumination layer being disposed adjacent to the support substrate. Illumination from the terminal edge of the illumination layer is directed into the optically confining region.

Devices for molecular biological analysis and diagnostics including waveguides

Apparatus and methods are provided for interacting light with particles, including but not limited to biological matter such as cells, in unique and highly useful ways. Optophoresis consists of subjecting particles to various optical forces, especially optical gradient forces, and more particularly moving optical gradient forces, so as to obtain useful results. In biology, this technology represents a practical approach to probing the inner workings of a living cell, preferably without any dyes, labels or other markers. In one aspect, a method is provided for separating particles by flowing the particles within a first constrained path, the first constrained path having an input and an output, and a sorting region, the sorting region coupling to a second constrained path, the second constrained path including an output, illuminating the sorting region with a moving optical gradient, characterized in that certain of the particles flow in a laminar manner between the first inlet and the output of the first constrained path, and selected particles are diverted from the first constrained path to the second constrained path under the force of the moving optical gradient.

Method for separation of particles

Apparatus and methods are provided for interacting light with particles, including but not limited to biological matter such as cells, in unique and highly useful ways. Optophoresis consists of subjecting particles to various optical forces, especially optical gradient forces, and more particularly moving optical gradient forces, so as to obtain useful results. In biology, this technology represents a practical approach to probing the inner workings of a living cell, preferably without any dyes, labels or other markers. In one aspect, a method is provided for interacting an optical gradient field in three dimensions with a particle by interfering two beams to generate a plurality of planar fronts, providing a plurality of particles in a medium, and moving the planar fronts relative to the particles, whereby the particles are separated at least in part based upon the dielectric constant of the particles.

Method of separating particles using an optical gradient

Apparatus and methods are provided for interacting light with particles, including but not limited to biological matter such as cells, in unique and highly useful ways. Optophoresis consists of subjecting particles to various optical forces, especially optical gradient forces, and more particularly moving optical gradient forces, so as to obtain useful results. In biology, this technology represents a practical approach to probing the inner workings of a living cell, preferably without any dyes, labels or other markers. The invention includes methods for separating particles in a medium where the particles having differing dielectric constants by providing a medium having a dielectric constant between the dielectric constants of the particles, subjecting the particles in the media to an optical gradient field, and separating the particles.

Methods of separating particles using an optical gradient

An electronic device for performing biological operations includes a support substrate and an array of microlocations disposed on the substrate. The array of microlocations include electronically addressable electrodes. A first collection electrode is disposed on the substrate and adjacent to a first side of the array of microlocations. A second collection electrode is disposed on the substrate and adjacent to a second side of the array of microlocations, the second side of the array being opposite of the first side such that the array of microlocations is disposed between the first and second collection electrodes. A flow cell is supported on the substrate. Preferably, the combined area of the collection electrodes is a substantial fraction, preferably at least 50% of the area of the footprint of the flow cell.

Electronic device for performing active biological operations and method of using same

A self-addressable, self-assembling microelectronic device is designed and fabricated to actively carry out and control multi-step and multiplex molecular biological reactions in microscopic formats. These reactions include nucleic acid hybridizations, antibody/antigen reactions, diagnostics, and biopolymer synthesis. The device can be fabricated using both microlithographic and micro-machining techniques. The device can electronically control the transport and attachment of specific binding entities to specific microlocations. The specific binding entities include molecular biological molecules such as nucleic acids and polypeptides. The device can subsequently control the transport and reaction of analytes or reactants at the addressed specific microlocations. The device is able to concentrate analytes and reactants, remove non-specifically bound molecules, provide stringency control for DNA hybridization reactions, and improve the detection of analytes. The device can be electronically replicated.

Method for enhancing the hybridization efficiency of target nucleic acids using a self-addressable, self-assembling microelectronic device

An electronic device for performing active biological operations includes a support substrate, a second substrate, a source of illumination, and an edge illumination layer. The support substrate includes first and second surfaces and a via between the first and second surfaces to permit fluid flow through the substrate. The second substrate includes a first surface that is adapted to be disposed in facing arrangement with the second surface of the first substrate. The second substrate includes an array of microlocations wherein the array is adapted to receive the fluid. A sealant is disposed between the second face of the support substrate and the first face of the second substrate. The device includes a source of illumination and an edge illumination layer having an input adapted to receive the illumination from the source, and an output adapted to direct the illumination to the array. The edge illumination layer is disposed adjacent to and between the support substrate and the second substrate. The illumination from the edge illumination layer is directed substantially parallel to the support substrate and second substrate and illuminates the array.

An optical detection system is adapted for detection of biological reactions. An excitation source illuminates a portion of an object to be examined, the portion preferably comprising one microlocation out of an array of microlocations. An intervening optical detection platform serves to direct the excitation radiation to the portion of the object to be illuminated. A detector receives the emitted radiation from the object to be examined, the detector being characterized in that the diameter of the region examined by the detector is the same as or smaller than the diameter of the illuminated region, and comprises less than the entire surface of the object to be examined, and most preferably images a whole or a part of a single microlocation. In operation, a microscopy system is formed in which the excitation radiation is substantially in focus at the surface of the object to be examined. In one aspect of this invention, the optical detection platform includes an excitation detector that measures reflected excitation radiation from the object to be examined. This information is compared to prestored information regarding the location of the microlocations and interstitial regions on the object to be examined, whereby alignment information is obtained. The excitation radiation may then be precisely directed to a given microlocation or portion thereof to perform the examining through the system.

Scanning optical detection system

Methods of manufacture and devices for performing active biological operations utilize various structures to advantageously collect and provide charged biological materials to an array of microlocations. In one embodiment, a device includes focusing electrodes to aid in the direction and transport of materials from a collection electrode to an array. Preferably, one or more intermediate transportation electrodes are utilized, most preferably of monotonically decreasing size between the collection electrode and the array, so as to reduce current density mismatches. In another aspect, a flow cell is utilized over devices to provide containment of solution containing materials to be analyzed. Preferably, the volume of the flow cell is more advantageously interrogated through use of relatively large collection and return electrodes, such as where the area of those electrodes relative to the footprint of the flowcell is at least 40%. In yet another embodiment, a first collection electrode is disposed adjacent an array, with a second collection electrode disposed on the at least an opposite portion of the array. Preferably, the combined area of the collection electrodes is a substantial fraction, preferably at least 50% of the area of the footprint of the flow cell. In yet another embodiment, a concentric ring design is provided. Various flip chip embodiments are disclosed.

Growing community of inventors

La Jolla, CA, United States of America

William Frank Butler