SYSTEMS, DEVICES, AND METHODS FOR ULTRA-SENSITIVE DETECTION OF MOLECULES OR PARTICLES

• US 20120196774A1
• Filed: 02/25/2011
• Published: 08/02/2012
• Est. Priority Date: 01/28/2011
• Status: Active Grant

• Current Assignee: Quanterix Corporation
• Original Assignee: Quanterix Corporation
• Inventors: Fournier, David, Kan, Cheuk, Lawson, John, Duffy, David C., Campbell, Todd, Rivnak, Andrew, Kagan, Michael
• Granted Patent: US 9,952,237 B2
• Time in Patent Office: Days
• US Class Current: 506/32
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2300/0803 Disc shape
• CPC Class Codes: B01L 2400/043 magnetic forces
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: G01N 2035/00287 movable lid/cover for sample or reaction tubes
• CPC Class Codes: G01N 35/00029 provided with flat sample substrates, e.g. slides G01N35/028 takes precedence
• CPC Class Codes: G01N 35/0098 involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C
• CPC Class Codes: G01N 35/1065 Multiple transfer devices

Abstract

Described are systems, devices, and methods which related to various aspects of assays for detecting and/or determining a measure of the concentration of analyte molecules or particles in a sample fluid. In some cases, the systems employ an assay consumable comprising a plurality of assay sites. The systems, devices, and/or methods, in some cases, are automated. In some cases, the systems, devices, and/or methods relate to inserting a plurality of beads into assay sites, sealing assay sites, imaging assay sites, or the like.

Claims

1. An apparatus for performing an assay, comprising:

an assay consumable handler configured to be operatively coupled to an assay consumable having a surface comprising a plurality of assay sites;

a sealer constructed and positioned to apply a sealing component to the surface of the assay consumable;

a sample loader configured to load an assay sample into at least a portion of the plurality of assay sites of the assay consumable;

an imaging system configured to acquire an image of at least a portion of the assay sites of the assay consumable containing assay sample; and

a computer implemented control system configured to automatically operate the sealer and receive information from the imaging system related to the image.

2. The apparatus of claim 1, further comprising the assay consumable operatively coupled to the assay consumable handler.

3. The apparatus of claim 2, wherein the assay consumable contains the assay sample and the assay sample comprises an unknown concentration of an analyte molecule.

4. The apparatus of claim 3, wherein the computer implemented control system is configured to determine a measure of the unknown concentration of the analyte molecule in the assay sample.

5. The apparatus of claim 1, wherein the computer implemented control system is configured to automatically operate the sample loader.

6. The apparatus of claim 1, wherein the sealing component comprises a film.

7. The apparatus of claim 1, wherein the sealer comprises a roller configured to contact and apply pressure to the sealing component during application to the surface of the assay consumable.

8. The apparatus of claim 6, wherein the film is formed of PDMS.

9-21. -21. (canceled)

22. An apparatus for sealing a plurality of assay sites, comprising:

an assay consumable handler configured to be operatively coupled to an assay consumable having a surface comprising a plurality of assay sites;

a sealer constructed and positioned to apply a sealing component to the surface of the assay consumable to form a plurality of sealed assay sites, wherein the contents of each sealed assay site is substantially isolated from the contents of each of the other plurality of sealed assay sites; and

a controller configured to automatically operate the sealer to apply the sealing component to the plurality of assay sites.

23-39. -39. (canceled)

40. An apparatus for inserting beads into assay sites on an assay consumable, comprising:

an assay consumable handler configured to be operatively coupled to an assay consumable having a surface comprising a plurality of assay sites;

a bead loader configured to insert individual beads into individual assay sites, such that each assay site containing a bead will contain no more than one bead; and

a controller configured to automatically operate the bead loader to insert individual beads into individual assay sites.

41-52. -52. (canceled)

53. An apparatus for performing an assay, comprising:

an assay consumable handler configured to be operatively coupled to an assay consumable having a surface comprising a plurality of assay sites;

a sample loader configured to load an assay sample containing analyte molecules or particles having an unknown concentration to be measured into at least a portion of the plurality of assay sites, such that a plurality of assay sites into which assay sample is loaded contain either zero or a single analyte molecule or particle;

a detector configured to interrogate at least a portion of the assay sites containing assay sample and determine a fraction of the plurality of assay sites interrogated that contain an analyte molecule or particle; and

a computer implemented system configured receive information from the detector and from the information determine a measure of the unknown concentration of the analyte molecules or particles in the assay sample.

54-72. -72. (canceled)

73. An apparatus for inserting beads into assay sites on an assay consumable, comprising:

an assay consumable handler configured to be operatively coupled to the assay consumable, wherein the assay consumable comprises a surface comprising a plurality of the assay sites;

a bead applicator configured to apply a plurality of magnetic beads to the surface of the assay consumable or place a plurality of magnetic beads in close proximity to the surface;

a bead loader comprising a magnetic field generator positioned adjacent to the assay consumable and configured to create relative motion between the magnetic beads and the assay sites; and

a controller configured to automatically operate the bead loader to create relative motion between the magnetic beads and the assay sites and insert beads into assay sites.

74-83. -83. (canceled)

84. An apparatus for removing excess beads from an assay consumable having a surface comprising a plurality of assay sites, comprising:

a assay consumable handler operatively coupled to the assay consumable, wherein the assay consumable comprises a plurality of beads, wherein a first portion of the beads are contained in the assay sites and a second portion of the beads are positioned on the surface of the assay consumable, but not contained within an assay site;

a wiper configured to remove substantially all of the second portion of beads from the surface; and

a controller configured to automatically operate the wiper to remove the second portion of the beads.

85-92. -92. (canceled)

93. An assay consumable, comprising:

a surface comprising a plurality of assay sites, wherein each of the assay sites has a volume between about 10 attoliters and about 50 picoliters; and

at least one channel formed in the surface at least partially surrounding the plurality of assay sites that is positioned and configured to collect excess assay sample liquid applied to the surface that overflows the assay sites.

94-97. -97. (canceled)

98. An automated method for forming a plurality of sealed assay sites for performing an assay, comprising:

operatively associating an assay consumable having a surface comprising a plurality of assay sites with a sealer apparatus comprising a sealer and a controller; and

applying a sealing component to the plurality of assay sites with the sealer apparatus such that a plurality of sealed assay sites are formed, wherein the contents of each sealed assay site is substantially isolated from the contents of each of the other plurality of sealed assay sites.

99-127. -127. (canceled)

128. A method for inserting beads into reaction vessels on an assay consumable, comprising:

generating a magnetic field in proximity to a surface of the assay consumable comprising a plurality of the reaction vessels such that a magnetic field vector of the magnetic field is directed from the surface towards a bottom of the reaction vessels and/or towards the perimeter of the surface;

delivering a plurality of magnetic beads in proximity to the surface; and

creating relative motion between the magnetic beads and the reaction vessels.

129-135. -135. (canceled)

136. A method for forming a plurality of sealed reaction vessels for performing an assay, comprising:

associating an assay consumable having a surface comprising a plurality of assay sites with a sealing component by applying the sealing component to the surface,

wherein the contents of each assay site are substantially isolated from the contents of each of the other plurality of assay sites following association of the sealing component without maintaining any pressure applied to the sealing component;

wherein

each of the assay sites has a volume between about 10 attoliters and about 50 picoliters.

137-142. -142. (canceled)

143. A method for forming a plurality of sealed reaction vessels for performing an assay, comprising:

associating an assay consumable having a surface comprising a plurality of assay sites with a sealing component by applying the sealing component to the surface of the assay consumable and applying pressure to the sealing component, wherein

the contents of each assay site are substantially isolated from the contents of each of the other plurality of assay sites following association of the sealing component with the assay consumable;

wherein the sealing component comprises a pressure-sensitive adhesive such that the pressure-sensitive adhesive is activated upon application of the pressure to the sealing component and the adhesive forms an adhesive bond between the sealing component and the surface of the assay consumable; and

wherein each of the assay sites has a volume between about 10 attoliters and about 50 picoliters.

144-146. -146. (canceled)

147. A method for forming a plurality of sealed assay sites for performing an assay, comprising:

providing an assay consumable having a surface comprising a plurality of assay sites wherein each of the assay sites has a volume between about 10 attoliters and about 50 picoliters; and

applying a liquid that is substantially immiscible with liquid contained in the plurality of assay sites to the plurality of assay sites such that a plurality of sealed assay sites are formed, wherein the contents of each sealed assay site is substantially isolated from the contents of each of the other plurality of sealed assay sites.

148. An apparatus for removing beads from a surface of an assay consumable, comprising:

a first magnet, wherein the first magnet is located adjacent to a surface of the assay consumable and is positioned opposite the surface comprising the plurality of assay sites;

a second magnet,

a third magnet, and

a metal object,

wherein the second magnet and third magnet are located adjacent the surface comprising the plurality of assay sites and such that the opposite poles of the second magnet and the third magnet are directed towards each other; and

wherein the metal object is positioned between the second magnet and the third magnet.

149-153. -153. (canceled)