Method and apparatus for detecting molecules

• US 10,222,333 B2
• Filed: 02/24/2010
• Issued: 03/05/2019
• Est. Priority Date: 03/23/2009
• Status: Active Grant

• Current Assignee: Geneseque AS
• Original Assignee: Geneseque AS
• Inventors: Kotsbak, Jarle
• Primary Examiner(s): Forman, Betty J
• Application Number:
  US13/258,219
• Publication Number:
  US 20120015833A1
• Time in Patent Office: 3,296 Days
• Field of Search: 359566
• CPC Class Codes: G01N 21/253 for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 21/6454 using an integrated detector array
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding

Abstract

The present disclosure is directed to a method of detecting a molecule attached to a surface comprising detecting a bead attached to the molecule. The method may be used for the detection of single molecules and the surface shape may be adapted to receive beads. The disclosure further describes an apparatus for detecting the presence of a molecule having a bead attached thereto comprising a surface adapted to detect a bead wherein the shape of the surface is configured to receive and retain a bead. Kits comprising the apparatus are provided and a use of the apparatus for detecting a bead or a molecule attached thereto is also described.

Claims

1. A method of detecting a single molecule attached to a surface with one or more light sensitive elements on or comprised within said surface wherein said surface is present in a photodetector without microlens and without filter layers, the method comprising:

illuminating said one or more light sensitive elements with a light source,

detecting a bead label attached to said single molecule

said bead label (i) has a diameter which is at least 30% of the width, breadth or diameter of said one or more light sensitive elements or (ii) is at least 1 μ

m in diameter,

said bead label and said surface are different entities,

said bead label is detected by a reduction in the amount of light received by said one or more light sensitive elements caused by the presence of said bead label which blocks light from reaching the one or more light sensitive elements, and

said single molecule is detected when the bead label is detected.

2. The method of claim 1, wherein said molecule is associated with said one or more light sensitive elements present in or on the surface.

3. The method of claim 2, wherein multiple molecules are associated with said one or more light sensitive elements but only one molecule type is associated with a light sensitive element.

4. The method of claim 1, wherein said surface is an image chip.

5. The method of claim 1, wherein said surface is coated with a layer of gold.

6. The method of claim 1, wherein said bead label is a magnetic bead.

7. The method of claim 1, wherein said single molecule attached to a surface is a target molecule, and wherein the method further comprises, prior to detecting, contacting the surface with a detector molecule, wherein said detector molecule is either attached to the bead label or is labelled with the bead label after addition to the surface.

8. The method of claim 1, wherein said bead label is directly detected by said one or more light sensitive elements.

9. The method of claim 1, wherein said single molecule attached to the surface is a target molecule and the bead is attached directly or indirectly to the target molecule.

10. The method of claim 1, wherein said single molecule attached to the surface is a target molecule and the target molecule is attached to the surface via a capture molecule which is attached to the surface.