Process for biosensor well formation

• US 9,322,062 B2
• Filed: 10/22/2014
• Issued: 04/26/2016
• Est. Priority Date: 10/23/2013
• Status: Active Grant

• Current Assignee: Genia Technologies, Inc. (Roche Holding AG), Roche Molecular Systems Inc (Roche Holding AG)
• Original Assignee: Genia Technologies, Inc. (Roche Holding AG)
• Inventors: Davis, Randall W., Tian, Hui, Foster, John, Chen, Roger, Liu, Edward Shian, Harada, Eric Takeshi, Aguirre, Anne, Trans, Andrew, Pollard, James, Cech, Cynthia, Yuan, Robert
• Primary Examiner(s): Ball, J. Christopher
• Application Number:
  US14/521,427
• Publication Number:
  US 20150153302A1
• Time in Patent Office: 552 Days
• US Class Current: 1/1
• CPC Class Codes: C12M 1/34 Measuring or testing with condition measuring or sensing means, e.g. colony counters
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode
• CPC Class Codes: C12Q 2565/631 being a biochannel or pore
• CPC Class Codes: G01N 27/3277 being a redox reaction, e.g. detection by cyclic voltammetry voltammetry per se G01N27/42, G01N27/48
• CPC Class Codes: G01N 27/3278 involving nanosized elements, e.g. nanogaps or nanoparticles nanopores G01N33/48721; magnetic beads G01N27/745
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: G01N 33/48721 Investigating individual macromolecules, e.g. by translocation through nanopores Coulter counters in general G01N15/12; fabrication methods for nanoscale apertures B81B1/00; sequencing of nucleic acids C12Q1/68

Abstract

A biochip for molecular detection and sensing is disclosed. The biochip includes a substrate. The biochip includes a plurality of discrete sites formed on the substrate having a density of greater than five hundred wells per square millimeter. Each discrete site includes sidewalls disposed on the substrate to form a well. Each discrete site includes an electrode disposed at the bottom of the well. In some embodiments, the wells are formed such that cross-talk between the wells is reduced. In some embodiments, the electrodes disposed at the bottom of the wells are organized into groups of electrodes, wherein each group of electrodes shares a common counter electrode. In some embodiments, the electrode disposed at the bottom of the well has a dedicated counter electrode. In some embodiments, surfaces of the sidewalls are silanized such that the surfaces facilitate the forming of a membrane in or adjacent to the well.

Claims

1. A biochip, comprising:

a substrate; and

a plurality of discrete sites formed on the substrate having a density of greater than five hundred wells per square millimeter, wherein each discrete site includes;

sidewalls disposed on the substrate to form a well, wherein the well is formed by depositing on the substrate a material having reactive oxide groups and etching the well into the material, and wherein the sidewalls have a hard, planar top surface for a membrane to be formed atop, wherein the membrane spans across and seals the well; and

an electrode disposed at the bottom of the well.

2. The biochip of claim 1, wherein the wells are formed such that cross-talk between the wells is reduced, including electrically isolating the electrodes from each other to reduce a signal from a nanopore or membrane formed atop a well to cross-talk with an electrode disposed at the bottom of a different well.

3. The biochip of claim 1, wherein the electrode disposed at the bottom of the well derives most of its signal from a nanopore or a membrane nearest to the electrode.

4. The biochip of claim 1, wherein the electrodes disposed at the bottom of the wells are organized into a plurality of groups of electrodes.

5. The biochip of claim 4, wherein each group of electrodes shares a common counter electrode.

6. The biochip of claim 1, wherein the electrode disposed at the bottom of the well has a dedicated counter electrode.

7. The biochip of claim 1, wherein surfaces of the sidewalls are silanized such that the surfaces facilitate the forming of a membrane in or adjacent to the well.

8. The biochip of claim 7, wherein surfaces of the sidewalls are hydrophobic such that the surfaces facilitate the forming of a hydrophobic membrane in or adjacent to the well.

9. The biochip of claim 7, wherein facilitating the forming of a membrane in or adjacent to the well comprises facilitating the adhering of the membrane to the silanized surfaces.

10. The biochip of claim 1, wherein surfaces of the sidewalls are silanized by covering the sidewalls with a layer of organofunctional alkoxysilane molecules.

11. The biochip of claim 10, wherein the layer of molecules is one molecule in thickness.