Methods for fabricating microarrays of biological samples

• US 6,110,426 A
• Filed: 12/30/1997
• Issued: 08/29/2000
• Est. Priority Date: 06/17/1994
• Status: Expired due to Term

• Current Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford University)
• Original Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford University)
• Inventors: Brown, Patrick O., Shalon, Tidhar Dari
• Primary Examiner(s): Marschel, Ardin H.
• Application Number:
  US09/001,027
• Time in Patent Office: 973 Days
• Field of Search: 422/50, 422/68.1, 435/6, 435/183.1, 436/501, 935/77, 935/78, 536/25.3
• US Class Current: 422/68.1
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00367 capillary
• CPC Class Codes: B01J 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/00475 Sheets
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00619 using hydrophilic or hydrophobic regions
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00641 the porous medium being continuous, e.g. porous oxide substrates
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00689 using computers
• CPC Class Codes: B01J 2219/00722 : Nucleotides
• CPC Class Codes: B01J 2219/00725 : Peptides
• CPC Class Codes: B01J 4/02 : for feeding measured , i.e. prescribed quantities of reagents
• CPC Class Codes: B01L 3/0244 : using pins
• CPC Class Codes: B01L 3/0248 : Prongs, quill pen type dispenser
• CPC Class Codes: B01L 3/0268 : using pulse dispensing or spraying, eg. inkjet type, piezo actuated ejection of droplets from capillaries
• CPC Class Codes: B01L 3/5088 : confining liquids at a location by surface tension, e.g. virtual wells on plates, wires B01L3/50857 takes precedence
• CPC Class Codes: C12Q 1/6809 : Methods for determination or identification of nucleic acids involving differential detection
• CPC Class Codes: C12Q 1/6837 : using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 2001/282 : with mapping; Identification of areas; Spatial correlated pattern
• CPC Class Codes: G01N 2035/00237 : Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 2035/1037 : Using surface tension, e.g. pins or wires
• CPC Class Codes: G01N 33/54366 : Apparatus specially adapted for solid-phase testing
• CPC Class Codes: Y10T 436/116664 : with automated titrator

Abstract

A method and apparatus for forming microarrays of biological samples on a support are disclosed. The method involves dispensing a known volume of a reagent at each selected array position, by tapping a capillary dispenser on the support under conditions effective to draw a defined volume of liquid onto the support. The apparatus is designed to produce a microarray of such regions in an automated fashion.

Claims

1. A method of forming a microarray of discrete analyte-assay regions on a solid support, where each discrete region in the microarray has a select(d, analyte-specific reagent, said method comprising,

(a) loading a solution of a selected analyte-specific reagent in a reagent-dispensing device having an elongate capillary channel adapted to hold a quantity of the reagent solution and having a tip region at which the channel is open-sided and the solution in the channel forms a meniscus,

(b) tapping the tip of the dispensing device against a solid support at a defined position on the surface, with an impulse effective to break the meniscus in the capillary channel and deposit a selected volume of solution on the surface, and

(c) repeating steps (a) and (b) until said microarray is formed.

3. The method of claim 1, wherein the reagents used to form the discrete regions in the microarray are distinct nucleic acid strands and wherein steps (a) and (b) are repeated until the microarray has about 1000 or more discrete regions of distinct nucleic acid strands per cm² of solid support.

4. The method of claim 1, which further includes, after performing steps (a) and (b), reloading the reagent-dispensing device with a new reagent solution by the steps of (i) dipping the capillary channel of the device in a wash solution, (ii) removing wash solution drawn into the capillary channel, and (iii) dipping the capillary channel into the new reagent solution.

2. The method of claim l, wherein the reagents used to form the discrete regions in the microarray are distinct nucleic acid strands and wherein steps (a) and (b) are repeated until the microarray has about 100 or more discrete regions of distinct nucleic acid strands per cm² of solid support.