Methods for screening substances in a microwell array

• US 6,893,877 B2
• Filed: 05/07/2001
• Issued: 05/17/2005
• Est. Priority Date: 01/12/1998
• Status: Expired due to Term

• Current Assignee: Massachusetts Institute of Technology
• Original Assignee: Massachusetts Institute of Technology
• Inventors: Kanigan, Tanya S., Brenan, Colin J. H., Hunter, Ian
• Primary Examiner(s): Soderquist, Arlen
• Application Number:
  US09/850,123
• Publication Number:
  US 20020001546A1
• Time in Patent Office: 1,471 Days
• Field of Search: 422/56, 422/58, 422/100, 422/82.05, 422/102, 422/82.09, 422/104, 356/244, 356/246, 436/165, 436/174, 436179-180, 359/599, 359/886, 362/318, 362/342, 362354-356
• US Class Current: 436/174
• CPC Class Codes: B01F 33/3021 the components to be mixed being combined in a single independent droplet, e.g. these droplets being divided by a non-miscible fluid or consisting of independent droplets
• CPC Class Codes: B01F 33/30351 using hydrophilic/hydrophobic surfaces
• 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 2219/00283 Reactor vessels with top opening
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00319 the blocks being mounted in stacked arrangements
• CPC Class Codes: B01J 2219/00351 Means for dispensing and evacuation of reagents
• CPC Class Codes: B01J 2219/0036 Nozzles
• CPC Class Codes: B01J 2219/00369 in multiple or parallel arrangements
• CPC Class Codes: B01J 2219/00373 Hollow needles
• CPC Class Codes: B01J 2219/00387 Applications using probes
• CPC Class Codes: B01J 2219/00389 Feeding through valves
• CPC Class Codes: B01J 2219/00423 using filtration, e.g. through porous frits
• CPC Class Codes: B01J 2219/0043 for direct application of reagents, e.g. through openings in a shutter
• CPC Class Codes: B01J 2219/00479 Means for mixing reactants or products in the reaction vessels
• CPC Class Codes: B01J 2219/00495 Means for heating or cooling the reaction vessels
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00511 Walls of reactor vessels
• CPC Class Codes: B01J 2219/00533 essentially rectangular
• CPC Class Codes: B01J 2219/00536 in the shape of disks
• CPC Class Codes: B01J 2219/00549 : 2-dimensional
• CPC Class Codes: B01J 2219/00585 : Parallel processes
• CPC Class Codes: B01J 2219/00587 : High throughput processes
• CPC Class Codes: B01J 2219/00596 : Solid-phase processes
• CPC Class Codes: B01J 2219/00648 : by the use of solid beads
• CPC Class Codes: B01J 2219/00659 : Two-dimensional arrays
• CPC Class Codes: B01J 2219/00686 : Automatic
• CPC Class Codes: B01L 2200/0642 : Filling fluids into wells by specific techniques
• CPC Class Codes: B01L 2300/0819 : Microarrays; Biochips
• CPC Class Codes: B01L 2300/0829 : Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/0845 : Filaments, strings, fibres, i.e. not hollow
• CPC Class Codes: B01L 2400/022 : droplet contacts the surface of the receptacle
• CPC Class Codes: B01L 3/0262 : using touch-off at substrate or container
• CPC Class Codes: B01L 3/0293 : for liquids
• CPC Class Codes: B01L 3/5025 : for parallel transport of multiple samples
• CPC Class Codes: B01L 3/50255 : Multi-well filtration
• CPC Class Codes: B01L 3/5085 : for multiple samples, e.g. microtitration plates
• CPC Class Codes: B01L 3/50857 : using arrays or bundles of open capillaries for holding samples
• CPC Class Codes: B01L 9/523 : for multisample carriers, e.g. used for microtitration plates
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: C40B 70/00 : Tags or labels specially adapted for combinatorial chemistry or libraries, e.g. fluorescent tags or bar codes
• 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 30/466 : with separation columns in parallel
• CPC Class Codes: G01N 30/6095 : Micromachined or nanomachined, e.g. micro- or nanosize
• CPC Class Codes: G01N 30/82 : Automatic means therefor
• CPC Class Codes: G01N 35/00069 : whereby the sample substrate is of the bio-disk type, i.e. having the format of an optical disk
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/25 : including sample preparation
• CPC Class Codes: Y10T 436/25625 : Dilution
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer
• CPC Class Codes: Y10T 83/0481 : Puncturing

Abstract

Methods for manufacturing and using an apparatus for manipulating and analyzing a large number of microscopic samples of a liquid or materials, including cells, in liquid suspension. Parallel through-holes are formed in a platen and loaded with a liquid. Loading may be performed in such a way as to create a gradient, with respect to the position of the through-holes, of the concentration of a particular substance or of another quantity. Mixing of the contents of through-holes may be obtained by bringing filled microwell arrays into contact with each other with registration of individual through-holes.

Claims

1. A method for loading a plurality of disparate sample containers, the sample containers comprising an integral structure, such that the concentration of a specified substance in each container is characterized by a gradient with respect to position of the container within the structure, the method comprising:

a. introducing a first liquid into the disparate containers; and

b. contacting the containers with a second liquid, the second liquid containing the specified substance, in such a manner that the degree of diffusion of the specified substance into the disparate containers differs in a controlled manner among the containers,

wherein contacting the containers with the second liquid includes differing a duration of contact with the second liquid as among the disparate containers.

2. A method for loading a plurality of disparate sample containers, the sample containers comprising an integral structure, such that the concentration of a specified substance in each container is characterized by a gradient with respect to position of the container within the structure, the method comprising:

a. introducing a first liquid into the disparate containers; and

b. contacting the containers with a second liquid, the second liquid containing the specified substance, in such a manner that the degree of diffusion of the specified substance into the disparate containers differs in a controlled manner among the containers,

wherein contacting the containers with the second liquid includes modulating the diffusion rate of the specified substance into the containers.

3. A method according to claim 2, further comprising the step of creating a gradient of the specified substance in the second liquid prior to contacting the containers with the second liquid.

4. A method according to claim 3, wherein the step of creating a gradient of the specified substance includes applying electrophoresis.

5. A method according to claim 2, wherein the step of modulating the diffusion rate includes contacting the containers with the second liquid through a membrane having a permeability variable with respect to position.

6. A method for loading a liquid into a plurality of through-hole platens, each platen having a plurality of through-holes, the method comprising:

a. stacking at least two substantially empty platens together in such an adjacent manner that at least one of the plurality of through-holes from each platen is registered with a through-hole of each other adjacent platen so as to form a registration set of through-holes unimpeded by any solid structure; and

b. transferring the liquid to form a continuous column of liquid in each registration set.

7. A method according to claim 6, wherein each platen is separated from each adjacent platen by an air gap.

8. A method according to claim 6, wherein the step of transferring liquid includes transporting the liquid through capillary tubes.

9. A method according to claim 6, wherein the step of transferring liquid includes transporting the liquid through at least one cannula.

10. A perforated platen having substantially parallel planar surfaces for manipulating distinct liquid samples, each sample having a volume less than 1 microliter, the platen comprising:

a. an inner layer of hydrophilic material;

b. two outer layers of hydrophobic material coupled to opposite sides of the inner layer;

c. a two-dimensional array of through-holes including, at least a first hole having a first liquid, and

d. at least one hole, adjacent to the first hole, containing a second liquid distinct from the first liquid, the through-holes each having a diameter less than 400 micrometers and a density of at least 1.6 through-holes per square millimeter,

wherein the two outer layers of hydrophobic material prevent outmigration of the first and second liquids.