Cell analysis in multi-through-hole testing plate

• US 20050059074A1
• Filed: 10/20/2004
• Published: 03/17/2005
• Est. Priority Date: 03/19/1999
• Status: Abandoned Application

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Schellenberger, Volker, Liu, Amy Deming
• Application Number:
  US10/969,104
• Publication Number:
  US 20050059074A1
• Time in Patent Office: Days
• US Class Current: 506/4
• 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 2219/00286 Reactor vessels with top and bottom openings
• 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 2219/0052 in the shape of elongated tubes
• CPC Class Codes: B01J 2219/00524 in the shape of fiber bundles
• CPC Class Codes: B01J 2219/00533 essentially rectangular
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00644 the porous medium being present in discrete locations, e.g. gel pads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00662 Two-dimensional arrays within two-dimensional arrays
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0809 rectangular shaped
• CPC Class Codes: B01L 2300/161 Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• 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: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 2035/00237 Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 35/1065 : Multiple transfer devices

Abstract

A spatially fixed library of mutated cells and a method for creating such a library. The library is created by creating a set of cells having a genetic diversity, causing the cells to multiply and express phenotypes, preparing a solution containing the cells diluted in a medium, populating a plurality of through-holes of a testing plate with the solution such that surface tension holds the solution in respective through-holes, and analyzing the phenotypes of the cells in the solution on a hole-by-hole basis. Cells expressing specified phenotypes may then be retrieved.

Claims

1. A method for creating a spatially fixed library of mutated cells, the method comprising:

a. creating a set of cells characterized by genetic diversity;

b. causing the cells to multiply and express phenotypes;

c. preparing a solution containing the cells diluted in a medium;

d. populating a plurality of through-holes of a testing plate with the solution such that surface tension holds the solution in respective through-holes; and

e. analyzing the phenotypes of the cells in the solution on a hole-by-hole basis.

2. The method for creating a spatially fixed library of mutated cells in accordance with claim 1, wherein the step of creating a set of cells characterized by genetic diversity includes exposing the cells to a mutagen.

3. The method of claim 1, wherein the step of populating the plurality of through-holes is performed prior to causing the cells to multiply and express phenotypes.

4. The method for creating a spatially fixed library of mutated cells in accordance with claim 1, wherein the step of introducing the set of cells in a medium into an array of through-holes includes delivering multiple cells to each of the through-holes of the plurality of through-holes.

5. The method for creating a spatially fixed library of mutated cells in accordance with claim 2, wherein the step of exposing the cells to a mutagen includes using at least one of ultraviolet and chemical mutagenesis.

6. The method for creating a spatially fixed library of mutated cells in accordance with claim 1, wherein the step of causing the cells to multiply and express phenotypes includes incubating the cells.

7. The method for creating a spatially fixed library of mutated cells in accordance with claim 1, wherein the step of analyzing the phenotypes includes optically analyzing contents of the through-holes.

8. The method for creating a spatially fixed library of mutated cells in accordance with claim 1, wherein the step of analyzing the phenotypes includes measuring fluorogenic groups liberated by hydrolysis of a substrate.

9. The method for creating a spatially fixed library of mutated cells in accordance with claim 1, wherein the step of analyzing the phenotypes includes measuring chromogenic groups liberated by hydrolysis of a substrate.

10. The method for creating a spatially fixed library of mutated cells in accordance with claim 1, wherein the step of analyzing the phenotypes includes optically imaging a plurality of through-holes of the testing plate.

11. The method for creating a spatially fixed library of mutated cells in accordance with claim 1, further comprising retrieving genetic determinants expressing a specified phenotype.

12. A method in accordance with claim 11, wherein the step of retrieving includes robotic extraction of contents of an identified through-hole.

13. A method in accordance with claim 11, wherein the step of retrieving includes non-robotic extraction of contents of an identified through-hole.

14. A library of genetic diversity comprising a set of mutated cells suspended in a medium, the medium retained by surface tension in a plurality of through-holes of a test plate, the cells induced to express phenotypes.

15. The library of genetic diversity of claim 14, wherein the medium additionally contains an analyte that produces a detectable characteristic.

16. The library of genetic diversity of claim 15, wherein said detectable characteristic is produced in an absorbance transcription assay.

17. The library of genetic diversity of claim 15, wherein said detectable characteristic is produced in a fluorescent transcription assay.

18. The library of genetic diversity of claim 15, wherein said detectable characteristic is produced in a fluorescent secreted enzyme assay.

19. The library of genetic diversity of claim 15, wherein said detectable characteristic is produced in a microorganism screening assay.