Optical fiber biosensor array comprising cell populations confined to microcavities

• US 6,210,910 B1
• Filed: 03/02/1998
• Issued: 04/03/2001
• Est. Priority Date: 03/02/1998
• Status: Expired due to Term

• Current Assignee: Tufts University
• Original Assignee: Trustees of Tufts College
• Inventors: Walt, David R., Taylor, Laura
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US09/033,462
• Time in Patent Office: 1,128 Days
• Field of Search: 385/12, 385/115, 385/123, 422/57, 422/58, 422/82.05, 422/82.08, 422/82.11, 435/7.2, 435/7.21, 435/287.1, 435/288.7, 435/808, 435/7.32, 436/518, 436/524, 436/527, 436/535, 436/164, 436/165, 436/172, 436/805
• US Class Current: 435/7.32
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00511 Walls of reactor vessels
• CPC Class Codes: B01J 2219/00524 in the shape of fiber bundles
• CPC Class Codes: B01J 2219/00585 Parallel 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 2219/00612 the surface being inorganic
• 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 2219/00628 Ionic
• 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 2219/00648 by the use of solid beads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00704 integrated with the reactor apparatus
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: G01N 21/253 : for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 33/54373 : involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: Y10S 385/901 : Illuminating or display apparatus
• CPC Class Codes: Y10S 435/808 : Optical sensing apparatus
• CPC Class Codes: Y10S 436/80 : Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 436/805 : Optical property

Abstract

A biosensor, sensor array, sensing method and sensing apparatus are provided in which individual cells or randomly mixed populations of cells, having unique response characteristics to chemical and biological materials, are deployed in a plurality of microwells formed at the distal end of individual fibers within a fiber optic array. The biosensor array utilizes an optically interrogatable encoding scheme for determining the identity and location of each cell type in the array and provides for simultaneous measurements of large numbers of individual cell responses to target analytes. The sensing method utilizes the unique ability of cell populations to respond to biologically significant compounds in a characteristic and detectable manner. The biosensor array and measurement method may be employed in the study of biologically active materials, in situ environmental monitoring, monitoring of a variety of bioprocesses, and for high throughput screening of large combinatorial chemical libraries.

Claims

1. A method for detecting the response of individual cells to at least one analyte of interest comprising:

providing a biosensor array comprising,

a fiber optic array comprising a plurality of individual fiber strands, each strand having a distal end and a proximal end and a core, said core diameter sized to accommodate a predetermined cell type, said core capable of being anisotropically etched, and a microwell formed on an end surface of the distal end of each of said strands, said microwell formed by anisotropic etching said core of each of said strands, said microwell sized to accommodate insertion of a single cell of said cell type;

inserting a plurality of cells in said microwells wherein each of said inserted cells in said microwells is optically coupled and in optical communication with one of said fiber strands;

optically coupling a detector means to said strands of said array, said detector means optically communicating with said microwells, said detector means capable of optically interrogating and detecting an optical response of said cells inserted in said microwells to at least one analyte of interest;

contacting the distal end of the array comprising said cells disposed in said microwells with a sample comprising said analyte of interest; and

measuring said optical response of said cells, said optical response measurement indicative of the presence or absence of at least one analyte.

2. The method of claim 1 further comprising illuminating said cells with excitation light energy.

3. The method of claim 1 further comprising capturing images of a plurality of detected optical responses from each of said cells, said images being characteristic of the presence or absence of at least one analyte.

4. The method of claim 1 wherein said cells are cell types selected from a group consisting of invertebrate neurons, mammalian neurons, adrenomedullary cells, lymphocytes, erythrocytes, E.coli bacteria, staphylococcus bacteria, myoblasts, neutrophils, erthroblasts, osteoblasts, chodrocytes, basophils, eosinophils, melanocytes, epithelial cells, epithedial cells, adipocytes, and naturally occuring or genetically engineered cells.

5. The method of claim 1 applied to monitoring cell metabolism changes in bioprocesses.

6. The method of claim 1 applied to monitoring of enzyme activity.

7. The method of claim 1 applied to in situ environmental monitoring of a material selected from a group consisting of heavy metals, metallic compounds, biohazardous materials and environmental pollutants.

8. The method of claim 1 applied to monitoring human, plant and animal pathology.

9. The method of claim 1 applied to screening combinatorial chemical libraries of large numbers of compounds.

10. The method of claim 1 further comprising encoding said cells with material selected from a group consisting of a dye, a fluorescent biomolecule, a fluorophore, a chromophore, and a chemiluscent or bioluminescent compound or material.

11. The method of claim 10 further comprising illuminating said encoded cells with excitation light energy.

12. The method of claim 10 further comprising indexing the location of individual cells in said array by emitted light energy produced by illuminating said encoded cells with excitation light energy.

13. A method for detecting the response of individual cells to at least one analyte of interest comprising:

a) contacting said analyte with a biosensor comprising;

i) a fiber optic array comprising a plurality of individual fiber strands, each strand having a distal end and proximal end;

ii) microwells formed on the distal ends of a plurality of said fiber strands; and

iii) a population of cells dispersed in said microwells; and

b) detecting a response of said cells to said analyte.

14. A method according to claim 13 wherein said cells are encoded.

15. A method according to claim 13 wherein said population of cells comprises at least a first subpopulation of a first cell type and a second subpopulation of a second cell type.

16. A method according to claim 15 wherein said first subpopulation comprises a first encoding moiety and said second subpopulation comprises a second encoding moiety.

17. A method according to claim 16 wherein at least one of said subpopulations of said encoded cells is encoded by attaching a dye to said cells.

18. A method according to claim 16 wherein at least one of said subpopulations of said encoded cells is encoded by the incorporation of a fluorophoric dye compound.

19. A method according to claim 16 wherein at least one of said subpopulations of said encoded cells is encoded by providing said moiety in the local cell environment.

20. A method according to claim 16 wherein at least one of said subpopulations of said encoded cells is genetically engineered to fluoresce.

21. A method according to claim 16 wherein at least one of said subpopulations of said encoded cells comprise a green fluorescent protein.

22. A method according to claim 16 wherein said first and second encoding moieties provide characteristic optical response signatures.

23. A method according to claim 16 further comprising detecting said first and second encoding moieties.

24. A method according to claim 16 wherein said cells are bacterial cells.

25. A method according to claim 16 wherein said cells are eukaryotic cells.

26. A method according to claim 25 wherein said eukaryotic cells are selected from yeast cells, plant cells or mammalian cells.

27. A method according to any one of claims 24, 25 or 26 wherein said cells are genetically engineered.

28. A method according to claim 13 wherein said microwells are coated with a biologically inert material.

29. A method according to claim 28 wherein said biologically inert material is selected from the group consisting of collagen, fibronectin, polylysine, polyethylene glycol and polystyrene.

30. A method according to claim 13 wherein said detecting is the detection of an optical response of said cells.

31. A method according to claim 30 further comprising capturing images of a plurality of detected optical responses from each of said cells, said images being characteristic of the presence or absence of said at least one analyte.