Optical fiber biosensor array comprising cell populations confined to microcavities

• US 6,667,159 B1
• Filed: 09/15/2000
• Issued: 12/23/2003
• Est. Priority Date: 03/02/1998
• Status: Expired due to Term

• Current Assignee: Trustees of Tufts College
• Original Assignee: Trustees of Tufts College
• Inventors: Taylor, Laura, Walt, David R.
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US09/663,510
• Time in Patent Office: 1,194 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, 435/287.2, 435/288.7, 435/808, 435/7.32, 436/164, 436/165, 436/172, 436/518, 436/524, 436/527, 436/535, 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 biosensor for detecting the response of individual cells to at least one analyte of interest 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;

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;

wherein each of said single cells in said microwells is optically coupled and in optical communication with one of said fiber strands.

2. The biosensor of claim 1 wherein said microwells have an internal diameter ranging from about 1 um to about 200 um, a depth ranging from about 0.25 um to about 200 um, and a volume ranging between about 1 fL to about 5 nanoliter.

3. The biosensor of claim 1 wherein the diameter of said core of said fiber strands ranges from about 1 um to about 200 um.

4. The biosensor of claim 1 wherein an interior surface of said microwell is coated with a thin film of a biologically compatible material selected from a group consisting of collagen, fibronectin, polylysine, polyethylene glycol, polystyrene, and metal.

5. The biosensor of claim 4 wherein said metal comprises gold, platinum or palladium.

6. The biosensor of claim 1 wherein an indicator compound is applied to an interior surface of said microwell for detecting cell responses to chemical or biological stimulation.

7. The biosensor of claim 1 wherein at least one cell type is inserted in at least one microwell, said cell type 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.

8. The biosensor of claim 7 wherein an indicator compound is attached or inserted into said cell.

9. The biosensor of claim 1 wherein a plurality of cells are inserted in a plurality of microwells.

10. The biosensor of claim 9 wherein said plurality of cells is comprised of at least two cell populations which are randomly mixed and inserted in said microwells.

11. The biosensor of claim 10 wherein each cell population is encoded with at least one optically interrogatable material selected from a group consisting of a fluorophore, a chromophore, and a chemiluminescent or bioluminescent compound or material.

12. An apparatus for detecting the response of individual cells to at least one analyte of interest comprising:

a) a biosensor array comprising

i) 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,

ii) 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,

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

iv) a detector optically coupled to a proximal end of said strands, said detector capable of detecting an optical response of said cells inserted in said microwells, said response being characteristic of the presence or absence of said analyte.

13. The apparatus of claim 12 further comprising an excitation light energy component coupled to said proximal end of said strands, said light energy component for illuminating said cells in said microwells.

14. The apparatus of claim 12 further comprising an image capturing system for capturing images of a plurality of detected optical responses from each individual cell, said images being characteristic of the presence or absence of said analyses.

15. The apparatus of claim 12 wherein said image capturing system comprises a CCD camera, a computer interface and a programmable computer.