PICOWELL CAPTURE DEVICES FOR ANALYSING SINGLE CELLS OR OTHER PARTICLES

• US 20120156675A1
• Filed: 06/09/2010
• Published: 06/21/2012
• Est. Priority Date: 06/09/2009
• Status: Abandoned Application

• Current Assignee: Oxford Gene Technology IP Limited (Sysmex Corporation)
• Original Assignee: Oxford Gene Technology IP Limited (Sysmex Corporation)
• Inventors: Lueerssen, Dietrich Wilhelm Karl, Malleo, Daniele
• Application Number:
  US13/377,211
• Publication Number:
  US 20120156675A1
• Time in Patent Office: Days
• US Class Current: 435/6.11
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/069 Absorbents; Gels to retain a fluid
• CPC Class Codes: B01L 2300/0822 Slides
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/0893 having a very large number of wells, microfabricated wells
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/50853 with covers or lids
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 23/20 Material Coatings immunocoatings C12M25/00
• CPC Class Codes: C12M 25/06 Plates; Walls; Drawers; Multilayer plates
• CPC Class Codes: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04

Abstract

A convenient way of isolating individual cells permits individual analysis of their contents. A capture support for individually capturing cells of interest comprises a first surface including at least one well sized to accommodate an individual cell, wherein the support is made of a differentially permeable material which permits transfer of a solvent and any low molecular weight species through the support from a second surface of the support to a well, but which is substantially impermeable to biopolymers. Single cells are captured, their contents are released, and the contents of individual cells are then analysed within a chamber containing suitable analytical components e.g. immobilised nucleic acid probes, immobilised antibodies, etc. Analysis of a single cell'"'"'s genome, transcriptome, proteome, etc. thus becomes possible.

Claims

1. A capture support for individually capturing cells of interest, comprising a first surface including at least one well sized to accommodate an individual cell, wherein the support is made of a differentially permeable material which permits movement of a solvent and any low molecular weight species through the support from a second surface of the support to a well, but which is substantially impermeable to biopolymers.

3. The capture support of claim 1, wherein at least one of the at least one wells is surrounded by a recess.

4. The capture support according to claim 3, wherein the recess is 0.1-5 μ

M deep.

5. The capture support according to claim 1, wherein the low molecular weight species have MW less than 1000 Da.

6. The capture support according to claim 1, wherein at least a portion of at least one well'"'"'s surface is coated with reagents to aid capture of the cells or particles.

7. The capture support according to claim 1, wherein at least a portion of at least one well'"'"'s surface is coated with one or more analytical reagents which permit analysis of biopolymers.

8. The capture support according to claim 1, wherein the capture support is made from a permeable polymer.

9. The capture support according to claim 8 wherein the permeable polymer is a polyacrylamide gel.

10. The capture support according to claim 1, wherein the at least one well is intersected by, or connected to, an open channel.

11. The capture support according to claim 1, wherein the capture support comprises a reservoir integral to the capture support.

12. A device for analysing the contents of an individual cell comprising (i) a capture support according to claim 1 and (ii) a lid for receiving the contents of a well.

13. The device according to claim 12, wherein at least a portion of the'"'"' lid is coated with analytical reagents.

14. The device according to claim 13, wherein the lid comprises recesses where the analytic reagents are located.

15. A process for analysing one or more cells of interest, comprising steps of:

capturing the cell(s) within well(s) of a capture support of claim 1;

sealing the well(s) with a lid to form one or more chamber(s) from which captured cells cannot exit;

releasing the contents of cell(s) such that they remain in the one or more chambers;

allowing the released contents to interact with one or more analytical component(s) within the one or more chambers, thereby permitting analysis of the contents.

16. A mould for the manufacture of a capture support according to claim 1, wherein the mould is capable of receiving material which solidifies within the mould.

17. A method for the manufacture of a capture support according to claim 1, said method comprising the steps of:

a) adding to a mould a material capable of solidifying;

b) incubating the mould under conditions which permit the material to solidify; and

c) separating the mould and the solidified material, to leave a capture support according to claim 1.

2. A capture support for individually capturing particles, comprising a first surface including at least one well with a diameter of less than 200 μ

m and depth of less than 200 μ

M, wherein the support is made of a differentially permeable material which permits movement of a solvent and any low molecular weight species through the support from a second surface of the support to a well, but which is substantially impermeable to biopolymers.