Device for magnetic immobilization of cells

• US 7,285,412 B2
• Filed: 02/28/2002
• Issued: 10/23/2007
• Est. Priority Date: 07/27/2001
• Status: Expired due to Fees

• Current Assignee: Surface Logix, Inc.
• Original Assignee: Surface Logix, Inc.
• Inventors: Kim, Enoch, Nussbaum, Michael, Raphel, Aaron, Casagrande, Rocco, Wang, Evelyn, Kirk, Gregory
• Primary Examiner(s): Naff; David M.
• Application Number:
  US10/084,063
• Publication Number:
  US 20030022370A1
• Time in Patent Office: 2,063 Days
• Field of Search: 435/4, 435/174, 435/177, 435/180
• US Class Current: 435/297.1
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00655 the compounds being bound to magnets embedded in or on the solid supports
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00743 Cells
• CPC Class Codes: B01L 2400/043 magnetic forces
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: B01L 3/5088 confining liquids at a location by surface tension, e.g. virtual wells on plates, wires B01L3/50857 takes precedence
• CPC Class Codes: C12M 33/00 Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus chemical or physical laboratory apparatus in general B01L, devices for taking cell samples A61B10/0045, withdrawing or distributing predetermined quantities of fluid B01L99/00
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 35/0098 involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C

Abstract

The present invention relates to an apparatus and methods that immobilize one or more cells associated with magnetic material on a substrate on which are located one or more magnetic receptacle(s). Alternatively, in another aspect the present invention, the device arrays cells associated with magnetic material on a substrate having a pattern of magnetic receptacles disposed thereon. The size of the magnetic receptacle(s) determines the number of target cells that it is capable of immobilizing. The size of the magnetic receptacle is defined by the strength of a localized magnetic field gradient. The localized magnetic field gradient may be derived from 1) permanent magnets embedded in the substrate or alternatively, the localized magnetic field gradient may be derived from an 2) external magnet whose strength is focused by objects of highly-permeable-magnetic material which create localized magnetic field gradients. The invention apparatus comprises a removable cell delivery device and a substrate, which has one or more magnetic receptacles disposed thereon.

Claims

1. A device for arraying a plurality of cells into discrete and predetermined locations for further experimentation, said device comprising a substrate having an essentially flat surface, wherein a plurality of magnets are contained in said substrate, wherein said plurality of magnets are arrayed in said substrate such that each magnet defines a localized magnetic field gradient to define a magnetic area, wherein said magnetic area is situated on the surface of the substrate in a predetermined location discrete from other magnetic areas to provide a plurality of discrete magnetic areas, wherein the plurality of magnetic areas defined by said plurality of magnets are disposed in a two-dimensional array on the substrate, wherein said localized magnetic field gradient immobilize one to about five cells within said each of said plurality of magnetic areas, wherein said cells are associated with magnetic material at the time that said cells are immobilized within said plurality of magnetic areas.

2. The device of claim 1 wherein said cells are hybridoma cells.

3. The device of claim 1 wherein the substrate is fabricated from a material selected from the group consisting of glass, urethane, rubber, molded plastic, polymethylmethacrylate, polycarbonate, polytetrafluoroethylene, polyvinylchloride, polydimethylsiloxane, and polysulfone.

4. The device of claim 1 further comprising a layer on top of said substrate wherein said layer has micro-gaps positioned over said magnetic areas.

5. The device of claim 1 further comprising a cell isolation device, wherein said cell isolation device comprises a membrane containing a plurality of wells that match the plurality of the magnetic areas, such that when said cell isolation device is placed on said substrate, said cell isolation device is capable of isolating said one to about 5 cells immobilized in one of said plurality of magnetic areas from other of said cells immobilized in said other of said plurality of magnetic areas arrayed within the cell isolation device.

6. The device of claim 5, wherein the wells of the cell isolation device are micro through-holes, wherein the micro through-holes are defined by inner walls of the membrane.

7. The device of claim 6, wherein the device further comprises a semi-permeable membrane opposite the substrate, wherein said semi-permeable membrane restricts cell movement between wells and is permeable to fluid.

8. The device of claim 7 wherein at least one of the walls of the micro through-holes are canted or perpendicular to the substrate.

9. The device of claim 5, wherein said plurality of magnetic areas further comprises immobilized cells, such that when the cell isolation device is placed on said substrate, said cells are capable of being transferred from said plurality of magnetic areas to said cell isolation device, and when the substrate is removed, the cells remain in the cell isolation device.

10. The device of claim 9 wherein said cells are capable of being transferred from said plurality of magnetic areas to said cell isolation device by centrifugal force.

11. The device of claim 1 wherein the substrate is coated with a hydrophobic agent.

12. The device of claim 11 wherein the hydrophobic agent is selected from the group consisting of teflon, perfluoronated plastic, polyethylene glycol, ethylene oxide-terminated trichlorosilane, and hydrophobic alkyltrichlorosilane.

13. The device of claim 1 wherein the substrate is coated with an anti-coagulant.

14. The device of claim 13 wherein the anti-coagulant is selected from the group consisting of heparin, heparin fragments, tissue-type plasminogen activator (tPA), urokinase (uPA), Hirudan, albumin, anti-platelet receptor GPIB antibodies, anti-platelet receptor GPIIb/IIIa antibodies, and anti-von Willebrand Factor (vWF) antibodies.

15. The device of claim 1 wherein at least one of the magnets is a permanent magnet.

16. The device of claim 1 wherein at least one of the magnets is made of highly-magnetically-permeable material.