System and method for three-dimensional in vitro flexible microelectrode array

Patent metadata

Field	Value
Publication number	US-11725170-B2
Application number	US-201916677328-A
Country	US
Kind code	B2
Filing date	Nov 7, 2019
Priority date	Nov 7, 2019
Publication date	Aug 15, 2023
Grant date	Aug 15, 2023

Abstract

The present disclosure relates to a biocompatible, in vitro probe system. The probe system may have a substrate and a culture well supported on the substrate. The culture well defines a three-dimensional volume for containing in vitro cultures of electroactive cells. The probe system has at least one probe subsystem supported on the substrate. The probe subsystem has at least one probe having an array of electrodes, with the probe being disposed within the culture well for in vitro electrically communicating with the electroactive cells. The probe subsystem is adapted to be interfaced to an external instrumentation/recording device.

First claim

What is claimed is:

1. A biocompatible, in vitro probe system comprising:
a substrate;
a culture well supported on the substrate and defining a three-dimensional volume for containing in vitro cultures of electroactive cells;
at least one probe subsystem supported on the substrate and including at least one probe having an array of electrodes, the probe being disposed within the culture well for in vitro electrically communicating with the electroactive cells, and adapted to be interfaced to an external electronic instrumentation/recording device;
the probe further including a first region, a second region and a hinge region including a void forming an engineered weakness for controllably weakening the probe at a selected location to aid in bending the probe during a manufacturing operation, the second region including the electrodes and forming a probe sensing body portion, and the first region being fixed to the substrate and adapted to be placed in electrical communication with the external electronic instrumentation/recording device; and
the electrodes being spaced elevationally apart along the second region at different distances from the first region.

2. The system of claim 1 , wherein the probe subsystem includes a plurality of the probes, with each one of the probes having a plurality of electrodes thereon.

3. The system of claim 1 , wherein the probe subsystem includes a plurality of the probes, with each said probe having a plurality of the electrodes thereon spaced apart from one another in a straight line.

4. The system of claim 3 , wherein each said probe of the probe subsystem includes a first region and a second region, with the second region including the plurality of electrodes, wherein each one of the second regions is disposed non-parallel to the first regions within the three-dimensional volume of the culture well.

5. The system of claim 3 , wherein the probes are arranged to extend non-parallel to the substrate, such that the probes collectively form an X-Y spatial arrangement of probes within the culture well, and such that the electrodes of the probes are arranged in X, Y and Z planes within the three-dimensional volume of the culture well.

6. The system of claim 5 , wherein the probes are arranged to extend generally orthogonal to the substrate, and parallel to one another, within the three-dimensional volume of the culture well.

7. The system of claim 1 , further comprising at least one electrical connection pad disposed on the substrate and in electrical communication with the probe subsystem.

8. The system of claim 1 , wherein the probe subsystem is used to apply an electrical signal to the electrode.

9. The system of claim 1 , wherein the electrode of the probe subsystem is used to receive electrical signals generated within the cultures of electroactive cells.

10. The system of claim 1 , wherein the probe subsystem is used to both apply electrical signals to the electrode and to receive electrical signals from the electrode.

11. The system of claim 1 , wherein the culture well is secured to an upper surface of the substrate by a biocompatible epoxy.

12. A biocompatible, in vitro probe system comprising:
a planar substrate;
a culture well secured to a surface of the substrate and defining a three-dimensional volume for containing in vitro cultures of electroactive cells;
at least one probe subsystem supported on the substrate and including a first region, a second region and a third region located between the first region and the second region;
the second region including a plurality of probes arranged in an X-Y plane extending non-parallel to the first region, and disposed inside the three-dimensional volume of the culture well;
at least a portion of the first region extending parallel to the substrate and out from the culture well;
the third region including a void forming an engineered weakness in the substrate to facilitate bending of the probe at the third region;
each of the probes including a plurality of elevationally spaced apart electrodes arranged at different distances from the first region that collectively form an in vitro, three-dimensional network of electrodes within the three-dimensional volume of the culture well; and
the probe subsystem including circuit traces extending from the first region into the second region and into electrical contact with the electrodes of each of the probes, for enabling an external electrical subsystem to electrically communicate with the electrodes on the probes.

13. The probe system of claim 12 , further comprising a plurality of independent electrical connection pads secured to the surface of the substrate, and in electrical communication with the circuit traces, the electrical connection pads being adapted for connection to the external electrical subsystem.

14. The probe system of claim 13 , wherein each one of the electrical connection pads is independently coupled to a single one of the circuit traces, to enable each one of the electrical connection pads to independently communicate with an associated one of the electrodes.

15. The probe system of claim 12 , wherein the probe subsystem is configured to at least one of:
apply an electrical signal to at least one of the electrodes; and
receive electrical signals from at least one of the electrodes.

16. The probe system of claim 12 , wherein the probes define a X-Y spatial arrangement of probes, with the electrodes defining a three-dimensional network of electrodes within the three-dimensional volume of the culture well.

17. A method for electrically communicating with a quantity of cultures of electroactive cells, comprising:
using a culture well to define a three-dimensional volume for containing in vitro cultures of electroactive cells;
using at least one probe subsystem having a portion with a plurality of probes extending into the culture wells, wherein each said probe includes a first region supported from a planar substrate, a second region extending non-parallel to the first region, and a third region separating the first region and the second region, the third region including a void to form an engineered weakness in the probe to permit the second region to be bent to extend non-parallel to the first region;
the second region including a plurality of elevationally spaced apart electrodes disposed at different distances from the first region, to form a three-dimensional network of electrodes within the three-dimensional volume of the culture well; and
using the electrodes to in vitro electrically communicate with the electroactive cells.

18. The method of claim 17 , further comprising interfacing the at least one probe subsystem to an external electrical subsystem for at least one of applying electrical signals to at least one of the electrodes, or receiving electrical signals from at least one of the electrodes.

Assignees

• L Livermore Nat Security Llc

Inventors

• Soscia David
• Enright Heather Ann
• Fischer Nicholas
• Lam Doris Mailie
• Tooker Angela C
• Triplett Michael
• Wheeler Elizabeth K

Classifications

• C12M23/12 Primary

  Well or multiwell plates (C12M25/04 takes precedence) · CPC title

• B01L3/5085 Primary

  for multiple samples, e.g. microtitration plates · CPC title

• C12M35/02

  Electrical or electromagnetic means, e.g. for electroporation or for cell fusion · CPC title

• C12N5/0068

  General culture methods using substrates (for specific animal cell type C12N5/06) · CPC title

• G01N27/30

  Electrodes, e.g. test electrodes; Half-cells (G01N27/414 takes precedence) · CPC title