Sensor array and apparatus for simultaneous observation of tissue electrophysiology, contractility, and growth

Patent metadata

Field	Value
Publication number	US-10914722-B2
Application number	US-201815901766-A
Country	US
Kind code	B2
Filing date	Feb 21, 2018
Priority date	Feb 24, 2017
Publication date	Feb 9, 2021
Grant date	Feb 9, 2021

Abstract

A sensor array for simultaneously measuring electrophysiological responses and contractility responses of a tissue includes: a substrate; a multi-electrode array (MEA) disposed in or on the substrate; and a plurality of interdigitized electrodes (IDEs) disposed in or on the substrate. The MEA and the IDEs are interpenetrating within a plane substantially parallel to an upper surface of the substrate. Systems for measuring such responses using the sensor array may also include contacts and/or connectors for interfacing with external control devices, electrodes, and cell culture components such as a chamber and lid. Fabrication techniques for making such systems include: forming a sensor array comprising an interpenetrating arrangement of IDEs and electrodes of a MEA in or on a substrate surface; forming a plurality of contacts for interfacing the system with one or more external devices in or on the substrate surface; and forming leads between the plurality of contacts and the sensor array.

First claim

What is claimed is:

1. A sensor array for simultaneously measuring electrophysiological responses and contractility responses of a tissue across two dimensions, the sensor array comprising:
a substrate;
a plurality of multi-electrode array structures forming a multi-electrode array (MEA) disposed in or on the substrate, wherein each MEA structure comprises a plurality of MEA electrodes aligned parallel to a first line; and
a plurality of interdigitized electrode (IDE) structures disposed in or on the substrate wherein each IDE structure comprises a plurality of IDEs aligned parallel to a second line; and
wherein the MEA and the IDEs are interpenetrating within a plane substantially parallel to an upper surface of the substrate;
wherein the sensor array is arranged according to an alternating pattern wherein one of the IDE structures is positioned between each adjacent pair of the MEA structures; and
wherein the sensor array comprises a rectangular grid defined within the plane substantially parallel to the upper surface of the substrate.

2. The sensor array as recited in claim 1 , wherein the rectangular grid defines a two-dimensional map of field potential-sensitive regions of the sensor array.

3. The sensor array as recited in claim 1 , wherein the plurality of IDEs are interleaved with electrodes of the MEA; and
wherein a longitudinal axis of each of the IDE structures is substantially parallel to a longitudinal axis of each of the MEA structures.

4. The sensor array as recited in claim 1 , wherein the IDEs are characterized by a substantially fractal geometry.

5. The sensor array as recited in claim 1 , wherein the MEA comprises sixteen electrodes arranged into four of the MEA structures, each MEA structure comprising four electrodes; and
wherein the sensor array comprises at least three, and no more than nine, IDEs arranged into up to five IDE structures each IDE structure comprising one to three of the IDEs.

6. The sensor array as recited in claim 1 , wherein leads of the IDEs and electrodes of the MEA each respectively comprise one or more materials selected from a group consisting of: titanium, platinum black, and carbon nanotubes.

7. The sensor array as recited in claim 1 , wherein the substrate comprises a material selected from a group consisting of: silicon, polydimethylsiloxane (PDMS); and
wherein the substrate comprises coating comprising a material selected from the group consisting of silicon oxide and silicon nitride.

8. The sensor array as recited in claim 1 , wherein the sensor array is characterized by an area less than or equal to an area of a single well of a standard, commercially available 96-well cell culture plate.

9. The sensor array as recited in claim 1 , wherein electrodes of the MEA structures and the IDEs are spatially separated by a distance greater than or equal to a mean diameter of an individual cell of the tissue.

10. The sensor array as recited in claim 1 , wherein each MEA structure comprises at least four electrodes of the MEA;
wherein the first line is parallel to a longitudinal axis of the MEA structures;
wherein the second line is parallel to a longitudinal axis of the IDE structures; and
wherein the first line is parallel to the second line.

11. The sensor array as recited in claim 1 , wherein electrodes of the MEA and the plurality of IDEs are arranged according to a checkerboard pattern.

12. The sensor array as recited in claim 1 , wherein the IDEs are characterized by a substantially linear geometry.

13. The sensor array as recited in claim 1 , wherein electrodes of the MEA and electrodes of the IDEs are spatially separated by a distance in a range from about 50 microns to about 100 microns.

14. The sensor array as recited in claim 1 , wherein the IDE structures are serially connected to one another.

15. The sensor array as recited in claim 1 , wherein the IDE structures are connected to one another in parallel.

16. A system for simultaneously measuring electrophysiological responses and contractility responses of the tissue, the system comprising:
the sensor array as recited in claim 1 ;
a first set of electrical connectors disposed in or on a first surface of the substrate, wherein the first set of electrical connectors are electrically coupled to the sensor array via a plurality of MEA leads;
a second set of electrical connectors disposed in or on a second surface of the substrate, the second surface being positioned along an opposing side of the substrate as the first surface, and wherein the second set of electrical connectors are electrically coupled to the sensor array via a plurality of IDE leads;
a cell culture chamber; and
an enclosure surrounding the sensor array, the first set of electrical connectors and the second set of electrical connectors, and the cell culture chamber.

17. The system as recited in claim 16 , wherein the enclosure comprises a faraday cage.

18. The system as recited in claim 16 , comprising an optical microscope and an objective lens positioned on opposing sides of the enclosure;
wherein the optical microscope illuminates tissue in the cell culture chamber; and
wherein the objective lens captures light from the illuminated tissue.

19. The system as recited in claim 16 , comprising an impedance analyzer and an electrical stimulator each electrically coupled to the sensor array via the second set of electrical connectors.

20. The system as recited in claim 19 , comprising a plurality of electrical stimulation electrodes electrically coupled to the electrical stimulator via the second set of electrical connectors.

21. The system as recited in claim 16 , comprising a multi-channel workstation electrically coupled to the sensor array via the first set of electrical connectors.

22. The system as recited in claim 21 , comprising an electrophysiological ground electrode and an electrophysiological stimulation electrode each electrically coupled to the multi-channel workstation via the first set of electrical connectors.

23. The system as recited in claim 16 , comprising a passivation layer formed in or on one or more regions of the upper surface of the substrate, wherein the upper surface of the substrate comprises:
the first set of electrical connectors;
the second set of electrical connectors;
one or more MEA leads connecting the MEA electrodes and the first set of electrical connectors; and/or
one or more IDE leads connecting the IDEs and the second set of electrical connectors.

24. A method, comprising: simultaneously measuring electrophysiological responses and contractility responses of a plurality of cells forming a tissue culture using the system as recited in claim 16 .

Assignees

• L Livermore Nat Security Llc

Inventors

• Qian Fang
• Huang Chao
• Ivanovskaya Anna Nikolaevna
• Kulp Kris
• Wheeler Elizabeth K

Classifications

• C12M35/02

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

• G01N33/4836 Primary

  using multielectrode arrays · CPC title

• C12M41/46

  of cellular or enzymatic activity or functionality, e.g. cell viability · CPC title

• C12M21/08

  for producing artificial tissue or for ex-vivo cultivation of tissue (prostheses A61F2/00, grafts A61L27/00) · CPC title