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A graphene three-dimensional microelectrode array chip, method and application thereof

A graphene and micro-electrode technology, which is applied in the field of electrophysiological detection of cells and tissues, can solve the problems of flexible cells or tissues in close contact, increase the specific surface area of ​​electrode sites, etc., and achieve excellent electrochemical stability and excellent electrical properties. Properties, the effect of high light transmittance

Active Publication Date: 2017-09-15
上海前瞻创新研究院有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In order to overcome the shortcoming that the electrode sites of the existing graphene microelectrode array chip plane cannot form close contact with flexible cells or tissues, and increase the specific surface area of ​​the electrode sites, the object of the present invention is to provide a graphene three-dimensional microelectrode array Chip, method and application thereof, specifically applicable to the detection of electrophysiological signals of cells and tissues

Method used

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  • A graphene three-dimensional microelectrode array chip, method and application thereof
  • A graphene three-dimensional microelectrode array chip, method and application thereof
  • A graphene three-dimensional microelectrode array chip, method and application thereof

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Effect test

Embodiment 1

[0015] The manufacturing process of graphene three-dimensional microelectrode chip is as follows: figure 2 As shown, the details are as follows:

[0016] (1) Clean the substrate: use Phiranha solution to clean the silicon wafer or quartz wafer or borosilicate glass wafer (such as Prex7740), then rinse with deionized water, blow dry with nitrogen, and treat with oxygen plasma for 5 minutes;

[0017] (2) Lift-off process to make the outer electrode leads and pins in the outer microelectrode array area: Spin-coat AZ4620 on the substrate, pattern it by photolithography, then sputter the metal layer (titanium / gold), and remove the photolithography with acetone Glue, leaving the metal layer. ( figure 2 .a)

[0018] (3) Production of three-dimensional microcolumn arrays: Spin-coat negative photoresist (SU8 3005 for silicon wafers, PI 7510 for quartz wafers and Prex7740 borosilicate glass), control the speed at 3000r / min, and after photolithography and development, cure to form ...

Embodiment 2

[0023] Graphene three-dimensional microelectrode performance testing method

[0024] The electrochemical impedance and cyclic voltammetry curve test methods are as follows:

[0025] (1) Gamry reference 600 electrochemical workstation was used to perform electrochemical characterization of microelectrodes using a three-electrode system.

[0026] (2) A plastic cavity is glued on the substrate with PDMS, and the microelectrode array area is located in the cavity. Inject PBS solution into the cavity.

[0027] (3) The graphene three-dimensional microelectrode site is used as the working electrode, the Ag / AgCl is used as the reference electrode, and the platinum wire is used as the counter electrode, and the three electrodes are immersed in the PBS solution described in (2).

[0028] (4) When performing electrochemical impedance analysis, the electrochemical workstation outputs a sinusoidal AC signal with a peak value of 50mV, and the frequency range is from 0.01HZ to 1MHz.

[00...

Embodiment 3

[0033] Cell culture and detection on a graphene three-dimensional microelectrode array chip

[0034] (1) Surface treatment of the graphene three-dimensional microelectrode array: before inoculating cells, the graphene three-dimensional microelectrode was soaked in 75% ethanol solution for one hour, and then dried under ultraviolet light; then poly-D-lysine (PDL) solution according to 2μg / cm 2 The concentration of 3D microelectrode array was dropped on the surface of the graphene three-dimensional microelectrode array, kept overnight at room temperature, washed three times with ultrapure water, and air-dried.

[0035] (2) Seeding cells on the surface of graphene three-dimensional microelectrode array: Digest conventionally cultured SH-SY5Y cells with digestion solution (0.25% trypsin, 0.02% EDTA solution), centrifuge, pipette, and dilute the cells at 1.0×10 7 cells / mm 3 The density is seeded on the graphene three-dimensional microelectrode array chip.

[0036] (3) Induced di...

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Abstract

The invention relates to a graphene three-dimensional microelectrode array chip, and a method and application thereof. The graphene three-dimensional microelectrode array chip is characterized in that a micro-column array is produced by utilizing negative photoresist, and a microelectrode array is produced by covering a single-layer graphene film on the microelectrode array; and the microelectrode array chip comprises two parts, namely a transparent graphene three-dimensional microelectrode array area and a peripheral gold electrode lead pin. A microelectrode site is a three-dimensional bulge. The three-dimensional microelectrode dome shaped (or hill shaped) microelectrode structure is beneficial for the rigid microelectrode site and the soft cell or tissue to form tight electric coupling, and the graphene has excellent electrical property, so that the electrophysiological detection sensitivity of the microelectrode array can be improved. In addition, the graphene three-dimensional microelectrode array on a transparent substrate is convenient for observation through an inverted microscope and application of various cell microscopic imaging methods, and can be combined with a micro-fluidic chip.

Description

technical field [0001] The invention relates to a graphene three-dimensional microelectrode array chip, a preparation method and an application thereof. The graphene three-dimensional microelectrode array chip provided by the invention can be used in the field of biomacromolecule, cell or tissue detection, especially in the detection of cells and tissues Electrophysiological testing. Background technique [0002] The microelectrode array chip is a sensor that integrates multiple micron-level electrodes on a chip for biological signal detection by using a micromachining process. The microelectrode array chip can simultaneously detect the electrical signals of biological macromolecules in the solution, detect the biological electrical signals of cells and tissues, or be used to electrically stimulate cells and tissues. [0003] At present, the electrode materials of microelectrode array chips are mostly metals such as gold and platinum, but their opaque nature cannot be compa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B81B1/00B81C1/00G01N27/00G01N33/483G01N33/487
CPCB81B1/00B81C1/00111G01N27/00G01N33/4836G01N33/48707G01N33/48728
Inventor 吴蕾唐琳金庆辉赵建龙
Owner 上海前瞻创新研究院有限公司
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