A microfluidic paper chip based on nano-gold modified screen-printed electrodes for online monitoring of intracellular dopamine

A technology of screen-printed electrodes and microfluidic paper chips, which is applied in measuring devices, material analysis through electromagnetic means, instruments, etc., can solve the problems of large sample volume, complicated operation, difficulty in integration and miniaturization, etc. Achieve high biocompatibility, low cost, and easy processing

Active Publication Date: 2020-07-03
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, traditional electrodes have disadvantages such as large amount of sample required, complex operation, difficulty in integration and miniaturization, which is not conducive to the realization of online detection at the cell level

Method used

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  • A microfluidic paper chip based on nano-gold modified screen-printed electrodes for online monitoring of intracellular dopamine
  • A microfluidic paper chip based on nano-gold modified screen-printed electrodes for online monitoring of intracellular dopamine
  • A microfluidic paper chip based on nano-gold modified screen-printed electrodes for online monitoring of intracellular dopamine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] (1) Wash the screen-printed carbon electrode with deionized water 3 times and blow dry with nitrogen. Drop 2.8 mmol / L chloroauric acid solution and 0.5 mol / L sulfuric acid solution on the surface of the screen-printed carbon electrode, and deposit gold nanoparticles on the screen-printed carbon electrode 4 by chronoamperometry. The surface is modified with gold nanoparticles. Working electrode 7 (such as figure 1 Shown). The screen-printed carbon electrode 4 is composed of a reference electrode 6, a working electrode 7 and a counter electrode 8. On the surface of the screen-printed carbon electrode 4, a reference electrode 6 is integrated on the side of the working electrode 7 and is located on the other side of the working electrode 7. The counter electrode 8 is integrated.

[0034] From figure 2 In A, it can be seen that the surface of the bare-screen-printed carbon electrode is relatively rough, which is mainly caused by micron-sized granular carbon powder; when the ...

Embodiment 2

[0042] The paper-based cell culture steps of a microfluidic paper chip based on nano-gold modified screen-printed electrodes for online monitoring of intracellular dopamine are as follows:

[0043] Take 100 uL density as 5×10 4 Cells / mL cell suspension was dropped on the filter paper surface of the paper-based microfluidic chip prepared in Example 1, and the entire paper-based microfluidic chip was placed in a petri dish, and then the petri dish was placed in 5wt% carbon dioxide. The place near the water in the 37°C incubator. Fluorescence imaging of cells cultured on paper chip for 1-4 days, see image 3 .

[0044] image 3 Middle A is a cell-free planted paper chip, with obvious paper fiber structure and no impurities; Figure 3B is a graph of cell growth after one day of culture, PC12 cells are evenly dispersed in the paper fiber, and the growth is good; image 3 Middle C is a graph showing the growth of cells after two days of culture. PC12 cells have grown well and increased in...

Embodiment 3

[0046] The linear relationship detection steps of a microfluidic paper chip for online monitoring of intracellular dopamine based on nano-gold modified screen-printed electrodes are as follows:

[0047] When the PC12 cells grow well, discard the culture medium, wash with phosphate buffered saline (PBS) three times, contact the cells with an appropriate amount of PBS solution (0.1 mol / L, pH 7.4), and place in an incubator to incubate for 5 minutes. Collect the PBS buffer in contact with PC12 cells, prepare a series of dopamine solutions with this solution, and place them on the paper-based microfluidic chip prepared in Example 1 in combination with the modified nano-gold screen printed electrode prepared in Example 1. As the working electrode, the differential pulse voltammetry is used for detection, and the results are as follows Figure 4 .

[0048] by Figure 4 It can be seen that the oxidation peak current response value of dopamine increases with the increase of dopamine concen...

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Abstract

The invention discloses a microfluidic paper chip for monitoring dopamine (DA) in cells online based on a nano-gold modified screen-printed electrode. Based on unique advantages of easiness for processing and low cost of a paper-based microfluidic chip, a novel technology for detecting the level of the dopamine released by pheochromocytoma cells (PC12 cells) is established by organically combininga flexible nano-gold modified electrochemical biosensor, so that online monitoring of the activity of the PC12 cells and Alzheimer's disease (AD) model cells is realized. According to the microfluidic paper chip, the DA releasing level of the AD model cells under the intervene of the PC12 cells, Abeta25-35 induced and constructed AD model cells, and curcumin is determined by adopting a differentpulse voltammetry; the microfluidic paper chip is successfully used for online monitoring and evaluation of normal PC12 cells and AD model cells and a novel technology can be provided for researches including medicine curative effect evaluation, new medicine development and the like of nerve-related diseases including the AD and the like.

Description

Technical field [0001] The invention relates to a microfluidic paper chip for online monitoring of intracellular dopamine based on nano-gold modified screen printing electrodes. The paper-based microfluidic chip detects intracellular DA and can be applied to the fields of early diagnosis and treatment of diseases, drug efficacy evaluation and the like . Background technique [0002] Dopamine is a catecholamine neurotransmitter secreted by nerve cells. It acts as a chemical messenger in the transmission of information between cells and can regulate human emotions and cognitive abilities, including thought, feeling, understanding, reasoning and so on. Studies have shown that neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, Huntington's disease, etc. are related to dopamine metabolism disorders. Therefore, the detection of dopamine molecular compounds at the cellular level is of great significance for the study of dopamine metabolic pathways. [0003] At ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327
CPCG01N27/327
Inventor 刘爱林刘萌萌李珊红郭子珍雷云彭花萍陈伟林新华
Owner FUJIAN MEDICAL UNIV
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