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A 3D printing device based on a bipolar electrode-light-emitting diode-photoresistor platform to detect hydrogen peroxide secretion from cells

A technology of 3D printing and cell secretion, which is applied in the direction of measuring devices, manufacturing auxiliary devices, and material analysis through electromagnetic means, which can solve the problems of restricting large-scale use, cumbersome steps, and expensive luminescent reagents, and achieve visualization and digitization. Avoid the interference of human factors, the effect of low price

Active Publication Date: 2021-08-03
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The successful output of visual signals by this system depends on the use of luminescent reagents. These luminescent reagents are expensive and cannot be reused, which will limit the large-scale use of this system to a certain extent.
The brightness change of the LED can sensitively reflect the concentration of the analyte, but only semi-quantitative detection can be realized by naked eye observation. If the colorimetric analysis of light bulbs with different brightness can be carried out, the experimental results will be more reliable, or quantitative detection can be realized, but this method The steps are cumbersome and require a certain amount of time for post-processing

Method used

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  • A 3D printing device based on a bipolar electrode-light-emitting diode-photoresistor platform to detect hydrogen peroxide secretion from cells
  • A 3D printing device based on a bipolar electrode-light-emitting diode-photoresistor platform to detect hydrogen peroxide secretion from cells
  • A 3D printing device based on a bipolar electrode-light-emitting diode-photoresistor platform to detect hydrogen peroxide secretion from cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] A BP-LED-E-LDR platform based on the detection of cell secreted H 2 o 2 The design and application of the LED-LDR fixture in the 3D printing device are as follows:

[0035] like figure 2 As shown in A, use the 3D printing drawing software to design the fixing device including the chute 11, the small slider 12 and the large slider 13, and then print the real object through the 3D printer. The 3D printer can choose rubber (rubber) soft elastic material , in the present invention, as long as the printed size and shape meet the requirements, that is, the printed chute 11, small slider 12 and large slider 13 conform to the experiment of the present invention, and the material is not the invention point of the present invention. Commercially available products can be used. Among them, the chute 11 is 150mm long, 25mm wide, and 20mm high. There are three slide rails with a width of 2.8mm inside. ×20mm), the two sides are closely attached to the front and rear surfaces thr...

Embodiment 2

[0041] A BP-LED-E-LDR platform based on the detection of cell secreted H 2 o 2 The 3D printed device under the optimal driving voltage of 4.1V, ΔR and cells in contact with different concentrations of H prepared in PBS 2 o 2 The linear relationship detection steps are as follows:

[0042] MCF-7 cells were cultured in culture flasks, and the medium was changed every two days. When the cells grow to 80-90% confluence, wash twice with PBS (0.01M, pH=7.4), then add 10mL of PBS, then put the culture bottle into the cell incubator and incubate for 5min, and finally collect the cells that have been in contact with the cells. PBS spare. PBS contacted with the above cells and standard H 2 o 2 Solution preparation with different concentration gradients including 1, 10 1 、10 1.5 、10 2 、10 2.5 、10 3 、10 3.5 、10 4 、10 5 、10 6 μM H 2 o 2 solution.

[0043] like figure 1 The entire detection device is constructed as shown, and the Pt electrodes 3 and 4 in the anode pool 1 ...

Embodiment 3

[0046] A BP-LED-E-LDR platform based on the detection of cell secreted H 2 o 2 Different numbers of MCF-7 cells in the 3D printed device secreted H 2 o 2 The detection method is as follows:

[0047] MCF-7 cells were cultured in culture flasks, and the medium was changed every two days. When the cells grow to 80-90% confluence, add 1 mL of trypsin to digest the cells. When the cells become round and fall off from the bottom of the culture bottle, add DMEM medium to stop the digestion, and draw part of the cell suspension for cell counting. Collect the cell suspension and centrifuge at 1000rpm for 5min in a centrifuge, discard the supernatant, then add 2.5mL PBS to resuspend the cell pellet, then add 20μL 10μg / mL PMA to stimulate the cells and place them in a cell culture incubator for incubation 5 min, then centrifuge and collect the supernatant and store it in the dark. Different numbers of cells collected (0, 2.5, 10, 15, 50, 100×10 5 Each / 2.5mL) PMA stimulation superna...

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Abstract

The invention discloses a 3D printing device for detecting hydrogen peroxide secreted by cells based on a bipolar electrode-light-emitting diode-photoresistor platform. Based on the characteristics of 3D printing that can realize personalized design and precise printing and the principle that isoelectronic redox reactions can occur at both ends of BPE for analyte detection, combined with the advantages of easy construction of BPE system, cheap and easy to obtain and reusable LED and LDR , constructed the BP‑LED‑E‑LDR platform for human breast cancer cells (MCF‑7) to secrete H 2 o 2 detection. Under optimal conditions, the present invention semi-quantitatively detects the concentration of the analyte in the sample pool through the visualization feature of BP-LED-E, and further quantitatively detects the H secreted by cells through LDR and digital multimeter. 2 o 2 It provides a new idea and a new platform for the preliminary judgment of cell growth and proliferation and tumor development and other related research fields.

Description

technical field [0001] The invention relates to a method based on the BP-LED-E-LDR platform to detect the secretion of hydrogen peroxide (H 2 o 2 ) 3D printing device, using DC power supply, Pt and GCE to build a BPE platform, and using easily available LED-LDR and digital multimeter as a visual signal output tool, and designed a fixed distance and angle between the LED-LDR. device, successfully detected H secreted by MCF-7 cells in different quantities under optimal conditions 2 o 2 , can be used in cell counting, judging cell growth status and tumor development process and other research fields. Background technique [0002] Bipolar electrochemistry takes place in an electrolyte solution containing two driving electrodes (driving electrode, DE) and a conductive object (ie bipolar electrode, BPE). Driven by a certain external voltage, the electrolyte solution between the two driving electrodes forms a gradient electric field. BPE located in the electrolyte solution is a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B29C64/393B29C64/295G01N27/416G01N27/12B33Y30/00B33Y50/02
CPCB33Y30/00B33Y50/02B29C64/295B29C64/393G01N27/12G01N27/416
Inventor 刘爱林郭子珍刘萌萌刘辉雷云
Owner FUJIAN MEDICAL UNIV
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