A cell electrochemical sensor based on 3D printing technology and its application
A 3D printing and electrochemical technology, which is applied in the field of cell electrochemical sensors, can solve the problems of poor detection accuracy and uneven distribution of sensors, and achieve the effects of small sample requirements, avoiding cross-contamination, and fast detection speed
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Embodiment 1
[0061] Example 1 Preparation of cell electrochemical sensor
[0062] A method for constructing cell electrochemical sensors based on 3D printing technology (such as figure 2 shown), including the following steps:
[0063] (1) Preparation of photosensitive methacryloyl gelatin (GelMA) solution: Take the LAP initiator and add it to DMEM cell culture medium to make the final concentration 0.5%, and then dissolve it in a water bath at 60°C for 30 minutes in the dark to obtain the dissolved solution. Standard initiator solution; add the GelMA material in the dissolved standard initiator solution, and dissolve it in the dark for 30 minutes in a water bath at 60° C., and shake it 3 times during the period to obtain the GelMA solution (mass concentration is 7.5%); then the obtained GelMA solution Use a 0.22μm sterile filter membrane to filter into a clean container for use.
[0064] (2) Preparation of cell / carbon nanofiber / GelMA composite hydrogel: add a certain amount of carbon na...
Embodiment 2
[0070] Example 2 Application of cell electrochemical sensor based on 3D printing technology
[0071] The electrochemical sensor after incubation obtained in Example 1 was evaluated for the cytotoxicity of deoxynivalenolium family toxins, specifically as follows:
[0072] (1) Drug stimulation: Remove the original culture medium on the working electrode, dilute the toxin standard substance with DMEM cell culture medium into a gradient concentration solution, and then take 150 μL DON and 15-ADON at 0.1, 0.2, 0.5, 1, 2 , 5ug / mL concentration range and DON+15-ADON toxin solution in the concentration range of 0.1+0.1, 0.2+0.2, 0.5+0.5, 1+1, 2+2, 5+5ug / mL are dropped on the printing work with cells On the electrode, place it in an incubator for 24 hours to measure the corresponding impedance value.
[0073] (2) Electrochemical signal value detection: use 150μL 2.5mM The PBS solution was used as the electrode reaction system, and the frequency range of the electrochemical impedance...
Embodiment 3
[0086] Embodiment 3 verification experiment
[0087] The CCK8 method was used to detect the cytotoxicity of DON, 15-ADON and DON+15-ADON alone and in combination: the density was 5×10 4 / mL lung cancer cells A549 were adherently inoculated into a 96-well plate, and after 24 hours of culture, the culture medium was removed, and 100 μL of the toxin solution at the same dose as in Example 2 was added. After 24 hours of toxin stimulation, aspirate the supernatant and add 100 μL of culture solution containing 10% CCK8 to each well, incubate at 37°C for 2 hours, then measure the absorbance value at 450 nm with a microplate reader, and calculate the inhibition rate of cell activity. The calculation method is as follows:
[0088]
[0089] Among them, OD 加药 : Absorbance value after toxin stimulation for 24h, OD 0加药 : Absorbance value after 24h stimulation without toxin, OD 空白 : Absorbance value of pure cell culture solution.
[0090] Depend on Figure 5 It can be seen that the ...
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