Gold-plated silicon nanocolumn electrodes and their applications
A technology of silicon nanometer and columnar electrodes, which is applied in the methods of stress-stimulated microbial growth, other methods of inserting foreign genetic materials, biochemical equipment and methods, etc. It can solve the problem of disordered nanowires on the electrode surface, unstable electroporation effect, and inability to Apply follow-up experiments and other issues to achieve the effect of reducing cell damage, increasing conductivity, and convenient and simple operation
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Embodiment 1
[0060] Example 1 Preparation of silicon nanocolumnar electrodes on gold-plated surface and construction of cell electroporation device
[0061] 1. Preparation of silicon nanocolumnar electrodes with gold-plated surface
[0062] (1) Fabrication of optical masks: Design four nanopillar array masks with different specifications. The material of the mask is glass, and the opaque position on the surface is covered with a chrome layer. The size of the mask is 127mm*127mm, such as Figure 1A As shown, the pattern covered by the chromium layer is a rectangular array, and the array specification is: a circle with a diameter of 1 μm is used as a structural unit, and the distance between the centers of the circles is respectively 5 μm, 10 μm, 15 μm and 20 μm. The size of each rectangular array is 10mm*10mm , the distance between each rectangular array is 3 mm, and the distance between rectangular array areas with different circle pitches is 4 mm.
[0063] (2) Photolithography: Take the s...
Embodiment 2
[0082] Example 2 Cell electroporation device for transfection of adherent cells
[0083] The cell electroporation device constructed in Example 1 was used to transfect adherent cells. In this example, adherent DC2.4 cells were electroporated to transfect pGL3-luc and adherent RAW264.7 macrophages were electroporated to transfect pGL3-luc Experiment as an example.
[0084] (1), electroporation and transfection pGL3-luc experiment of adherent DC 2.4 cells, the transfection method includes the following steps:
[0085] (1) placing the gold-plated nanocolumn silicon chip electrode in a 6-well plate;
[0086] (2) After DC 2.4 cells were digested with trypsin, the medium was added to stop the digestion, the cells were blown down gently with a pipette, the cells were transferred to a 15mL centrifuge tube, centrifuged at 1000g for 5min, and resuspended in 1640 medium, Ensure that the final cell density is 4*10 7cells / mL, take 100 μL of cells and drop them on the silicon wafer elect...
Embodiment 3
[0120] Example 3 Cell electroporation device transfection suspension cells
[0121] The cell electroporation device constructed in Example 1 was used to transfect suspension cells. In this example, the suspension Jurkat cells were electroporated to transfect pGL3-luc and mRNA as an example.
[0122] 1. Suspended Jurkat cells were electroporated to transfect pGL3-luc experiment. The transfection method includes the following steps:
[0123] (1) Take out a certain amount of Jurkat cells, centrifuge at 1000g for 5min, discard the supernatant, resuspend the cells with 1-2mL of PBS, count with a cell counter, centrifuge again at 1000g for 5min, discard the supernatant, and use BTX for commercial electroporation resuspended in liquid, the final cell size is 4*10 7 a / mL;
[0124] (2) Add the pGL3-luc plasmid to be transfected to the cell suspension to ensure that the final concentration of the plasmid is 0.08 μg / μL;
[0125] (3) Assemble the cell electroporation device according t...
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