Automatic high-throughput single cell capture method based on liquid droplet microfluidic chip
A microfluidic chip, single-cell technology, applied in the intersection of microfluidic technology and cell biology, can solve the problems of cell damage, high cost, inconvenience of wide application, etc. wide range of effects
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Embodiment 1
[0039] A method for preparing a high-throughput microfluidic chip that automatically captures single cells is as follows:
[0040] (1) Preparation of SU-8 template for microfluidic chip with high-throughput automatic capture of single cells
[0041] The microfluidic chip was prepared by photolithography and etching methods to prepare the SU-8 template with channel part protrusions; first, the thickness of the first layer of SU-8 glue was 100 μm, pre-baked at 95°C for 20 minutes, and the temperature was naturally lowered, and the mask was placed on the SU-8 template. -8 glue plate, UV exposure for 30s, 95°C post-baking for 20min, and natural cooling; secondly, based on the foundation of the first layer of SU-8 glue, the thickness of the second layer of SU-8 glue is 150μm, and 95°C for 30min before baking. Cool down naturally, put the mask on the SU-8 glue plate, expose to ultraviolet light for 40s, bake at 95°C for 40min, and cool down naturally; finally, use ethyl lactate to d...
Embodiment 2
[0045] The density of single cell suspension is 5×10 4 cells / mL, the chip automatically captures single-cell experiments
[0046] The microfluidic chip prepared above was soaked in 75% ethanol, sterilized by ultraviolet irradiation for 1 h, the outlet of the liquid flow channel was blocked, and placed in a vacuum incubator to evacuate for 10 min. breast cancer cells (MCF-7) single cell suspension in 5×10 4 The cell density of cells / mL is added to the chip inlet. At this time, the flow velocity of the single-cell suspension is 0.027m / s. When all the droplet generation units are filled with the single-cell suspension, the outlet of the liquid flow channel is opened, and the The excess liquid in the medium was drained, and after the cells adhered to the bottom surface of the chip, fresh high-glucose DMEM medium was added and cultured in a 37°C incubator. The distribution of single cells in the chip is shown in Figure 4 shown. The cell density was 5 x 10 4 When the cells / mL ...
Embodiment 3
[0048] The density of single cell suspension is 5×10 5 cells / mL, the chip automatically captures single-cell experiments
[0049] The microfluidic chip prepared above was soaked in 75% ethanol, sterilized by ultraviolet irradiation for 1 h, the outlet of the liquid flow channel was blocked, and placed in a vacuum incubator to evacuate for 10 min. breast cancer cells (MCF-7) single cell suspension in 5×10 5 The cell density of cells / mL is added to the chip inlet. At this time, the flow velocity of the single-cell suspension is 0.027m / s. When all the droplet generation units are filled with the single-cell suspension, the outlet of the liquid flow channel is opened, and the The excess liquid in the medium was drained, and after the cells adhered to the bottom surface of the chip, fresh high-glucose DMEM medium was added and cultured in a 37°C incubator. The distribution of single cells in the chip is shown in Figure 5 shown. The cell density was 5 x 10 5 When the cells / mL ...
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