Device for capturing circulating tumor cells by combining functionalized microspheres with filtering chip and application of device
A tumor cell and chip technology, applied in the field of modern medicine, can solve the problems of low efficiency, unfavorable hysteresis of tissue biopsy, high cost, etc., and achieve the effect of improving the capture ability
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Embodiment 1
[0073] Synthesis of Example 1 Nano Zinc Oxide Modified Microspheres
[0074] Polystyrene microspheres and SiO 2 The steps of synthesizing nano zinc oxide on the surface of microspheres are as follows:
[0075] (1) Cleaning of the microspheres, pipette 200 μl of polystyrene microspheres with a diameter of 23 μm and silica microspheres of 13 μm, and wash with ultrapure water for 3 times, 10 min each time.
[0076] (2) Microsphere pretreatment: place the cleaned microspheres on the surface of a slide and wash them on a plasma cleaner for 180 s.
[0077] (3) Adhere the treated microspheres obtained in step (1) to the seed layer, shake well in the seed layer for 10 seconds, centrifuge, put them back into the seed solution and repeat once, centrifuge the microspheres and rinse with ultrapure water Wash once.
[0078] (4) Put the microspheres covering the seed crystal layer into a 1ml EP tube, and put them in the growth solution at 95°C to synthesize 4H. In this experiment, silic...
experiment example 2
[0083] Preparation of Experimental Example 2 Filter Chip
[0084] (1) Acetone, ethanol, and water were used to ultrasonically clean the 400-micron low-resistance silicon wafer, each time for 10 minutes;
[0085] (2) After drying, the silicon surface is oxidized into silicon dioxide oxide, the thickness of silicon dioxide is 2 μm, the glue is applied by a glue machine to 1.5 μm, the ultraviolet lithography is 5s, the silicon dioxide is etched by RIE, and the deep silicon is etched by STS Machine dry etching silicon 65 microns, acetone degum.
[0086] (3) Keep the etching template obtained in (2) as clean and flat as possible. After cleaning with ethanol and experimental ultrapure water, silanize it for 10 minutes.
[0087] (4) 10g of polydimethylsiloxane was mixed evenly at a ratio of 1:10, poured on the surface of the silicon template, vacuumed for 30 minutes, transferred to a 95-degree hot plate and heated for 60 minutes.
[0088] (5) Use a scalpel to cut the PDMS template ...
Embodiment 4
[0099] The performance test of cell sorting and enrichment in the cell line of embodiment 4
[0100] (1) Four tumor cell lines MCF-7, Bxpc-3, panc-1, and HeLa are used as cell models in Filter CHIP, and the ratio of microspheres to tumor cells is 10 2 , to test the capture efficiency of circulating tumor cells at different flow rates.
[0101] The results of this experiment are as Figure 5 As shown, the capture efficiency of circulating tumor cells at a flow rate of 1ml.h -1 to 3ml.h -1 In the range of , as the speed increases, the capture efficiency does not decrease but increases. This is because the flow rate is too low, and a stable turbulent fluid cannot travel inside the chip. The capture efficiency of tumor cells is limited, and the cells in the fluid with a low flow rate cannot be effectively trapped. After the flow rate was greater than 3ml.h-1, the cell capture efficiency decreased.
[0102] (2) The purity of circulating tumor cells captured by microfluidics is ...
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