Circulation tumor cell automatic separation and purification micro-fluidic chip and separation and purification method thereof
A microfluidic chip and automatic separation technology, applied in the field of biomedicine, can solve problems affecting CTCs protein and molecular research, achieve the effect of accurate critical separation size, avoid cell damage, and avoid false results
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Embodiment 1
[0040] This embodiment provides a microfluidic chip 10 for automatic separation and purification of circulating tumor cells.
[0041] see figure 1 As shown, a microfluidic chip 10 for automatic separation and purification of circulating tumor cells includes a separation and purification chamber 100 , a separation column 200 , a microcolumn 300 and a target cell collection chamber 600 . The microfluidic chip 10 for automatic separation and purification of circulating tumor cells is denoted as microfluidic chip A for automatic separation and purification of circulating tumor cells.
[0042] Automatic Separation and Purification of Circulating Tumor Cells The separation and purification chamber 100 of the microfluidic chip 10 is formed by bonding a bottom sheet and an upper cover. The material of the bottom sheet includes but not limited to: glass, PDMS, silicon material, etc.; the material of the upper cover includes but not limited to: glass, PDMS, silicon material, etc. The ...
Embodiment 2
[0059] Using the microfluidic chip A for the automatic separation and purification of circulating tumor cells in Example 1 to detect clinical samples
[0060] In this example, the microfluidic chip A for automatic separation and purification of circulating tumor cells in Example 1 was used to detect peripheral blood samples (numbered 1-20) of 20 tumor patients.
[0061] 1. Reagent formula:
[0062] (1) Preparation of blocking solution: minimum essential medium (MEM) contains 15% polysucrose, 0.2% Pluronic and 5% bovine serum albumin (BSA).
[0063] (2) Hank's balanced salt solution:
[0064]
[0065] 2. Specific detection steps:
[0066] (1) Closed: The separation and purification chamber of the microfluidic chip 10 for automatic separation and purification of circulating tumor cells is composed of a bottom sheet of PDMS and an upper cover glass sheet, in order to prevent non-specific contact between cells and PDMS and the glass surface of the separation and purification ...
Embodiment 3
[0075] Using the microfluidic chip A for the automatic separation and purification of circulating tumor cells in Example 1 to detect cell lines
[0076] 1. Sample preparation
[0077] In order to prove that the microfluidic chip A for automatic separation and purification of circulating tumor cells of the present invention has a good capture rate, can effectively detect tumor cells in the sample, and remove normal blood cells in the sample at the same time, the number of tumor cells in this example has been Known samples were taken as examples for testing. In this embodiment, the epithelial cell line MCF-10A, the mesenchymal tumor cell line U118, the epithelial-mesenchymal mixed lung cancer cell line PC-9 and the negative control CCRF-HSB-2 lymphoblastoid cells were selected for the experiment. Those skilled in the art As long as you know the name of the cell line, you can buy it. Use a cell counter to take a certain number of the above-mentioned cells, add them to 5 mL of n...
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