Manufacture method of microfluidic chip with channel inner wall coated with Parylene
A technology of microfluidic chip and manufacturing method, which is applied in the direction of coating, microstructure technology, gaseous chemical plating, etc., can solve the problems that complex channel network coating cannot be carried out, and achieve shortening coating time, reducing dosage, and improving The effect of utilization
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Embodiment 1
[0034] A microfluidic droplet digital PCR chip is manufactured by using the chip manufacturing method provided by the invention, and the chip is used for digital PCR reaction. figure 1 It is a flow chart of the method for fabricating a PDMS microfluidic chip coated with Parylene C provided in the embodiment of the present invention. figure 2 It is the design drawing of the droplet digital PCR chip in the embodiment of the present invention.
[0035] In this embodiment, the mold of the channel layer of the chip is made on a silicon wafer by multi-layer soft lithography technology, the material used to make the chip is PDMS, and the coated Parylene films are all Parylene C films. Production process such as figure 1 Shown: First, spin-coat 100 μm SU-8 negative photoresist on a clean silicon wafer 1, and after pre-baking at 95°C, use the film plate containing the droplet digital PCR chip channel structure as a mask for selective UV exposure 30s, copy the graphics on the mask pl...
Embodiment 2
[0040] Utilize the original method of first sealing and then coating the parylene film to manufacture the chip, the manufacturing process of the chip is as follows: use the photoresist mold among the embodiment 1, take PDMS as material by molding method, make the chip channel layer 5, used The conditions are the same as in Example 1. Seal the chip channel layer 5 with the glass after plasma treatment. The chip after sealing is coated with Parylene C thin film, and the amount of coating material is also 6.64g. At this time, the coating of Parylene C on the inner wall of the channel relies on the diffusion of chlorinated p-xylene monomer into the chip through the inlet and outlet, and the coating distance is limited, and only the channel near the inlet and outlet is coated with Parylene C.
[0041] figure 2 The droplet digital PCR chip consists of two parts: droplet generation and droplet storage. The droplet generating part includes an oil inlet 9, a PCR reaction solution i...
Embodiment 3
[0044] The chips fabricated by the two methods were used for digital PCR reaction respectively. The 10ul PCR reaction solution consists of the following components: 5ul 3D Digital PCR Master Mix, 0.5ul Gene Expression Assay (18S), 0.5ul 2% Tween-20, 2ul nuclease-free water, 2ul plasmid template. The composition of the solidifiable oil used to generate the droplets was: 69.2 wt% Silicone Oil PMX-200-10cSt, 12 wt% 5225C Formulation Aid, 12 wt% Bis-19, 6 wt% RTV615-B, 0.8% SM3110 Catalyst. The PCR reaction solution enters the chip from the injection port 10 at a flow rate of 100ul / h, and the oil enters the chip from the injection port 9 at a flow rate of 280ul / h. The PCR reaction solution and the oil meet at the droplet generation point 11 to generate uniform water-in-oil droplets. After the droplets are generated, they enter the buffer pool 12. The flow rate of the droplets in the buffer pool slows down, and they enter the liquid storage pool evenly in two ways. 13. A uniform...
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