Fetal cell capturing module, application method thereof, micro-fluidic chip for fetal cell capturing and application method of micro-fluidic chip for fetal cell capturing
A microfluidic chip and fetal cell technology, applied in the field of cell capture, can solve the problems of low capture efficiency, high cost and low purity of fetal cell analysis
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Embodiment 1
[0112] Example 1 Experimental Example of Magnetic Beads + Disulfide Bond Modified Antibody
[0113] 1.1 Experimental method Magnetic beads are selected from Thermo Fisher Company, Dynabeads TM MyOne TM CarboxylicAcid, the product number is 65012, refer to the instruction manual for the method of use, the specific display is as follows: after taking out the magnetic beads, shake and mix well, take 20 μL of magnetic beads and wash them three times with 15mM MES buffer pH 6.0, add 1-(3-dimethylaminopropyl) - 3-Ethylcarbodiimide hydrochloride solution (see instructions for concentration), react for 30 minutes, magnetically separate, wash three times with 15mM MES buffer pH 6.0, add 3-mercapto-2-propanamine, react for 30 minutes, magnetically separate, Wash with 15mM MES buffer pH 6.0 three times, then add 0.01% (mass fraction) of m-pyridyl disulfide polyethylene glycol succinimide valerate (polyethylene glycol molecular weight is 2000) solution, react for 30 minutes, Magnetic se...
Embodiment 2
[0118] Example 2 Preparation of microfluidic chip for fetal cell capture
[0119] see figure 1 structure to make a microfluidic chip. The mask plate of the chip is a silicon-based chip containing su-8 photoresist channels obtained by ultraviolet lithography. Dimethylsiloxane (PDMS) prepolymer is poured into the chip, and after pumping, heating, and degassing The PDMS channel layer containing microfluidic channels can be obtained by four steps of molding and drilling. The PDMS channel layer and the glass slide were bonded into a complete chip using a plasma cleaner (Harrick, model: PDC-002). The slide glass is preferably a sailboat brand 25.4×76.2 mm glass slide without frosted edges.
[0120] The chip is provided with two sample inlets: inlet (1) and inlet (2) and one sample outlet: outlet (3). Between the sample inlet and the sample outlet is a fluid microchannel filled with microarrays. Such as figure 1 shown. In this example, the sample inlet and the sample outlet are...
Embodiment 3
[0128] Example 3 Detecting the Capture Efficiency of Microfluidic Chips to Fetal Cells
[0129] 3.1 Experimental method
[0130] Take the microfluidic chip obtained by the recognition molecule coupling method 2 in Example 2, and use 3% bovine serum albumin solution as the blocking solution to block the chip for 30 minutes.
[0131] Add cultureable cell lines to 1mL healthy human peripheral blood to simulate the peripheral blood environment of pregnant women, (the cultureable fetal cell line is JEG-3 (human choriocarcinoma cell line), which was purchased from the Shanghai Institute of Biological Sciences, Chinese Academy of Sciences Cell Resource Center, Cat. No. TCHu195). Specifically: the number of JEG-3 cells is about 100, and the number of white blood cells is 9.63×10 6 cells / mL, red blood cells 3.98×10 9 samples / mL, the obtained blood was passed through the injection port (1) through the syringe pump, and at the same time, the buffer solution was passed through the inje...
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