Brain malignant tumor cell combined quantitative detection card, preparation method thereof, and using method thereof
A technology for quantitative detection of brain malignant tumors, applied in the field of microfluidic technology and immunofluorescence microsphere immunochromatography, can solve the problems that can only be applied to the detection of middle and late stages of cancer, cannot be accurately judged, and has high technical requirements, achieving accurate Effects of controlling reaction time, improving detection accuracy, and avoiding environmental pollution
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Embodiment 1
[0066] A combined quantitative detection card for brain malignant tumor cells, including a bottom card and a face card, the bottom card is provided with: a sample loading area, a binding area, a microchannel, a time valve, a detection area and a collection area, and the sample loading area Located at one end of the microchannel, the binding zone is connected to one or more detection zones through the microchannel and a time valve, and is located at the other end of the microchannel, and the detection zone is connected to the collection zone through the microchannel; multiple A sample loading chamber, the sample loading chamber is a cylindrical microchannel, the bottom of which is directly connected to the binding area.
[0067] There are two binding areas, namely binding area A and binding area B; the binding area A is connected to the detection areas A1 and A2 through a microchannel and a time valve; the binding area B is connected to the detection area through a microchannel ...
Embodiment 2
[0087] A method for preparing a combined quantitative detection card for brain malignant tumor cells, comprising the following steps:
[0088] (1) Preparation of bottom card:
[0089] (1-1) Drawing: draw the mask layout through the drawing software according to the design of the microchannel pattern;
[0090] (1-2) Mask: After the mask layout is made, print it on a transparent film to make a photolithography mask plate, and use this mask plate to make a silicon wafer mold through a photolithography process;
[0091] (1-3) Photolithography: plate a layer of barrier layer on the surface of the substrate, and then evenly throw a layer of photosensitive material-photoresist on the barrier layer with a glue machine; prepare the required film on the photomask Channel pattern; cover the photomask on the substrate, irradiate the substrate coated with photoresist with ultraviolet light, and the photoresist undergoes photochemical reaction; remove the exposed photoresist by chemical me...
Embodiment 3
[0099] The method for using the brain malignant tumor cell combined quantitative detection card comprises the following steps:
[0100] (a) The aqueous solution A containing dextran, trehalose and gelatin was added to the aqueous solution B containing polyethylene glycol to form a fibril network, and then 1 wt% egg white lysozyme was added for heat incubation at 60°C, the fibrils The network is transformed into mature fibrils, and finally the serum standard is added to it, and the mature fibrils are used to wrap the serum standard, and water-in-water micro-droplets with a diameter of 8 μm can be prepared and prepared into a series of concentrations of more than 5 , use several detection cards of the same batch to detect the standard solution of each concentration, take the fluorescence intensity of the detection area as the ordinate, and the concentration of the serum standard solution as the abscissa, draw a standard curve and save the standard curve in the multi-color fluores...
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