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

Inactive Publication Date: 2019-05-14
周辉
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, using serum identification, due to individual differences, some brain malignant tumors lack highly specific markers and cannot be effectively detected, and their reliability is poor; medical imaging scans can non-invasively and quickly monitor the patient's brain malignant tumors. However, even with the most advanced and accurate imaging detection methods, the detection diameter of brain malignant tumors should be at least 2-3 mm, that is, brain malignant tumors smaller than this diameter range are often not found through imaging examinations, so they can only be used in Detection of advanced cancer; histopathological sections are often identified only by puncture or postoperative brain malignant tumor tissue, and the technical requirements are high, and it cannot be obtained repeatedly, for a long time or at any time. In addition, this method also has certain advantages. Traumatic and cannot accurately determine whether metastases have occurred

Method used

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  • Brain malignant tumor cell combined quantitative detection card, preparation method thereof, and using method thereof
  • Brain malignant tumor cell combined quantitative detection card, preparation method thereof, and using method thereof

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a brain malignant tumor cell combined quantitative detection card, a preparation method thereof, and a using method thereof. The brain malignant tumor cell combined quantitative detection card comprises a bottom card and a surface card; the bottom card is provided with a sample adding zone, bonding zones, microchannels, time valves, detection zones, and collection zones, wherein the sample adding zone is positioned at one end of the microchannels, the bonding zones are connected with the one or more detection zones through the microchannels and the time valves and are positioned at the other end of the microchannels, and the detection zones are connected with the collection zones through the microchannels; a plurality of sample adding cavities are formed in the sample adding zone, the sample adding cavities are cylindrical microchannels, and the bottoms of the sample adding cavities are directly connected to the bonding zones. According to the brain malignant tumor cell combined quantitative detection card, the preparation method thereof, and the using method thereof,the flow rate and the flow quantity of a to-be-tested sample are controlled by the microfluidic technology, brain malignant tumor cells with the same marker enter the detection zones through a microchannel, so that mutual interference is avoided, thus the error is reduced, and reaction sensitivity is increased; and the to-be-tested sample is prepared into water-in-water micro-droplets, so that the detection limit is increased.

Description

technical field [0001] The invention relates to the technical field of the combination of microfluidic technology and immunofluorescence microsphere immunochromatography technology, in particular to a combined quantitative detection card for brain malignant tumor cells and its preparation method and use method. Background technique [0002] In recent years, the incidence of intracranial tumors has been on the rise. According to statistics, intracranial tumors account for about 5% of systemic tumors and 70% of childhood tumors, while 20-30% of other malignant tumors will eventually transfer to the brain. Due to its expansive and infiltrating growth, once it occupies a certain space in the brain, no matter whether its nature is benign or malignant, it will inevitably increase intracranial pressure, compress brain tissue, cause central nervous system damage, and endanger the patient's life. Tumors that grow in the brain are collectively called brain tumors, including primary br...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/58G01N33/533G01N33/574
Inventor 周辉
Owner 周辉
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