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Enzyme chip based on quantum dot fluorescence detection, preparation method and application

A fluorescence detection, quantum dot technology, applied in fluorescence/phosphorescence, analysis by chemical reaction of materials, material excitation analysis, etc., can solve problems such as detection of enzyme-based disease markers that have not been reported.

Inactive Publication Date: 2012-03-21
TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the reports of quantum dot fluorescence detection have been increasing in recent years, the research on the detection of enzyme disease markers has not been reported.

Method used

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  • Enzyme chip based on quantum dot fluorescence detection, preparation method and application
  • Enzyme chip based on quantum dot fluorescence detection, preparation method and application
  • Enzyme chip based on quantum dot fluorescence detection, preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] 1. Preparation of cadmium telluride quantum dots (prepared with reference to CN 200810101429.4),

[0064] (1) Add cadmium nitrate and thioglycolic acid in sequence in deionized water, so that the molar ratio of cadmium nitrate and thioglycolic acid is 1:50. After the complexation reaction is completed, add sodium hydride telluride equimolar to cadmium nitrate to obtain water-soluble quantum point precursor solution, the final concentration of cadmium ion is 10 -5 M;

[0065] (2) Add the precursor solution obtained in step (1) to an ultrasonic atomizer. After ultrasonic atomization, the precursor solution exists in the form of submicron droplets to obtain the precursor solution of water-soluble quantum dots. mist;

[0066] (3) Feed in nitrogen, the flow rate of nitrogen gas is 5L / min, and the droplets of the precursor solution obtained in step (2) are brought into the temperature-stabilized reaction device, and the droplets of the precursor solution are heated (the tem...

Embodiment 2

[0079] 1. The preparation of cadmium telluride quantum dots is the same as in Example 1.

[0080] 2. The substrate of the enzyme chip was fabricated by mask photolithography technology as in Example 1, and the side length of the prepared substrate microchamber was 30 mm.

[0081] 3. Using an enzyme chip to detect the activity of lactate dehydrogenase:

[0082] (1) Preparation of the agar solution: dissolve the agar in water to obtain the agar solution, the concentration of the agar is 5% by weight, and the water temperature is 70°C. Inject agar solution into each microchamber on the substrate made in step 2, cool to room temperature, and form a layer of agar gel at the bottom of the microchamber;

[0083] (2) Prepare the quantum dot solution: disperse the cadmium telluride quantum dots in 0.01 mol / liter phosphate buffer (pH 6.8) to obtain the cadmium telluride quantum dot solution, and the telluride in the cadmium telluride quantum dot solution The molar concentration of cad...

Embodiment 3

[0090] 1. Preparation of cadmium telluride quantum dots (prepared with reference to CN 200810101429.4)

[0091] (1) Add cadmium nitrate and thioglycolic acid sequentially in deionized water, so that the molar ratio of cadmium nitrate and thioglycolic acid is 1:3. After the complexation reaction is completed, add sodium hydride telluride equivalent to cadmium nitrate to obtain water-soluble quantum point precursor solution, the final concentration of cadmium ions was 5×10 -2 M;

[0092] (2) Add the precursor solution obtained in step (1) to an ultrasonic atomizer. After ultrasonic atomization, the precursor solution exists in the form of submicron droplets to obtain the precursor solution of water-soluble quantum dots. mist;

[0093] (3) Feed nitrogen, the flow rate of nitrogen is 3L / min, the droplets of the precursor solution obtained in step (2) are brought into the temperature-stabilized reaction device, and the droplets of the precursor solution are heated (the temperatur...

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Abstract

The invention relates to an enzyme chip based on quantum dot fluorescence detection, a preparation method and application, which manufactures substrates of the enzyme chip through a mask photoetching. A plurality of mini-chambers are obtained on the substrates. A reaction system formed by a water-solubility quantum dot, enzyme oligomer and coenzyme or formed by the water-solubility quantum dot, enzyme and the coenzyme is loaded in each mini-chamber which is used for detection and arranged on the substrates provided with the plurality of mini-chambers. Or one reaction system formed by the water-solubility quantum dot, the enzyme oligomer and the coenzyme or formed by the water-solubility quantum dot, the enzyme and the coenzyme is respectively loaded in different mini-chambers which are arranged on the substrate and used for detection. By means of the reaction of the enzyme to be detected and the enzyme oligomer or the reaction of the enzyme and the enzyme oligomer to be detected, fluorescence strength of the water-solubility quantum dot changes. High-selectivity quantitative detection of activity of the enzyme to be detected and / or density of the enzyme oligomer to be detected canbe achieved through detection of the quantity of the change of the fluorescence strength of the water-solubility quantum dot.

Description

technical field [0001] The invention belongs to the field of biological detection, in particular to an enzyme chip based on quantum dot fluorescence multi-enzyme joint detection, an enzyme chip based on quantum dot fluorescence detection of enzyme reaction substrates, an enzyme chip based on quantum dot fluorescence detection of enzyme activity and the above enzyme chip preparation methods and applications. Background technique [0002] Among the many early rapid detection methods, the biochip is the most rapidly developed detection device in recent years and is considered to have the most industrialization prospects. It has very obvious advantages, such as integration, parallelism, rapid detection and so on. Enzyme chip is a kind of biological chip, which is detected by the binding reaction of enzyme molecules and corresponding substrates, so it has the advantages of high specificity, low cost, safe operation, and convenient on-site determination. Because many physiologica...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N21/75
Inventor 唐芳琼任湘菱杨柳青
Owner TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI
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