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A multi-channel particle detection device and detection method for detecting micron particles

A technology for particle detection and micron particles, which is applied in measuring devices, individual particle analysis, particle and sedimentation analysis, etc., can solve the problems that the chip cannot be reused, consumes a lot of time and energy, and the counting result is inaccurate, etc., to achieve fully automatic operation, reduce work intensity, and reduce the effect of detection cost

Active Publication Date: 2021-04-06
WUHAN LIFE ORIGIN BIOTECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] At present, liquid particle counters have a certain application basis in the fields of food safety, clinical diagnosis, and biological detection. For example, the cell counting plate commonly used in biological detection includes a sample pool with a counting grid. Cell inlet, when in use, inject the culture solution with cells into the sample pool until the cells are evenly distributed and then count. However, because the depth of the sample pool is greater than the size of the cells, the cells will appear layered and superimposed in the sample pool , leading to inaccurate counting results
[0003] In order to improve the accuracy of the counting results, flow cytometer counting, microfluidic chip counting and image recognition counting are usually used at present. However, the chips used in the flow cytometer cannot be reused, resulting in high measurement costs. Chip counting and image recognition counting instruments are expensive, limiting their use
[0004] At the same time, when the above method is used, only one sample can be detected at a time. When the number of samples is large, it takes a lot of time and energy, and the detection efficiency is low.

Method used

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  • A multi-channel particle detection device and detection method for detecting micron particles
  • A multi-channel particle detection device and detection method for detecting micron particles
  • A multi-channel particle detection device and detection method for detecting micron particles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 Using the "double-antibody sandwich method" to detect the biomarker procalcitonin (hereinafter referred to as PCT)

[0070] S1. Preparation of MNPs-Ab1 (magnetic nanoparticles-capture antibody) conjugates, wherein the magnetic nanoparticles have a particle size of 180nm

[0071] S101. Take 200 μL of COOH-MNPs with a concentration of 10 mg / mL and a particle size of 180 nm, wash twice with deionized water, resuspend with 500 μL of deionized water, add 30 μL of EDC with a concentration of 10 mg / mL and 15 μL of a concentration of 10 mg / mL of NHS was activated at room temperature for 15-20 minutes.

[0072] S102. Remove excess EDC and NHS by magnetic separation, resuspend with 500 μL PBS with pH=7.4 to obtain MNPs suspension, dilute Ab1 that recognizes PCT to 1 mg / mL with PBS, and add 100 μL of 1 mg / mL Ab1 to the MNPs suspension After shaking and reacting at room temperature for 2 hours, add 200 μL of 3% BSA solution, shake and react at room temperature for 30 mi...

Embodiment 2

[0082] Example 2 Realized the detection of veterinary drug residue ractopamine (Rac) in pig urine by competitive immunoassay reaction

[0083] S1. Preparation of magnetic nanoparticles-ractopamine antibody (MNPs-Ab) conjugates, the magnetic nanoparticles are COOH-MNPs with a particle size of 1 μm

[0084] Take 200 μL of COOH-MNPs with a concentration of 10 mg / mL and a particle size of 1 μm, wash them twice with pure water, magnetically separate them, resuspend them with 500 μL of pure water, add 50 μL of EDC with a concentration of 10 mg / mL and 25 μL of EDC with a concentration of 10 mg / mL 1 mL of NHS, activated at room temperature for 15-20 minutes, then magnetically separated to remove excess EDC and NHS, resuspended with 500 μL of PBS with pH = 7.4, added 0.2 mg of ractopamine antibody, shaken at room temperature for 2 hours, then added 200 μL 3% BSA solution in the reaction solution, shaken at room temperature for 30 minutes to block, magnetically separated and removed the...

Embodiment 3

[0090] Example 3 Detection of Salmonella in milk by DNA double-strand hybridization and magnetic separation

[0091] S1. Preparation of "PS microsphere-detection probe" conjugate, the particle size of PS microsphere is 3 μm

[0092] Take 300 μL of NH with a concentration of 10 mg / mL and a particle size of 3 μm 2 -PS microspheres were washed twice with deionized water, centrifuged and washed with NaHCO 3 Resuspend in PBS buffer with a concentration of 10 mM, add 292 μg 4-(N-maleimidomethyl)cyclohexane-1-carboxysulfosuccinimide sodium salt (Sulfo-SMCC) and react at room temperature Activation was carried out for 4 hours. After activation, the PS microspheres were washed 3 times with PBS and set aside.

[0093] Add 125 mM dithiothreitol to PBS buffer containing 10 mM NaOH, then add 50 nM thiol-modified oligonucleotide probes, react at room temperature for 2 h, add Sulfo-SMCC activated PS microspheres, After reacting for 12 hours, wash 4-5 times with PBS buffer solution to obta...

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Abstract

The invention discloses a multi-channel particle detection device and detection method for detecting micron particles, and relates to the fields of food safety, in vitro diagnosis and environmental monitoring. The device includes a box body and a lifting mechanism, and the bottom of the box body is provided with several small holes Test tubes, a number of sample pools corresponding to small-hole test tubes are arranged under the box body, particle inlets are opened on the small-hole test tubes, first electrodes are arranged inside the small-hole test tubes, and second electrodes are arranged outside the small-hole test tubes. The test tube can realize simultaneous detection of multiple samples; the sample cell is equipped with electrolyte and particles to be identified, each particle to be identified includes insulating microspheres, and the surface of the insulating microspheres is modified with specific recognition molecules, which are used for Specifically binds to the corresponding analyte; analytes include biomarkers, drug residues and / or bacteria. The invention can improve detection efficiency and reduce detection cost.

Description

technical field [0001] The invention relates to the fields of food safety, in vitro diagnosis and environmental monitoring, in particular to a multi-channel particle detection device and a detection method for detecting micron particles. Background technique [0002] At present, liquid particle counters have a certain application basis in the fields of food safety, clinical diagnosis, and biological detection. For example, the cell counting plate commonly used in biological detection includes a sample pool with a counting grid. Cell inlet, when in use, inject the culture solution with cells into the sample pool until the cells are evenly distributed and then count. However, because the depth of the sample pool is greater than the size of the cells, the cells will appear layered and superimposed in the sample pool , leading to inaccurate counting results. [0003] In order to improve the accuracy of the counting results, flow cytometer counting, microfluidic chip counting an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N15/12G01N33/543G01N33/569
CPCG01N15/12G01N33/54326G01N33/56911G01N2015/1006G01N2015/1024G01N2015/1029G01N2015/135
Inventor 陈翊平黄汉英王知龙冯婉娴
Owner WUHAN LIFE ORIGIN BIOTECH LTD
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