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High-throughput detection method for metal ions and metal nanoparticles based on single cell enrichment and single cell mass spectrometry

A single-cell and cell-based technology, applied in the field of high-throughput detection, can solve problems such as high detection limit, consumption of 100mL algae solution, and complex processing

Active Publication Date: 2020-11-24
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In the process of realizing the present invention, the applicant found that the design of the above-mentioned patent 201910246159.4 has the following limitations: (1) the cell sample needs to be made into a single-cell array before it can be detected, and the sample pretreatment is more complicated; (2) although the combined isotope dilution method, but due to the use of LA-ICP-MS as a detector, its detection throughput is low; (3) for single-cell samples, its detection limit is high, at the fg / cell level, it is difficult to achieve some trace or super Detection of Trace Samples
In the process of realizing the present invention, the applicant found that the design of the above-mentioned patent 201910246159.4 has the following limitations: (1) The sample consumption is large, and it needs to consume tens or even hundreds of milliliters of samples in the process of realizing metal adsorption. Not applicable; (2) The required biomass is large, and each sample needs to consume 100mL algae solution; (3) The detection limit of this method is high, and the uniformity of the sample is high, otherwise the repeatability of the sample detection is poor

Method used

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  • High-throughput detection method for metal ions and metal nanoparticles based on single cell enrichment and single cell mass spectrometry
  • High-throughput detection method for metal ions and metal nanoparticles based on single cell enrichment and single cell mass spectrometry
  • High-throughput detection method for metal ions and metal nanoparticles based on single cell enrichment and single cell mass spectrometry

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Embodiment 1

[0109] Embodiment 1, the detection of trace metal ions or metal nanoparticles of the present invention

[0110] 1. Culture of Tetrahymena thermophila

[0111] Thermophila Tetrahymena B2086, inoculated into 5 mL SPP medium at 3000 cells / mL, cultured in a shaker at 28 °C and 135 rpm / min for 24 h, until the cell density increased to 2×10 5 cells / mL or so. The formula of SPP medium is: add 20g to 1L deionized water Peptone, 2g glucose, 1g yeast extract, 0.03g ferric citrate, sterilized at 120°C for 20min, and then added 1% (v / v) double antibody.

[0112] 2. Metal or metal nanoparticle exposure

[0113] Gold nanoparticles (AuNPs, diameter, 5nm), stock solution 1g / L, serially diluted to 100ng / L, 500ng / L, 1μg / L, 5μg / L, 10μg / L working solution. Take 2 mL of the above-mentioned Tetrahymena suspension, add 20 μL of the above-mentioned working solution respectively, and expose Tetrahymena to 1, 5, 10, 50 or 100 ng / L of gold nanoparticles (AuNPs, diameter, 5 nm) respectively, and add...

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Abstract

The invention discloses a high-throughput detection method for metal ions and metal nanoparticles based on single cell enrichment and single cell mass spectrometry. The invention provides a method fordetecting the metal ions and metal nanoparticles. The method comprises the following steps: absorbing and enriching the metal ions and the metal nanoparticles by utilizing cells or unicellular organisms; marking the treated cells or single-cell organisms by using a metal isotope A, wherein the metal isotope A is other metal isotopes except a to-be-detected metal element and can be used for marking survival and dead cells or single-cell organisms at the same time; and then carrying out single-cell mass spectrometry flow detection to obtain an intracellular signal intensity value of the to-be-detected metal element, and further calculating to obtain the content of the to-be-detected metal element in a to-be-detected sample. The sample pretreatment is simple, and no harmful reagent is introduced; the demand on samples is very small; the flux is high; the sensitivity is high; cell-related metal signals may be differentiated.

Description

technical field [0001] The invention relates to the field of detection of metal ions and metal nanoparticles, in particular to a high-throughput detection method of metal ions and metal nanoparticles based on single-cell enrichment and single-cell mass spectrometry. Background technique [0002] Atomic absorption spectrometry (AAS), inductively coupled plasma-atomic emission spectrometry (ICP-AES) and inductively coupled plasma mass spectrometry (ICP-MS) are currently commonly used elemental analysis methods. For the detection of metal nanoparticles, the commonly used methods are laser ablation-inductively coupled plasma mass spectrometry (LA-ICP-MS), single particle / single cell-inductively coupled plasma mass spectrometry (SP / SC-ICP-MS) Wait. The AAS analysis method needs to change the lamp source for the determination of different elements, and only one element can be determined at a time, and multiple elements cannot be analyzed at the same time, and the efficiency of sa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N15/10G01N33/569G01N33/58
CPCG01N15/1031G01N33/56905G01N33/58G01N2015/1006
Inventor 曲广波吴琪史建波江桂斌
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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