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A method for analyzing the content of silver nanoparticles in single cells

A silver nanoparticle and nanoparticle technology, which is applied in individual particle analysis, particle and sedimentation analysis, measurement devices, etc., can solve the problems of difficult accurate quantification, low single-cell analysis throughput, and lack of single-cell standard substances.

Active Publication Date: 2021-05-28
INST OF HIGH ENERGY PHYSICS CHINESE ACAD OF SCI
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Problems solved by technology

However, the following bottlenecks need to be solved urgently: 1) There is a lack of suitable single-cell reference materials, and due to the influence of matrix effects and element fractionation, it is difficult to quantify accurately: 2) The throughput of single-cell analysis based on LA-ICP-MS is low

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  • A method for analyzing the content of silver nanoparticles in single cells
  • A method for analyzing the content of silver nanoparticles in single cells
  • A method for analyzing the content of silver nanoparticles in single cells

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Embodiment 1

[0039] (1) Preparation of single-cell samples: RAW264.7 cells were cultured in high-glucose DMEM / F12 medium with 10% fetal bovine serum and 1% penicillin-streptomycin double antibody solution, at 37°C and 90% humidity and 5% CO 2 , 95% air in a constant temperature incubator. When the RAW264.7 cells were cultured to 80-90% coverage in the culture flask, they were digested and passaged with 0.25% trypsin. After 24 hours of adherent growth, 2 mg / L of 20nm silver nanoparticles solution was added and incubated for 12 hours. After RAW264.7 cells were incubated with silver nanoparticles, the cells were first washed with phosphate buffered saline (PBS). Subsequently, the cells were collected after being digested with 0.25% trypsin and centrifuged at 1200 rpm for 3 min. Then, centrifuge and wash three times with 0.9% NaCl solution, collect the cells, add culture medium again to suspend to form a single cell suspension, and count the cell concentration with a cell counting plate.

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Abstract

The invention discloses a method for analyzing the content of silver nanoparticles in a single cell, comprising: a) treating a single cell array containing silver particles with an isotope diluent to obtain a single cell array containing the isotope diluent; b) adopting laser ablation ‑Inductively Coupled Plasma Mass Spectrometry (LA‑ICP‑MS) Determination of Single Cells in Arrays 109 Ag / 107 Ag ratio, according to the 109 Ag / 107 The ratio of Ag was calculated to obtain the content of silver nanoparticles in single cells. The invention provides a process and mechanism for studying the interaction between cells and nanomaterials at the single-cell level. The isotope dilution method combined with LA-ICP-MS is used to quantitatively analyze the content of silver nanoparticles in single cells, and the quantitative analysis results are accurate. High analytical throughput.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to an analysis method for silver nanoparticles in single cells. Background technique [0002] Inductively coupled plasma mass spectrometry (ICP-MS method) is a mass spectrometry elemental analysis method using plasma as the ion source, and is mainly used for simultaneous determination of multiple elements. The working principle of ICP-MS is that after the sample is introduced into the atomization system by the carrier gas (argon gas) for atomization, it enters the central area of ​​the plasma in the form of aerosol, and is desolvated, vaporized, dissociated and ionized in a high temperature and inert atmosphere. , converted into positively charged positive ions. Positive ions enter the mass analyzer through the ion collection system, and the mass analyzer separates them according to the mass-to-charge ratio, and then determines the content of the corresponding elements in the sa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N15/10
Inventor 王萌郑令娜汪冰陈汉清丰伟悦
Owner INST OF HIGH ENERGY PHYSICS CHINESE ACAD OF SCI