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A method for the absolute quantification of gold nanorods in single cells

An absolute quantitative and detection method technology, which is applied in the direction of measuring devices, particle and sedimentation analysis, individual particle analysis, etc., can solve problems such as insufficient convincing power, inability to detect actual cell uptake, and failure to realize absolute quantitative detection of a single nanoparticle.

Active Publication Date: 2021-02-26
INST OF HIGH ENERGY PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

However, many research methods on the metabolism of nanomaterials in animals mainly focus on the tissue level, such as fluorescent labeling methods, which cannot detect the actual uptake of cells; and non-quantitative detection methods such as TEM may cause deviations due to instrumental and human reasons. Lack of strength
[0004] At present, at the level of single-cell detection, the existing related experiments only stop at the quantitative detection of the element content in a single cell, which is a relatively quantitative detection method, and has not realized the absolute quantitative detection of a single nanoparticle in a single cell.

Method used

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  • A method for the absolute quantification of gold nanorods in single cells
  • A method for the absolute quantification of gold nanorods in single cells
  • A method for the absolute quantification of gold nanorods in single cells

Examples

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Effect test

Embodiment 1

[0035]Example 1: Absolute quantitative detection method of AuNRs in single cells

[0036](1) Production of standard sample plates

[0037]First, a layer of 200nm thick polymethylmethacrylate (PMMA) is modified on the surface of the ITO conductive glass and heated at 180°C; then using photoresist technology, a series of diameters of 5μm are obtained under the action of the developer. 、A sample pit array with a depth of 200nm, marked with panes and letters around the dot matrix. Observe under an inverted microscope, the observation results are as followsfigure 2 Shown.

[0038]Then drop the AuNRs solution after ultrasound on the surface of the conductive glass, wait for the solution to dry, and wash off the PMMA layer on the glass with acetone. Finally, a series of sample dot arrays containing AuNRs can be obtained. Observe under SEM. The observation results are as followsimage 3 Shown.

[0039](2) Preparation of standard curve for absolute quantitative detection of AuNRs

[0040]Under SEM, the numb...

Embodiment 2

[0045]Example 2: Application of the method for absolute quantitative detection of single-cell AuNRs in in vivo animal experiments (1) Preparation of standard sample plate and standard curve for absolute quantitative detection of AuNRs

[0046]Same as steps (1) and (2) in Example 1.

[0047](2) Separation of mouse liver parenchymal cells and kupffer cells

[0048]Gold nanorods with different concentrations and different modifications were injected into the tail vein of mice, mouse hepatocytes were separated by cardiac retrograde perfusion, hepatocytes were separated by 50g low-speed centrifugation, and kupffer cells were separated by magnetic bead sorting.

[0049](3) Determination of the number of gold nanorods in mouse single cells

[0050]Spread the two kinds of cells separated in step 2 on a polydimethylfluorosilane (PDMS) cell microplate with a pore size of 20μm to make them single-cell distribution, and use LA-ICP-MS to detect each cell The signal value of Au element and the number of gold na...

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Abstract

The invention provides an absolute quantitative detection method for gold nanorods in single cells. The method comprises the following steps of firstly preparing a standard sample plate by using photoresist technology to obtain a series of sample pit arrays containing AuNRs, performing counting statistics on the AuNRs in the sample pits, simultaneously determining the content of Au in the corresponding sample pits, establishing an absolute quantitative detection standard of the AuNRs according to the correspondence between the content of Au and the number of AuNRs, finally distributing the cells exposed to AuNRs in single cells, measuring the content of Au in each cell, and counting the number of AuNRs swallowed in each cell according to an absolute quantitative detection standard curve ofAuNRs. The method can be used to compare the ability of different liver cells to ingest gold nanorods, to create models of different concentrations and different modifications of gold nanorods distributed in mouse liver cells, and to study the metabolic behavior of animals at the cellular level. At the same time, the method can also be applied to other absolute quantitative detection of nanoparticles which are easy to count under SEM.

Description

Technical field[0001]The invention belongs to the technical field of nanomaterial detection, and relates to an absolute quantitative detection method of gold nanorods in single cells.Background technique[0002]The development of nanotechnology has prompted the birth of a batch of nanomaterials with unique properties of light, electricity, and magnetism. Due to their superior performance, nanomaterials have huge application prospects in biomedical diagnosis, treatment and drug delivery. Noble metal nanomaterials, especially gold nanoparticles (AuNPs) of various shapes have become a research hotspot in recent years. Due to their size and morphology, easy control synthesis, high chemical stability, easy surface modification, and unique optoelectronic properties, they are Bioimaging, biosensing, cancer diagnosis, therapeutic applications and drug delivery have broad application prospects. For example, Chinese patent application CN201410370072.5 discloses a nuclide-Cherenkov luminescence-...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N15/10
Inventor 何潇罗文赫
Owner INST OF HIGH ENERGY PHYSICS CHINESE ACAD OF SCI
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