A silicon nanowire array chip method for detecting cell metabolites and lipids

A silicon nanowire array and cell metabolism technology, applied in the field of mass spectrometry detection, can solve the problems of single detection object, poor detection repeatability, and small detection molecular weight range, and achieve the effect of wide application range, simple operation and excellent small molecule detection ability.

Active Publication Date: 2021-05-11
HANGZHOU WELL HEALTHCARE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these array targets have the ability to detect small molecules at multiple sites, they have high cost, single detection object, small molecular weight range and poor detection repeatability.
Therefore, the above techniques have not been applied in the detection of metabolites in cell samples and actual clinical samples

Method used

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  • A silicon nanowire array chip method for detecting cell metabolites and lipids
  • A silicon nanowire array chip method for detecting cell metabolites and lipids
  • A silicon nanowire array chip method for detecting cell metabolites and lipids

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 3

[0061] Concrete method embodiment 3 is:

[0062] a, The silicon nanowire array chip and the corresponding perforated PDMS film are bonded to each other after oxygen plasma treatment to obtain a porous cell incubation chip with the silicon nanowire array as the bottom of the hole and PDMS as the wall of the hole;

[0063] b, transfer the cells into the wells of the cell incubation chip at an appropriate concentration, and incubate for an appropriate time to make the cells completely adhere to the silicon nanowires at the bottom;

[0064] c, the cells in each well are washed with a cleaning agent, the PDMS channel membrane on the surface of the silicon nanowire array chip is removed, and the silicon nanowire array chip with cells adhered to the array surface is completely dried for mass spectrometry detection.

[0065] These three embodiments are verified in Experiment 1 below.

[0066] In step 3, the acquired cell mass spectrum is analyzed in combination with a statistical ana...

Embodiment 1

[0078] Example 1, the specific steps of direct detection of cell metabolites and lipids in the suspension based on the mass spectrometry of the silicon nanowire array chip are as follows: the collected cells are washed several times with PBS and counted, and after completion, they are diluted to 10^5- 10^8 / mL cell suspension solution, the obtained cell suspension is directly used as the sample to be tested, and 1-5 μL is dropped onto the sample point on the silicon nanoarray chip, and after natural drying, the chip is fixed on the mass spectrometer target plate, Metabolic molecular spectrum information is obtained by detecting in positive or negative ion mode.

Embodiment 2

[0079] Example 2, based on the mass spectrometry of the silicon nanowire array chip, the specific steps of directly detecting the metabolites and lipids of the adherent cells on the chip surface are: a piece of PDMS membrane with the same size as the silicon nanowire chip and a thickness of 3-6mm is corresponding to the chip The silicon nanowire array chip and the corresponding perforated PDMS film are bonded to each other after oxygen plasma treatment to obtain a porous cell incubation chip with the silicon nanowire array as the bottom of the hole and PDMS as the wall of the hole. . The cells are transferred into the wells of the cell incubation chip at an appropriate concentration, and the cells are completely adhered to the silicon nanowires at the bottom after incubation for 12-24 hours. Use PBS to wash the cells in each well, peel off the PDMS channel membrane on the surface of the silicon nanowire array chip, and obtain a silicon nanowire array chip with cells attached t...

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Abstract

The invention discloses a method for detecting cell metabolites and lipids by a silicon nanowire array chip. The invention obtains a silicon nanowire array chip with multiple sites through photolithographic patterning and metal-assisted etching, and then in multiple surface modification on the basis of silicon nanowire array chip at the site; the obtained chip integrates superior desorption ability, ionization ability and high-throughput detection ability of matrix-free mass spectrometry, which simplifies the pretreatment and processing of complex samples. The detection process; and by establishing a spectrogram analysis model, the method can simultaneously detect the cell subtypes of single cell samples, mixed cell samples, xenograft tumor tissue samples, and cell samples after drug action. The operation is simple and the application range is wide.

Description

technical field [0001] The invention relates to the field of mass spectrometry detection, in particular to a silicon nanowire array chip detection method for cell metabolites and lipids. Background technique [0002] Metabolomics is one of the omics that has received widespread attention in clinical and scientific research fields. As a downstream product of genomics and proteomics, it reflects the real-time metabolic changes of cells. Existing clinical research results have shown that many diseases such as cancer, Parkinson's, Alzheimer's, obesity, etc. are related to the metabolic disorder of focal cells. Therefore, the detection and analysis of cell metabolites can help discover potential disease markers, which is expected to be applied For drug screening and to promote the development of precision medicine. Matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS), as a new type of soft ionization mass spectrometry, has the advantages of high throughput, h...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/626H01L21/027H01L21/48
CPCG01N27/628H01L21/0274H01L21/4828
Inventor 邬建敏王涛陈晓明
Owner HANGZHOU WELL HEALTHCARE TECH CO LTD
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