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In-vivo lipid three-dimensional mass spectrum imaging method based on integral zebra fish model

A mass spectrometry imaging, zebrafish technology, applied in the field of mass spectrometry detection, can solve problems such as spleen enlargement, mental retardation, dystonia, etc., and achieve the effect of high chemical specificity and high integrity

Pending Publication Date: 2020-12-18
GUANGDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are some applications of three-dimensional MALDI mass spectrometry imaging, but they are limited to the study of a single organ or tissue, and many diseases involve multiple tissues or organs, such as NPC1 disease
The disease is a lysosomal lipid storage disease of autosomal recessive inheritance, which accounts for 95% of NPC cases. The abnormal lipid metabolites in individuals are studied, but the disease will involve multiple organs such as internal organs and the nervous system, and patients will have symptoms such as enlarged spleen, dystonia, ataxia, and mental retardation

Method used

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  • In-vivo lipid three-dimensional mass spectrum imaging method based on integral zebra fish model
  • In-vivo lipid three-dimensional mass spectrum imaging method based on integral zebra fish model
  • In-vivo lipid three-dimensional mass spectrum imaging method based on integral zebra fish model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Acquisition of Whole Zebrafish Mass Spectrometry Imaging Data

[0049] Sample preparation for frozen sections: NPC1 - / - After the adult zebrafish in group and WT group were euthanized, the whole zebrafish was embedded with embedding agent, and placed in a -80°C refrigerator until it solidified; after that, frozen section samples were prepared and placed in a cryostat at -20°C Take a tissue section sample with a thickness of 14 μm every 56 μm and adhere to the ITO plate, NPC1 - / - 20 slices were taken from the control group and the WT group respectively. Place the ITO plate with the tissue slice attached in the vacuum drying oven for 30 min before depositing the NEDC matrix, and each plate needs about 6 mL.

[0050] Matrix deposition instrument: matrix deposition instrument (Imageprep, Bruker Company). Deposition conditions: 35% of spray power; modulation 20%; deposition time 1.0s; incubation time 15s; drying time 60s.

[0051] Mass Spectrometry Instrument: ...

Embodiment 2 3

[0052] Embodiment 2 Construction of three-dimensional MALDI mass spectrometry image

[0053] The mass spectrometry imaging data was imported into SCiLS Lab 2016b (Bremen, Germany) Premium 3D for processing. By adjusting the x and y axes, 20 tissue slices could be accurately stacked according to the original biological appearance, making it approximately represent the whole animal model. Normalization was performed with the total ion current method, the m / z interval was set to ±0.1Da, and all MALDI mass spectrometry images were processed using a weak denoising method, and then the mass spectrometry images of lipids in the original position of the tissue were constructed.

[0054] Such as figure 2 Shown: figure 2 A. figure 2 B. figure 2 C are two-dimensional MALDI mass spectrometry images distributed in the spine (m / z 452.324), brain (m / z 790.854) and intestinal tract (m / z 867.534). The largest sections of the three organs are located in the 11th slice , on the 17th and ...

Embodiment 3

[0055] Example 3 Screening of differential lipids in the zebrafish NPC1 disease model

[0056] Embedded NPC1 after euthanasia - / - The whole zebrafish of the group and the WT group, after obtaining the frozen section tissue samples and carrying out matrix deposition, according to the method of embodiment 1, respectively to NPC1 - / - MALDI mass spectrometry imaging was performed on all sliced ​​samples of the WT group and the WT group, and then the three-dimensional MALDI patterns were reconstructed according to the method in Example 2. Fifteen kinds of differential lipid m / z were screened, and the detected differential m / z values ​​were entered into the lipidomics mass spectrometry database Lipidsearch4.0 for searching, and compared with the secondary fragment information to determine the differential m / z The molecular formula corresponding to the z value and the name of the lipid corresponding to the molecular formula. The search conditions are: the molecular weight deviation...

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Abstract

The invention belongs to the technical field of mass spectrum detection, and discloses an in-vivo lipid three-dimensional mass spectrum imaging method based on an integral zebra fish model. The invention establishes the in-vivo lipid three-dimensional mass spectrum imaging method based on the integral zebra fish model, the method is applied to the research of a zebra fish C1 type Niemann-Pick disease model, and the result shows that fifteen lipids with obvious spatial distribution difference exist in different organs of zebra fish. According to the three-dimensional MALDI mass spectrum imagingmethod established by the invention, complex sample pretreatment is not needed, various compounds in a tissue sample can be detected at the same time, and the integrity of selected tissues and a multi-dimensional visual effect can be ensured, so that the spatial distribution difference between an experimental group and a control group can be visually and comprehensively obtained; the method has the advantages of high integrity and high chemical specificity.

Description

technical field [0001] The invention relates to the technical field of mass spectrometry detection, in particular to a method based on three-dimensional mass spectrometry imaging of lipids in a whole zebrafish model. Background technique [0002] MALDI mass spectrometry imaging technology is a new technology for visual analysis and detection. It combines the component identification ability of mass spectrometry with computer-aided image visualization technology. By using different matrices at the tissue, cell or subcellular level, it can be realized. In situ imaging analysis of molecular compounds, peptides, proteins, nucleic acids, lipids and other molecules and their metabolites. This method can realize in situ qualitative and quantitative analysis of various molecules in tissue samples without complicated pretreatment, and draw their two-dimensional or three-dimensional spatial distribution maps, which has the advantages of high sensitivity, high analysis speed and high c...

Claims

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Application Information

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IPC IPC(8): G01N27/62G01N1/04G01N1/28
CPCG01N27/628G01N1/04G01N1/286G01N2001/2866
Inventor 梁晓萍曹胜西
Owner GUANGDONG UNIV OF TECH
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