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Biochemical sample in-situ processing device and mass spectrometry imaging method for phosphorylated proteins

An in-situ processing and biochemical processing technology, applied in the field of analytical chemistry, to achieve the effect of clear imaging, convenient use, promotion of establishment and development, and popularization and application.

Active Publication Date: 2020-04-28
INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, there is no report on mass spectrometry imaging of phosphorylated proteins, so the mass spectrometry imaging analysis technology for phosphorylated proteins has strong practical significance

Method used

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  • Biochemical sample in-situ processing device and mass spectrometry imaging method for phosphorylated proteins
  • Biochemical sample in-situ processing device and mass spectrometry imaging method for phosphorylated proteins
  • Biochemical sample in-situ processing device and mass spectrometry imaging method for phosphorylated proteins

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Experimental program
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Effect test

Embodiment 1

[0057] (1) Transfer the frozen section of biological tissue to the pretreated ITO glass slide, dry it in vacuum, transfer and fix it on the slide holder, and fix the matching double-layer magnet with strong magnetic field strength;

[0058] (2) Use a matrix sprayer to spray magnetic nanomaterials that can bond with -COOH; the biochemical treatment tank is prepared to hold various solutions required for biochemical treatment, including 70% ethanol and 100% ethanol for washing off fat Classes and other non-amino acid small molecules;

[0059] (3) Dry in vacuum, spray the matrix with a matrix sprayer, and detect by mass spectrometry imaging. A good imaging effect of amino acid mass spectrometry has been achieved, such as Figure 5 shown.

Embodiment 2

[0061] (1) Prepare biological tissue slices cut with surface protease, prepare frozen slices by a frozen section method, spray trypsin solution, incubate for 2 hours, and obtain biological tissue frozen slices after vacuum drying.

[0062] (2) Place the frozen section of biological tissue after protease digestion on the slide glass 3 of the biochemical sample in situ processing device provided by the present invention, and spray the surface of the frozen section of biological tissue at a concentration of 0.01ug / ul-100ug / ul ferric oxide suspension 5-250ul and incubate for 15min;

[0063] (3) Place the treated biological tissue frozen section in a magnetic field with vertical magnetic field lines for adsorption, and the magnetic ferric iron tetroxide suspension is combined with the phosphorylated peptide in the enzyme-cleaved peptide on the surface of the biological tissue frozen section , so that the phosphorylated peptides are fixed in place by magnetic adsorption;

[0064] ...

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Abstract

The invention discloses an in-situ processing device for biochemical samples and a mass spectrometry imaging method for phosphorylated proteins, and relates to the technical field of analytical chemistry. The device for in-situ processing of biochemical samples includes: a biochemical treatment tank, a glass slide holder arranged in the biochemical treatment tank, and a slide glass and a double-layer magnet arranged in the slide glass holder, the double-layer magnet The magnetic field lines of the method pass through the glass slide vertically; the phosphorylated protein spectrum imaging method utilizes ferric oxide suspension to adsorb and immobilize phosphorylated peptides, and then mass spectrometry can be performed. The invention uses magnetic field technology and magnetic materials to process biochemical samples in situ, and is applied to mass spectrum imaging of phosphorylated proteins.

Description

technical field [0001] The invention relates to the technical field of analytical chemistry. More specifically, it relates to an in-situ processing device for biochemical samples and a mass spectrometry imaging method for phosphorylated proteins. Background technique [0002] The processing of biochemical molecules on the surface of biological tissue samples is a mode of sample processing, which is widely used in the sample pretreatment process of various biological tissue surface analysis, such as staining of animal tissue surfaces, immunohistochemistry, transmission electron microscopy, cryo-electron microscopy and other samples surface treatment etc. For simple surface biochemical treatments, such as staining and immunohistochemistry, biochemical molecules only undergo simple non-covalent adsorption or covalent bonding on the sample surface, and spatial displacement is not easy to occur. However, in the more complicated and multi-step biochemical treatment process of th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/28G01N27/62
CPCG01N1/28G01N27/62
Inventor 刘科辉娄新徽霍雨萌侯国丽
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI
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