A treatment method for protein in situ enzyme cleavage
A processing method and in-situ processing technology, applied in the field of analytical chemistry, can solve the problems of small mass detection range of mass spectrometry, protein molecular displacement, and affecting the imaging effect of mass spectrometry, etc.
Active Publication Date: 2020-04-28
INST OF ZOOLOGY CHINESE ACAD OF SCI
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Problems solved by technology
The reported sample preparation for protein spectrum imaging either directly detects the mass of protein molecules on the surface of tissue sections. Due to the small detection range of mass spectrometry, this method can only detect the distribution of a small number of proteins with small masses; This method needs to digest the protein in the solution state, which will cause the displacement of the protein molecule in the spatial distribution and leave the original position before the digestion, which will affect the effect of mass spectrometry imaging.
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Embodiment 1
[0051] Example 1 Using the processing method of the present invention for mass spectrometry imaging sample preparation of kidney tissue slices
[0052] (1) Prepare mouse kidney tissue sections, and prepare frozen sections by a frozen section method. Spread 20ul of poly-lysine solution evenly on the surface of the glass slide, and dry it at 80°C to prevent the slide from falling off during the experimental operation. Transfer cryosections to glass slides.
[0053] (2) Synthesis of NHS-EDC-styrene magnetic microspheres: MES is morpholineethanesulfonic acid, first use 25mMMES50mg / ml of pH=5.0 to configure EDC (1-(3-dimethylaminopropyl)-3- Ethylcarbodiimide hydrochloride) and NHS (N-hydroxysuccinimide) solutions, and then react 20ul of 0.5% W / V magnetic microspheres with a prepared 50mg / ml EDC solution, and then use Prepared 50mg / ml NHS solution for activation.
[0054] (3) The glass slide with tissue slices is fixed on the biochemical sample in situ processing device, so that ...
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Abstract
The invention discloses a processing method for protein in-situ digestion, and relates to the technical field of analytical chemistry. The processing method comprises the following steps of: fixing aglass slide with a tissue section on a biochemical sample in-situ processing device, wherein the biochemical sample in-situ processing device comprises a magnet, and magnetic lines of force of the magnet vertically penetrate through the glass slide; spraying the magnetic microspheres capable of bonding protein to the surface of the tissue slice, and incubating at room temperature; washing the tissue slice with 70% ethanol for 1-2 times, washing the tissue slice with 96% ethanol for 15 s, and vacuum drying; and spraying the protease solution on the surface of the tissue slice, and carrying outvacuum drying after full in-situ enzyme digestion. The method is suitable for sample preparation of in-situ mass spectrometry imaging of the surface protein of the biological tissue section.
Description
technical field [0001] The invention relates to the technical field of analytical chemistry. More specifically, it relates to a treatment method for in situ enzyme cleavage of proteins. Background technique [0002] Immunohistochemistry is generally used for the imaging of biological macromolecular proteins. With the development of mass spectrometry technology, there have been a few reports on the research progress of protein mass spectrometry imaging technology. The reported sample preparation for protein spectrum imaging either directly detects the mass of protein molecules on the surface of tissue sections. Due to the small detection range of mass spectrometry, this method can only detect the distribution of a small number of proteins with small masses; This method needs to digest the protein in the solution state, which will cause the displacement of the protein molecule in the spatial distribution and leave the original position before the digestion, which will affect ...
Claims
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IPC IPC(8): G01N33/68G01N1/28
CPCG01N1/2813G01N1/286G01N33/6848G01N2001/2873
Inventor 刘科辉娄新徽霍雨萌孟青
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI