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Microorganism single-cell metabonomics analysis method

A metabolomics and analysis method technology, applied in the field of preprocessing and data acquisition of microbial single cell metabolomics, can solve the problems of limited application, low data acquisition efficiency, insufficient single cell positioning accuracy, etc. Efficient results for single-cell data acquisition

Pending Publication Date: 2022-02-08
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] High-throughput, high-precision and efficient single-cell data acquisition is crucial for the capture of microbial metabolic heterogeneity. However, there is no precedent for the development of single-cell metabolomics technology in the field of microorganisms using mass spectrometry imaging technology, which is suitable for microbial The scale matrix application strategy is not yet perfect, the accuracy of single cell positioning is still insufficient, and the data collection efficiency is low. These factors greatly limit the application of single cell metabolomics technology in the field of microorganisms

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 The preprocessing and data acquisition method of Chlamydomonas reinhardtii single-cell metabolomics based on MALDI mass spectrometry includes the following steps:

[0031] 1. Surface arraying of indium tin oxide conductive glass sheets: use laser etching technology to prepare arrays of indium tin oxide conductive glass sheets based on positioning-guided patterns;

[0032] 2. Cleaning: During the experiment, take out 2 mL of the cultured TAP medium containing Chlamydomonas reinhardtii into a 2 mL centrifuge tube, centrifuge at 3000 rpm for 5 minutes, discard the supernatant, and add ultrapure water to wash;

[0033] 3. Dilution: gradiently dilute the washed cell liquid and check by microscope to ensure that each microliter of cell liquid contains 100-200 single cells;

[0034] 4. Pre-preparation: Manually add 0.5uL of cell solution to the center of the laser-etched dot matrix, and then place it on an ultra-clean workbench to dry to complete the preparation of t...

Embodiment 2

[0039] Example 2: Preprocessing and data acquisition method of Vibrio single-cell metabolomics based on MALDI mass spectrometry

[0040] Contains the following steps:

[0041] 1. Surface arraying of indium tin oxide conductive glass sheets: use laser etching technology to prepare arrays of indium tin oxide conductive glass sheets based on positioning-guided patterns;

[0042] 2. Cleaning: During the experiment, take out 2 mL of the cultured TAP medium containing Chlamydomonas reinhardtii into a 2 mL centrifuge tube, centrifuge at 3000 rpm for 5 minutes, discard the supernatant, and add normal saline to wash;

[0043] 3. Dilution: gradiently dilute the washed cell liquid and check by microscope to ensure that each microliter of cell liquid contains 100-200 single cells;

[0044] 4. Pre-preparation: Manually add 0.5uL of cell solution to the center of the laser-etched dot matrix, and then place it on an ultra-clean workbench to dry to complete the preparation of the cell chip. ...

Embodiment 3

[0049] Example 3: The preprocessing and data acquisition method of Saccharomyces cerevisiae single-cell metabolomics based on MALDI mass spectrometry comprises the following steps:

[0050] 1. Surface arraying of indium tin oxide conductive glass sheets: use laser etching technology to prepare arrays of indium tin oxide conductive glass sheets based on positioning-guided patterns;

[0051] 2. Cleaning: During the experiment, take out 2 mL of the cultured TAP medium containing Chlamydomonas reinhardtii into a 2 mL centrifuge tube, centrifuge at 3000 rpm for 5 minutes, discard the supernatant, and add ultrapure water to wash;

[0052] 3. Dilution: gradiently dilute the washed cell liquid and check by microscope to ensure that each microliter of cell liquid contains 100-200 single cells;

[0053] 4. Pre-preparation: Manually add 0.5uL of cell solution to the center of the laser-etched dot matrix, and then place it on an ultra-clean workbench to dry to complete the preparation of th...

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Abstract

The invention discloses a microorganism single-cell metabonomics analysis method based on MALDI mass spectrometry. The method specifically comprises the following steps: arraying the surface of a conductive slide, centrifugally washing a microbe cell suspension in double distilled water, carrying out gradient dilution after microbe washing, dropwise adding the diluted cell liquid to a specified position of the arrayed slide and drying, carrying out MALDI mass spectrometry operation and the like. The method overcomes the defects of complex pretreatment of microbial single cells, poor repeatability between data points and inaccurate data acquisition image positioning at present, and has the advantages of simple steps, high background repeatability, accurate positioning, high data acquisition speed and high sensitivity.

Description

technical field [0001] The invention relates to the technical field of mass spectrometry detection, in particular to a method for preprocessing and data acquisition of microbial single-cell metabolomics based on MALDI mass spectrometry. Background technique [0002] The field of single-cell metabolomics research is at the forefront of systems biology and analytical chemistry. This method can reveal the causes of metabolic heterogeneity at the single-cell level of microorganisms, establish precise metabolic models at the single-cell level, and explore the stability of intestinal microorganisms. In addition, single-cell metabolomics technology has great application potential in cancer target screening and precision medicine. [0003] At present, the mass spectrometers used in the research and development of single-cell metabolomics technology mainly include matrix-assisted laser desorption ionization (Matrix-Assisted Laser Desorption / Ionization, MALDI), secondary ion mass spec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/64
CPCG01N27/64
Inventor 陶飞孟宣霖许平
Owner SHANGHAI JIAO TONG UNIV
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