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Detecting method for unicellular mass spectrum flow cytometry

A flow cytometry and single-cell technology, applied in the field of biological inspection or detection, can solve the problems of detection efficiency not meeting high-throughput detection, cumbersome operation, interference, etc.

Inactive Publication Date: 2018-03-13
SHANGHAI JIAO TONG UNIV
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Problems solved by technology

[0004] Using this technology, it is possible to simultaneously accurately measure multiple protein DNA and mRNA markers in a single cell, but the detection efficiency of mass cytometry is still far from meeting the high-throughput detection in actual clinical practice; at the same time, during the labeling process, The antibody of each sample needs to be labeled separately, and the operation is cumbersome; moreover, the instrument detection signal drift during detection, the pretreatment of samples between batches before detection and the difference in labeling efficiency will inevitably interfere with the experiment

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  • Detecting method for unicellular mass spectrum flow cytometry
  • Detecting method for unicellular mass spectrum flow cytometry
  • Detecting method for unicellular mass spectrum flow cytometry

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Embodiment 1

[0058] The preparation method of this Barcoding kit comprises the following steps:

[0059] Table 1 Barcoding kit ratio scheme

[0060]

[0061]

[0062] 1) Take 5 μL of three kinds of heavy metal chloride solutions, mark the polymer (polymer), and obtain the polymer of chelating three kinds of specific metals;

[0063] 2) Use metal-labeled polymers to label three anti-CD45 antibody solutions of the same concentration, use an ultra-micro spectrophotometer Nanodrop instrument to detect the concentration of the three labeled antibodies, and use antibody stabilization for the three labeled antibodies Buffer (antibody stable diluent) was diluted to the same concentration;

[0064] 3) Take 1 μL of the three labeled antibodies and dissolve them in 10 μL and 100 μL of Cell Staining Buffer solution (cell staining buffer, CSB) respectively, dilute to the original concentration of 1% and 10% of the antibody solution, and prepare the concentration respectively Three gradient anti...

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Abstract

The invention discloses a detecting method for unicellular mass spectrum flow cytometry. A novel encoding method of an isotope metal concentration ratio marking antibody is applied, metals with different ratios is used, thus a new label combination is structured; thus a detecting channel of the mass spectrum flow cytometry is expanded by a few metal label number, a Barcoding technology which moreefficient than the existed technology is developed.

Description

technical field [0001] The present invention belongs to the field of biotechnology, and in particular the present invention relates to biological inspection or detection, using cell Barcoding technology to pre-mark cells before single-cell mass flow cytometry, and to analyze the group to which each cell belongs after mass spectrometry method. Background technique [0002] Mass cytometry (Mass Cytometry) is a flow cytometry technique that uses the principle of mass spectrometry to perform multi-parameter detection of single cells. It inherits the high-speed analysis characteristics of traditional flow cytometry, and has the high-resolution capability of mass spectrometry detection, which is a new development direction of flow cytometry. It can perform fine immunophenotyping and signaling pathway analysis on complex samples; automatically group immune cells and perform detailed analysis on the functional polymorphism of immune cells; it can perform fine subgroup analysis on c...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N15/14
CPCG01N15/1434
Inventor 蒙泓宇丁显廷张婷张璐璐
Owner SHANGHAI JIAO TONG UNIV
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