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Method for analyzing single cell inclusion based on micro flow-controlled chip

A technology of a microfluidic chip and an analysis method, which is applied in the field of analysis of single-cell contents, and can solve the problems of time-consuming capillary electrophoresis technology and the like

Inactive Publication Date: 2004-06-30
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a method for analyzing the contents of a single cell using a microfluidic chip as an operating platform. This method can analyze nucleic acids, proteins, amino acids, sugars and other substances in a single cell individually, and can also analyze in parallel. The above substances, while the method has considerable throughput, overcomes the time-consuming problem of capillary electrophoresis

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0018] See attached figure 1 Chip structure diagram. The running buffer is 100mM TBE, the cells are PC12 (rat adrenal pheocytoma), and the concentration is about 10 5 individual / mL. Cell lysate was 1% SDS. The dye used was YOYO-I (0.3uM). The laser wavelength is 488nm. Cells are lysed in the channel (9) between the lysis tank and the waste liquid tank, and the released RNA is negatively charged, so it is detected at the detection point I. When detecting positively charged species, it can be detected at detection point II. The detected RNA is attached figure 2 shown.

Embodiment 2

[0020] See attached figure 1 The structure diagram of the chip is shown. The running buffer is 100mM TBE, the cells are PC12 (rat adrenal pheocytoma), and the concentration is about 10 5 individual / mL. Cell lysate was 1% SDS. The sieving agent was 1% HPMC (50cp). The laser wavelength is 488nm. Apply a voltage of 500V to the cell fluid pool, running buffer pool, and cell lysate pool, and ground the waste pool. Detection at detection point I. The detected RNA is attached image 3 shown.

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Abstract

The invention is a micro-flow control chip-based single-cell endosome analysis, made on a chip platform. First, exert voltage on cell pool and earth cell waste liquor one, the electric power drives the cells to flow from the cell pool to the cell waste liquor one, observe by microscope, and when the cells reach the exit of the detecting channel, make the electrodes of the two pools suspend; at the same time, exert high voltage on run buffer liquor pool and cell cracking liquor pool, and earth waste liquor pool; in this voltage mode, ensure the cells, the cell cracking liquor, screening agent, and run buffer liquor to enter separating channel in order to complete cracking and separating single cell; and detect the endosome on two ends of the channel. It can make single or parallel analysis on the endosome.

Description

Technical field: [0001] The invention relates to an analysis method for single cell content, and in particular provides an analysis method based on a microfluidic chip. Background technique: [0002] The traditional method of analyzing cell inclusions is based on a large number of cells to obtain the total value of a certain inclusion (such as protein, nucleic acid, etc.), and the ratio of this value to the number of cells is extrapolated to the content of the substance in a single cell. For relatively homogeneous cell populations, this method is acceptable, but in other cases, such as the early stage of some serious diseases, only the composition of individual cells changes, at this time, the traditional method will average out. The specific change. Therefore, analysis at the single cell level is helpful for early diagnosis of diseases. In addition, single-cell analysis is relatively accurate in information acquisition compared with traditional methods; it is possible to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/00C12Q1/68G01N33/68
Inventor 盖宏伟刘晓君戴忠鹏马银法林炳承
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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