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Continuous unicellular inclusion analyzing method based on microflow control chip platform

A technology of microfluidic chips and analysis methods, applied in biochemical equipment and methods, measurement/testing of microorganisms, etc., can solve problems such as complex operation of capillary electrophoresis technology

Inactive Publication Date: 2004-11-24
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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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. Amino acids, sugars and other substances, or parallel analysis of the above substances, at the same time, the method has considerable throughput, which overcomes the shortcomings of complex operation of capillary electrophoresis technology

Method used

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Embodiment 1

[0024] See attached drawing 1 chip structure diagram. The running buffer is 10mM PBS, the cells are carp red blood cells, the concentration is about 10 5 individual / mL. Cell lysate is 0.7% SDS. The dye used was YOYO-I (1 uM). The laser wavelength is 488nm. Cells are lysed in the channel between the buffer pool and the waste pool, and the released nucleic acids are negatively charged. If the buffer contains only PBS, the detection point is at Figure 1B. When the buffer contains PBS and HPMC, the detection point is at the accompanying drawing 1A

[0025] Shown in accompanying drawing 8 is the nucleic acid detection figure of carp red blood cell. The running buffer is 10mMPBS, with a concentration of about 10 7 individual / mL. Cell lysate is 0.7% SDS. The laser wavelength is 488nm. The electric field strength in the channels of the buffer pool and the waste pool is 120V / cm, and the detection point is detected at Figure 1B.

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Abstract

The continuous unicellular inclusion analyzing method based on microflow control chip platform features that in the cross passage microflow control chip as operation platform, after the buffering liquid with cell dividing agent and screening agent is added into buffering pool and the cell liquid is added into the cell pool, voltage is applied between the buffering pool and the waste liquid pool, so that single cell is introduced into the horizontal passage to contact with the cell dividing agent and divided and the released inclusion migrates directionally under the action of electric field to complete the electrophoresis. The method of the present invention may be used in continuous single cell sampling and cracking, and in analyzing intercellular nucleic acid, protein, amino acid, saccharide and other matters in relatively high flux and relatively simple process.

Description

Technical field: [0001] The invention mainly relates to an analysis method for realizing continuous single cell sampling, lysing and inclusion detection on a microfluidic chip platform. 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; i...

Claims

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Application Information

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IPC IPC(8): C12Q1/02
Inventor 盖宏伟於林芬马银法林炳承
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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