Microfluid system used for unicell ultrasonic gene introduction, and introduction method thereof

A microfluidic system, gene introduction technology, applied in other methods of inserting foreign genetic materials, treatment of microorganisms with electricity/wave energy, methods of stress stimulation of microbial growth, etc., can solve problems such as difficult interpretation of cell analysis data, and achieve saving Raw materials, high reliability, and the effect of increasing work efficiency

Active Publication Date: 2016-05-11
HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the heterogeneity of the response of each cell and the different life or metabolic cycle, the result of this averaging often makes the analysis data of cells difficult to interpret

Method used

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  • Microfluid system used for unicell ultrasonic gene introduction, and introduction method thereof
  • Microfluid system used for unicell ultrasonic gene introduction, and introduction method thereof
  • Microfluid system used for unicell ultrasonic gene introduction, and introduction method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Such as figure 1 , figure 2 As shown, the microfluidic system for single-cell ultrasonic gene delivery is made of a microfluidic pipeline 7 on a PMMA substrate 6 by a hot pressing process, and an ultrasonic transducer 4 is provided on a polyimide substrate 8 by a MEMS process;

[0031] The microfluidic pipeline is an integrated three-section structure, the front section is "Y" shaped, the middle section is serpentine, and the rear section is linear; the two ports in the front section of the "Y" shape are respectively provided with inlets for the cell fluid to be treated 1 and the target gene solution inlet 2, a mixing chamber 3 is provided at the intersection of the two branches of the "Y" front section, and a treated cell liquid outlet 5 is provided at the port of the straight rear section; PMMA substrate 6 and polyimide substrate 8 are bonded and fixed together, and the ultrasonic transducer 4 is positioned above the serpentine middle section of the microfluidic pip...

Embodiment 2

[0041]This example is used to illustrate how to use the above-mentioned microfluidic system to realize single-cell ultrasonic gene transfer:

[0042] First, at the same time, the cell solution to be treated is injected into the microfluidic pipeline from the inlet of the cell solution to be treated through a syringe pump, and the target gene solution is injected into the microfluidic pipeline from the target gene solution inlet;

[0043] Then, the cell fluid to be treated and the target gene solution converge at the mixing chamber of the microfluidic pipeline, and mix evenly, and enter the serpentine middle section; turn on the corresponding ultrasonic transducer, generate ultrasonic waves and act on the cells 10 to be treated, and Ultrasonic micropores are generated on the cell surface, and the target gene enters the cells to be treated through the ultrasonic micropores to complete the introduction of the target gene 11;

[0044] Finally, the treated cells enter the linear re...

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Abstract

The invention discloses a microfluid system for unicell ultrasonic gene introduction, and an introduction method thereof. The microfluid system is characterized in that a PPMA base body is provided with a microfluid pipeline through a hot pressing technology, and a polyimide substrate is provided with an ultrasonic transducer with different frequencies through an MEMS technology; and the PMMA base body and the polyimide substrate are bonded and fixed together, and the ultrasonic transducer is positioned above the microfluid pipeline. When the microfluid system is used, a cell sap to be treated and a target gene solution are injected to the microfluid pipeline and are uniformly mixed, the ultrasonic transducer is started, and the target gene goes through ultrasonic micropores and is introduced to cells to be treated when the ultrasonic micropores are generated on the surface of the cells to be treated. The microfluid system can accurately realize unicell ultrasonic gene delivery without cell size and kind restriction or change of cell survival environment, and has the advantages of high introduction success rate, device micromation, rapid and simple operate, high precision and high reliability.

Description

technical field [0001] The invention relates to a device and a method for introducing genes into biological cells, in particular to a device and a method for single-cell ultrasonic precise gene introduction. Background technique [0002] Since the 1990s, with the rapid development of ultrasonic medicine and the deepening of theoretical research on ultrasonic cavitation, ultrasonic gene delivery technology has gradually emerged. Ultrasonic gene delivery is a new type of cell membrane perforation technology developed recently, which has been applied to cell transfection of DNA and drugs. When ultrasonic waves of a certain intensity propagate in the liquid, bubbles are formed, and when the bubbles burst, local high temperature, high pressure and shock waves are formed, which is the so-called cavitation effect. Using the cavitation effect, in a liquid environment, nano-scale micro-perforations can be produced on the cell membrane, and gene fragments in the liquid around the cel...

Claims

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Application Information

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IPC IPC(8): C12M1/42C12N13/00C12N15/87
CPCB01L3/5027B01L2300/0861B01L2400/0439C12N13/00C12N15/87
Inventor 尤晖张萍林荣辉张瑞黄哲杨明鹏李小军王晓飞
Owner HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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