Microinjection method and device

a micro-injection and device technology, applied in the field of micro-injection devices, can solve the problems of difficult injection of a substance into a specific cell, certain limit regarding the reduction of external diameter, and complex operation, and achieve the effect of reducing the invasiveness of the cell

Inactive Publication Date: 2007-04-19
OLYMPUS CORP +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

[0006] It is an object of the present invention to solve the aforementioned problems of the prior art techniques. In other words, it is an object of the present invention to provide a method for introducing a physiologically active substance such as a gene into a cell, which introduces a physiologically active substance such as any given gene into any given cell in a view under a microscope, while significantly reducing inva

Problems solved by technology

However, it has been difficult to inject a substance into a specific cell.
However, since a hollow glass capillary has been used as a needle to be injected, there has been a certain limit regarding reduction in the external diameter thereof.
Thus, these conventional methods have been problematic in that a cell bursts or suffers fatal damage when a needle is injected therein, or in that op

Method used

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Examples

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example 1

[0099] Using the device shown in FIG. 1, DNA was introduced into nerve cells that were cultured in a petri dish for culture.

[0100] As nerve cells, PC12. cells (nervous system clone cells isolated from rat adrenal medulla pheochromocytoma) were used. As a medium, DMEM (Dulbecco's Modified Eagle Medium) containing 10% fetal bovine serum (FBS) was used. The culturing was carried out at 37° C. in 5% CO2. As DNA, a recombinant expression vector containing NGF receptor gene was used, and 1 μg / ml DNA solution was used.

[0101] The needle used for the device shown in FIG. 1 composed of the carbon nanotube shown in FIG. 11, which had a diameter of 50 nm and a length of 3 μm.

[0102] First, the needle was immersed in the DNA solution, so as to allow DNA to attach to the surface thereof. Thereafter, the needle was inserted into the nucleus of a nerve cell, and then released the DNA therein. After completion of the introduction of the DNA, the nerve cell was continuously cultured. Even 3 days la...

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Abstract

An object of the present invention is to provide a method for introducing a physiologically active substance such as a gene into a cell, which introduces a physiologically active substance such as any given gene into any given cell in a view under a microscope, while significantly reducing invasiveness to the cell, and a device used for the above method. The present invention provides a method for introducing a physiologically active substance into a cell, which comprises: allowing a physiologically active substance to attach around a needle having a diameter of 500 nm or less, provided that it is able to be inserted into a cell; and inserting the above-described needle into the cell; and a microinjection device for carrying out the aforementioned method.

Description

TECHNICAL FIELD [0001] The present invention relates to a method for introducing a physiologically active substance into cells and a microinjection device used for the above method. BACKGROUND ART [0002] Examples of a technique of introducing gene DNA into cultured cells or the like may include the calcium precipitation method, the lipid transfer method, the viral vector method, electroporation, the gene gun method, and the microinjection method. In the above methods other than the microinjection method, DNA is introduced in cells at a certain probability, and thus it is impossible to introduce DNA into only a specific cell. On the other hand, the microinjection method has been problematic in that since the diameter of the edge of a glass pipette is approximately 1 μm, cells are easily damaged when such a glass pipette is inserted into the cell nucleus thereof. In addition, when different genes are introduced into multiple cells, the same number of pipettes as that of genes should b...

Claims

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Application Information

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IPC IPC(8): C12N15/00C12M1/00C12M3/00
CPCB82Y30/00C12M35/00
Inventor MIYAWAKI, ATSUSHIIMABAYASHI, HIROYUKIKARAKI, SACHIKO
Owner OLYMPUS CORP
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