Microinjection process for introducing an injection substance pa

Details Microinjection process for introducing an injection substance pa Microinjection process for introducing an injection substance pa

1999-06-03

2000-05-16

Ketter, James

Chemistry: molecular biology and microbiology

Process of mutation, cell fusion, or genetic modification

Introduction of a polynucleotide molecule into or...

4352851, C12N 1564

Patent

active

060636297

ABSTRACT:
The invention relates to a microinjection process for introducing an injection substance, particularly genetic material, into procaryotic and eucaryotic cells, as well as cell compartments of the latter (plastids, cell nuclei). It is disadvantageous in the prior art that the cell is damaged by the glass pipette. This problem is obviated by the invention in that the nanopipette (10), which has an external diameter of 0.05 to 0.2 .mu.m, an internal diameter of 0.1 to 1.5 mm and a tip diameter of 0.025 to 0.3 .mu.m is used and is filled with the injection substance (1) and a heat-expandable substance or substance mixture and the capillary of the nano-pipette (10) is then sealed with an adhesive, the pipette tip, with the aid of a microscope and a micromanipulator, is stuck into the desired plastids, bacterium or cell compartment/cell nucleus and the nanopipette is heated by means of a regulatable heater (12) until the injection substance passes out of the pipette tip at an outflow rate of up to 1 femtoliter per second and enters the plastids, bacterium or cell compartment/cell nucleus, whose diameter is 1 to 20 .mu.m.
The invention also relates to the correspondingly filled nanopipette, which is heatable by a regulatable heater for temperature-controlled injection purposes.

REFERENCES:
patent: 3720354 (1973-03-01), Drummond et al.
patent: 4625677 (1986-12-01), Neher
patent: 5225750 (1993-07-01), Higuchi et al.
Patent Abstracts of Japan, Higuchi Ryuichi, "Dispensing Apparatus" 05133851, May 28, 1993.

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