A novel method of DNA transfection by laser microbeam cell surgery

  title={A novel method of DNA transfection by laser microbeam cell surgery},
  author={Motowo Tsukakoshi and Shun-ichi Kurata and Yoshio C O Rikagaku Ken Nomiya and Yoji Ikawa and Takahiro Kasuya},
  journal={Applied Physics B},
A new technique is presented to incorporate exogeneous gene materials (DNA) into cells with a microbeam irradiation from an uv pulsed laser. A frequency-multiplied Nd:YAG laser, 355 m wavelength, 5 ns pulse duration, punches a self-healing hole of submicrometer aperture in cell membrane under selected irradiation conditions. It takes a fraction of a second for the aperture to close, long enough to allow the foreign DNA, contained in the medium, to slip into the cell. The method offers a clear… 
Laser-Based Gene Transfection and Gene Therapy
This review covers four different laser-assisted transfection techniques and their advantages and disadvantages and suggests Universality towards various cell lines is possibly the main advantage of laser- assisted optoporation in comparison with presently existing methods of cell transfections.
Dielectric microsphere mediated transfection using a femtosecond laser.
An enhanced optical field generated under polystyrene microspheres of 1000 nm diameter excited by a femtosecond laser pulse has advantages both in high throughput of many cell treatments and precise processing of minute areas on cell membranes.
Optical forces in biophotonics: transfection and cell sorting
Lately, several groups successfully used ultrashort laser pulses to selectively permeabilize the membrane of living cells to achieve transport of foreign molecules, like DNA, into the cells. For
Femtosecond optical transfection of cells: viability and efficiency.
Contrary to recent literature, in which 100% efficiency is claimed, this measure of efficiency accounts for all irradiated cells, including those lost as a result of laser treatment, thereby providing a true biological measure of the technique.
Femtosecond optical transfection of individual mammalian cells
The entire protocol takes ∼5 d to complete, from the initial setup of the femtosecond optical transfection system to the final stage of fluorescence imaging to assay for successful expression of the gene of interest.
Nano particles insertion into individual mammalian cells using optical tweezers
Transfection is the process of introducing DNA into cells so that the introduced DNA will function and produce proteins. This technique is useful to study the function of various DNA sequences and in
Gold nanoparticle mediated cell manipulation using fs and ps laser pulses for cell perforation and transfection
Manipulation of cells requires the delivery of membrane-impermeable substances like genetic materials or proteins into the cytoplasm. Thus delivery of molecules over the cell membrane barrier is one
Optical transfection of mammalian cells
This work is investigating the introduction and subsequent expression of foreign DNA into living mammalian cells by laser-assisted photoporation with a femtosecond-pulsed titanium sapphire laser at 800 nm, in cells that are adherent.
Single cell optical transfection
This discussion explores a procedure called optical injection, where a laser field transiently increases the membrane permeability to allow species to be internalized and provides a forecast of future applications of this rapidly developing and exciting technology.
In vitro study for laser gene transfer in BHK-21 fibroblast cell line
The present work applied the introduced here home-made laser setup with its synchronized parameters to introduce pBK-CMV phagemid, containing LacZ and neomycin resistance (neor )genes into BHK-21 fibroblast cell line to introduce safe, efficient and less damaging modality for gene transfer in animal cells.


Laser microsurgery in cell and developmental biology.
New applications of laser microbeam irradiation to cell and developmental biology include a new instrument with a tunable wavelength (217- to 800-nanometer) laser microbeam and a wide range of
A microsurgical methodology for human cells in vitro: evolution and applications.
A methodology for micromanipulating human cells of normal and malignant origin, in vitro, has evolved from the study of about 2000 HeLa, ERK (a subline of HeLa cells), and human embryonic lung cells
A high-sensitivity laser microprobe mass analyzer
A laser microprobe mass analyzer has been developed. It is intended for application in biomedical and physiological research. A frequency-doubled ruby laser is focussed through an incident light
Mechanisms of DNA uptake by mammalian cells: fate of exogenously added DNA monitored by the use of fluorescent dyes.
It is found that DNA forms a tight complex with Ca Pi and that DNA in this complex is resistant to nucleases present in serum or added externally and upon the concentration of DNA in the complex.
Expression of a bacterial gene in mammalian cells.
Transfection of cultured monkey kidney cells with recombinant DNA constructed with a cloned Escherichia coli gene that codes for xanthine-guanine phosphoribosyltransferase and several different SV40
Transforming activity of DNA of chemically transformed and normal cells
It seems that endogenous transforming genes of uninfected cells can be activated and efficiently transmitted by transfection, consistent with the hypothesis that normal cells contain genes that are capable of inducing transformation if expressed at abnormal levels.