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1 July 2002 Laser-assisted optoporation of single cells
Herbert Schneckenburger, Anita Hendinger, Reinhard Sailer, Wolfgang S. L. Strauss, Marco Lyttek, Michael Schmitt
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Journal of Biomedical Optics, Vol. 7, Issue 3, (July 2002). https://doi.org/10.1117/1.1485758
The plasma membrane of Chinese hamster ovary cells was made permeable using the focused beam of an argon ion laser (488 nm) and phenol red as a light absorbing dye. Small circular dark spots on the cell surface appeared immediately after laser irradiation and disappeared within about 5 min. They were related to transient changes in membrane properties, which could be visualized using the fluorescent marker laurdan, and were probably due to a local increase in temperature. According to a colony forming assay, cell viability was maintained by using light doses up to 2.5 MJ/cm2 applied for 1 s. In addition to measurements of the efflux of the cytoplasmic marker calcein, cell transfection using a green fluorescent protein (GFP) coding plasmid was studied: brightly fluorescent GFP with an emission maximum around 510 nm was observed within part of the cells after 24 h. The transfection rates after laser irradiation were around 30% for younger subcultures and less than 10% for aging cells. This may be due to age dependent changes in the phase transition of membrane lipids from gel phase to liquid crystalline phase. High transfection rates, visual control and universality towards various cell lines are possibly the main advantages of laser-assisted optoporation in comparison with presently existing methods of cell transfection.
©(2002) Society of Photo-Optical Instrumentation Engineers (SPIE)
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Herbert Schneckenburger, Anita Hendinger, Reinhard Sailer, Wolfgang S. L. Strauss, Marco Lyttek, and Michael Schmitt "Laser-assisted optoporation of single cells," Journal of Biomedical Optics 7(3), (1 July 2002). https://doi.org/10.1117/1.1485758
Published: 1 July 2002
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Cited by 101 scholarly publications and 7 patents.
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KEYWORDS
Luminescence
Laser irradiation
Green fluorescent protein
Argon ion lasers
Linear filtering
Optical tweezers
Laser cutting
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