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Nanoprocessing of biological targets with nanojoule near infrared femtosecond laser pulses

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Abstract

Nanoprocessing of living cells and tissues including targeted transfection of cells is an important technique for nanosurgery, gene therapy and related biomedical applications. We delineate how high-intensity (1012 W/cm2) near-infrared (NIR) 80 MHz nanojoule femtosecond laser pulses of an ultracompact non-amplified Ti:sapphire laser system can create highly localised membrane perforations within a minute focal volume, enabling noninvasive direct transfection of mammalian cells with DNA. We suspended Chinese hamster ovarian (CHO), rat kangaroo kidney epithelial (PtK2) and rat fibroblast cells in 0.5 ml culture medium in a sterile miniaturised cell chamber (JenLab GmbH, Jena, Germany) containing 0.2 µg plasmid DNA vector pEGFP-N1 (4.7 kb), which codes for green fluorescent protein (GFP). The NIR laser beam was introduced into a femtosecond laser scanning microscope (JenLab GmbH, Jena, Germany) and focussed on the edge of the cell membrane of a target cell for 16 ms.

© 2003 Optical Society of America

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