Violet Diode-Assisted Photoporation and Transfection of Cells

Various methods for transfecting molecules such as DNA, RNA, proteins, or drugs with high efficiency and low toxicity have been implemented and optimized for many different cell types. These include widely used techniques such as chemical transfection (lipid-based techniques), the use of viral vectors and electroporation. To date, laserassisted transfection of cells has relied on cumbersome Argon ion lasers or large, high-power lasers capable of achieving femtosecond pulses (titanium sapphire), with a few exceptions. The work reported in this article, and also recently published in Optics Express, shows how the successful transfection and creation of stable cell lines is achieved using a compact, relatively inexpensive, violet diode laser (Toptica or Nichia, at 405 nm wavelength). Turovets et al. use an argon fluoride excimer laser at 193 nm to transfect plant cells. Shirahata et al. transfect mammalian cells using a 355nm pulsed neodymium-yttrium-aluminum garnet (Nd:YAG) with pulse duration of 10ns, and Mohanty et al. also use a pulsed Nd:YAG laser of wavelength 1064 nm and pulse duration of 17 ns. Sagi et al. also transfect mammalian cells using a Holmium:YAG laser (2.1 mm wavelength). These wavelengths were all used to create pores in the plasma membrane or to change the permeability of the plasma membrane through a variety of effects such as heating, absorption, photochemical effects, or the creation of reactive oxygen species. 1,2