Single cell optical transfection

Abstract

The plasma membrane of a eukaryotic cell is impermeable to most hydrophilic substances, yet the insertion of these materials into cells is an extremely important and universal requirement for the cell biologist. To address this need, many transfection techniques have been developed including viral, lipoplex, polyplex, capillary microinjection, gene gun and electroporation. The current discussion explores a procedure called optical injection, where a laser field transiently increases the membrane permeability to allow species to be internalized. If the internalized substance is a nucleic acid, such as DNA, RNA or small interfering RNA (siRNA), then the process is called optical transfection. This contactless, aseptic, single cell transfection method provides a key nanosurgical tool to the microscopist-the intracellular delivery of reagents and single nanoscopic objects. The experimental possibilities enabled by this technology are only beginning to be realized. A review of optical transfection is presented, along with a forecast of future applications of this rapidly developing and exciting technology.

Spatially, Temporally, and Quantitatively Controlled Delivery of Broad Range of Molecules into Selected Cells through Plasmonic Nanotubes

“…A step behind this direction is represented by ultrafast optoporation that is an alternative to the classical methods of intracellular delivery. [ 11,12 ] Very recently, optoporation was also achieved in the microsecond regime by generating microbubbles oscillating in size. [ 13 ] In this case, the induced shear stress creates transient pores in the cell membrane thus enabling delivery of molecules.…”

Dynamics of laser-induced bubble pairs

“…Assuming that the momentum of the fluid motion is largely contained in the respective jet flows, it is to be expected that the relation m jet1 v jet1 = m jet2 v jet2 holds because of conservation of momentum, where m jeti and v jeti denote the mass and velocity of the jet from bubble i (i = 1, 2), respectively. This implies the possibility of achieving a combination of a massive and slow jet opposed by a thin and fast jet that could be useful for various applications, for instance micropumping and opto-injection for cell transfection Stevenson et al 2010). The use of laser-based methods for introducing genetic materials or other substances into living cells has become a very active area of research, and various techniques for opto-poration of the cell membrane have been developed (Tirlapur & König 2002;Vogel et al 2005;Baumgart et al 2008;Stevenson et al 2010;Antkowiak et al 2013a,b;Davis et al 2013).…”

Enhancement and optimization of plasmid expression in femtosecond optical transfection

“…According to this definition, the value of efficiency is not corrected for the proliferation of cells during the 48 hours. The previously reported values of efficiency for femtosecond cellular transfection for different cell lines typically lies in the range of 40-60% [1]. The efficiency of transfection depends on a number of physical parameters such as beam spot size, quality, timing of dose, accuracy of poration, which can be optimized [9][10][11].…”

“…Optically assisted transfection is a process in which a highly localised laser beam alters the permeability of the plasma membrane of a cell, thereby allowing the entry of extracellular nucleic acids such as DNA, RNA or si-RNA into the cell [1]. Cell transfection is used as a tool for studying and elucidating the basic cellular processes and genotypic changes inside the cells.…”

“…Cell transfection is used as a tool for studying and elucidating the basic cellular processes and genotypic changes inside the cells. The ability to permeate the lipid cell membrane in a sterile and relatively non-invasive manner makes this technique a powerful tool when compared to other established transfection methods such as viral vectors, chemical transfection, electroporation, especially in the field of drug delivery and gene therapy [1].…”