Jun 2, 2014
Protein injection by plasmonic laser treatment
The cellular membrane represents the ultimate barrier for the delivery of therapeutic molecules into living cells. A variety of methods are available to cross it, but all have different disadvantages. A new technique utilizing the interaction of gold nanoparticles (AuNPs) with laser pulses, termed gold nanoparticle mediated (GNOME) laser transfection, adds versatility to the common techniques. It has been proven to efficiently deliver fluorescent molecules, siRNA, and Morpholino oligomers. Reporting here in Nanotechnology, researchers present its applicability to the injection of functional proteins into living cells. The viability of the cells is conserved while simultaneously reaching high throughput.
The working procedure of GNOME laser transfection consists of the incubation of cells with spherical AuNPs for 3 hours. The particles sediment on the cell monolayer and attach non-specifically to the cellular membrane. The molecules of interest are added to the culture medium. In the last step, the sample is irradiated by a weakly focused scanning laser beam. The spatial highly localized AuNP-laser interaction provides transient membrane permeabilization via heat generation. In order to facilitate the applicability of GNOME laser transfection for routine usage, an automated and encased set-up is constructed.
Clear transfection time
Using GNOME laser transfection, a green fluorescent protein (GFP) is injected into ZMTH3 cells. Furthermore, the selective induction of apoptosis via Caspase 3 delivery is demonstrated. Compared with other protein transfection techniques, GNOME laser transfection has a well defined time of transfection point, which allows temporal investigations of the protein signaling and trafficking kinetics.
More information can be found in the journal Nanotechnology 25 245101.
About the author
Dag Heinemann and Stefan Kalies are research scientists in the department of Biomedical Optics at the Laser Zentrum Hannover e.V., Germany. Their research is focused on the laser-mediated manipulation of living cells. The team of the department carries out interdisciplinary research on biomedical laser applications and the respective optical technologies, works together with industrial partners on developing applications in this field, and uses this know-how in university teaching and academic education and training.