Characterization of nanoparticle mediated laser transfection by femtosecond laser pulses for applications in molecular medicine

Background: In molecular medicine, the manipulation of cells is prerequisite to evaluate genes as therapeutic targets or to transfect cells to develop cell therapeutic strategies. To achieve these purposes it is essential that given transfection techniques are capable of handling high cell numbers in reasonable time spans. To fulfill this demand, an alternative nanoparticle mediated laser transfection method is presented herein. The fs-laser excitation of cell-adhered gold nanoparticles evokes localized membrane permeabilization and enables an inflow of extracellular molecules into cells. Results: The parameters for an efficient and gentle cell manipulation are evaluated in detail. Efficiencies of 90% with a cell viability of 93% were achieved for siRNA transfection. The proof for a molecular medical approach is demonstrated by highly efficient knock down of the oncogene HMGA2 in a rapidly proliferating prostate carcinoma in vitro model using siRNA. Additionally, investigations concerning the initial perforation mechanism are conducted. Next to theoretical simulations, the laser induced effects are experimentally investigated by spectrometric and microscopic analysis. The results indicate that near field effects are the initial mechanism of membrane permeabilization. Conclusion: This methodical approach combined with an automated setup, allows a high throughput targeting of several 100,000 cells within seconds, providing an excellent tool for in vitro applications in molecular medicine. NIR fs lasers are characterized by specific advantages when compared to lasers employing longer (ps/ns) pulses in the visible regime. The NIR fs pulses generate low thermal impact while allowing high penetration depths into tissue. Therefore fs lasers could be used for prospective in vivo applications.

Compact discs as versatile cost-effective substrates for releasable nanopatterned aluminium films

We demonstrate that standard polycarbonate compact disk surfaces can provide unique adhesion to Al films that is both strong enough to permit Al film nanopatterning and weak enough to allow easy nanopatterned Al film detachment using Scotch tape. Transferred Al nanohole arrays on Scotch tape exhibit excellent optical and plasmonic performance.

Boron nitride nanosheet-veiled gold nanoparticles for surface-enhanced raman scattering

Atomically thin boron nitride (BN) nanosheets have many properties desirable for surface-enhanced Raman spectroscopy (SERS). BN nanosheets have a strong surface adsorption capability toward airborne hydrocarbon and aromatic molecules. For maximized adsorption area and hence SERS sensitivity, atomically thin BN nanosheet-covered gold nanoparticles have been prepared for the first time. When placed on top of metal nanoparticles, atomically thin BN nanosheets closely follow their contours so that the plasmonic hot spots are retained. Electrically insulating BN nanosheets also act as a barrier layer to eliminate metal-induced disturbances in SERS. Moreover, the SERS substrates veiled by BN nanosheets show an outstanding reusability in the long term. As a result, the sensitivity, reproducibility, and reusability of SERS substrates can be greatly improved. We also demonstrate that large BN nanosheets produced by chemical vapor deposition can be used to scale up the proposed SERS substrate for practical applications.

Investigation of conformationally switchable biomolecular ligands at aqueous metallic interfaces.

An atomistic understanding was garnered through large scale molecular simulations, for the adsorption and interaction of reconfigurable hybrid biomolecule components at different aqueous metallic interfaces. This would allow for the development of future hybrid biomolecular ligands that could be used to synthesise novel and tuneable materials with unique electromagnetic properties.