Optical transfection is a promising technique for the delivery of foreign genetic material into cells by transiently changing the permeability of the cell membrane. Of the different optical light sources that have been used, femtosecond laser based transfection has been one of the most effective methods for optical transfection which is generally implemented using a free space bulk optical setup. In conventional optical transfection methods the foreign genetic material to be transfected is homogenously mixed in the medium. Here we report the first realization of an integrated optical transfection system which can achieve transfection along with localized drug delivery by combining a microlens fiber based optical transfection system with a micro-capillary based microfluidic system. A fiber based illumination system is also incorporated in the system in order to achieve visual identification of the cell boundaries during transfection. A novel fabrication method is devised to obtain easy and inexpensive fabrication of microlensed fibers, which can be used for femtosecond optical transfection. This fabrication method offers the flexibility to fabricate a microlens which can focus ultra-short laser pulses at a near infrared wavelength to a small focal spot (~3 µm) whilst keeping a relatively large working distance (~20 µm). The transfection efficiency of the integrated system with localized plasmid DNA delivery, is approximately 50%, and is therefore comparable to that of a standard free space transfection system. Also the use of integrated system for localized gene delivery resulted in a reduction of the required amount of DNA for transfection. The miniaturized, integrated design opens a range of exciting experimental possibilities, including the dosing of tissue slices, targeted drug delivery, and targeted gene therapy in vivo.

Integrated optical transfection system using a microlens fiber combined with microfluidic gene delivery

3D Photoluminescent Nanostructures Containing Quantum Dots Fabricated by Two-Photon Polymerization: Influence of Quantum Dots on the Spatial Resolution of Laser Writing

In numerous applications of optical scanning microscopy, a reference tapered fiber lens with high symmetry at sub-wavelength scale remains a challenge. Here, we demonstrate the ability to manufacture it with a wide range of geometry control, either for the length from several hundred nanometers to several hundred microns, or for the curvature radius from several tens of nanometers to several microns on the endface of a single mode fiber. On this basis, a scanning optical microscope has been developed, which allows for fast characterization of various sub-wavelength tapered fiber lenses. Focal position and depth of microlenses with different geometries have been determined to be ranged from several hundreds of nanometers to several microns. FDTD calculations are consistent with experimental results.

Rapid fabrication of micro-nanometric tapered fiber lens and characterization by a novel scanning optical microscope with submicron resolution.

Shaped optical fiber tips have recently attracted a lot of interest for photonic jet light focusing due to their easy manipulation to scan a sample. However, lensed optical fibers are not new. This study analyzes how fiber tip parameters can be used to control focusing properties. Our study shows that the configurations to generate a photonic jet (PJ) can clearly be distinguished from more classical-lensed fibers focusing. PJ is a highly concentrated, propagative light beam, with a full width at half maximum (FWHM) that can be lower than the diffraction limit. According to the simulations, the PJs are obtained when light is coupled in the guide fundamental mode and when the base diameter of the microlens is close to the core diameter. For single mode fibers or fibers with a low number of modes, long tips with a relatively sharp shape achieve PJ with smaller widths. On the contrary, when the base diameter of the microlens is larger than the fiber core, the focus point tends to move away from the external surface of the fiber and has a larger width. In other words, the optical system (fiber/microlens) behaves in this case like a classical-lensed fiber with a larger focus spot size. The results of this study can be used as guidelines for the tailored fabrication of shaped optical fiber tips according to the targeted application.

Photonic Jet-Shaped Optical Fiber Tips versus Lensed Fibers

We present a novel route to directly integrate an array of microlenses at the extremity of an optical fiber bundle The method is based on photopolymerization at the end of the fiber The method is based on the control of exposure dose and volume of the deposited droplet of photopolymerizable formulation Optical properties of the integrated microlenses are discussed on the basis of FDTD calculations

Integration of polymer microlens array at fiber bundle extremity by photopolymerization.

We report the high speed scanning submicronic microscopy (SSM) using a low cost polymer microlens integrated at the extremity of an optical fiber. These microlenses are fabricated by a free-radical photopolymerization method. Using a polymer microlens with a radius of curvature of 250 nm, a sub-micrometric gold pattern is imaged experimentally by SSM. Different distances between the tip and the sample are used with a high scanning speed of 200 cm/s. In particular, metallic absorption contrasts are described with an optical spatial resolution of 250 nm at the wavelength of 532 nm. Moreover, finite-difference time-domain (FDTD) simulations concerning the focal lengths of microlenses with different geometries and heights support the experimental data.

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P. Renaud Goud

Papers

High speed sub-micrometric microscopy using optical polymer microlens