Welding of PMMA by a femtosecond fiber laser.

Developing versatile joining techniques to weld transparent materials on a micrometer scale is of great importance in a growing number of applications, especially for the fabrication and assembly of biomedical devices. In this paper, we report on fs-laser microwelding of two transparent layers of polymethyl methacrylate (PMMA) based on nonlinear absorption and localized heat accumulation at high repetition rates. A fiber CPA laser system was used delivering 650-fs pulses at 1030 nm with repetition rates in the MHz regime. The laser-induced modifications produced by the focused beam into the bulk PMMA were firstly investigated, trying to find a suitable set of process parameters generating continuous and localized melting. Results have been evaluated based on existing heat accumulation models. Then, we have successfully laser welded two 1-mm-thick PMMA layers in a lap-joint configuration. Sealing of the sample was demonstrated through static and dynamic leakage tests. This fs-laser micro-welding process does not need any pre-processing of the samples or any intermediate absorbing layer. Furthermore, it offers several advantages compared to other joining techniques, because it prevents contamination and thermal distortion of the samples, thus being extremely interesting for application in direct laser fabrication of microfluidic devices.

Editorial for the Special Issue on New Trends and Applications in Femtosecond Laser Micromachining.

Femtosecond laser micromachining is becoming an established fabrication technique for transparent material processing in three dimensions [...].

Fabrication of binary Fresnel lenses in PMMA by femtosecond laser surface ablation.

We report on the fabrication of binary Fresnel lenses by femtosecond laser surface ablation of poly(methyl methacrylate) (PMMA) substrates. Tight focusing of the laser pulses produced a minimum ablated feature size of 600 nm, enabling the creation of lenses with numerical apertures as high as 0.5 and focal lengths ranging from 500 µm to 5 mm. A precise control of the ablation depth allowed the achievement of a 30% focusing efficiency, close to the maximum theoretical value for this kind of lenses.

High-resolution electrophoretic separation and integrated-waveguide excitation of fluorescent DNA molecules in a lab on a chip.

By applying integrated-waveguide laser excitation to an optofluidic chip, fluorescently labeled DNA molecules of 12 or 17 different sizes are separated by CE with high operating speed and low sample consumption of approximately 600 pL. When detecting the fluorescence signals of migrating DNA molecules with a PMT, the LOD is as low as 2.1 pM. In the diagnostically relevant size range (approximately 150-1000 base-pairs) the molecules are separated with reproducibly high sizing accuracy (> 99%) and the plug broadening follows Poissonian statistics. Variation of the power dependence of migration time on base-pair size--probably with temperature and condition of the sieving gel matrix--indicates that the capillary migration cannot be described by a simple physical law. Integrated-waveguide excitation of a 12-microm narrow microfluidic segment provides a spatio-temporal resolution that would, in principle, allow for a 20-fold better accuracy than the currently supported by state-of-the-art electrophoretic separation in microchips, thereby demonstrating the potential of this integrated optical approach to fulfill the resolution demands of future electrophoretic microchips.

Femtosecond Laser-Micromachining of Glass Micro-Chip for High Order Harmonic Generation in Gases.

We report on the application of femtosecond laser micromachining to the fabrication of complex glass microdevices, for high-order harmonic generation in gas. The three-dimensional capabilities and extreme flexibility of femtosecond laser micromachining allow us to achieve accurate control of gas density inside the micrometer interaction channel. This device gives a considerable increase in harmonics' generation efficiency if compared with traditional harmonic generation in gas jets. We propose different chip geometries that allow the control of the gas density and driving field intensity inside the interaction channel to achieve quasi phase-matching conditions in the harmonic generation process. We believe that these glass micro-devices will pave the way to future downscaling of high-order harmonic generation beamlines.

Integration of femtosecond laser written optical waveguides in a lab-on-chip.

We use direct femtosecond laser writing to integrate optical waveguides into a commercial fused silica lab-on-chip (LOC). We fabricate high quality waveguides intersecting the microfluidic channels and use them to optically address with high spatial selectivity their content. Fluorescence from the photoexcited volume is efficiently collected at a 90 degrees angle by a high numerical aperture fiber, resulting in a compact and portable setup. Our approach is quite powerful because it allows the integration of photonic functionalities, by simple post-processing, into commercial LOCs, fabricated with standard techniques. By taking advantage of the unique three-dimensional capabilities of femtosecond laser writing, more complex functionalities, such as splitters or Mach-Zehnder interferometers, can be implemented.

Femtosecond Laser Microfabrication of an Integrated Device for Optical Release and Sensing of Bioactive Compounds.

Flash photolysis of caged compounds is one of the most powerful approaches to investigate the dynamic response of living cells. Monolithically integrated devices suitable for optical uncaging are in great demand since they greatly simplify the experiments and allow their automation. Here we demonstrate the fabrication of an integrated bio-photonic device for the optical release of caged compounds. Such a device is fabricated using femtosecond laser micromachining of a glass substrate. More in detail, femtosecond lasers are used both to cut the substrate in order to create a pit for cell growth and to inscribe optical waveguides for spatially selective uncaging of the compounds present in the culture medium. The operation of this monolithic bio-photonic device is tested using both free and caged fluorescent compounds to probe its capability of multipoint release and optical sensing. Application of this device to the study of neuronal network activity can be envisaged.

Particle Manipulation by Optical Forces in Microfluidic Devices.

Since the pioneering work of Ashkin and coworkers, back in 1970, optical manipulation gained an increasing interest among the scientific community. Indeed, the advantages and the possibilities of this technique are unsubtle, allowing for the manipulation of small particles with a broad spectrum of dimensions (nanometers to micrometers size), with no physical contact and without affecting the sample viability. Thus, optical manipulation rapidly found a large set of applications in different fields, such as cell biology, biophysics, and genetics. Moreover, large benefits followed the combination of optical manipulation and microfluidic channels, adding to optical manipulation the advantages of microfluidics, such as a continuous sample replacement and therefore high throughput and automatic sample processing. In this work, we will discuss the state of the art of these optofluidic devices, where optical manipulation is used in combination with microfluidic devices. We will distinguish on the optical method implemented and three main categories will be presented and explored: (i) a single highly focused beam used to manipulate the sample, (ii) one or more diverging beams imping on the sample, or (iii) evanescent wave based manipulation.

Investigation of temperature effect on cell mechanics by optofluidic microchips.

Here we present the results of a study concerning the effect of temperature on cell mechanical properties. Two different optofluidic microchips with external temperature control are used to investigate the temperature-induced changes of highly metastatic human melanoma cells (A375MC2) in the range of ~0 - 35 °C. By means of an integrated optical stretcher, we observe that cells' optical deformability is strongly enhanced by increasing cell and buffer-fluid temperature. This finding is supported by the results obtained from a second device, which probes the cells' ability to be squeezed through a constriction. Measured data demonstrate a marked dependence of cell mechanical properties on temperature, thus highlighting the importance of including a proper temperature-control system in the experimental apparatus.

Straightforward 3D hydrodynamic focusing in femtosecond laser fabricated microfluidic channels.

We report on the use of femtosecond laser irradiation followed by chemical etching as a microfabrication tool for innovative microfluidic networks that implement hydrodynamic focusing. The capability of our microfabrication technology to interconnect microchannels in three dimensions was exploited to demonstrate 2D hydrodynamic focusing, either in the horizontal or in the vertical plane, and full 3D hydrodynamic focusing. In all cases only two inlets were required, one for the sample and one for the sheath flows. Fluidic characterization of all devices was provided. In addition, taking advantage of the possibility to write optical waveguides using the same technology, a monolithic cell counter based on 3D hydrodynamic focusing and integrated optical detection was validated. Counting rates up to 5000 cells s(-1) were achieved in this very compact device, where focusing and counting operations were implemented in less than 1 mm(3). Integration of this hydrodynamic focusing module into several devices fabricated by the same technology as optical cell stretchers and cell sorters is envisaged.

Modulation-frequency encoded multi-color fluorescent DNA analysis in an optofluidic chip.

We introduce a principle of parallel optical processing to an optofluidic lab-on-a-chip. During electrophoretic separation, the ultra-low limit of detection achieved with our set-up allows us to record fluorescence from covalently end-labeled DNA molecules. Different sets of exclusively color-labeled DNA fragments-otherwise rendered indistinguishable by spatio-temporal coincidence-are traced back to their origin by modulation-frequency-encoded multi-wavelength laser excitation, fluorescence detection with a single ultrasensitive, albeit color-blind photomultiplier, and Fourier analysis decoding. As a proof of principle, fragments obtained by multiplex ligation-dependent probe amplification from independent human genomic segments, associated with genetic predispositions to breast cancer and anemia, are simultaneously analyzed.