Nanochannels in Fused Silica through NaOH Etching Assisted by Femtosecond Laser Irradiation.

Sodium hydroxide (NaOH) is increasingly drawing attention as a highly selective etchant for femtosecond laser-modified fused silica. Unprecedented etching contrasts between the irradiated and pristine areas have enabled the fabrication of hollow, high-aspect-ratio structures in the bulk of the material, overcoming the micrometer threshold as the minimum feature size. In this work, we systematically study the effect of NaOH solutions under different etching conditions (etchant concentration, temperature, and etching time) on the tracks created by tightly focused femtosecond laser pulses to assess the best practices for the fabrication of hollow nanostructures in bulk fused silica.

Lab-on-Chip Systems for Cell Sorting: Main Features and Advantages of Inertial Focusing in Spiral Microchannels.

Inertial focusing-based Lab-on-Chip systems represent a promising technology for cell sorting in various applications, thanks to their alignment with the ASSURED criteria recommended by the World Health Organization: Affordable, Sensitive, Specific, User-friendly, Rapid and Robust, Equipment-free, and Delivered. Inertial focusing techniques using spiral microchannels offer a rapid, portable, and easy-to-prototype solution for cell sorting. Various microfluidic devices have been investigated in the literature to understand how hydrodynamic forces influence particle focusing in spiral microchannels.

Methods and Protocol for Single-Cell Motility Assays under Topological Constraints.

The extracellular environment plays a crucial role in many physiological and pathological processes involving cell motility, such as metastatic invasion in cancer development, by heavily impacting the migration strategies adopted by the cells. The study of how mechanical constraints affect the dynamics of cell migration may be relevant to gain more insight into such processes, and it may prove to be a powerful tool in the hands of biologists. In this chapter, we describe the methods used to investigate the ability of neoplastic cells to migrate through narrowing, rigid microstructures upon chemoattractant stimulation.

label-free tissue histology through a microstructured imaging window.

Tissue histopathology, based on hematoxylin and eosin (H&E) staining of thin tissue slices, is the gold standard for the evaluation of the immune reaction to the implant of a biomaterial. It is based on lengthy and costly procedures that do not allow longitudinal studies. The use of non-linear excitation microscopy , largely label-free, has the potential to overcome these limitations.

Microfluidic Lab-on-a-Chip for Studies of Cell Migration under Spatial Confinement.

Understanding cell migration is a key step in unraveling many physiological phenomena and predicting several pathologies, such as cancer metastasis. In particular, confinement has been proven to be a key factor in the cellular migration strategy choice. As our insight in the field improves, new tools are needed in order to empower biologists' analysis capabilities.

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 [...

Rapid Prototyping of 3D Biochips for Cell Motility Studies Using Two-Photon Polymerization.

The study of cellular migration dynamics and strategies plays a relevant role in the understanding of both physiological and pathological processes. An important example could be the link between cancer cell motility and tumor evolution into metastatic stage. These strategies can be strongly influenced by the extracellular environment and the consequent mechanical constrains.

Effects of Thermal Annealing on Femtosecond Laser Micromachined Glass Surfaces.

Femtosecond laser micromachining (FLM) of fused silica allows for the realization of three-dimensional embedded optical elements and microchannels with micrometric feature size. The performances of these components are strongly affected by the machined surface quality and residual roughness. The polishing of 3D buried structures in glass was demonstrated using different thermal annealing processes, but precise control of the residual roughness obtained with this technique is still missing.

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.

Generation of deep ultraviolet sub-2-fs pulses.

We demonstrate the generation of few-cycle deep ultraviolet pulses via frequency upconversion of 5-fs near-infrared pulses in argon using a laser-fabricated gas cell. The measured spectrum extends from 210 to 340 nm, corresponding to a transform-limited pulse duration of 1.45 fs.

Disposable Optical Stretcher Fabricated by Microinjection Moulding.

Microinjection moulding combined with the use of removable inserts is one of the most promising manufacturing processes for microfluidic devices, such as lab-on-chip, that have the potential to revolutionize the healthcare and diagnosis systems. In this work, we have designed, fabricated and tested a compact and disposable plastic optical stretcher. To produce the mould inserts, two micro manufacturing technologies have been used.

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.

Rapid Prototyping of Plastic Lab-on-a-Chip by Femtosecond Laser Micromachining and Removable Insert Microinjection Molding.

We have introduced a new hybrid fabrication method for lab-on-a-chip devices through the combination of femtosecond laser micromachining and removable insert micro-injection molding. This method is particularly suited for the fast prototyping of new devices, while maintaining a competitive low cost. To demonstrate the effectiveness of our approach, we designed, fabricated, and tested a completely integrated flow cytometer coupled to a portable media device.

Editorial for the Special Issue on Ultrafast Laser Fabrication for Lab-on-a-Chip.

Ultrafast laser microfabrication is a very powerful method for producing integrated devices in transparent materials [1].[..

Microfluidic Based Optical Microscopes on Chip.

Last decade's advancements in optofluidics allowed obtaining an ever increasing integration of different functionalities in lab on chip devices to culture, analyze, and manipulate single cells and entire biological specimens. Despite the importance of optical imaging for biological sample monitoring in microfluidics, imaging is traditionally achieved by placing microfluidics channels in standard bench-top optical microscopes. Recently, the development of either integrated optical elements or lensless imaging methods allowed optical imaging techniques to be implemented in lab on chip systems, thus increasing their automation, compactness, and portability.

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.

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.

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.

Femtosecond laser microstructuring for polymeric lab-on-chips.

This paper provides an overview of femtosecond laser microfabrication in polymeric materials, with emphasis on lab-on-chip applications. Due to the nonlinear interaction of femtosecond laser pulses with polymers, laser-induced modifications are localized to the focal volume, enabling high resolution patterning in 3D. Femtosecond laser microfabrication offers unmatched versatility in fabricating surface microchannels and diffractive optics by means of laser ablation, buried optical waveguides and micro-optics through refractive index modification and complex 3D microstructures in photoresists by two-photon polymerization.

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.

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.

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.

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.

Fluorescence monitoring of microchip capillary electrophoresis separation with monolithically integrated waveguides.

Using femtosecond laser writing, optical waveguides were monolithically integrated into a commercial microfluidic lab-on-a-chip device, with the waveguides intersecting a microfluidic channel. Continuous-wave laser excitation through these optical waveguides confines the excitation window to a width of 12 microm, enabling high-resolution monitoring of the passage of different types of fluorescent analytes when migrating and being separated in the microfluidic channel by microchip capillary electrophoresis. Furthermore, we demonstrate on-chip-integrated waveguide excitation and detection of a biologically relevant species, fluorescently labeled DNA molecules, during microchip capillary electrophoresis.

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.