High throughput clogging free microfluidic particle filter by femtosecond laser micromachining.

In recent decades, driven by the needs of industry and medicine, researchers have been investigating how to remove carefully from the main flow microscopic particles or clusters of them. Among all the approaches proposed, crossflow filtration is one of the most attractive as it provides a non-destructive, label-free and in-flow sorting method. In general, the separation performance shows capture and separation efficiencies ranging from 70% up to 100%. However, the maximum flow rate achievable (µL/min) is still orders of magnitude away from those suitable for clinical or industrial applications mainly due to the low stiffness of the materials typically used. In this work, we propose an innovative hydrodynamic-crossflow hybrid filter geometry, buried in a fused silica substrate by means of the femtosecond laser irradiation followed by chemical etching technique. The material high stiffness combined with the accuracy of our manufacturing technique allows the 3D fabrication of non-deformable channels with higher aspect ratio posts, while keeping the overall device dimensions compact. The filter performance has been validated through experiments with both Newtonian (water-based solution of microbeads) and non-Newtonian fluids (blood), achieving separation efficiencies of up to 94% and large particles recovery rates of 100%, even at very high flow rates (mL/h).

Optofluidic Flow Cytometer with In-Plane Spherical Mirror for Signal Enhancement.

Statistical analysis of the properties of single microparticles, such as cells, bacteria or plastic slivers, has attracted increasing interest in recent years. In this regard, field flow cytometry is considered the gold standard technique, but commercially available instruments are bulky, expensive, and not suitable for use in point-of-care (PoC) testing. Microfluidic flow cytometers, on the other hand, are small, cheap and can be used for on-site analyses. However, in order to detect small particles, they require complex geometries and the aid of external optical components. To overcome these limitations, here, we present an opto-fluidic flow cytometer with an integrated 3D in-plane spherical mirror for enhanced optical signal collection. As a result, the signal-to-noise ratio is increased by a factor of six, enabling the detection of particle sizes down to 1.5 µm. The proposed optofluidic detection scheme enables the simultaneous collection of particle fluorescence and scattering using a single optical fiber, which is crucial to easily distinguishing particle populations with different optical properties. The devices have been fully characterized using fluorescent polystyrene beads of different sizes. As a proof of concept for potential real-world applications, signals from fluorescent HEK cells and Escherichia coli bacteria were analyzed.

3D in-plane integrated micro reflectors enhancing signal capture in lab on a chip applications.

The integration of micro-optics in lab on a chip (LOCs) devices is crucial both for increasing the solid angle of acquisition and reducing the optical losses, aiming at improving the signal-to-noise ratio (SNR). In this work, we present the thriving combination of femtosecond laser irradiation followed by chemical etching (FLICE) technique with CO2 laser polishing and inkjet printing to fabricate in-plane, 3D off-axis reflectors, featuring ultra-high optical quality (RMS ∼3 nm), fully integrated on fused silica substrates. Such micro-optic elements can be used both in the excitation path, focusing an incoming beam in 3D, and in the acquisition branch, harvesting the optical signal coming from a specific point in space. The flexibility of the manufacturing process allows the realization of micro-optics with several sizes, shapes and their integration with photonic circuits and microfluidic networks.

Photocell-Based Optofluidic Device for Clogging-Free Cell Transit Time Measurements.

Measuring the transit time of a cell forced through a bottleneck is one of the most widely used techniques for the study of cell deformability in flow. It in turn provides an accessible and rapid way of obtaining crucial information regarding cell physiology. Many techniques are currently being investigated to reliably retrieve this time, but their translation to diagnostic-oriented devices is often hampered by their complexity, lack of robustness, and the bulky external equipment required. Herein, we demonstrate the benefits of coupling microfluidics with an optical method, like photocells, to measure the transit time. We exploit the femtosecond laser irradiation followed by chemical etching (FLICE) fabrication technique to build a monolithic 3D device capable of detecting cells flowing through a 3D non-deformable constriction which is fully buried in a fused silica substrate. We validated our chip by measuring the transit times of pristine breast cancer cells (MCF-7) and MCF-7 cells treated with Latrunculin A, a drug typically used to increase their deformability. A difference in transit times can be assessed without the need for complex external instrumentation and/or demanding computational efforts. The high throughput (4000-10,000 cells/min), ease of use, and clogging-free operation of our device bring this approach much closer to real scenarios.

Simplified 3D hydrodynamic flow focusing for lab-on-chip single particle study.

Accurately control of the position of a fluid and particle within lab-on-a-chip platform is a critical prerequisite for many downstream analysis processes, such as detection, trapping and separation, moving the sensing at the single-particle level. With the development of microfluidic fabrication technology, particle/cell focusing has shifted from two to three dimensions. 3D hydrodynamic focusing, which sorts and aligns the incoming cloud of particles so that they pass through the interrogation area one by one, enables new possibilities and breakthroughs in the single-cell analysis system. Despite the excellent results shown in literature, there is still a lack of a device that can simultaneously fulfilling the requirements of high throughput, compactness, high integrability, and ease of use operation to become a widely accepted work center for biomedical research and clinical applications. Here, we proposed a unique 3D flow focusing microfluidic device buried in fused silica substrate that potentially combines all this advantages. By designing a sample channel suspended inside a larger buffer channel, manufactured by exploiting the laser-assisted micromachine technique, a not size-dependent focusing capability is shown. A spatially and temporally stable central flow of a mixture of 15 µm and 6 µm PS particles to a 1 µm PS microsphere solution has been obtained with high accuracy. Finally, to test the achievable focusing resolution, the chip was tested for the detection of Escherichia Coli bacteria in water solution as proof of concept of biological application.

Bubble dynamics and speed of jets for needle-free injections produced by thermocavitation.

Significance: The number of injections administered has increased dramatically worldwide due to vaccination campaigns following the COVID-19 pandemic, creating a problem of disposing of syringes and needles. Accidental needle sticks occur among medical and cleaning staff, exposing them to highly contagious diseases, such as hepatitis and human immunodeficiency virus. In addition, needle phobia may prevent adequate treatment. To overcome these problems, we propose a needle-free injector based on thermocavitation. Aim: Experimentally study the dynamics of vapor bubbles produced by thermocavitation inside a fully buried 3D fused silica chamber and the resulting high-speed jets emerging through a small nozzle made at the top of it. The injected volume can range from ∼0.1 to 2 µL per shot. We also demonstrate that these jets have the ability to penetrate agar skin phantoms and ex-vivo porcine skin. Approach: Through the use of a high-speed camera, the dynamics of liquid jets ejected from a microfluidic device were studied. Thermocavitation bubbles are generated by a continuous wave laser (1064 nm). The 3D chamber was fabricated by ultra-short pulse laser-assisted chemical etching. Penetration tests are conducted using agar gels (1%, 1.25%, 1.5%, 1.75%, and 2% concentrations) and porcine tissue as a model for human skin. Result: High-speed camera video analysis showed that the average maximum bubble wall speed is about 10 to 25 m/s for almost any combination of pump laser parameters; however, a clever design of the chamber and nozzle enables one to obtain jets with an average speed of ∼70 m/s. The expelled volume per shot (0.1 to 2 µl) can be controlled by the pump laser intensity. Our injector can deliver up to 20 shots before chamber refill. Penetration of jets into agar of different concentrations and ex-vivo porcine skin is demonstrated. Conclusions: The needle-free injectors based on thermocavitation may hold promise for commercial development, due to their cost and compactness.

Battery-free fully integrated microfluidic light source for portable lab-on-a-chip applications.

Integrating a light source inside a Lab-on-a-Chip (LOC) platform has always been as challenging as much as an appealing task. Besides the manufacturing issues, one of the most limiting aspects is due to the need for an energy source to feed the light emission. A solution independent of external energy sources can be given by Chemiluminescence (CL): a well-known chemical phenomenon in which light emission is achieved because of a chemical reaction. Here we present the fabrication and the characterization of a chemiluminescent light source, fully integrated on a microfluidic platform by means of the direct writing technique known as Femtosecond Laser Micromachining. The key advantage is the possibility to insert within LOC devices light sources with complete placement freedom in 3D, wide flexibility of the emitting source geometry and no external feeding energy. The characterization is carried out by investigating the effect of confining a chemiluminescent rubrene-based reaction in small volumes and the inject pressures impact on the emission spectra. Moreover, exploiting microfluidics principles, it's possible to move from the typical flash-type CL emission to a prolonged one (several hours). This allows to disengage bulky, external light sources, adding an extra step on the road to real device portability.