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Multiphoton polymerization using upconversion nanoparticles for adaptive high-resolution 3D printing
Lensless two-photon imaging through a multicore fiber with coherence-gated digital phase conjugation
Single micro-droplets of diameter 70-80 μm were produced on demand with inks of viscosity 0.6-9 mPa.s with good controllability and reproducibility, thus enabling to print two-dimensional patterns with a precision of 13 μm. Furthermore, the primary droplet produced with the most viscous fluid was about 66% of the capillary diameter. Preliminary results also showed that the process is linearly scalable to narrower capillaries (100-200 μm), thus paving the way for a compact laser-assisted inkjet printer. A possible application of the device would be additive manufacturing as the printed patterns could be consequently cured.
Here, we demonstrate two-photon excitation imaging of fluorescent beads through a multimode optical fiber. We show that our method maintains the advantages of two-photon excitation microscopy compared to single-photon excitation such as reduced photo-bleaching, deeper penetration depth and sectioning capability. Our method is based on time-gated digital phase conjugation, which allows the generation of focused pulses on the other side of a multimode fiber. To acquire an image, the focused femtosecond pulse is scanned in a three-dimensional mesh, producing two-photon excitation on each spatial location of the sample. By collecting the fluorescence through the fiber, a 3D two-photon image is reconstructed.
We present a method based on digital phase conjugation to selectively excite in a multimode fiber specific optical fiber modes that follow similar optical paths as they travel through the fiber. The excited modes interfere constructively at the fiber output generating an ultrashort spatially focused pulse. The excitation of a limited number of modes following similar optical paths limits modal dispersion, allowing the transmission of the ultrashort pulse. We have experimentally demonstrated the delivery of a focused spot of pulse width equal to 500 fs through a 30 cm, 200 micrometer core step index multimode fiber. The results of this study show that two-photon imaging capability can be added to ultra-thin lensless endoscopy using commercial multimode fibers.
However, at the device level, surgical endoscopic probes are required. We present a miniaturized light probe where a photoactive material can be 1) mixed, pressurized and injected 2) photopolymerized or photoactivated and 3) monitored during the chemical reaction. The device enables surgeries to be conducted through a hole smaller than 1 mm in diameter.
Beside basic injection mechanics, the tool consists of an optical fiber guiding the light required for photopolymerization and for chemical analysis. Combining photorheology and fluorescence spectroscopy, the current state of the photopolymerization is inferred and monitored in real time. Biocompatible and highly tuneable Poly-Ethylene-Glycol (PEG) hydrogels were used as the injection material.
The device was tested on a model for intervertebral disc replacement. Gels were successfully implanted into a bovine caudal model and mechanically tested in-vitro during two weeks. The photopolymerized gel was evaluated at the tissue level (adherence and mechanical properties of the implant), at the cellular level (biocompatibility and cytotoxicity) and ergonomic level (sterilization procedure and feasibility study).
This paper covers the monitoring aspect of the device.
Photopolymerizable hydrogels for implants: Monte-Carlo modeling and experimental in vitro validation
For applications with restricted physical access and illumination time, such as an Nucleus Pulposus replacement, photopolymerization of volumes with a large volume/illumination-area ratio becomes highly challenging. During polymerization, the material’s absorption and scattering coefficients change and directly influence local polymerization rates. By understanding and controlling such polymerization patterns, local material properties can be engineered (e.g. elastic modulus, swelling ratio), to match the set of mechanical requirements for the implant. Thus, it is essential to better understand and model photopolymerization reactions.
Experiments were conducted by polymerizing a hydrogel in a column-like volume using an optical fiber for light delivery. Quantitative scattering and absorption values as well as monomer conversion rates of the hydrogel sample were validated using a newly established Monte Carlo model for photopolymerization. The results were used to study and predict 3D polymerization patterns for different illumination configurations. In particular, we show an example of a lumbar intervertebral disc replacement where the jelly core of the intervertebral disc (Nucleus Pulposus) is replaced by an in situ photopolymerized hydrogel.
The results provide insights for the development of novel endoscopic light-scattering polymerization probes paving the way for a new generation of implantable hydrogels.
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