Two-photon Polymerization Research Articles

Two-photon polymerization for random rough surface preparation

A typical tool to characterize diffuse materials is the determination of the scattering and absorption coefficients. Most of these evaluation methods consider optically smooth surfaces between the investigated and the surrounding medium. However, realistic surfaces generally show a distinct roughness, which influences the reflected and transmitted light of the investigated material. Hence, it is necessary to understand the optical behavior of these rough surfaces. We present a method for the preparation of such optically rough surfaces using two-photon polymerization. The properties of these rough surfaces can be precisely adjusted within a range of a few hundred nanometers. Additionally, an easy-to-use preparation method is shown to compensate for reflections from the backside of the used substrate. Hence, these surfaces can be used for measuring the reflectance leading to a better understanding of the scattering behavior of rough surfaces and their influence on the determination of the optical properties of turbid media.

Two-photon polymerization based reusable master template to fabricate polymer microneedles for drug delivery

Microneedle patches have been widely used in a minimally invasive manner for various drug delivery applications. However, for developing these microneedle patches, master molds are required, which are generally made of metal and are very expensive. Two-photon polymerization (2PP) technique can be used for fabricating microneedles more precisely and at a much lower cost. This study reports a novel strategy for developing microneedle master templates using the 2PP method. The main advantage of this technique is that there is no requirement for post-processing after laser writing, and that for the fabrication of polydimethylsiloxane (PDMS) molds, harsh chemical treatments such as silanization are not required. This is a one-step process for manufacturing of microneedle templates which allows easy replication of negative PDMS molds. This is done by adding resin to the master-template and annealing at a specific temperature, thereby making the PDMS peel-off easy and allowing re-use of the master template multiple times. Using this PDMS mold, two types of polyvinyl alcohol (PVA)-rhodamine (RD) microneedle patches were developed, namely, dissolving (D-PVA) and hydrogel (H-PVA) patches and were characterized using suitable techniques. This technique is affordable, efficient and does not require post-processing for development of microneedle templates required for drug delivery applications.•Two photon polymerization can be used for cost-effective fabrication of polymer microneedles for transdermal drug delivery.•Post-processing or surface-modification procedures are not required for these master templates.•Using a simple annealing step, the master template becomes reusable and robust for peeling off polymers like PDMS.

Smart Nematic Liquid Crystal Polymers for Micromachining Advances.

The miniaturization of tools is an important step in human evolution to create faster devices as well as precise micromachines. Studies around this topic have allowed the creation of small-scale objects capable of a wide range of deformation to achieve complex tasks. Molecular arrangements have been investigated through liquid crystal polymer (LCP) to program such a movement. Smart polymers and hereby liquid crystal matrices are materials of interest for their easy structuration properties and their response to external stimuli. However, up until very recently, their employment at the microscale was mainly limited to 2D structuration. Among the numerous issues, one concerns the ability to 3D structure the material while controlling the molecular orientation during the polymerization process. This review aims to report recent efforts focused on the microstructuration of LCP, in particular those dealing with 3D microfabrication via two-photon polymerization (TPP). Indeed, the latter has revolutionized the production of 3D complex micro-objects and is nowadays recognized as the gold standard for 3D micro-printing. After a short introduction highlighting the interest in micromachines, some basic principles of liquid crystals are recalled from the molecular aspect to their implementation. Finally, the possibilities offered by TPP as well as the way to monitor the motion into the fabricated microrobots are highlighted.

Gold Metallization of Hybrid Organic-Inorganic Polymer Microstructures 3d Printed by Two-Photon Polymerization

Gold Metallization of Hybrid Organic-Inorganic Polymer Microstructures 3d Printed by Two-Photon Polymerization

Fabrication of Plano-convex Microlenses using Two-Photon Polymerization for Bioimaging with Non-Linear Excitation Microscopy

A recent challenge in bioimaging is the observation and imaging of vital, thick, and complex tissues in real time and in non-invasive mode. In the last decade, non-linear excitation microscopy showed several advantages for in-vivo imaging compared to conventional confocal techniques. Nevertheless, deep tissue imaging remains challenging, especially for thick media, due to spherical aberrations induced on focused beams by the tissue. A low numerical aperture objective lens coupled to high dioptric power microlenses, implanted in the tissue, can be beneficial for the reduction of optical aberrations. In this context, we fabricated a system of plano-convex microlenses and microscaffolds on a single chip by means of two-photon polymerization), to be used for non-linear imaging of biological specimens.

Biocompatible polymers with tunable mechanical properties and conductive functionality on two-photon 3D printing.

Two-photon polymerization (TPP)-based 3D printing technology utilizes the two-photon absorption process of near-infrared radiation, enabling the fabrication of micro- and nano-scale three-dimensional structures with extremely high resolution. It has been widely applied in scientific fields closely related to living organisms, such as tissue engineering, drug delivery, and biosensors. Nevertheless, the existing photoresist materials have poor mechanical tunability and are hardly able to be doped with functional materials, resulting in constraints on the preparation of functional devices with micro-nano structures. In this paper, TPP printable polymer formulas with good mechanical tunability, high resolution, strong functional scalability, and excellent biocompatibility are proposed, by using the synergistic effects of a hydroxyl group-containing photocurable resin prepolymer, UV acrylate monomer, long-chain hydrophilic crosslinking monomer and photo-initiator. This can ensure the printability and help to improve the flexibility of the printed polymer, thereby solving the problem the photosensitive materials suitable for two-photon 3D printing in previous research had in balancing the formability and flexibility. The results of nanoindenter analysis showed that the Young's modulus of the printed structure can be adjusted between 0.3 GPa and 1.43 GPa, realizing mechanical tunability. Also, complex structures, such as micro-scaffold structures and high aspect ratio hollow microneedles were printed to explore the structural stability as well as the feasibility of biodevice application. Meanwhile, the proposed polymer formula can be functionalized to be conductive by doping with functional nanomaterial MXene. Finally, the biocompatibility of the proposed polymer formula was studied by culturing with human normal lung epithelial cells. The results indicated a good potential for biodevice applications.

An optically fabricated gradient nanochannel array to access the translocation dynamics of T4-phage DNA through nanoconfinement.

It has been widely recognized that nanostructures in natural biological materials play important roles in regulating life machinery. Even though nanofabrication techniques such as two-photon polymerization (TPP) provide sub-100 nm fabrication resolution, it remains technologically challenging to produce 3D nanoscale features modeling the complexity in vivo. We herein demonstrate that a nanochannel array carrying different sizes and nanostructures with gradually transitioning dimensions can be easily produced on a slightly tilted nano-stage. Using the gradient nanochannel array, we systematically investigate the factors affecting the dynamics of DNA translocation through nanoconfinement, including the size of biomolecules and geometrical features of the physical environment, which resembles the selectivity of nanopores in the cell membrane. It is observed that T4-phage DNA shows distinctive conformational transition dynamics during translocation through nanochannels driven by electric field or flow, and the deformation energy required for DNA to enter the nanochannels depends on both chemical environmental conditions, i.e., the ionic strength regulating DNA persistence length and nanochannel dimension. In the electric field, DNA repeatedly gets stretched and compressed during its migration through the nanochannel, reflected by elevated fluctuation in extension, which is substantially greater than the thermal fluctuation. However, driven by flow, DNA remains stretched during translocation through nanochannels, and shows variances in extension of merely a few hundred nanometers. These results indicate that the optically fabricated gradient nanochannel array is a suitable platform for optimizing the experimental conditions for biomedical applications such as gene mapping, and verify that production of complex three dimensional (3D) nanostructures can be greatly simplified by including slight inclination during TPP fabrication.

Buoyant magnetic milliswimmers reveal design rules for optimizing microswimmer performance.

Magnetically-actuated swimming microrobots are an emerging tool for navigating and manipulating materials in confined spaces. Recent work has demonstrated that it is possible to build such systems at the micro and nanoscales using polymer microspheres, magnetic particles and DNA nanotechnology. However, while these materials enable an unprecedented ability to build at small scales, such systems often demonstrate significant polydispersity resulting from both the material variations and the assembly process itself. This variability makes it difficult to predict, let alone optimize, the direction or magnitude of microswimmer velocity from design parameters such as link shape or aspect ratio. To isolate questions of a swimmer's design from variations in its physical dimensions, we present a novel experimental platform using two-photon polymerization to build a two-link, buoyant milliswimmer with a fully customizable shape and integrated flexible linker (the swimmer is underactuated, enabling asymmetric cyclic motion and net translation). Our approach enables us to control both swimming direction and repeatability of swimmer performance. These studies provide ground truth data revealing that neither the first order nor second order models currently capture the key features of milliswimmer performance. We therefore use our experimental platform to develop design guidelines for tuning the swimming speeds, and we identify the following three approaches for increasing speed: (1) tuning the actuation frequency for a fixed aspect ratio, (2) adjusting the aspect ratio given a desired range of operating frequencies, and (3) using the weaker value of linker stiffness from among the values that we tested, while still maintaining a robust connection between the links. We also find experimentally that spherical two-link swimmers with dissimilar link diameters achieve net velocities comparable to swimmers with cylindrical links, but that two-link spherical swimmers of equal diameter do not.

Advances in Structural Color Research Based on Two-Photon Polymerization 3D Printing Technology

Advances in Structural Color Research Based on Two-Photon Polymerization 3D Printing Technology

Two-photon polymerization of femtosecond high-order Bessel beams with aberration correction

Two-photon polymerization of femtosecond high-order Bessel beams with aberration correction

Complex illumination system for fast interferometric measurements

Freeform metrology is an enabling technology for today’s research and advanced manufacturing. The Tilted Wave Interferometer is a full field measurement system for fast and flexible measurements. It is based on an off-axis illumination scheme based on a microlens array. In this contribution, we present a novel illumination system for the tilted wave interferometer, that allows to reduce the measurement time by a factor of four using parallelization based on wavelength multiplexing. Here we present a design solution that utilizes the flexibility of 3D-printing. The microlenses are realized as multi-order diffractive optical elements, providing a high efficiency compared to colorfilter based realizations. To boost the light efficiency of the novel illumination system further, a field lens functionality is added to the system by adding individual micro-prisms to each microlens. The system is manufactured by the use of grayscale two-photon polymerisation.

Two-Photon Polymerization: Fundamentals, Materials, and Chemical Modification Strategies.

Two-photon polymerization (TPP) has become a premier state-of-the-art method for microscale fabrication of bespoke polymeric devices and surfaces. With applications ranging from the production of optical, drug delivery, tissue engineering, and microfluidic devices, TPP has grown immensely in the past two decades. Significantly, the field has expanded from standard acrylate- and epoxy-based photoresists to custom formulated monomers designed to change the hydrophilicity, surface chemistry, mechanical properties, and more of the resulting structures. This review explains the essentials of TPP, from its initial conception through to standard operating principles and advanced chemical modification strategies for TPP materials. At the outset, the fundamental chemistries of radical and cationic polymerization are described, along with strategies used to tailor mechanical and functional properties. This review then describes TPP systems and introduces an array of commonly used photoresists including hard polyacrylic resins, soft hydrogel acrylic esters, epoxides, and organic/inorganic hybrid materials. Specific examples of each class-including chemically modified photoresists-are described to inform the understanding of their applications to the fields of tissue-engineering scaffolds, micromedical, optical, and drug delivery devices.

Entering a New Dimension in Powder Processing for Advanced Ceramics Shaping.

Filigree structures can be manufactured via two-photon polymerization (2PP) operating in the regime of nonlinear light absorption. For the first time, it is possible to apply this technique to the powder processing of ceramic structures with a feature size in the range of the critical defect sizes responsible for brittle fracture and, thus, affecting fracture toughness of high-performance ceramics. In this way, tailoring of advanced properties can be achieved already in the shaping process. Traditionally, 2PP relies on transparent polymerizable resins, which are diametrically opposed to the usually completely opaque ceramic resins and slurries. Here a transparent and photocurable suspension of nanoparticles (resin) with very high mass fractions of yttria-stabilized zirconia particles (YSZ) is presented. Due to the extremely well-dispersed nanoparticles, scattering of light can be effectively suppressed at the process-relevant wavelength of 800nm. Sintered ceramic structures with a resolution of down to 500nm are obtained. Even at reduced densities of 1-4g cm-3 , the resulting compressive strength with 4.5GPa is equivalent or even exceeding bulk monolithic yttria-stabilized zirconia. A ceramic metamaterial is born, where the mechanical properties of yttria-stabilized zirconia are altered by changing geometrical parameters, and gives access to a new class of ceramic materials.

Mechanical Control of the Optical Bandgap in One-Dimensional Photonic Crystals.

Over the last several years, two-photon polymerization has been a popular fabrication approach for photonic crystals due to its high spatial resolution. One-dimensional photonic crystals with photonic bandgap reflectivities over 90% have been demonstrated for the infrared spectral range. With the success of these structures, methods which can provide tunability of the photonic bandgap are being explored. In this study, we demonstrate the use of mechanical flexures in the design of one-dimensional photonic crystals fabricated by two-photon polymerization for the first time. Experimental results show that these photonic crystals provide active mechanically induced spectral control of the photonic bandgap. An analysis of the mechanical behavior of the photonic crystal is presented and elastic behavior is observed. These results suggest that one-dimensional photonic crystals with mechanical flexures can successfully function as opto-mechanical structures.

A Power Compensation Strategy for Achieving Homogeneous Microstructures for 4D Printing Shape-Adaptive PNIPAM Hydrogels: Out-of-Plane Variations.

In the last decade, 3D printing has attracted significant attention and has resulted in benefits to many research areas. Advances in 3D printing with smart materials at the microscale, such as hydrogels and liquid crystalline polymers, have enabled 4D printing and various applications in microrobots, micro-actuators, and tissue engineering. However, the material absorption of the laser power and the aberrations of the laser light spot will introduce a decay in the polymerization degree along the height direction, and the solution to this problem has not been reported yet. In this paper, a compensation strategy for the laser power is proposed to achieve homogeneous and high aspect ratio hydrogel structures at the microscale along the out-of-plane direction. Linear approximations for the power decay curve are adopted for height steps, discretizing the final high aspect ratio structures. The strategy is achieved experimentally with hydrogel structures fabricated by two-photon polymerization. Moreover, characterizations have been conducted to verify the homogeneity of the printed microstructures. Finally, the saturation of material property is investigated by an indirect 3D deformation method. The proposed strategy is proved to be effective and can be explored for other hydrogel materials showing significant deformation. Furthermore, the strategy for out-of-plane variations provides a critical technique to achieve 4D-printed homogeneous shape-adaptive hydrogels for further applications.

Modeling of two-photon polymerization in the strong-pulse regime

This paper reports a study of two-photon polymerization induced by femtosecond laser pulses having microjoule pulse energy and kilohertz repetition rates. Light-matter interaction and polymerization kinetics are modeled in highly confined spatiotemporal scales. The model employs a non-diffractive Bessel beam, considers the effects of temperature-dependent species diffusions, and regards propagation and termination kinetic constants as functions of double-bond conversion. The model is validated by comparing the size of features predicted from simulations to those generated experimentally. The model is used to investigate how the time and energy required to create a single volume element ("voxel") change under various conditions of irradiation. The results show that polymerizing a single voxel requires a minimum exposure time that is constant across a range of irradiation conditions, and is largely determined by the chemical kinetics. In the regime where the pulse energy is low (< 10 μ J), it is more energy-efficient to use fewer pulses having higher energy within the same total exposure time. However, this trend reverses in the regime where the pulse energy is high (10 μ J - 30 μ J), because radical-radical recombination becomes significant, which wastes absorbed energy. This work advances the understanding of two-photon polymerization in the strong-pulse regime and is a step toward increasing throughput to a level suitable for industrial applications. • Two-photon polymerization is induced by few and strong femtosecond laser pulses • A Bessel beam is utilized to rapidly fabricates high aspect ratio structures • Polymerizing a single voxel requires a minimum exposure time • It is more energy-efficient to use fewer pulses when the pulse energy is low (<10 μ J) • Radical-radical recombination is severe when the pulse energy is high (>10 μ J)

Microscale 3D Printing and Tuning of Cellulose Nanocrystals Reinforced Polymer Nanocomposites.

The increasing demand for functional materials and an efficient use of sustainable resources makes the search for new material systems an ever growing endeavor. With this respect, architected (meta-)materials attract considerable interest. Their fabrication at the micro- and nanoscale, however, remains a challenge, especially for composites with highly different phases and unmodified reinforcement fillers. This study demonstrates that it is possible to create a non-cytotoxic nanocomposite ink reinforced by a sustainable phase, cellulose nanocrystals (CNCs), to print and tune complex 3D architectures using two-photon polymerization, thus, advancing the state of knowledge toward the microscale. Micro-compression, high-res scanning electron microscopy, (polarised) Raman spectroscopy, and composite modeling are used to study the structure-property relationships. A 100% stiffness increase is observed already at 4.5 wt% CNC while reaching a high photo-polymerization degree of ≈80% for both neat polymers and CNC-composites. Polarized Raman and the Halpin-Tsai composite-model suggest a random CNC orientation within the polymer matrix. The microscale approach can be used to tune arbitrary small scale CNC-reinforced polymer-composites with comparable feature sizes. The new insights pave the way for future applications where the 3D printing of small structures is essential to improve performances of tissue-scaffolds, extend bio-electronics applications or tailor microscale energy-absorptiondevices.

Direct laser writing breaking diffraction barrier based on two-focus parallel peripheral-photoinhibition lithography

Direct laser writing (DLW) enables arbitrary three-dimensional nanofabrication. However, the diffraction limit poses a major obstacle for realizing nanometer-scale features. Furthermore, it is challenging to improve the fabrication efficiency using the currently prevalent single-focal-spot systems, which cannot perform high-throughput lithography. To overcome these challenges, a parallel peripheral-photoinhibition lithography system with a sub-40-nm two-dimensional feature size and a sub-20-nm suspended line width was developed in our study, based on two-photon polymerization DLW. The lithography efficiency of the developed system is twice that of conventional systems for both uniform and complex structures. The proposed system facilitates the realization of portable DLW with a higher resolution and throughput.

Design, Fabrication, and Application of Mini-Scaffolds for Cell Components in Tissue Engineering

The concept of "lockyballs" or interlockable mini-scaffolds fabricated by two-photon polymerization from biodegradable polymers for the encagement of tissue spheroids and their delivery into the desired location in the human body has been recently introduced. In order to improve control of delivery, positioning, and assembly of mini-scaffolds with tissue spheroids inside, they must be functionalized. This review describes the design, fabrication, and functionalization of mini-scaffolds as well as perspectives on their application in tissue engineering for precisely controlled cell and mini-tissue delivery and patterning. The development of functionalized mini-scaffolds advances the original concept of "lockyballs" and opens exciting new prospectives for mini-scaffolds' applications in tissue engineering and regenerative medicine and their eventual clinical translation.

A Review on Stimuli-Actuated 3D Micro/Nanostructures for Tissue Engineering and the Potential of Laser-Direct Writing via Two-Photon Polymerization for Structure Fabrication.

In this review, we present the most recent and relevant research that has been done regarding the fabrication of 3D micro/nanostructures for tissue engineering applications. First, we make an overview of 3D micro/nanostructures that act as backbone constructs where the seeded cells can attach, proliferate and differentiate towards the formation of new tissue. Then, we describe the fabrication of 3D micro/nanostructures that are able to control the cellular processes leading to faster tissue regeneration, by actuation using topographical, mechanical, chemical, electric or magnetic stimuli. An in-depth analysis of the actuation of the 3D micro/nanostructures using each of the above-mentioned stimuli for controlling the behavior of the seeded cells is provided. For each type of stimulus, a particular recent application is presented and discussed, such as controlling the cell proliferation and avoiding the formation of a necrotic core (topographic stimulation), controlling the cell adhesion (nanostructuring), supporting the cell differentiation via nuclei deformation (mechanical stimulation), improving the osteogenesis (chemical and magnetic stimulation), controlled drug-delivery systems (electric stimulation) and fastening tissue formation (magnetic stimulation). The existing techniques used for the fabrication of such stimuli-actuated 3D micro/nanostructures, are briefly summarized. Special attention is dedicated to structures' fabrication using laser-assisted technologies. The performances of stimuli-actuated 3D micro/nanostructures fabricated by laser-direct writing via two-photon polymerization are particularly emphasized.