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Two-Photon Polymerization: Functionalized Microstructures, Micro-Resonators, and Bio-Scaffolds

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Two-Photon Polymerization: Functionalized Microstructures, Micro-Resonators, and Bio-Scaffolds polymers Review Two-Photon Polymerization: Functionalized Microstructures, Micro-Resonators, and Bio-Scaffolds Adriano J. G. Otuka 1,*,†, Nathália B. Tomazio 1,2,†, Kelly T. Paula 1 and Cleber R. Mendonça 1,* 1 Photonics Group, São Carlos Institute of Physics, University of São Paulo, São Carlos 13566-590, SP, Brazil; [email protected] (N.B.T.); [email protected] (K.T.P.) 2 Device Research Laboratory, “Gleb Wataghin” Institute of Physics, University of Campinas, Campinas 13083-859, SP, Brazil * Correspondence: [email protected] (A.J.G.O.); [email protected] (C.R.M.) † Contributed equally to this work as the first author. Abstract: The direct laser writing technique based on two-photon polymerization (TPP) has evolved considerably over the past two decades. Its remarkable characteristics, such as 3D capability, sub- diffraction resolution, material flexibility, and gentle processing conditions, have made it suitable for several applications in photonics and biosciences. In this review, we present an overview of the progress of TPP towards the fabrication of functionalized microstructures, whispering gallery mode (WGM) microresonators, and microenvironments for culturing microorganisms. We also describe the key physical-chemical fundamentals underlying the technique, the typical experimental setups, and the different materials employed for TPP. Keywords: direct laser writing; ultrashort laser pulses; two-photon polymerization; functional microdevices; whispering gallery mode microresonators; scaffolds for biological applications Citation: Otuka, A.J.G.; Tomazio, N.B.; Paula, K.T.; Mendonça, C.R. Two-Photon Polymerization: 1. Introduction Functionalized Microstructures, Micro-Resonators, and Bio-Scaffolds. The processing of materials by ultrashort lasers, which allows high precision-patterning Polymers 2021, 13, 1994. https:// of 2D and 3D micro/nanostructures, has become a cornerstone technology for a wide range doi.org/10.3390/polym13121994 of areas related to academic research and engineering [1]. An ultrashort laser-based fabrica- tion technique that has been widely used is direct laser writing via two-photon polymeriza- Academic Editor: Lech Sznitko tion (TPP) [2,3]. As opposed to the most conventional micro/nanofabrication techniques, TPP is a maskless patterning tool and does not require harsh processing conditions nor a Received: 30 April 2021 cleanroom facility [4]. Furthermore, this technique is not limited to the processing materials Accepted: 3 June 2021 in the powder form, such as selective laser sintering (SLS) [5–8], another laser fabrica- Published: 18 June 2021 tion method that allows sculpting macroscopic structures with micrometric resolution. TPP leverages the nonlinear nature of two-photon absorption to fabricate 3D polymeric Publisher’s Note: MDPI stays neutral microstructures with arbitrary geometries and sub-diffraction features [9]. Moreover, a with regard to jurisdictional claims in large variety of materials can be employed as photoresist, or be incorporated into it, which published maps and institutional affil- allows customizing the microstructures’ physical, chemical or biological properties to meet iations. specific applications [10,11]. For example, improvements in the spatial resolution and the amount of dry-related shrinkage undergone by the microstructures can be accomplished by varying the photoresist components proportion [1,12–14]. The TPP technique has found applications in numerous fields, including photon- Copyright: © 2021 by the authors. ics [15–19], biosciences [20–22], and micromechanical systems [23–25]. Precise photonic Licensee MDPI, Basel, Switzerland. building blocks, such as optical microresonators [26], photonic crystals [27], and waveg- This article is an open access article uides [28], as well as photonic wire bondings [29] and free-form coupling elements [16], distributed under the terms and have been successfully fabricated by TPP. Active photonic structures have also been fab- conditions of the Creative Commons ricated [30,31]. In the field of biological sciences, TPP has mainly been used to create Attribution (CC BY) license (https:// microscaffolds suitable for culturing cells, tissues, and microorganisms [21,32,33], and creativecommons.org/licenses/by/ microneedles [22] and porous microstructures [20] for drug delivery. TPP has also been 4.0/). Polymers 2021, 13, 1994. https://doi.org/10.3390/polym13121994 https://www.mdpi.com/journal/polymers Polymers 2021, 13, x FOR PEER REVIEW 2 of 31 croscaffolds suitable for culturing cells, tissues, and microorganisms [21,32,33], and mi- croneedles [22] and porous microstructures [20] for drug delivery. TPP has also been a remarkable tool for fabricating optically driven micromachines that allows grabbing or Polymers 2021probing, 13, 1994 nanoscale objects [34]. Microstructures containing movable parts, such as nanot- 2 of 30 weezers [23], microgears [24], and micropump setups [35], have been demonstrated. This paper summarizes the advances of TPP towards three of its major applications: functionalized microstructures,a remarkable tool whispering for fabricating gallery optically mode driven (WGM micromachines) microresonators, that allows and grabbing microscaffolds foror culturing probing nanoscalemicroorganisms. objects [34 The]. Microstructuresfavorable properties containing of TPP, movable such parts, as such as fine-tuning of the nanotweezersstructure dimensions, [23], microgears the integration [24], and micropump of the fabricated setups [35 structures], have been into demonstrated. a range of substrates, andThis its paper material summarizes flexibility, the advanceshave pushed of TPP forward towards the three design of its majorand po- applications: tential of applicationsfunctionalized of WGM microresonators. microstructures, whispering Moreover, gallery the material mode (WGM) flexibility microresonators, of TPP, and along with its 3D microscaffoldscapability and for gentle culturing processing microorganisms. conditions, The has favorable made it propertieswell suited of for TPP, such as fine-tuning of the structure dimensions, the integration of the fabricated structures into the fabrication of bioactive microscaffolds with spatially varying properties. a range of substrates, and its material flexibility, have pushed forward the design and The further sectionspotential of of the applications present review of WGM are microresonators. organized as follows: Moreover, In theSection material 2, we flexibility of briefly describe theTPP, physical along with-chemical its 3D capability fundamentals and gentle underlying processing the conditions, TPP technique. has made We it well suited subsequently showfor the the typical fabrication experimental of bioactive setups microscaffolds and different with spatially materials varying employed properties. in TPP. Section 3 providesThe an further overview sections of the of general the present applications review are of organized TPP. Emphas as follows:is is given In Section 2, we to the fabrication brieflyof WGM describe microresonators the physical-chemical and 3D microscaffolds fundamentals underlying to study microorgan- the TPP technique. We ism’s development.subsequently Finally, Section show the 4 presents typical experimental the overall setupsconclusions and different and prospects materials employedof in TPP in the near future.TPP. Section 3 provides an overview of the general applications of TPP. Emphasis is given to the fabrication of WGM microresonators and 3D microscaffolds to study microorganism’s development. Finally, Section4 presents the overall conclusions and prospects of TPP in 2. Two-Photon Polymerization (TPP) Fundamentals the near future. 2.1. Two-Photon Absorption 2. Two-Photon Polymerization (TPP) Fundamentals The spatial confinement of TPP, which allows fabricating 3D microstructures with 2.1. Two-Photon Absorption arbitrary geometry and submicrometric features, arises from the nonlinear nature of the two-photon absorptionThe (TPA) spatial phenomenon. confinement ofThe TPP, latter which refers allows to fabricatingan electronic 3D microstructurestransition with arbitrary geometry and submicrometric features, arises from the nonlinear nature of the excited by the combined action of two photons [2,36], such that the sum of the energy of two-photon absorption (TPA) phenomenon. The latter refers to an electronic transition the individual photonsexcited must by the be combined resonant action with ofthe two transition photons [energy2,36], such [37] that. When the sum the ofpho- the energy of tons have the samethe energy, individual as photonsindicated must in be Figure resonant 1b, with the theprocess transition is called energy degenerated [37]. When the photons [38]. TPA can alsohave be observed the same energy, with photons as indicated of different in Figure1 b,energies the process since is calledthe sum degenerated of their [ 38]. TPA energies matches canthe alsoenergy be observedgap. with photons of different energies since the sum of their energies In a quantummatches mechanics the energy framework, gap. TPA is described by considering the existence of a virtual intermediateIn astate, quantum which mechanics takes part framework, in the electronic TPA is described transition by [36,39] considering. In this the existence picture, a transitionof ato virtual a real intermediate state is possible state, only which if takesthe time part interval in the electronic between transition the two- [36,39]. In this picture, a transition to a
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