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Corneal fibroblasts maintain homeostasis of the corneal stroma by mediating the synthesis and degradation of extracellular collagen, and these actions are promoted by transforming growth factor-β (TGF-β) and interleukin-1β (IL-1β), respectively. The cornea is densely innervated with sensory nerve fibers that are not only responsible for sensation but also required for physiological processes such as tear secretion and wound healing. Loss or dysfunction of corneal nerves thus impairs corneal epithelial wound healing and can lead to neurotrophic keratopathy. The sensory neurotransmitter substance P (SP) promotes corneal epithelial wound healing by enhancing the stimulatory effects of growth factors and fibronectin. We have now investigated the role of SP in collagen metabolism mediated by human corneal fibroblasts in culture. Although SP alone had no effect on collagen synthesis or degradation by these cells, it promoted the stimulatory effect of TGF-β on collagen type I synthesis without affecting that of IL-1β on the expression of matrix metalloproteinase-1. This effect of SP on TGF-β-induced collagen synthesis was accompanied by activation of p38 mitogen-activated protein kinase (MAPK) signaling and was attenuated by pharmacological inhibition of p38 or of the neurokinin-1 receptor. Our results thus implicate SP as a modulator of TGF-β-induced collagen type I synthesis by human corneal fibroblasts, and they suggest that loss of this function may contribute to the development of neurotrophic keratopathy. NEW & NOTEWORTHY: This study investigates the role of substance P (SP) in collagen metabolism mediated by human corneal fibroblasts in culture. We found that, although SP alone had no effect on collagen synthesis or degradation by corneal fibroblasts, it promoted the stimulatory effect of transforming growth factor-β on collagen type I synthesis without affecting that of interleukin-1β on the expression of matrix metalloproteinase-1. [ABSTRACT FROM AUTHOR]

Advanced nucleic acid amplification techniques require separating test samples into numerous units (digitalization), which enables higher sensitivity and accuracy than traditional methods. Reagent partition tools are commercially available, but are usually complex and expensive, hindering mass adoption. Herein, the fabrication of a large‐scale micro‐through‐hole array on glass substrates for an advanced digital nucleic acid amplification technique (NAAT) is demonstrated. To meet the requirement of hole quantities (over tens of thousands) for valid nucleic acid statistics, ultrafast Bessel pulses with continuous translation of the glass substrate are applied. A single‐shot Bessel pulse can lead to a single hole, and 100 holes can be produced per second (scalable to thousands per second). Experiments using the fabricated chips show that the performance meets the NAAT requirements. This work provides a highly efficient and low‐cost scheme for reagent partitioning that promotes the wide accessibility of advanced digital NAATs as well as a variety of other applications. [ABSTRACT FROM AUTHOR]

Live imaging of living cells in 3D micro/nano‐environment is important for advanced biological studies. An issue for super‐resolution imaging is refractive index mismatch between the materials used for imaging platform and medium containing cells (water). Fluoropolymer CYTOP is the ideal platform material due to its refractive index close to that of water. Herein, femtosecond laser (220 fs) is exploited to ablate CYTOP at three laser wavelengths of 257, 515, and 1030 nm. Linear power dependences of the ablation rate obtained for all wavelengths indicate that ablation mechanisms of CYTOP are attributed to the production of fragmented CYTOPs (frag‐CYTOPs) inside the film to induce efficient light absorption for subsequent laser pulses. The frag‐CYTOPs can effectively absorb UV–Vis light, whereas those left behind generate green fluorescence. Importantly, the shortest wavelength of 257 nm offers the best ablation quality in terms of surface smoothness and low fluorescence with the highest ablation rate. We elucidate that the refractive‐index matching of CYTOP/water is effective for clear imaging of shapes/fluorescence of cells on 3D structures, e.g., HeLa cells on V‐shaped trenches. The fabrication of a microscopic 3D fluorescence code tag embedded in the CYTOP films is further demonstrated. [ABSTRACT FROM AUTHOR]

Fabrication of nanostructures with a feature size much smaller than the laser wavelength is challenging due to the optical diffraction limit. It's well known that the irradiation of polarized ultrafast laser generates periodic nanostructures, so called laser-induced periodic surface structures (LIPSS). Owing to the modulated field, the surface is periodically ablated to form specific patterns, which can be used for some photonic applications including surface-enhanced Raman scattering (SERS). In this paper, we investigate the morphologies of LIPSS on ZnO substrates by mask-less ultrafast laser processing. By adjusting the laser processing parameters, including fluence, pulse number, polarization, and pulse duration, the homogenous nanostrip array and nanopillar array are created. Furthermore, by adjusting the laser fluence, a single nanogroove with a width of ∼20 nm and a single nanocavity with a diameter of ∼24 nm are created. The gold nanoparticles are then coated on the ZnO nanopillar array for SERS application. We found that the concentration of defects in ZnO substrate is increased by the laser irradiation, which is beneficial for SERS performances to achieve an enhancement factor of SERS as high as 2.28 × 107. [ABSTRACT FROM AUTHOR]

Electrochemical surface‐enhanced Raman scattering (EC‐SERS) is a promising technique for the diagnosis of trace amounts of neurotransmitters, because it can elucidate neurotransmitters' behavior on electrodes to deduce their functions in the human body. However, the current EC‐SERS devices need several tens of milliliters of analyte solution to collect enough signal for analysis. Miniaturization of EC‐SERS devices is crucial for the early diagnosis of disease and point‐of‐care testing. Herein, a new type of EC‐SERS sensor based on 3D microfluidic chips for the analysis of neurotransmitters in ultrasmall volumes is proposed. The microfluidic EC‐SERS chip is fabricated by a ship‐in‐a‐bottle technique based on hybrid laser processing. The working electrode is modified using silver/zinc oxide materials, enabling the formation of a unique "candy apple" structure. To assess the fabricated microfluidic EC‐SERS chips, ascorbic acid is analyzed using the ingenious microfluidic EC‐SERS chips to elucidate its redox reaction by EC‐SERS spectroscopy. Significantly, a sub‐10 μL volume of analyte solution is sufficient for EC‐SERS analysis, which is several orders smaller in volume than the requirements of current EC‐SERS devices. The unprecedented microfluidic EC‐SERS chips fabricated by the ship‐in‐a‐bottle integration technique can be used in portable and smart analyzers for next‐generation biomedicines and catalysts. [ABSTRACT FROM AUTHOR]

Photonic chips have been recognized as a promising platform for information technology. On‐chip optical interferometers as fundamental building blocks of photonic chips are under extensive research with both conventional and novel fabrication techniques. Among them, recently, femtosecond laser has been attracting a great deal of attention as a powerful tool for directly writing micro‐optical devices in various materials due to its flexibility and three‐dimensional fabrication capability. This article gives a review on the working principles and recent achievements of on‐chip waveguides and interferometers fabricated by femtosecond laser, showing their potential applications in various scenarios. [ABSTRACT FROM AUTHOR]

Corneal fibroblasts are embedded within an extracellular matrix composed largely of collagen type 1, proteoglycans, and other proteins in the corneal stroma, and their morphology and function are subject to continuous regulation by collagen. During wound healing and in various pathological conditions, corneal fibroblasts differentiate into myofibroblasts characterized by the expression of a-smooth muscle actin (α-SMA). Endo180, also known as urokinase-type plasminogen activator (uPA) receptor-associated protein (uPARAP), is a collagen receptor. Here we investigated whether targeting of Endo180 and the uPA receptor (uPAR) by uPA might play a role in the regulation of a-SMA expression by culturing corneal fibroblasts derived from uPA-deficient (uPA-/-) or wild-type (uPA +/+) mice in a collagen gel or on plastic. The expression of α-SMA was upregulated, the amounts of full-length Endo180 and uPAR were increased, and the levels of both transforming growth factor-β (TGF-β) expression and Smad3 phosphorylation were higher in uPA-/- corneal fibroblasts compared with uPA +/+ cells under the collagen gel culture condition. Antibodies to Endo180 inhibited these effects of uPA deficiency on α-SMA and TGF-β expression, whereas a TGF-β signaling inhibitor blocked the effects on Smad3 phosphorylation and α-SMA expression. Our results suggest that uPA deficiency might promote the interaction between collagen and Endo180 and thereby increase α-SMA expression in a manner dependent on TGF-β signaling. Expression of α-SMA is thus negatively regulated by uPA through targeting of Endo180 and uPAR. [ABSTRACT FROM AUTHOR]

Surface-enhanced Raman scattering (SERS) enables trace-detection for biosensing and environmental monitoring. Optimized enhancement of SERS can be achieved when the energy of the localized surface plasmon resonance (LSPR) is close to the energy of the Raman excitation wavelength. The LSPR can be tuned using a plasmonic superstructure array with controlled periods. In this paper, we develop a new technique based on laser near-field reduction to fabricate a superstructure array, which provides distinct features in the formation of periodic structures with hollow nanoclusters and flexible control of the LSPR in fewer steps than current techniques. Fabrication involves irradiation of a continuous wave laser or femtosecond laser onto a monolayer of self-assembled silica microspheres to grow silver nanoparticles along the silica microsphere surfaces by laser near-field reduction. The LSPR of superstructure array can be flexibly tuned to match the Raman excitation wavelengths from the visible to the infrared regions using different diameters of silica microspheres. The unique nanostructure formed can contribute to an increase in the sensitivity of SERS sensing. The fabricated superstructure array thus offers superior characteristics for the quantitative analysis of fluorescent perfluorooctanoic acid with a wide detection range from 11 ppb to 400 ppm. [ABSTRACT FROM AUTHOR]

Background: This case report describes the surgical outcome in a patient with congenital X-linked retinoschisis (CXLRS) and the results of proteomic analysis of surgically extracted samples from both vitreous and intraschisis cavities by mass spectrometry.Case Presentation: A 3-month-old boy presented with extensive retinoschisis involving macula and retinal periphery in both eyes. Genetic analysis confirmed retinoschisin 1 mutation (c.554C > T), and an electroretinogram showed significant reduction of b-wave and decreased cone and rod responses, which led to a diagnosis of CXLRS. By performing pars plana vitrectomy, including inner wall retinectomy, clear visual axes with stable retinal conditions and functional vision in both eyes were obtained during the 4 years of follow-up. Proteomic analysis of surgically retrieved fluid from the intraschisis cavity revealed a higher expression of interphotoreceptor retinoid-binding protein (IRBP) than that from the vitreous humor. However, both samples showed equal levels of albumin, transferrin, and pigment epithelium-derived factor.Conclusions: Cellular adhesive imperfection in CXLRS may cause IRBP diffusion from the interphotoreceptor matrix, resulting in the strong expression of IRBP in the intraschisis cavity. An impaired retinoid cycle caused by an absence of IRBP in the retina may potentially underlie the pathology of CXLRS. [ABSTRACT FROM AUTHOR]

In the last decades, research and development of microfluidics have made extraordinary progress, since they have revolutionized the biological and chemical fields as a backbone of lab-on-a-chip systems. Further advancement pushes to miniaturize the architectures to nanoscale in terms of both the sizes and the fluid dynamics for some specific applications including investigation of biological sub-cellular aspects and chemical analysis with much improved detection limits. In particular, nano-scale channels offer new opportunities for tests at single cell or even molecular levels. Thus, nanofluidics, which is a microfluidic system involving channels with nanometer dimensions typically smaller than several hundred nm, has been proposed as an ideal platform for investigating fundamental molecular events at the cell-extracellular milieu interface, biological sensing, and more recently for studying cancer cell migration in a space much narrower than the cell size. In addition, nanofluidics can be used for sample manipulation in analytical chemistry, such as sample injections, separation, purifications or for quantitative and qualitative determinations. Among the nanofabrication technologies, ultrafast laser manufacturing is a promising tool for fabrication of nanofluidics due to its flexibility, versatility, high fabrication resolution and three dimensional (3D) fabrication capability. In this paper, we review the technological advancements of nanofluidic systems, with emphasis on fabrication methods, in particular ultrafast laser manufacturing. We present the challenges for issues concerning channel sizes and fluid dynamics, and introduce the applications in physics, biology, chemistry and engineering with future prospects. [ABSTRACT FROM AUTHOR]

The mechanical strength of hydrogel microstructures is crucial for obtaining the desired flexibility, robustness, and biocompatibility for various applications such as cell scaffolds and soft microrobots. In this study, we demonstrate the fabrication of microstructures composed of cellulose nanofibers (CNFs) and poly(ethylene glycol) diacrylate (PEGDA) hydrogels by multiphoton polymerization. The stress of the fabricated microstructure during tensile testing increased with an increase in the CNF concentration, indicating that the mechanical strength of the microstructure was enhanced by using CNFs as fillers. Moreover, the swelling ratio of the microstructure increased with increasing CNF concentration in the PEGDA hydrogel. Our results show the potential of the technique for the microfabrication of advanced cell scaffolds and soft microrobots with the desired mechanical strength. [ABSTRACT FROM AUTHOR]

Various material processing techniques have been proposed for fabrication of smart surfaces that can modulate cellular behavior and address specific clinical issues. Among them, laser-based technologies have attracted growing interest due to processing versatility. Latest development of ultrashort pulse lasers with pulse widths from several tens of femtoseconds (fs) to several picoseconds (ps) allows clean microfabrication of a variety of materials at micro- and nanoscale both at surface and in volume. In this study, we addressed the possibility of 3D microfabrication of photosensitive glass (PG) by high repetition rate ps laser-assisted etching (PLAE) to improve the fabrication efficiency for the development of useful tools to be used for specific biological applications. Microfluidic structures fabricated by PLAE should provide the flow aspects, 3D characteristics, and possibility of producing functional structures to achieve the biologically relevant microenvironments. Specifically, the microfluidic structures could induce cellular chemotaxis over extended periods in diffusion-based gradient media. More importantly, the 3D characteristics could reproduce capillaries for in vitro testing of relevant organ models. Single cell trapping and analysis by using the fabricated microfluidic structures are also essential for understanding individual cell behavior within the same population. To this end, this paper demonstrates: (1) generation of 3D structures in glass volume or on surface for fabrication of microfluidic channels, (2) subtractive 3D surface patterning to create patterned molds in a controlled manor for casting polydimethylsiloxane (PDMS) structures and developing single cell microchambers, and (3) designing glass photo-masks to be used for sequel additive patterning of biocompatible nanomaterials with controlled shapes, sizes, and periodicity. Mesenchymal stem cells grown on laser-processed glass surfaces revealed no sign of cytotoxicity, while a collagen thin coating improved cellular adhesion. [ABSTRACT FROM AUTHOR]

A new 3D nanofluidic biochip for the study of cancer cell migration and invasion is proposed. In this design, femtosecond laser‐assisted etching is applied to create embedded microfluidic channels, with a base thickness of less than 100 µm for high‐resolution imaging using inverted microscopes. The glass deformation is thermally controlled during fabrication to create pillar‐like formations separated by narrow constricted channels with widths of less than 1 µm spanning lengths of more than tens of microns, mimicking the 3D intravasation–extravasation configuration. Time‐lapse microscopy is used to observe the behavior of prostate cancer (PC3) cells in chemoattractant‐free media over long time intervals as the cells invade the narrow spaces. The PC3 cells are observed to be capable of breaching the fabricated submicrometric intravasation‐like barriers while retaining their viability and proliferation activity. The cells are further able to penetrate the extravasation‐like confining spaces, confirming their dynamic adaptability as they pass through constricted channels with volumes much less than that of the cell nucleus. [ABSTRACT FROM AUTHOR]

Purpose. To report a case of corneal perforation, in a patient with a history of herpetic keratitis, during combination chemotherapy including cetuximab. Case. We report the case of a 71-year-old man who was diagnosed with a hypopharyngeal carcinoma and received radiation therapy combined with cetuximab, the epidermal growth factor receptor (EGFR) inhibitor monoclonal antibody. He was referred to us because of ocular hyperemia and corneal perforation in his left eye. In spite of conservative therapy, his corneal perforation was exacerbated, with iris incarceration into the wound site and exposure to the surface of the cornea. He therefore discontinued treatment with the combination chemotherapy and underwent lamellar keratoplasty using a preserved donor cornea. After treatment with cetuximab resumed, there was no recurrence of the corneal perforation. Conclusion. We have presented the first case of cetuximab-related corneal perforation in a patient who had a history of recurrent herpetic keratitis. EGFR inhibitors, such as cetuximab, can induce corneal perforation in cases with a history of herpetic stromal keratitis. [ABSTRACT FROM AUTHOR]

Hybrid micro/nanostructures and light-absorbing pigment of melanin are two key factors for the excellent antireflective properties of the butterfly wings of Ornithoptera goliath. In this work, femtosecond laser ablation of metals in an organic solvent (FLAMOS) was used to generate hybird micro/nanostructured cross-linked ultrahigh/low spatial frequency laser-induced periodic surface structures (UHSFLs/LSFLs), which well mimic the antireflective architecture of Ornithoptera goliath. The simultaneously deposited carbon or newly formed transition metal carbide (TMC) plays the same role of light-absorbing pigment of melanin on Ornithoptera goliath. A total of 13 transition metals and one transition-metal alloy (Ti, V, Nb, Ta, Mo, W, Fe, Pd, Pt, Ni, Au, Ag, Cu, and CuZn) were systematically investigated by FLAMOS to demonstrate the material-dependent formation of cross-linked hybrid UHSFL/LSFL nanostructures and metal carbonization. Hybrid UHSFLs/LSFLs were only found to be achievable when using group IVB–VIB transition metals (e.g., Ti, V, Nb, Ta, Mo, and W), while LSFLs and hole-rich microstructures were obtained with group VIII (e.g., Fe, Pd, Pt, and Ni) and IB/IB–IIB (e.g., Au, Ag, Cu, and CuZn) transition metals or alloy. This work represents the first-ever demonstration that the orientations of UHSFLs are perpendicular to the curvatures of LSFLs because of Marangoni bursting rather than parallel to the direction of light polarization, as is commonly thought to be the case. The surface chemistry and exceptional multioptical properties of structured metasurfaces were assessed, including thermally resistant antireflectance in the UV–near-infrared (NIR) range, enhanced selective absorbance in the NIR–mid-IR (MIR) range, and polarization-dependent NIR–MIR reflectance. Synergistic effects from TMCs/carbon networks and surface nanostructures resulted in the reflectance of Fe, CuZn, Ni, Ta, Mo, Ti, V, Nb, and W metasurfaces below 1% in the UV–NIR range (less than 0.2% over the entire UV–NIR range for Ni, Ti, V, and W metasurfaces). [ABSTRACT FROM AUTHOR]

We report on experimental study on chemical etch selectivity of fused silica irradiated by femtosecond laser with either linear or circular polarization in a wide range of pulse energies. The relationships between the etch rates and pulse energies are obtained for different polarization states, which can be divided into three different regions. A drop of the etch rate for high pulse energy region is observed and the underlying mechanism is discussed. The advantage of using circularly polarized laser is justified owing to its unique capability of providing a 3D isotropic etch rate. [ABSTRACT FROM AUTHOR]

We investigate the mechanism of laser-induced plasma-assisted ablation (LIPAA), by which high-quality and high-efficiency ablation of transparent materials, such as glass, can be performed with a single conventional pulsed laser. The laser-induced plasma induces transient absorption of the laser beam (532 nm) by the glass substrate. The origin of the transient absorption is electron excitation by ions with kinetic energy more than approximately 10 eV in the plasma, which is observed by measuring transient polarization change in the glass substrate applied with a high external pulsed electric field during the plasma-assisted electron excitation (plasma-conductivity measurement). A possible mechanism of LIPAA is proposed based on the results obtained. [ABSTRACT FROM AUTHOR]

A high-intensity femtosecond (fs) laser can fabricate complicated three-dimensional microstructures inside photosensitive glass with high spatial resolution. In this work, the mechanism of the photoreaction of the photosensitive glass to the infrared fs laser is investigated. We examine the photoinduced electron excitation process on the basis of the determination of the critical dose and a change of the optical-absorption spectrum after the fs laser irradiation. The photoreaction mechanism is discussed in comparison with the case of an ultraviolet nanosecond laser irradiation. Finally, the successive interband electron excitation through defect levels by multiphoton absorption is proposed. [ABSTRACT FROM AUTHOR]

Nitridation of InP is performed by KrF excimer laser irradiation in an NH[sub 3] ambient. The N-In and N-P bonds are formed in the irradiated area in proportion to the number of laser pulses. The x-ray photoelectron spectroscopy spectra of the nitrided samples contain a small concentration of oxide components after aging in an air atmosphere than nonirradiated samples. [ABSTRACT FROM AUTHOR]

Abstract: A novel all‐femtosecond‐laser‐processing technique is proposed for the fabrication of 2D periodic metal nanostructures inside 3D glass microfluidic channels, which have applications to real‐time surface‐enhanced Raman spectroscopy (SERS). In the present study, 3D glass microfluidic channels are fabricated by femtosecond‐laser‐assisted wet etching. This is followed by the space‐selective formation of Cu‐Ag layered thin films inside the microfluidic structure via femtosecond laser direct writing ablation and electroless metal plating. The Cu‐Ag films are subsequently nanostructured by irradiation with linearly polarized beams to form periodic surface structures. This work demonstrates that a double exposure to laser beams having orthogonal polarization directions can generate arrays of layered Cu‐Ag nanodots with dimensions as small as 25% of the laser wavelength. The resulting SERS microchip is able to detect Rhodamine 6G, exhibiting an enhancement factor of 7.3 × 108 in conjunction with a relative standard deviation of 8.88%. This 3D microfluidic chip is also found to be capable of the real‐time SERS detection of Cd2+ ions at concentrations as low as 10 ppb in the presence of crystal violet. This technique shows significant promise for the fabrication of high performance microfluidic SERS platforms for the real‐time sensing of toxic substances with ultrahigh sensitivity. [ABSTRACT FROM AUTHOR]

Purpose. To report a case of persistent corneal epithelial defect that had occurred after a trigeminal nerve block. Case Presentation. A 75-year-old female had suffered from postherpetic neuralgia for 8 years. She underwent Gasserian ganglion block surgery and noticed declining visual acuity in the right eye on the following day. She presented with severe hyperemia and corneal epithelial defects in the right eye and experienced remarkable reduction of sensitivity in the right cornea. She was diagnosed with neurotrophic keratopathy. Ofloxacin eye ointment and rebamipide ophthalmic suspension ameliorated the corneal epithelial defects but superficial punctate keratopathy, corneal superficial neovascularization, and Descemet’s fold persisted. Although the epithelial defects occasionally recurred, the corneal sensation and epithelial defects, Descemet’s fold, and corneal superficial neovascularization all improved around 5 months after trigeminal nerve block. The HRT II Rostock Cornea Module (RCM) could not detect any corneal subbasal nerve fibers at postoperative 4 months; however, it could detect them at postoperative 6 months. Conclusions. As the nerve block effect wore off, the corneal subbasal nerve fibers slowly regenerated. As the corneal sensation improved, the corneal epithelial defects and superficial neovascularization also improved. The HRT II RCM appeared useful for observing loss and regeneration of the corneal subbasal nerve fibers. [ABSTRACT FROM AUTHOR]

Abstract: Development of tunable microlenses by taking advantage of the physical adaptability of fluids is one of the challenges of optofluidic techniques, since it offers many applications in biochips, consumer electronics, and medical engineering. Current optofluidic tuning methods using electrowetting or pneumatic pressure typically suffer from high complexity involving external electromechanical actuating devices and limited tuning performance. In this paper, a novel and simple tuning method is proposed that changes the liquid refractive index in an optofluidic channel while leaving the shape of the microlens unchanged. To create an optofluidic microlens with high robustness and optical performance, built‐in microlenses are fabricated inside 3D glass microfluidic channels by optimized single‐operation wet etching assisted by a femtosecond laser. Tuning of focusing properties is demonstrated by filling the channel with media having different indices. Continuous tuning over a wide range (more than threefold tunability for both focal length and focal spot size) is also achieved by pumping sucrose solutions with different concentrations into the microchip channels. Reversible tuning is experimentally verified, indicating intriguing properties of the all‐glass optofluidic microchip. Both the proposed tuning method and the all‐glass architecture with built‐in microlens offer great potential toward numerous applications, including microfluidic adaptive imaging and biomedical sensing. [ABSTRACT FROM AUTHOR]

The extremely high peak intensity associated with ultrashort pulse width of femtosecond laser allows us to induce nonlinear interaction such as multiphoton absorption and tunneling ionization with materials that are transparent to the laser wavelength. More importantly, focusing the femtosecond laser beam inside the transparent materials confines the nonlinear interaction only within the focal volume, enabling three-dimensional (3D) micro- and nanofabrication. This 3D capability offers three different schemes, which involve undeformative, subtractive, and additive processing. The undeformative processing preforms internal refractive index modification to construct optical microcomponents including optical waveguides. Subtractive processing can realize the direct fabrication of 3D microfluidics, micromechanics, microelectronics, and photonic microcomponents in glass. Additive processing represented by two-photon polymerization enables the fabrication of 3D polymer micro- and nanostructures for photonic and microfluidic devices. These different schemes can be integrated to realize more functional microdevices including lab-on-a-chip devices, which are miniaturized laboratories that can perform reaction, detection, analysis, separation, and synthesis of biochemical materials with high efficiency, high speed, high sensitivity, low reagent consumption, and low waste production. This review paper describes the principles and applications of femtosecond laser 3D micro- and nanofabrication for lab-on-a-chip applications. A hybrid technique that promises to enhance functionality of lab-on-a-chip devices is also introduced. [ABSTRACT FROM AUTHOR]

Microarray technology provides an excellent platform for biomedical and biochemical research including basic scientific studies, drug discovery, and diagnostics. Here, we develop a novel method referred to as real-time two-photon lithography in a controlled flow in which femtosecond laser two-photon lithography is performed in situ in the sequential mode stopping and flowing the flow of liquid resin containing microparticles to achieve 100% trapping on a one-bead-to-one-trap basis. Polydisperse particles can be all trapped to form a desired array by freely designing trap structures, resulting in an unprecedentedly high capture efficiency of ∼100%. No persistent pressure is needed after trapping which reduces the complexity of the system. In addition, trapping of particle-cluster arrays with a controlled number of particles is also achieved via this method. The trapped particles inside the microchip are successfully applied as microlenses for high quality imaging. The present technology marks an essential step towards a versatile platform for the integration of bead-based assays and paves the way for developing innovative microfluidics, optofluidics, micro-optics and single-cell analysis devices. [ABSTRACT FROM AUTHOR]

Microfluidic devices and biochips offer miniaturized laboratories for the separation, reaction, and analysis of biochemical materials with high sensitivity and low reagent consumption. The integration of functional or biomimetic elements further functionalizes microfluidic devices for more complex biological studies. The recently proposed ship-in-a-bottle integration based on laser direct writing allows the construction of microcomponents made of photosensitive polymer inside closed microfluidic structures. Here, we expand this technology to integrate proteinaceous two-dimensional (2D) and three-dimensional (3D) microstructures with the aid of photo-induced cross-linking into glass microchannels. The concept is demonstrated with bovine serum albumin and enhanced green fluorescent protein, each mixed with photoinitiator (Sodium 4-[2-(4-Morpholino) benzoyl-2-dimethylamino] butylbenzenesulfonate). Unlike the polymer integration, fabrication over the entire channel cross-section is challenging. Two proteins are integrated into the same channel to demonstrate multi-protein patterning. Using 50% w/w glycerol solvent instead of 100% water achieves almost the same fabrication resolution for in-channel fabrication as on-surface fabrication due to the improved refractive index matching, enabling the fabrication of 3D microstructures. A glycerol-water solvent also reduces the risk of drying samples. We believe this technology can integrate diverse proteins to contribute to the versatility of microfluidics. [ABSTRACT FROM AUTHOR]

Background: Graft detachment is a complication of non-Descemet stripping automated endothelial keratoplasty (nDSAEK). We report a case of spontaneous reattachment of an extensively dislocated graft after nDSAEK.Case Presentation: A 54-year-old male underwent penetrating keratoplasty (PKP) for keratoconus in his left eye in 2001. Following graft opacity due to rejection, a second PKP was implemented in May 2014. The graft was kept in good condition after the reoperation and yet, visual acuity (VA) declined due to cataract. PEA+IOL was then performed in May 2015. Because edema appeared in the graft 6 months after the PEA+IOL, nDSAEK was carried out in May 2016. Although the donor graft well attached immediately after the nDSAEK, the graft was almost completely dislocated 3 h later except a temporal part. Air was reinjected into the anterior chamber on the following day and the detachment was resolved. Despite of the treatment, about 1/5 of the graft remained detached and the detachment deteriorated to 3/4 of the graft 9 days later. Because the patient could not decide whether to undergo another operation immediately, we decided to follow him up first and found that the partially detached graft reattached spontaneously 1 month later during the follow-up. Although the cornea had a mild edema remaining in the superior temporal area, his BCVA improved to 1.0. Three months later, the graft remained in position and the cornea kept its transparency.Conclusions: Spontaneous reattachment was observed during the follow-up in a case that had shown a comparatively extensive graft dislocation after nDSAEK. [ABSTRACT FROM AUTHOR]

Purpose: To report a case of neurotrophic keratopathy associated with nasopharyngeal carcinoma. Case Report: A 59-year-old man who had been diagnosed with a nasopharyngeal carcinoma was referred to the authors because of visual disturbance and pain in his right eye. Slit-lamp examination revealed a corneal epithelial defect and corneal stromal edema surrounding the epithelial defect area in his right eye. Magnetic resonance imaging showed a mass in his cavernous sinus, which was identified as nasopharyngeal carcinoma (NPC). We diagnosed neurotrophic keratopathy associated with NPC and initiated treatment with preservative-free artificial tears, antibiotic eye drops, fibronectin, a therapeutic contact lens, and amniotic membrane transplantation. However, the persistent corneal epithelial defect was unresponsive to these treatments. Conclusion: Neurotrophic keratopathy secondary to NPC is thought to be rare. We presented a case of neurotrophic keratopathy associated with cavernous sinus metastasis of an NPC. The development of new and more effective treatments for this refractory disease is anticipated. [ABSTRACT FROM AUTHOR]

The unique characteristics of ultrafast lasers have rapidly revolutionized materials processing after their first demonstration in 1987. The ultrashort pulse width of the laser suppresses heat diffusion to the surroundings of the processed region, which minimizes the formation of a heat-affected zone and thereby enables ultrahigh precision micro- and nanofabrication of various materials. In addition, the extremely high peak intensity can induce nonlinear multiphoton absorption, which extends the diversity of materials that can be processed to transparent materials such as glass. Nonlinear multiphoton absorption enables three-dimensional (3D) micro- and nanofabrication by irradiation with tightly focused femtosecond laser pulses inside transparent materials. Thus, ultrafast lasers are currently widely used for both fundamental research and practical applications. This review presents progress in ultrafast laser processing, including micromachining, surface micro- and nanostructuring, nanoablation, and 3D and volume processing. Advanced technologies that promise to enhance the performance of ultrafast laser processing, such as hybrid additive and subtractive processing, and shaped beam processing are discussed. Commercial and industrial applications of ultrafast laser processing are also introduced. Finally, future prospects of the technology are given with a summary. [ABSTRACT FROM AUTHOR]

Bio-imaging, fluoropolymer CYTOP, fluorescence generation, femtosecond laser ablation, refractive index matching Femtosecond Laser Direct-Writing Ablation of Transparent Fluoropolymer Toward Super-Resolution Imaging of Cell Movements (Adv. Keywords: bio-imaging; fluoropolymer CYTOP; fluorescence generation; femtosecond laser ablation; refractive index matching EN bio-imaging fluoropolymer CYTOP fluorescence generation femtosecond laser ablation refractive index matching 1 1 1 06/13/23 20230609 NES 230609 B Laser Ablation b In article 2202048, Kazunari Ozasa, Koji Sugioka, and co-workers demonstrate that 3D structures of CYTOP, a remarkably efficient substance for observing cells in 3D due to its high transparency and refractive index matching that of water, are successfully fabricated using fs-laser direct-writing ablation. [Extracted from the article]

Ultrafast laser synthesis and processing of materials is a burgeoning field that is still in its infancy. This article and the theme articles in this issue review recent developments in the fundamental physics of ultrafast laser–solid interactions as well as the state of our understanding of ultrafast laser-driven surface morphology, modification of transparent media, 3D photo-polymerization and additive fabrication, spallation of graphene, and biological interactions. Also reviewed is the current state of emerging commercial high average power lasers, central to the widespread adoption of ultrafast laser synthesis and processing of materials. It is remarkable that ultrafast lasers with 20 to 600 femtosecond pulse duration can have such a dramatic impact on materials. As we learn more about the fundamental mechanisms that drive the ultrafast laser-material response, even more applications are anticipated to emerge. This revolutionary approach to materials synthesis and processing has already spawned several commercial technologies and promises to create many more in the near future. [ABSTRACT FROM AUTHOR]

To report the light- and dark-adapted perimetric findings in a patient with multiple evanescent white dot syndrome (MEWDS). The patient was a 25-year-old Japanese woman who underwent comprehensive ophthalmological examinations including measurements of the visual acuity, dilated ophthalmoscopy, Goldmann kinetic perimetry, electroretinography (ERG), indocyanine green fundus angiography (ICGA), and optical coherence tomography (OCT). Kinetic perimetry was performed under light- and dark-adapted conditions. The patient was diagnosed with MEWDS by the fundus and visual field findings, and the ICGA abnormalities. Light-adapted perimetry showed an enlargement of the blind spot; however, the size of the blind spot was normalized with dark-adaptation. Amplitude of cone ERG was more reduced than that of rod ERG in the affected eye. The OCT images showed multiple disruptions of the ellipsoid and interdigitation zones. These abnormalities were still present 9 months after the onset although the fundus appeared normal. These findings indicate a persistent cone-dominated dysfunction in a patient with MEWDS. [ABSTRACT FROM AUTHOR]

Microfluidic frameworks known as micro-total-analysis-systems or lab-on-a-chip have become versatile tools in cell biology research, since functional biochips are able to streamline dynamic observations of various cells. Glass or polymers are generally used as the substrate due to their high transparency, chemical stability and cost-effectiveness. However, these materials are not well suited for the microscopic observation of cell migration at the fluid boundary due to the refractive index mismatch between the medium and the biochip material. For this reason, we have developed a new method of fabricating three-dimensional (3D) microfluidic chips made of the low refractive index fluoric polymer CYTOP. This novel fabrication procedure involves the use of a femtosecond laser for direct writing, followed by wet etching with a dilute fluorinated solvent and annealing, to create high-quality 3D microfluidic chips inside a polymer substrate. A microfluidic chip made in this manner enabled us to more clearly observe the flagellum motion of a Dinoflagellate moving in circles near the fluid surface compared to the observations possible using conventional microfluidic chips. We believe that CYTOP microfluidic chips made using this new method may allow more detailed analysis of various cell migrations near solid boundaries. [ABSTRACT FROM AUTHOR]

We present the fabrication of a waveguide-like structure in a polydimethylsiloxane (PDMS) polymer substrate using a femtosecond laser to study the mechanism of symbiosis between filamentous cyanobacteria, Phormidium, and a seedling root. While symbiosis occurring underground promotes the growth of vegetable seedlings, the details of the mechanism remain unclear. Understanding the mechanisms of Phormidium gliding to the seedling root will facilitate improving the mat formation of Phormidium, which will lead to increased vegetable production. We assumed a symbiosis mechanism in which sunlight propagates through the seedling root and is scattered underground to guide the Phormidium gliding. Once attached to the root, Phormidium uses the scattered light for photosynthesis. Photosynthetic products, in turn, promote an increase in Phormidium mat formation and vegetable growth. To verify this assumption, the optical characteristics of the seedling root were investigated. A waveguide-like structure with the same optical characteristics of the root was subsequently fabricated by femtosecond laser in PDMS polymer to assess the light illumination effect on Phormidium behavior. [ABSTRACT FROM AUTHOR]

Optimal design and fabrication of novel devices for high-performance optofluidic applications is a key issue for the development of advanced lab-on-a-chip systems. Parallel cell counting with a high success rate and simple mode of operation is a challenging goal. Current cell-counting methods, using optical waveguides or flow cytometry, typically require a precise coupling of the probe light and involve complex operations. In the present paper, a novel multifunctional cell counting microdevice is designed. It uses a center-pass optofluidic microlens array (MLA) consisting of seven microlenses and an M-shaped confining wall with 9 μm-diameter apertures. The device can be fabricated in a three-dimensional microchannel by ship-in-a-bottle femtosecond laser integration based on two-photon polymerization with optimized experimental parameters. Each microlens produces approximately the same intensity at the focal positions (within ±5%) under white-light illumination, while the confining wall restricts 6∼8 μm-width cells to passing through the edges of two adjacent microlenses because the aperture opens toward their centers. The device demonstrates coupling-free parallel cell counting with a 100% success rate by monitoring the optical intensity variations at each spot. As a result, this method features both easy operation and high performance. Furthermore, the confining wall can filter deformed cells having 15 μm width. [ABSTRACT FROM AUTHOR]