Sign in to use this feature.

Years

Between: -

Subjects

remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline

Journals

Article Types

Countries / Regions

Search Results (60)

Search Parameters:
Keywords = two-photon laser microscopy

Order results
Result details
Results per page
Select all
Export citation of selected articles as:
24 pages, 5439 KB  
Review
Review on the Application of Optoelectronic and Photonic Technologies in the Modernization of Traditional Chinese Medicine
by Yihan Huang, Li Zou, Junwei Hu, Huaqi Liu, Shula Chen, Xiaoyan Yi, Ouying Chen and Liancheng Wang
Photonics 2026, 13(7), 628; https://doi.org/10.3390/photonics13070628 - 29 Jun 2026
Viewed by 286
Abstract
The modernization of traditional Chinese medicine (TCM) is significantly impeded by the elusive material basis of its meridian system and by a lack of objective, quantitative diagnostic standards. Recent breakthroughs in photonic technologies and optoelectronic chips offer transformative paradigms to address these systemic [...] Read more.
The modernization of traditional Chinese medicine (TCM) is significantly impeded by the elusive material basis of its meridian system and by a lack of objective, quantitative diagnostic standards. Recent breakthroughs in photonic technologies and optoelectronic chips offer transformative paradigms to address these systemic bottlenecks. This review systematically evaluates the complete academic and engineering chain of “Photonic TCM,” spanning fundamental mechanisms, optical diagnostics, advanced therapeutics, and core chip-level technologies. Specifically, we analyze how ultra-weak photon emission (UPE), two-photon microscopy, and infrared thermography can objectify meridian dynamics and acupuncture pathways. For clinical translation, laser acupuncture has emerged as a robust, non-invasive modality for managing disorders such as chronic pain and insomnia, supported by cumulative evidence-based data. At the device level, vertical-cavity surface-emitting laser (VCSEL)-based photonic computing chips enable ultrafast herbal medicine recognition, while flexible optoelectronics and lab-on-a-chip systems lay the technical groundwork for wearable neuromodulation. Crucially, this review concludes that the Photonic TCM paradigm is transitioning from isolated clinical validation to integrated engineering implementation. We identify biological tissue scattering and parameter heterogeneities as the primary bottlenecks. To navigate these challenges, we propose that the field’s future should converge toward edge-computing-driven wearable closed-loop systems and multi-dimensional optical big data ecosystems. Ultimately, these technological trajectories will steer TCM from an empirical discipline toward a data-driven, precise, and standardized medical science. Full article
(This article belongs to the Special Issue Light-Based Technologies in Biophotonics)
Show Figures

Figure 1

14 pages, 5149 KB  
Article
Two Theoretical Model Comparisons for Calculating the Optical Propagation Loss of Silicon-on-Insulator Waveguides
by Mingqi Bi, Degui Sun, Yu Lin, Yuxiong Li, Peng Yu, Zihao Yu, Yue Sun, Shuning Guo, Lijun Guo and Miao Yu
Coatings 2026, 16(3), 323; https://doi.org/10.3390/coatings16030323 - 6 Mar 2026
Viewed by 985
Abstract
Silicon photonic integrated circuit (Si-PIC) components/devices based on silicon-on-insulator (SOI) waveguides have become critical components in modern optoelectronic information systems. This investigation systematically examines optical propagation losses (OPLs) induced by the sidewall roughness (SWR) of a waveguide through comparative analysis of two scattering-loss [...] Read more.
Silicon photonic integrated circuit (Si-PIC) components/devices based on silicon-on-insulator (SOI) waveguides have become critical components in modern optoelectronic information systems. This investigation systematically examines optical propagation losses (OPLs) induced by the sidewall roughness (SWR) of a waveguide through comparative analysis of two scattering-loss theoretical frameworks: the SWR-improved Payne–Lacey (P-L) three-dimensional (3-D) formalism and Hörmann’s 3-D perturbation model. Crucially, our computational results identify SWR = 10 nm as the convergence threshold where both models exhibit consistent OPL predictions across waveguide architectures. Single-mode SOI rib waveguides with 0.5 µm high ribs on 2.0 µm silicon film and a 2.0 μm BOX layer were designed and fabricated using the classic ICP-RIE technique. Furthermore, SWRs of 28 nm were obtained with confocal laser scanning microscopy for SOI waveguides, leading to OPLs of 2.66 and 2.67 dB/cm for TE and TM modes, respectively, from the 2-D SWR-enhanced P-L model, and 1.7 and 1.9 dB/cm, respectively, from the Hörmann 3-D model. Finally, the average experimental result of OPL for the same waveguide was 2.61 dB/cm, showing a strong agreement with the numerical values of the SWR-improved P-L 3-D formalism, providing a robust framework for optimizing industrial-grade SOI waveguide-based PIC devices/components. Full article
Show Figures

Figure 1

26 pages, 2284 KB  
Review
Key Methodologies in Characterizing the Multi-Scale Structures of Gluten Proteins in Dough: A Comparative Review
by Feifei Su, Yiyuan Zou, Zehua Zhang, Zhiling Tang, Haoran Luo, Fayin Ye and Guohua Zhao
Biomolecules 2026, 16(3), 382; https://doi.org/10.3390/biom16030382 - 3 Mar 2026
Viewed by 846
Abstract
Gluten proteins are key components in wheat flour that determine the formation of dough and the quality of flour-based products. Upon hydration and mixing, gluten proteins undergo complex structural transformations to form a gluten network, exhibiting a hierarchical multi-scale structure spanning molecular, aggregate, [...] Read more.
Gluten proteins are key components in wheat flour that determine the formation of dough and the quality of flour-based products. Upon hydration and mixing, gluten proteins undergo complex structural transformations to form a gluten network, exhibiting a hierarchical multi-scale structure spanning molecular, aggregate, and network scales. Due to the extreme complexity of gluten proteins, accurately characterizing their multi-scale structures remains challenging, requiring the combined application of multiple techniques, which are still relatively limited and thus warrant further exploration. Therefore, this review presents the principles, operational details, and result presentations of current techniques at different structural scales, including electrophoresis, high-performance liquid chromatography, proteomics, Fourier transform infrared spectroscopy, and Fourier transform Raman spectroscopy at the molecular scale; size-exclusion chromatography, asymmetrical flow field-flow fractionation, dynamic light scattering, multi-angle light scattering, differential refractive index, and ultraviolet absorbance at the aggregate scale; and confocal laser scanning microscopy, scanning electron microscopy, confocal Raman microscopy, and two-photon excitation microscopy at the network scale, among others. It further compares the advantages and disadvantages of similar techniques, facilitating their scenario-based selective utilization. Finally, it outlines the ongoing challenges and future perspectives for the development and application of techniques for the multi-scale structural characterization of gluten proteins. Full article
(This article belongs to the Section Biomacromolecules: Proteins, Nucleic Acids and Carbohydrates)
Show Figures

Figure 1

18 pages, 5438 KB  
Article
Ultrafast NIR kHz and GHz Burst Laser Micro-Structuring of Polyimide Films
by Shuai Wang, Chiara Mischo, Walter Perrie, Jose Rajendran, Amin Ibrahim, Yin Tang, Patricia Scully, Dave Atkinson, Yue Tang, Matthew Bilton, Richard Potter, Laura Corner, Geoff Dearden and Stuart Edwardson
Photonics 2026, 13(2), 179; https://doi.org/10.3390/photonics13020179 - 11 Feb 2026
Viewed by 981
Abstract
An ultrafast laser system combined with an optical delay line allowed ablation and in-scription at 1 kHz and 1 GHz pulse burst within transparent polyimide films. The two-photon-induced absorption results in clean surface ablation, while inscription results in polymer decomposition, creating carbonised regions [...] Read more.
An ultrafast laser system combined with an optical delay line allowed ablation and in-scription at 1 kHz and 1 GHz pulse burst within transparent polyimide films. The two-photon-induced absorption results in clean surface ablation, while inscription results in polymer decomposition, creating carbonised regions within the polymer. Three pulse bursts at 1 GHz increased the observed coupling to the material significantly. Modified regions (with linewidths down to a few microns) were investigated using optical microscopy, white light interferometry, SEM and Raman spectroscopy, supporting the increasing carbon density relative to the pristine polymer. As depth of field was only a few microns at high NA, 3D micro-structuring was achieved. Polymer decomposition produces gaseous products, resulting in internal stress and thus affecting inscription fidelity. An inscribed subsurface electrode with dimensions of 5 mm × 0.3 mm × 3 μm connected to conducting vias had a resistance of R = 10.6 ± 0.2 kΩ, along with resistivity of ρ ~ 0.19 Ω cm; hence, it had DC conductivity, σ ~ 5.3 Scm−1. This conductivity is similar to that of bulk graphite and could well form the basis of future flexible sensors, demonstrating single-step 3D subsurface inscription of carbon or laser-induced graphene structures. Full article
(This article belongs to the Special Issue Ultrafast Optics: From Fundamental Science to Applications)
Show Figures

Graphical abstract

11 pages, 1849 KB  
Article
Miniaturized Multicolor Femtosecond Laser Based on Quartz-Encapsulated Nonlinear Frequency Conversion
by Bosong Yu, Siying Wang, Aimin Wang, Yizhou Liu and Lishuang Feng
Photonics 2025, 12(9), 836; https://doi.org/10.3390/photonics12090836 - 22 Aug 2025
Viewed by 4129
Abstract
Ultrafast lasers operating at 740 nm and 820 nm have attracted widespread attention as two-photon light sources for the detection of biological metabolism. Here, we report on a solid-like quartz-encapsulated femtosecond laser with a repetition rate of 80 MHz, delivering 740 nm and [...] Read more.
Ultrafast lasers operating at 740 nm and 820 nm have attracted widespread attention as two-photon light sources for the detection of biological metabolism. Here, we report on a solid-like quartz-encapsulated femtosecond laser with a repetition rate of 80 MHz, delivering 740 nm and 820 nm femtosecond laser pulses. This home-built laser system was realized by employing an erbium-doped 1560 nm fiber laser as the fundamental laser source. A quartz-encapsulated nonlinear frequency conversion stage, consisting of a second-harmonic generation (SHG) stage and self-phase modulation (SPM)-based nonlinear spectral broadening stage, was utilized to deliver 30 mW, 53.7 fs, 740 nm laser pulses and the 15 mW, 60.8 fs, 820 nm laser pulses. Further imaging capabilities of both wavelengths were validated using a custom-built inverted two-photon microscope. Clear imaging results were obtained from mouse kidney sections and pollen samples by collecting the corresponding fluorescence signals. The achieved results demonstrate the great potential of this laser source for advanced two-photon microscopy in metabolic detection. Full article
(This article belongs to the Special Issue Advances in Solid-State Laser Technology and Applications)
Show Figures

Figure 1

24 pages, 19590 KB  
Review
Multiphoton Tomography in Cosmetic Research
by Karsten König and Aisada König
Cosmetics 2025, 12(2), 44; https://doi.org/10.3390/cosmetics12020044 - 4 Mar 2025
Cited by 7 | Viewed by 5528
Abstract
Background: Multiphoton tomography (MPT) is a femtosecond laser imaging technique that enables high-resolution virtual biopsies of human skin. It provides a non-invasive method for analyzing cellular metabolism, structural changes, and responses to cosmetic products, providing insights into cell–cosmetic interactions. This review explores the [...] Read more.
Background: Multiphoton tomography (MPT) is a femtosecond laser imaging technique that enables high-resolution virtual biopsies of human skin. It provides a non-invasive method for analyzing cellular metabolism, structural changes, and responses to cosmetic products, providing insights into cell–cosmetic interactions. This review explores the principles, historical development, and key applications of MPT in cosmetic research. Methods: The latest MPT device combines five modalities: (i) two-photon fluorescence: visualizes cells, elastin, and cosmetic ingredients; (ii) second harmonic generation (SHG): maps the collagen network; (iii) fluorescence lifetime imaging (FLIM): differentiates eumelanin from pheomelanin and evaluates the impact of cosmetics on cellular metabolic activity; (iv) reflectance confocal microscopy (RCM): images cell membranes and cosmetic particles; and (v) white LED imaging for dermoscopy. Results: MPT enables in-depth examination of extracellular matrix changes, cellular metabolism, and melanin production. It identifies skin responses to cosmetic products and tracks the intratissue distribution of sunscreen nanoparticles, nano- and microplastics, and other cosmetic components. Quantitative measurements, such as the elastin-to-collagen ratio, provide insights into anti-aging effects. Conclusions: MPT is a powerful in vivo imaging tool for the cosmetic industry. Its superior resolution and metabolic information facilitate the evaluation of product efficacy and support the development of personalized skincare solutions. Full article
Show Figures

Graphical abstract

12 pages, 2518 KB  
Article
In Situ Multiphysical Metrology for Photonic Wire Bonding by Two-Photon Polymerization
by Yu Lei, Wentao Sun, Xiaolong Huang, Yan Wang, Jinling Gao, Xiaopei Li, Rulei Xiao and Biwei Deng
Materials 2024, 17(21), 5297; https://doi.org/10.3390/ma17215297 - 31 Oct 2024
Cited by 4 | Viewed by 3015
Abstract
Femtosecond laser two-photon polymerization (TPP) technology, known for its high precision and its ability to fabricate arbitrary 3D structures, has been widely applied in the production of various micro/nano optical devices, achieving significant advancements, particularly in the field of photonic wire bonding (PWB) [...] Read more.
Femtosecond laser two-photon polymerization (TPP) technology, known for its high precision and its ability to fabricate arbitrary 3D structures, has been widely applied in the production of various micro/nano optical devices, achieving significant advancements, particularly in the field of photonic wire bonding (PWB) for optical interconnects. Currently, research on optimizing both the optical loss and production reliability of polymeric photonic wires is still in its early stages. One of the key challenges is that inadequate metrology methods cannot meet the demand for multiphysical measurements in practical scenarios. This study utilizes novel in situ scanning electron microscopy (SEM) to monitor the working PWBs fabricated by TPP technology at the microscale. Optical and mechanical measurements are made simultaneously to evaluate the production qualities and to study the multiphysical coupling effects of PWBs. The results reveal that photonic wires with larger local curvature radii are more prone to plastic failure, while those with smaller local curvature radii recover elastically. Furthermore, larger cross-sectional dimensions contribute dominantly to the improved mechanical robustness. The optical-loss deterioration of the elastically deformed photonic wire is only temporary, and can be fully recovered when the load is removed. After further optimization based on the results of multiphysical metrology, the PWBs fabricated in this work achieve a minimum insertion loss of 0.6 dB. In this study, the multiphysical analysis of PWBs carried out by in situ SEM metrology offers a novel perspective for optimizing the design and performance of microscale polymeric waveguides, which could potentially promote the mass production reliability of TPP technology in the field of chip-level optical interconnection. Full article
(This article belongs to the Special Issue Advances in Laser Processing of Materials)
Show Figures

Figure 1

9 pages, 5195 KB  
Article
Advancing Atomic Force Microscopy: Design of Innovative IP-Dip Polymer Cantilevers and Their Exemplary Fabrication via 3D Laser Microprinting
by Peter Gaso, Daniel Jandura, Sergii Bulatov, Dusan Pudis and Matej Goraus
Coatings 2024, 14(7), 841; https://doi.org/10.3390/coatings14070841 - 4 Jul 2024
Cited by 4 | Viewed by 3425
Abstract
This paper presents the design and fabrication of new types of polymer-based cantilevers for atomic force microscopy. The design and fabrication are aimed at the capability of 3D laser microprinting technology based on two-photon polymerization on a standard silicon substrate. IP-Dip commercial material [...] Read more.
This paper presents the design and fabrication of new types of polymer-based cantilevers for atomic force microscopy. The design and fabrication are aimed at the capability of 3D laser microprinting technology based on two-photon polymerization on a standard silicon substrate. IP-Dip commercial material from the Nanoscribe company was used for the fabrication of the designed cantilevers. The fabricated microprinted cantilevers facilitate precise manipulation at the nanoscopic scale, which is essential for studying nanomaterials’ mechanical, electrical, and optical properties. The cantilevers’ flexibility allows for the integration of functional elements such as piezoelectric layers and optical fibers, enabling combined measurements of multiple physical parameters. Various cantilever geometries, including rectangular and V-shaped, are examined, and their resonance frequencies are calculated. The experimental process involves preparing the cantilevers on a silicon substrate and coating them with aluminum for enhanced reflectivity and conductivity. Scanning electron microscope analysis documents the precise form of prepared polymer cantilevers. The functionality of the probes is validated by scanning a step-height standard grating. This study demonstrates the versatility and precision of the fabricated cantilevers, showcasing their potential for large-area scans, living cell investigation, and diverse nanotechnology applications. Full article
Show Figures

Figure 1

12 pages, 3930 KB  
Article
Nanosecond Laser Fabrication of Dammann Grating-like Structure on Glass for Bessel-Beam Array Generation
by Prasenjit Praharaj and Manoj Kumar Bhuyan
Photonics 2024, 11(5), 473; https://doi.org/10.3390/photonics11050473 - 18 May 2024
Cited by 3 | Viewed by 2664
Abstract
The generation of optical beam arrays with prospective uses within the realms of microscopy, photonics, non-linear optics, and material processing often requires Dammann gratings. Here, we report the direct fabrication of one- and two-dimensional Dammann grating-like structures on soda lime glass using a [...] Read more.
The generation of optical beam arrays with prospective uses within the realms of microscopy, photonics, non-linear optics, and material processing often requires Dammann gratings. Here, we report the direct fabrication of one- and two-dimensional Dammann grating-like structures on soda lime glass using a nanosecond pulsed laser beam with a 1064 nm wavelength. Using the fabricated grating, an axicon lens, and an optical magnification system, we propose a scheme of generation of a diverging array of zero-order Bessel beams with a sub-micron-size central core, extending longitudinally over several hundred microns. Two different grating fabrication strategies are also proposed to control the number of Bessel beams in an array. It was demonstrated that Bessel beams of 12 degrees conical half-angle in an array of up to [5 × 5] dimensions can be generated using a suitable combination of Dammann grating, axicon lens and focusing optics. Full article
(This article belongs to the Special Issue Laser Processing and Modification of Materials)
Show Figures

Figure 1

11 pages, 6829 KB  
Communication
A 20 MHz Repetition Rate, Sub-Picosecond Ti–Sapphire Laser for Fiber Delivery in Nonlinear Microscopy of the Skin
by Ádám Krolopp, Luca Fésűs, Gergely Szipőcs, Norbert Wikonkál and Róbert Szipőcs
Life 2024, 14(2), 231; https://doi.org/10.3390/life14020231 - 7 Feb 2024
Cited by 4 | Viewed by 2293
Abstract
Nonlinear microscopy (NM) enables us to investigate the morphology or monitor the physiological processes of the skin through the use of ultrafast lasers. Fiber (or fiber-coupled) lasers are of great interest because they can easily be combined with a handheld, scanning nonlinear microscope. [...] Read more.
Nonlinear microscopy (NM) enables us to investigate the morphology or monitor the physiological processes of the skin through the use of ultrafast lasers. Fiber (or fiber-coupled) lasers are of great interest because they can easily be combined with a handheld, scanning nonlinear microscope. This latter feature greatly increases the utility of NM for pre-clinical applications and in vivo tissue imaging. Here, we present a fiber-coupled, sub-ps Ti–sapphire laser system being optimized for in vivo, stain-free, 3D imaging of skin alterations with a low thermal load of the skin. The laser is pumped by a low-cost, 2.1 W, 532 nm pump laser and delivers 0.5–1 ps, high-peak-power pulses at a ~20 MHz repetition rate. The spectral bandwidth of the laser is below 2 nm, which results in a low sensitivity for dispersion during fiber delivery. The reduction in the peak intensity due to the increased pulse duration is compensated by the lower repetition rate of our laser. In our proof-of-concept imaging experiments, a ~1.8 m long, commercial hollow-core photonic bandgap fiber was used for fiber delivery. Fresh and frozen skin biopsies of different skin alterations (e.g., adult hemangioma, basal cell cancer) and an unaffected control were used for high-quality, two-photon excitation fluorescence microscopy (2PEF) and second-harmonic generation (SHG) z-stack (3D) imaging. Full article
(This article belongs to the Special Issue Non-invasive Skin Imaging Development and Applications)
Show Figures

Figure 1

18 pages, 5728 KB  
Article
Simultaneous Two- and Three-Photon Deep Imaging of Autofluorescence in Bacterial Communities
by Alma Fernández, Anton Classen, Nityakalyani Josyula, James T. Florence, Alexei V. Sokolov, Marlan O. Scully, Paul Straight and Aart J. Verhoef
Sensors 2024, 24(2), 667; https://doi.org/10.3390/s24020667 - 20 Jan 2024
Cited by 6 | Viewed by 3928
Abstract
The intrinsic fluorescence of bacterial samples has a proven potential for label-free bacterial characterization, monitoring bacterial metabolic functions, and as a mechanism for tracking the transport of relevant components through vesicles. The reduced scattering and axial confinement of the excitation offered by multiphoton [...] Read more.
The intrinsic fluorescence of bacterial samples has a proven potential for label-free bacterial characterization, monitoring bacterial metabolic functions, and as a mechanism for tracking the transport of relevant components through vesicles. The reduced scattering and axial confinement of the excitation offered by multiphoton imaging can be used to overcome some of the limitations of single-photon excitation (e.g., scattering and out-of-plane photobleaching) to the imaging of bacterial communities. In this work, we demonstrate in vivo multi-photon microscopy imaging of Streptomyces bacterial communities, based on the excitation of blue endogenous fluorophores, using an ultrafast Yb-fiber laser amplifier. Its parameters, such as the pulse energy, duration, wavelength, and repetition rate, enable in vivo multicolor imaging with a single source through the simultaneous two- and three-photon excitation of different fluorophores. Three-photon excitation at 1040 nm allows fluorophores with blue and green emission spectra to be addressed (and their corresponding ultraviolet and blue single-photon excitation wavelengths, respectively), and two-photon excitation at the same wavelength allows fluorophores with yellow, orange, or red emission spectra to be addressed (and their corresponding green, yellow, and orange single-photon excitation wavelengths). We demonstrate that three-photon excitation allows imaging over a depth range of more than 6 effective attenuation lengths to take place, corresponding to an 800 micrometer depth of imaging, in samples with a high density of fluorescent structures. Full article
(This article belongs to the Special Issue Recent Advances in Biophotonics Sensors)
Show Figures

Figure 1

23 pages, 6863 KB  
Review
Advances in Ultrafast Fiber Lasers for Multiphoton Microscopy in Neuroscience
by Thulasi Srinivasan and Murat Yildirim
Photonics 2023, 10(12), 1307; https://doi.org/10.3390/photonics10121307 - 26 Nov 2023
Cited by 19 | Viewed by 8299
Abstract
Multiphoton microscopy (MPM) has emerged as a vital tool in neuroscience, enabling deeper imaging with a broader field of view, as well as faster and sub-cellular resolution. Recent innovations in ultrafast fiber laser technology have revolutionized MPM applications in living brains, offering advantages [...] Read more.
Multiphoton microscopy (MPM) has emerged as a vital tool in neuroscience, enabling deeper imaging with a broader field of view, as well as faster and sub-cellular resolution. Recent innovations in ultrafast fiber laser technology have revolutionized MPM applications in living brains, offering advantages like cost-effectiveness and user-friendliness. In this review, we explore the progress in ultrafast fiber laser technology, focusing on its integration into MPM for neuroscience research. We also examine the utility of femtosecond fiber lasers in fluorescence and label-free two- and three-photon microscopy applications within the field. Furthermore, we delve into future possibilities, including next-generation fiber laser designs, novel laser characteristics, and their potential for achieving high spatial and temporal resolution imaging. We also discuss the integration of fiber lasers with implanted microscopes, opening doors for clinical and fundamental neuroscience investigations. Full article
(This article belongs to the Special Issue Recent Advances in Multiphoton Microscopy)
Show Figures

Figure 1

11 pages, 10802 KB  
Article
Simultaneous 3D Construction and Imaging of Plant Cells Using Plasmonic Nanoprobe-Assisted Multimodal Nonlinear Optical Microscopy
by Kun Liu, Yutian Lei and Dawei Li
Nanomaterials 2023, 13(19), 2626; https://doi.org/10.3390/nano13192626 - 23 Sep 2023
Cited by 3 | Viewed by 2196
Abstract
Nonlinear optical (NLO) imaging has emerged as a promising plant cell imaging technique due to its large optical penetration, inherent 3D spatial resolution, and reduced photodamage; exogenous nanoprobes are usually needed for nonsignal target cell analysis. Here, we report in vivo, simultaneous 3D [...] Read more.
Nonlinear optical (NLO) imaging has emerged as a promising plant cell imaging technique due to its large optical penetration, inherent 3D spatial resolution, and reduced photodamage; exogenous nanoprobes are usually needed for nonsignal target cell analysis. Here, we report in vivo, simultaneous 3D labeling and imaging of potato cell structures using plasmonic nanoprobe-assisted multimodal NLO microscopy. Experimental results show that the complete cell structure can be imaged via the combination of second-harmonic generation (SHG) and two-photon luminescence (TPL) when noble metal silver or gold ions are added. In contrast, without the noble metal ion solution, no NLO signals from the cell wall were acquired. The mechanism can be attributed to noble metal nanoprobes with strong nonlinear optical responses formed along the cell walls via a femtosecond laser scan. During the SHG-TPL imaging process, noble metal ions that crossed the cell wall were rapidly reduced to plasmonic nanoparticles with the fs laser and selectively anchored onto both sides of the cell wall, thereby leading to simultaneous 3D labeling and imaging of the potato cells. Compared with the traditional labeling technique that needs in vitro nanoprobe fabrication and cell labeling, our approach allows for one-step, in vivo labeling of plant cells, thus providing a rapid, cost-effective method for cellular structure construction and imaging. Full article
(This article belongs to the Special Issue Nonlinear Optics in Low-Dimensional Nanomaterials)
Show Figures

Figure 1

35 pages, 7519 KB  
Review
Optical Methods for Non-Invasive Determination of Skin Penetration: Current Trends, Advances, Possibilities, Prospects, and Translation into In Vivo Human Studies
by Maxim E. Darvin
Pharmaceutics 2023, 15(9), 2272; https://doi.org/10.3390/pharmaceutics15092272 - 3 Sep 2023
Cited by 46 | Viewed by 8624
Abstract
Information on the penetration depth, pathways, metabolization, storage of vehicles, active pharmaceutical ingredients (APIs), and functional cosmetic ingredients (FCIs) of topically applied formulations or contaminants (substances) in skin is of great importance for understanding their interaction with skin targets, treatment efficacy, and risk [...] Read more.
Information on the penetration depth, pathways, metabolization, storage of vehicles, active pharmaceutical ingredients (APIs), and functional cosmetic ingredients (FCIs) of topically applied formulations or contaminants (substances) in skin is of great importance for understanding their interaction with skin targets, treatment efficacy, and risk assessment—a challenging task in dermatology, cosmetology, and pharmacy. Non-invasive methods for the qualitative and quantitative visualization of substances in skin in vivo are favored and limited to optical imaging and spectroscopic methods such as fluorescence/reflectance confocal laser scanning microscopy (CLSM); two-photon tomography (2PT) combined with autofluorescence (2PT-AF), fluorescence lifetime imaging (2PT-FLIM), second-harmonic generation (SHG), coherent anti-Stokes Raman scattering (CARS), and reflectance confocal microscopy (2PT-RCM); three-photon tomography (3PT); confocal Raman micro-spectroscopy (CRM); surface-enhanced Raman scattering (SERS) micro-spectroscopy; stimulated Raman scattering (SRS) microscopy; and optical coherence tomography (OCT). This review summarizes the state of the art in the use of the CLSM, 2PT, 3PT, CRM, SERS, SRS, and OCT optical methods to study skin penetration in vivo non-invasively (302 references). The advantages, limitations, possibilities, and prospects of the reviewed optical methods are comprehensively discussed. The ex vivo studies discussed are potentially translatable into in vivo measurements. The requirements for the optical properties of substances to determine their penetration into skin by certain methods are highlighted. Full article
Show Figures

Graphical abstract

14 pages, 2234 KB  
Article
Impact of the Protein Environment on Two-Photon Absorption Cross-Sections of the GFP Chromophore Anion Resolved at the XMCQDPT2 Level of Theory
by Vladislav R. Aslopovsky, Andrei V. Scherbinin, Nadezhda N. Kleshchina and Anastasia V. Bochenkova
Int. J. Mol. Sci. 2023, 24(14), 11266; https://doi.org/10.3390/ijms241411266 - 10 Jul 2023
Cited by 7 | Viewed by 3603
Abstract
The search for fluorescent proteins with large two-photon absorption (TPA) cross-sections and improved brightness is required for their efficient use in bioimaging. Here, we explored the impact of a single-point mutation close to the anionic form of the GFP chromophore on its TPA [...] Read more.
The search for fluorescent proteins with large two-photon absorption (TPA) cross-sections and improved brightness is required for their efficient use in bioimaging. Here, we explored the impact of a single-point mutation close to the anionic form of the GFP chromophore on its TPA activity. We considered the lowest-energy transition of EGFP and its modification EGFP T203I. We focused on a methodology for obtaining reliable TPA cross-sections for mutated proteins, based on conformational sampling using molecular dynamics simulations and a high-level XMCQDPT2-based QM/MM approach. We also studied the numerical convergence of the sum-over-states formalism and provide direct evidence for the applicability of the two-level model for calculating TPA cross-sections in EGFP. The calculated values were found to be very sensitive to changes in the permanent dipole moments between the ground and excited states and highly tunable by internal electric field of the protein environment. In the case of the GFP chromophore anion, even a single hydrogen bond was shown to be capable of drastically increasing the TPA cross-section. Such high tunability of the nonlinear photophysical properties of the chromophore anions can be used for the rational design of brighter fluorescent proteins for bioimaging using two-photon laser scanning microscopy. Full article
(This article belongs to the Topic Theoretical, Quantum and Computational Chemistry)
Show Figures

Figure 1

Back to TopTop