Non-invasive probing of dynamic ion migration in light-emitting electrochemical cells by an advanced nanoscale confocal microscope.

In this study, we firstly propose an optical approach to investigate the ion profile of organic films in light-emitting electrochemical cells (LECs) without any invasive sputtering processes. In contrast to previous literatures, this pure optical strategy allows us to record clear and non-destructive ion profile images in the (Ru(dtb-bpy)3(PF6)2) consisted organic layer without interferences of complex collisions from the bombardment of secondary sputter induced ions in a conventional time-of-flight secondary ion mass spectrometry. By using the advanced position sensitive detector (PSD)-based Nanoscale Confocal Microscope, ion distribution profiles were successfully acquired based on the observation of nanoscale optical path length difference by measuring the refractive-index variation while the thickness of the LEC layer was fixed. Dynamic time-dependent ion profile displayed clear ion migration process under a 100 V applied bias at two ends of the LEC. This technique opens up a new avenue towards the future investigations of ion distributions inside organic/inorganic materials, Li-ion batteries, or micro-fluid channels without damaging the materials or disturbing the device operation.

Correlation between the optical absorption and twisted angle of bilayer graphene observed by high-resolution reflectance confocal laser microscopy.

We report a systematic study of the optical absorption of twisted bilayer graphene (tBLG) across a large range of twist angles from 0° to 30° using a high-resolution reflectance confocal laser microscopy (RCLM) system. The high-quality single crystalline tBLG was synthesized via the efficient plasma enhanced chemical vapor deposition techniques without the need of active heating. The sensitivity of acquired images from the RCLM were better than conventional optical microscopes. Although the highest spatial resolution of RCLM is still lower than scanning electron microscopes, it possesses the advantages of beam-damage and vacuum free. Moreover, the high intensity-resolution (sensitivity) images firstly allowed us to distinguish the slight absorption differences and analyze the correlation between the optical absorption and twisted angle of tBLG after data processing procedures. A maximum absorption (minimum transmission) was observed at the stacking angle of tBLG from 10° to 20°, indicating the interplay between the laser and the electron/hole van-Hove singularities when tBLG oriented around the critical angle (θc∼13°). The twisted angle correlated optical absorption paves an alternative way not only to visibly identify the interlayer orientation of tBLG but also to reflect the characterization of the interlayer coupling via its band structure.

Label-Free Characterization of Collagen Crosslinking in Bone-Engineered Materials Using Nonlinear Optical Microscopy.

Engineered biomaterials provide unique functions to overcome the bottlenecks seen in biomedicine. Hence, a technique for rapid and routine tests of collagen is required, in which the test items commonly include molecular weight, crosslinking degree, purity, and sterilization induced structural change. Among them, the crosslinking degree mainly influences collagen properties. In this study, second harmonic generation (SHG) and coherent anti-Stokes Raman scattering (CARS) microscopy are used in combination to explore the collagen structure at molecular and macromolecular scales. These measured parameters are applied for the classification and quantification among the different collagen scaffolds, which were verified by other conventional methods. It is demonstrated that the crosslinking status can be analyzed from SHG images and presented as the coherency of collagen organization that is correlated with the mechanical properties. Also, the comparative analyses of SHG signal and relative CARS signal of amide III band at 1,240 cm−1 to δCH2 band at 1,450 cm−1 of these samples provide information regarding the variation of the molecular structure during a crosslinking process, thus serving as nonlinear optical signatures to indicate a successful crosslinking.

Imaging of nanoscale birefringence using polarization-resolved chromatic confocal microscopy.

We demonstrate a homebuilt confocal microscope with ∼60 nm axial resolution to visualize the optical path length (OPL) of liquid crystals (LCs) inside a 2-domain alignment LC cell. Since the microscope is sensitive to light polarization, it is capable of determining LC orientation by accounting for the OPL variation, ΔOPL. The resolution of birefringence depends on the measured ΔOPL from two cross-polarized channel detections, of which the concept is different from other polarization-resolved optical imaging techniques, but is relatively simple in optical layout and analysis. The different orientations of LCs and the voltage-dependent LC rotation properties in the 2-domain LC cell are monitored and analyzed. Additionally, the complicated LC orientation distribution at the junction of the two domains with different alignments can be clearly observed. It shows great possibilities of examining tissue birefringence related to disease progression and tiny birefringence variation of electro-optical materials under an external field, which are hardly resolved by conventional optical imaging techniques.

Wave-vector-encoded nonlinear endomicroscopy.

Based on a rigid square fiber for wave vector delivery, we present a novel (to the best of our knowledge) wave-vector-encoded nonlinear-optical endomicroscopy (WENE). WENE overcomes three tangled issues, including femtosecond pulse broadening induced signal degradation, complexity of packaging miniaturized scanners in the distal end, and pixel-like images, which cannot be fully addressed by current distal scanning nonlinear endomicroscopy (NE) or fiber-bundle-based proximal scanning NE. Due to the advantages of its simplicity in overall configuration and package in the distal end, the capability of addressing the issue of pulse broadening, and offering continuous wave vector delivery, the demonstrated WENE shows great promise for future basic research on biomedical processes and minimally invasive utilization for clinical diagnosis.

Label-free characterization of collagen fibers in cancerous esophagus tissues using ratiometric nonlinear optical microscopy.

IMPACT STATEMENT: The issue of classifying esophageal cancer at various developmental stages is crucial for determining the optimized treatment protocol for the patients, as well as the prognosis. Precision improvement in staging esophageal cancer keeps seeking quantitative and analytical imaging methods that could augment histopathological techniques. In this work, we used nonlinear optical microscopy for ratiometric analysis on the intrinsic signal of two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) from single collagen fibers only in submucosa of esophageal squamous cell carcinoma (ESCC). The blind tests of TPEF/SHG and forward (F)/backward (B) SHG were demonstrated to compare with the histology conclusion. The discussion of sensitivity and specificity was provided via statistical comparison between the four stages of esophageal cancer. To the best of our knowledge, this is the first study of using these two ratios in combination for staging ESCC.

Ultrahigh-resolution optical coherence tomography/angiography with an economic and compact supercontinuum laser.

In this study, a Q-switch pumped supercontinuum laser (QS-SCL) is used as a light source for in vivo imaging via ultrahigh-resolution optical coherence tomography and angiography (UHR-OCT/OCTA). For this purpose, an OCT system based on a spectral-domain detection scheme is constructed, and a spectrometer with a spectral range of 635 - 875 nm is designed. The effective full-width at half maximum of spectrum covers 150 nm, and the corresponding axial and transverse resolutions are 2 and 10 µm in air, respectively. The relative intensity noise of the QS-SCL and mode-locked SCL is quantitatively compared. Furthermore, a special processing algorithm is developed to eliminate the intrinsic noise of QS-SCL. This work demonstrates that QS-SCLs can effectively reduce the cost and size of UHR-OCT/OCTA instruments, making clinical applications feasible.

Tuning the Color Temperature of White-Light-Emitting Electrochemical Cells by Laser-Scanning Perovskite-Nanocrystal Color Conversion Layers.

The development of white-light-emitting electrochemical cells (LECs) has attracted great attention owing to their numerous advantages. Recently, perovskite materials have also shown many outstanding optoelectronic properties in light absorption and emission, and hence they are suitable for serving as the color conversion layers (CCLs) in solid-state white-light-emitting diodes (LEDs). Here, white LECs were fabricated by integrating non-doped blue-green LECs with CCLs made of a single composition of perovskite nanocrystal (NCs). Moreover, the correlated color temperatures (CCTs) of the white LECs can be tuned by modifying the optical properties of the perovskite NCs, in the same way as so as the color conversion properties of CCLs are tuned, through laser scan. By controlling the laser power, scanning number, and duty cycle of the scanned grating patterns on perovskite-NC CCLs, the CCTs of the white LECs can be tuned from 2502 K to nearly 4300 K. Since this method is much different from that used with conventional CCLs, which use multiple compositions of perovskite NCs to produce white light, the inherent anion-exchange issue of perovskite NCs can be avoided.

In situ high-resolution thermal microscopy on integrated circuits.

The miniaturization of metal tracks in integrated circuits (ICs) can cause abnormal heat dissipation, resulting in electrostatic discharge, overvoltage breakdown, and other unwanted issues. Unfortunately, locating areas of abnormal heat dissipation is limited either by the spatial resolution or imaging acquisition speed of current thermal analytical techniques. A rapid, non-contact approach to the thermal imaging of ICs with sub-µm resolution could help to alleviate this issue. In this work, based on the intensity of the temperature-dependent two-photon fluorescence (TPF) of Rhodamine 6G (R6G) material, we developed a novel fast and non-invasive thermal microscopy with a sub-µm resolution. Its application to the location of hotspots that may evolve into thermally induced defects in ICs was also demonstrated. To the best of our knowledge, this is the first study to present high-resolution 2D thermal microscopic images of ICs, showing the generation, propagation, and distribution of heat during its operation. According to the demonstrated results, this scheme has considerable potential for future in situ hotspot analysis during the optimization stage of IC development.

Widely wavelength tunable fast intensity-modulated light source for biophotonic applications.

High modulation depth, fast (megahertz to gigahertz), intensity-modulated light sources of various wavelengths within the 0.7-1.35 µm bio-penetration window are highly desirable for many biophotonic diagnosis systems. In this Letter, we present a novel scheme of a wavelength tunable, ultra-broadband light source which simply consists of a pump laser, a nonlinear fiber, and demodulation circuits. The working wavelength range of the light source is from 0.7 to 1.35 µm which covers a vast part of the bio-penetration window, and its modulation frequencies extends from tens of megahertz to gigahertz. The performances of the proposed light source in either working wavelength range or modulation frequency bandwidth are much superior to any typical laser diodes or solid state lasers currently employed in the frequency-domain or other biophotonic utilization. The wide applicability of this novel light source in diverse biophotonic applications can be observed from our carefully designed diffused optical spectroscopy phantom measurement.

Non-invasive image-guided laser microsurgery by a dual-wavelength fiber laser and an integrated fiber-optic multi-modal system.

A new approach to non-invasive image-guided laser micro-treatment is demonstrated by a dual-wavelength fiber laser source and an integrated fiber-based multi-modal system. The fiber-based source, operated in 1.55 and 1.2 µm simultaneously, was directly connected to an integrated fiber-based multi-modal system for imaging and laser micro-treatment at the same time. The 1.2 µm radiations, within the 1.2-1.35 µm bio-penetration window of skin, were utilized for spectral domain optical coherence tomography imaging. The 1.55 µm radiations, highly absorptive to waters, were utilized for laser microsurgery. The new approach, which is simple in configuration and accurately controls the positions and exposure time of the laser microsurgery, shows great promises for future clinical applications.

Simple approach to three-color two-photon microscopy by a fiber-optic wavelength convertor.

A simple approach to multi-color two-photon microscopy of the red, green, and blue fluorescent indicators was reported based on an ultra-compact 1.03-µm femtosecond laser and a nonlinear fiber. Inside the nonlinear fiber, the 1.03-µm laser pulses were simultaneously blue-shifted to 0.6~0.8 µm and red-shifted to 1.2~1.4 µm region by the Cherenkov radiation and fiber Raman gain effects. The wavelength-shifted 0.6~0.8 µm and 1.2~1.4 µm radiations were co-propagated with the residual non-converted 1.03-µm pulses inside the same nonlinear fiber to form a fiber-output three-color femtosecond source. The application of the multi-wavelength sources on multi-color two-photon fluorescence microscopy were also demonstrated. Overall, due to simple system configuration, convenient wavelength conversion, easy wavelength tunability within the entire 0.7~1.35 µm bio-penetration window and less requirement for high power and bulky light sources, the simple approach to multi-color two-photon microscopy could be widely applicable as an easily implemented and excellent research tool for future biomedical and possibly even clinical applications.

Spiral surface plasmon modes inside metallic nanoholes.

Spiral surface plasmon (SSP) modes that propagate inside a silver (Ag) nanohole are investigated by performing both simulations and theoretical analyses. The SSP modes are formed by a linear combination of two rotating SP eigenmodes of the Ag nanohole in the fast-wave branch. Inside a uniform Ag nanohole, the handedness and the number of strands of the SSP modes are determined by both the component SP eigenmodes and their rotation directions. The spiral pitch of the SSP mode increases with the nanohole radius for a fixed wavelength and is inversely related to the incident wavelength for a fixed nanohole radius. Inside a tapered Ag nanohole, the spiral pitch decreases with the reduction of nanohole radius. However, the azimuth-integrated field energy density increases to a maximum value and then falls. For a tapered Ag-clad fiber capped by a tapered Ag nanorod, the SSP mode reverses its handedness when it passes through the fiber-nanorod interface. Furthermore, using this composite structure, the field energy density of SSP mode that arrives at the tip of the tapered nanorod is largely increased.

Optical coherence tomography-guided laser microsurgery for blood coagulation with continuous-wave laser diode.

Blood coagulation is the clotting and subsequent dissolution of the clot following repair to the damaged tissue. However, inducing blood coagulation is difficult for some patients with homeostasis dysfunction or during surgery. In this study, we proposed a method to develop an integrated system that combines optical coherence tomography (OCT) and laser microsurgery for blood coagulation. Also, an algorithm for positioning of the treatment location from OCT images was developed. With OCT scanning, 2D/3D OCT images and angiography of tissue can be obtained simultaneously, enabling to noninvasively reconstruct the morphological and microvascular structures for real-time monitoring of changes in biological tissues during laser microsurgery. Instead of high-cost pulsed lasers, continuous-wave laser diodes (CW-LDs) with the central wavelengths of 450 nm and 532 nm are used for blood coagulation, corresponding to higher absorption coefficients of oxyhemoglobin and deoxyhemoglobin. Experimental results showed that the location of laser exposure can be accurately controlled with the proposed approach of imaging-based feedback positioning. Moreover, blood coagulation can be efficiently induced by CW-LDs and the coagulation process can be monitored in real-time with OCT. This technology enables to potentially provide accurate positioning for laser microsurgery and control the laser exposure to avoid extra damage by real-time OCT imaging.

Non-invasive biopsy of doped-ions inside optical substrate by modified two-photon microscopy.

Doped-ion based optical elements play key roles in optical signal processes, including amplification, absorption, wavelength-filtering, lighting, and polarizing plate. Non-invasively mapping the spatial distribution of the ion concentrations in these optical elements is highly desirable either during the fabrication process or to determine their optical qualities. In this work, we applied modified two-photon fluorescence (m-TPF) microscopy to trace the ion-distributions deep inside the optical elements. For demonstration purposes, polyvinyl alcohol (PVA) polymer films inside polarizing plates are taken as an example, where the spatial distributions of Iodine-dyed ions were measured by the m-TPF microscope in a fast and non-invasive way. The durability of the polarizer films can be distinguished from the axial distribution of the Iodine-dyed ions, without the need to perform a biopsy. This proposed method and demonstrated results show great potential for monitoring the spatial distributions of doped-ions in the optical elements quickly and non-destructively, which would be of great benefit in both scientific research and industrial applications.

Novel wearable and wireless ring-type pulse oximeter with multi-detectors.

The pulse oximeter is a popular instrument to monitor the arterial oxygen saturation (SPO2). Although a fingertip-type pulse oximeter is the mainstream one on the market at present, it is still inconvenient for long-term monitoring, in particular, with respect to motion. Therefore, the development of a wearable pulse oximeter, such as a finger base-type pulse oximeter, can effectively solve the above issue. However, the tissue structure of the finger base is complex, and there is lack of detailed information on the effect of the light source and detector placement on measuring SPO2. In this study, the practicability of a ring-type pulse oximeter with a multi-detector was investigated by optical human tissue simulation. The optimal design of a ring-type pulse oximeter that can provide the best efficiency of measuring SPO2 was discussed. The efficiency of ring-type pulse oximeters with a single detector and a multi-detector was also discussed. Finally, a wearable and wireless ring-type pulse oximeter was also implemented to validate the simulation results and was compared with the commercial fingertip-type pulse oximeter.

High power NIR fiber-optic femtosecond Cherenkov radiation and its application on nonlinear light microscopy.

We reported a record high power (>250 mW) and compact near-infrared fiber-optic femtosecond Cherenkov radiation source and its new application on nonlinear light microscopy for the first time (to our best knowledge). The high power femtosecond Cherenkov radiation was generated by 1.03 µm femtosecond pulses from a portable diode-pumped laser and a photonic crystal fiber as a compact, flexible, and highly efficient wavelength convertor. Sectioned nonlinear light microscopy images from mouse brain blood vessel network and rat tail tendon were then performed by the demonstrated light source. Due to the advantages of its high average output power (>250 mW), high pulse energy (>4 nJ), excellent wavelength conversion efficiency (>40%), compactness, simplicity in configuration, and turn-key operation, the demonstrated femtosecond Cherenkov radiation source could thus be widely applicable as an alternative excitation source to mode-locked Ti:Sapphire lasers for future clinical nonlinear microscopy or other applications requiring synchronized multi-wavelength light sources.

Higher-order modulations of fs laser pulses for GHz frequency domain photon migration system.

Except the fundamental modulation frequency, by higher-order-harmonic modulations of mode-locked laser pulses and a simple frequency demodulation circuit, a novel approach to GHz frequency-domain-photon-migration (FDPM) system was reported. With this novel approach, a wide-band modulation frequency comb is available without any external modulation devices and the only electronics to extract the optical attenuation and phase properties at a selected modulation frequency in FDPM systems are good mixers and lock-in devices. This approach greatly expands the frequency range that could be achieved by conventional FDPM systems and suggests that our system could extract much more information from biological tissues than the conventional FDPM systems. Moreover, this demonstration will be beneficial for discerning the minute change of tissue properties.

Simultaneous 0.8, 1.0, and 1.3 µm multispectral and common-path broadband source for optical coherence tomography.

Simultaneous multispectral generation in 0.8, 1.0, and 1.3 µm wavelength ranges by efficient energy conversions of 1.0 µm wavelength femtosecond pulses through a nonlinear fiber was reported. The output spectral range of this multispectral light source was composed of 0.6-0.9 µm blue-shifted Cherenkov radiation (CR), 1.0 µm residual pump, and 1.1-1.7 µm red-shifted soliton self-frequency shift (SSFS) with more than 1 mW/nm power-spectral densities. Output characteristics of the multispectral light source were then quantitatively analyzed and the central wavelengths of CR and SSFS emissions can be further easily adjusted by changing the input power into wavelength conversion fiber. Example spectral-domain optical coherence tomography (OCT) images of an IR card and finger skin were also performed with the demonstrated source. Due to the advantages of its simplicity, easily operated, and wavelength tunability, the reported multispectral source could be widely applicable for various spectroscopic OCT applications.

Observation of spontaneous polarization misalignments in periodically poled crystals using second-harmonic generation microscopy.

Periodically poled crystal (PPC) is a key component for nonlinear optical applications. Its poling quality relies largely on successful domain inversion and the alignment of spontaneous polarization (SP) vectors in each domain. Here we report the unexpected observation of bulk second harmonic generation (SHG) in PPC when excitation propagating along its optical axis. Based on its tensorial nature, SHG is highly sensitive to the orientation of SP, and therefore the misalignment of SP in each domain of PPC can be revealed noninvasively by SHG microscopy. This nonlinear imaging modality provides optical sectioning capability with 3D sub-micrometer resolution, so it will be useful for in situ investigation of poling quality in PPC.