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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

A sub-100 fs self-starting Cr:forsterite laser generating 1.4 W output power.

Without cavity dumping or external amplification, we report a femtosecond Cr:forsterite laser with a 1.4 W output power and 2 W in continuous wave (CW) operated with a crystal temperature of 267 K. In the femtosecond regime, the oscillator generates Kerr-lens-mode-locked 84 fs pulses with a repetition rate of 85 MHz, corresponding to a high 16.5 nJ pulse energy directly from a single Cr:forsterite resonator. This intense femtosecond Cr:forsterite laser is ideal to pump varieties of high power fiber light sources and could be thus ideal for many biological and spectroscopy applications.

Miniaturized video-rate epi-third-harmonic-generation fiber-microscope.

With a micro-electro-mechanical system (MEMS) mirror, we successfully developed a miniaturized epi-third-harmonic-generation (epi-THG) fiber-microscope with a video frame rate (31 Hz), which was designed for in vivo optical biopsy of human skin. With a large-mode-area (LMA) photonic crystal fiber (PCF) and a regular microscopic objective, the nonlinear distortion of the ultrafast pulses delivery could be much reduced while still achieving a 0.4 microm lateral resolution for epi-THG signals. In vivo real time virtual biopsy of the Asian skin with a video rate (31 Hz) and a sub-micron resolution was obtained. The result indicates that this miniaturized system was compact enough for the least invasive hand-held clinical use.

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.

Miniaturized multiphoton microscope with a 24Hz frame-rate.

With miniaturized tube lenses and a micro-electro-mechanical system (MEMS) mirror, we constructed a miniaturized multiphoton microscope system. Through a two-dimensional asynchronous scanning of the MEMS mirror, 24Hz frame rate can be realized. With a high numerical aperture objective, sub-micron resolution can also be achieved at the same time.