Photonic-Crystal-Fiber-Coupled, Hand-Held, Polarization-Resolved Second-Harmonic-Generation Microscope for In Vivo Visualization of Dermal Collagen Fibers in Human Skin

Second-harmonic-generation (SHG) microscopy is a powerful tool for in vivo monitoring of collagen fibers in human skin. Furthermore, polarization-resolved SHG microscopy can provide additional insights regarding the direction of collagen fibers, i.e., collagen fiber orientation. However, their practical use in the dermatological field is still limited due to the bulky and complicated setup. In this paper, we constructed a photonic-crystal-fiber-coupled, hand-held polarization-resolved SHG microscope for in vivo monitoring of collagen fibers in human skin. Fiber delivery of ultrashort pulse light was achieved without significant change of the linear polarization by a large-mode-area photonic-crystal-fiber whereas the SHG microscopy setup was enclosed into a hand-held probe head. The combination of PCF with the hand-held probe head largely enhances the flexibility of measurement sites in the human skin

Dual terahertz comb spectroscopy with a single free-running fibre laser

Dual THz comb spectroscopy has the potential to be used as universal THz spectroscopy with high spectral resolution, high spectral accuracy, and broad spectral coverage; however, the requirement for dual stabilized femtosecond lasers hampers its versatility due to the bulky size, high complexity, and high cost. We here report the first demonstration of dual THz comb spectroscopy using a single free-running fibre laser. By tuning the cavity-loss-dependent gain profile with an intracavity Lyot filter together with precise management of the cavity length and dispersion, dual-wavelength pulsed light beams with slightly detuned repetition frequencies are generated in a single laser cavity. Due to sharing of the same cavity, such pulsed light beams suffer from common-mode fluctuation of the repetition frequency, and hence the corresponding frequency difference between them is passively stable around a few hundred hertz within millihertz fluctuation. This considerably stable frequency difference enables dual THz comb spectroscopy with a single free-running fibre laser. While greatly reducing the size, complexity, and cost of the laser source by use of a single free-running fibre laser, the dual THz comb spectroscopy system maintains a spectral bandwidth and dynamic range of spectral power comparable to a system equipped with dual stabilized fibre lasers, and can be effectively applied to high-precision spectroscopy of acetonitrile gas at atmospheric pressure. The demonstrated results indicate that this system is an attractive solution for practical applications of not only THz spectroscopy but also THz-pulse-based measurements.Comment: 29 pages, 7 figure

Nanometer-precision surface metrology of millimeter-size stepped objects using full-cascade-linked synthetic-wavelength digital holography using a line-by-line full-mode-extracted optical frequency comb

Digital holography (DH) is a powerful tool for surface profilometry of objects with sub-wavelength precision. In this article, we demonstrate full-cascade-linked synthetic-wavelength DH (FCL-SW-DH) for nanometer-precision surface metrology of millimeter-size stepped objects. 300 modes of optical frequency comb (OFC) with different wavelengths are sequentially extracted at a step of mode spacing from a 10GHz-spacing, 3.72THz-spanning electro-optic modulator OFC (EOM-OFC). The resulting 299 synthetic wavelengths and a single optical wavelength are used to generate a fine-step wide-range cascade link covering within a wavelength range of 1.54 um to 29.7 mm. We determine the 0.1000mm-stepped surface with axial uncertainty of 6.1 nm within the maximum axial range of 14.85 mm.Comment: 22 pages, 6 figure

Evaluation of the histological and mechanical features of tendon healing in a rabbit model

Objectives This study aimed to evaluate the histological and mechanical features of tendon healing in a rabbit model with second-harmonic-generation (SHG) imaging and tensile testing. Materials and Methods A total of eight male Japanese white rabbits were used for this study. The flexor digitorum tendons in their right leg were sharply transected, and then were repaired by intratendinous stitching. At four weeks post-operatively, the rabbits were killed and the flexor digitorum tendons in both right and left legs were excised and used as specimens for tendon healing (n = 8) and control (n = 8), respectively. Each specimen was examined by SHG imaging, followed by tensile testing, and the results of the two testing modalities were assessed for correlation. Results While the SHG light intensity of the healing tendon samples was significantly lower than that of the uninjured tendon samples, 2D Fourier transform SHG images showed a clear difference in collagen fibre structure between the uninjured and the healing samples, and among the healing samples. The mean intensity of the SHG image showed a moderate correlation (R2 = 0.37) with Young’s modulus obtained from the tensile testing. Conclusion Our results indicate that SHG microscopy may be a potential indicator of tendon healing

Advances in 5-ALA-PDD of gastric cancer

Photodynamic diagnosis based on 5-aminolevulinic acid-induced protoporphyrin IX has been clinically applied in many fields based upon its evidenced efficacy and adequate safety. In order to establish a personalized medicine approach for treating gastric cancer patients, rapid intraoperative detection of malignant lesions has become important. Feasibility of photodynamic diagnosis using 5-aminolevulinic acid for gastric cancer patients has been investigated, especially for the detection of peritoneal dissemination and lymph node metastasis. This method enables intraoperative real-time fluorescence detection of peritoneal dissemination, exhibiting higher sensitivity than white light observation without histopathological examination. The method also enables detection of metastatic foci within excised lymph nodes, exhibiting a diagnostic accuracy comparable to that of a current molecular diagnostics technique. Although several complicating issues still need to be resolved, such as the effect of tissue autofluorescence and the insufficient depth penetration of excitation light, this simple and rapid method has the potential to become a useful diagnostic tool for gastric cancer, as well as urinary bladder cancer and glioma

Application of Scan-less Two-Dimensional Confocal Microscopy Based on a Combination of Confocal Slit With Wavelength/Space Conversion

Confocal laser microscope (CLM) has been widely used in the fields of the non-contact surface topography, biomedical imaging, and other applications, because the confocality gives two-dimensional (2D) optical-sectioning or three-dimensional (3D) imaging capability with the depth selectivity. Combination of line-focused CLM with one-dimensional (1D) spectral encoding CLM enables us to obtain the 2D confocal image without the need for the mechanical scanning. So-called scan-less 2D CLM is a unique imaging modality, however, there are no attempts to apply for practical application. In this paper, we constructed scan-less 2D CLM with the image acquisition time of 0.23 ms, the lateral resolution of 1.2 µm, the depth resolution of 2.4 µm, and apply it for different kinds of application to evaluate its practical potential

Photo-Induced Cell Damage Analysis for Single- and Multifocus Coherent Anti-Stokes Raman Scattering Microscopy

In this study, we investigated photo-induced damage to living cells during single-and multifocus excitations for coherent anti-Stokes Raman scattering (CARS) imaging. A near-infrared pulsed laser (709 nm) was used to induce cell damage. We compared the photo-induced cell damage in the single- and the multifocus excitation schemes with the condition to obtain the same CARS signal in the same frame rate. For the evaluation of cell viability, we employed 4', 6-diamidino-2-phenylindole (DAPI) fluorophores that predominantly stained the damaged cells. One-and two-photon fluorescence of DAPI fluorophores were, respectively, excited by an ultraviolet light source and the same near-infrared light source and were monitored to evaluate the cell viability during near-infrared pulsed laser irradiation. We found lower uptake of DAPI fluorophores into HeLa cells during the multifocus excitation compared with the single- focus excitation scheme in both the one- and the two-photon fluorescence examinations. This indicates a reduction of photo-induced cell damage in the multifocus excitation. Our findings suggested that the multifocus excitation scheme is expected to be suitable for CARS microscopy in terms of minimal invasiveness