Multimodal and multiplex spectral imaging of rat cornea ex vivo using a white‐light laser source

Abstract

We applied our multimodal nonlinear spectral imaging microscope to the measurement of rat cornea. We successfully obtained multiple nonlinear signals of coherent anti‐Stokes Raman scattering (CARS), third‐order sum frequency generation (TSFG), and second harmonic generation (SHG). Depending on the nonlinear optical processes, the cornea tissue was visualized with different image contrast mechanism simultaneously. Due to white‐light laser excitation, multiplex CARS and TSFG spectra were obtained. Combined multimodal and spectral analysis clearly elucidated the layered structure of rat cornea with molecular structural information. This study indicates that our multimodal nonlinear spectral microscope is a promising bioimaging method for tissue study.

Modern trends in biophotonics for clinical diagnosis and therapy to solve unmet clinical needs

“…Results from bulk bone are shown in Figure . Another group used a white light laser source to obtain multiplex CARS and third‐order sum frequency generation spectra . This combined multimodal and spectral analysis elucidated the layered structure of rat cornea with molecular and structural information.…”

Ultra-multiplex CARS spectroscopic imaging with 1-millisecond pixel dwell time

“…Owing to the high peak power of the laser source, we are not only able to obtain the CARS signal but also other signals resulting from different nonlinear optical processes such as second harmonic generation (SHG), third harmonic generation (THG), and two-photon excitation fluorescence (TPEF), thereby enabling multi-modal nonlinear optical (or multi-photon) imaging [27][28][29]. We previously used multi-modal nonlinear optical imaging to successfully elucidate that the Rootletin filaments [30] of the rootlets in the retina are SHG-active [31].…”

Multimodal Imaging of Living Cells with Multiplex Coherent Anti-stokes Raman Scattering (CARS), Third-order Sum Frequency Generation (TSFG) and Two-photon Excitation Fluorescence (TPEF) Using a Nanosecond White-light Laser Source

“…So far, CARS, second harmonic generation (SHG), third harmonic generation (THG), and third-order sum frequency generation (TSFG) imaging have been reported by our group. 19,20 In the present study, we have extended our microscopic system further by launching another multiphoton channel, two-photon excitation fluorescence (TPEF).…”