| Laser scanning confocal microscopy (LSCM) is widely used in biological science. In most applications, however, the laser is used only as a monochromatic excitation source for fluorescence. Many aspects of laser technology and its scattering property are not fully used. Conventional LSCM measures on-axis scattered light only, neglecting the large angle scattering that contains information of a target that would not be easily obtained by on-axis detection.;The objective of this work was to develop a novel optical microscopy technique based on two key features: laser multi-spectral confocal imaging and large-angle azimuthal scatter diffuse imaging. The two features combined give a spatial-spectral discrimination capability that can enhance contrast and spatial resolution not available from current optical microscopy. The goal was to contribute a new tool to biomedical, analytical, sensing, and metrological applications.;This work describes the principle of laser multi-spectral diffuse scatter confocal microscopy. Theoretical calculations and image simulations based on Finite Difference Time Domain (FDTD) were explored and provided a theoretical basis and a guide for the experiments. Simulated confocal images and large-angle scatter diffuse images of several targets were obtained and discussed. To the best of my knowledge, this specific work has not been done by other researchers. The results show that large angle scatter imaging has the ability to differentiate between targets that are not easily obtained only from confocal imaging. The microscopic imaging system was based on a system architecture configured similarly to an optical wireless network that allows simultaneous multiple communication connections among any number of transmitters and receivers. The entire imaging system, including control program, optical components, layout, and various other components, was optimized. Confocal images and large angle scatter diffuse images were obtained, and they demonstrated high dynamic range and high image contrast of the imaging system. The experimental results were found to be promising, with an ability to uncover and interpret features not easily achieved by conventional confocal microscopic methods.;The combination of confocal method and large scattering angle imaging can achieve more information about objects than one of them separately. This research proves that Laser Multi-Spectral Diffuse Scatter Confocal Microscopy is a novel technology that can achieve more information about targets and can be widely used for biomedical, analytical, sensing. and metrological applications. |
