The Development Of Multimodality Reflectance And Fluorescence Confocal Microscope

Confocal laser scanning microscopy has been widely accepted in dermatological research, due to its advantages in non-invasive, real time, high resolution in vivo imaging. Conventional confocal laser scanning microscopy can be divided into two modes: reflectance mode and fluorescence mode. The reflectance mode demonstrates tissue structural information based on the different refractive indices among the various tissue microstructures. The fluorescence mode can detect tissue pathological information with the excitation of exogenous fluorescent dye to produce contrast. However, up to now most researchers have been focusing on either reflectance or fluorescence confocal imaging separately which can only provide either structural or pathological information of tissue. This greatly affected the accuracy of dermatological diagnosis by lack of enough diagnostic information.In order to improve diagnostic performance of dermatosis efficiently, we developed a new design of noninvasive simultaneous multimodality fluorescence and reflectance confocal microscope which combines the structural imaging and pathological imaging. Moreover, in the confocal system the fluorescence imaging is aim to autofluorescence substance such as nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) which exclude the toxic affection from the introduction of exogenous fluorescent dyes. With both imaging modality acquired simultaneously, the combined image revealed the morphological and pathological information of the suspicious site, yielding a possibility of precancer diagnosis of skin.The study contents in this thesis are demonstrated as follows:1 The hardware design of multimodality reflectance and fluorescence confocal microscope, including reflectance and fluorescence optical system, scanning system and detecting system.2 The software design of multimodality reflectance and fluorescence confocal microscope, including scan mode algorithm design, acquisition mode algorithm design and image reconstruction algorithm design.3 In vivo applications in skin tissue imaging by multimodality reflectance and fluorescence confocal microscope, including epidermis tissue imaging in mouse skin and melanoma imaging diagnosis.