Optical microscope is one of the most popular imaging methods to find out metabolism and structural information of a cell for bio-medical researches. Among the many optical microscope technologies, Confocal microscope and Two-photon microscope is the most popular microscope for biomedical imaging. Because a confocal microscope is available for both reflectance and fluorescence mode, it can be used in various appli-cations to obtain the structural information of the sample. The Two-photon microscope has better penetration depth compare to confocal microscope. By using this better penetration depth, it is possible to obtain images of cell that is deeper from the surface.
While the Confocal and Two-photon microscope (TPM) is only available to obtain structural infor-mation, there are many other microscopes that are available to obtain chemical or biological information form the sample. The fluorescence lifetime imaging microscope (FLIM) is the most popular imaging technique to obtain bio-chemical information of the sample. Because the lifetime of the fluorescence is various depending on the sample environment like pH, ion and Oxygen concentration, dynamic interaction between two molecules, it is possible to predict the causality of the cell interaction and development by analyzing the lifetime changes of the sample. The Hyper Spectral Imaging (HSI) is another important method to obtain chemical information changes of the sample. The HSI is a spectral analyzing method to calculate the reflectance of the sample and predict what the sample is and analyze the property changes of the sample by comparing the reflectance spectrum of the sample.
But those microscopes are available only for single modality which means that a microscope can measure single information from the sample. To overcome this barrier, multi-modal microscope had been proposed that combining several of microscopes into one system to obtain the same image with different modality. The multi-modal microscope can be used to find out unknown parameter of the sample by analyzing many information obtained using each modality of the multi-modal microscope.
Although there are many kind of imaging microscope technologies, only four of microscopes which were mentioned above will be treated as candidates to be integrated to Multi-modal microscope. Because Confocal microscope and TPM is available to obtain structural information and FLIM and HIS are available to obtain chemical information, the Multi-modal microscope can obtain much more information from the sample and analyze the sample interaction and development more precisely compare to single modality mi-croscope.
There were some researches on developing multi-modal microscope in recent years, but most of those researches were just sharing the relay optics for scanning microscope. And most of those multi-modal micro-scopes were table-top system that is not available for practical user like pathologist because the system inte-gration to work at multi-modality is difficult for practical reasons. I propose a kinematic mounting mechanism for simple and stable mode switching. The kinematic coupling is a very popular method to ensure high in-position stability for high precision machine. Each modality can be switched easily by changing filter and galvano mirror position using proposed kinematic coupling mechanism.
Although there were several of studies to make multi-modal microscope itself in several modality, there were rare studies to find out the correlation between the image acquisition modes like confocal and FLIM. Both of two imaging microscopes are already used for biomedical imaging researches, but it is not considered where those imaging modes have correlations to each other.
In this research, I propose the optimal design method to make multi-modal microscope that has four modalities of Confocal, TPM, FLIM and HSI. And analyze the performance of proposed Multi-modal micro-scope to prove that the multi-modal microscope work well to obtain various imaging information from the sample. In the next step, I analyze each modality and compare the image acquisition condition case by case to find out the correlation of each modality. These data-processing methods will be used to increase the accuracy of disease diagnosis or predict what is happening inside of the cell during their development.
The multi-modal microscope is becoming very powerful imaging tool recently, and many of studies are showing the possibility of further research finding out unknown parameters. The multi-modal microscope is expected to become one of the most promising imaging techniques for bio-medial researches.
멀티모달 현미경은 다양한 모달리티를 이용하여 시편으로부터 다양한 영상정보를 획득한 후 이를 복합적으로 분석하여, 기존의 현미경보다 더욱 많은 정보를 얻어낼 수 있는 복합 현미경을 말한다. 본 연구에서는 기존의 광학현미경에 공초점 현미경, 이광자 현미경, 형광수명시간 현미경, 스펙트럴 이미징 현미경을 결합함으로써 시편으로부터 다양한 영상을 획득하여 이를 이미징화 하고 분석할 수 있는 현미경을 설계 및 제작하고, 제작된 현미경을 바이오 메디컬 이미징에 적용함 으로써 정상 및 암 조직을 구분하고 동맥경화 혈관에 대해서 정상과 동맥경화의 차이점을 밝혀내는 연구를 수행하였다. 공초점 현미경은 시편의 3차원 이미징 및 고 분해능 영상획득이 가능하다는 장점이 있으며, 이광자 현미경은 깊은 투과깊이, 형광수명시간 현미경의 시편의 화학적 정보, 스펙트럴 이미징은 시편의 특성변화를 측정할 수 있으므로 제안된 모달리티를 통합하면 시편의 형태적인 정보뿐만 아니라 시편의 화학적 정보도 획득할 수 있는 장점을 가지고 있다. 제안된 멀티모달 현미경의 설계 및 제작을 위해서 고정밀 위치결정기구인 kinematic mounting을 이용하여 광학 기기의 위치를 변경하여 모달리티를 변경할 수 있도록 만들어 주었다. 또 이광자 현미경 및 형광수명시간 현미경의 성능향상을 위하여 negative chirping 기구를 설계 및 제작하여 멀티모달 현미경에 추가하였으며, 스펙트럴 이미징의 성능이 최대가 될 수 있도록 분광광학계의 설계 및 제작을 수행하였다. 제작된 멀티모달 현미경을 이용하여 대자암 및 동맥경화 혈관을 이미징 하였고, 이미징 된 결과를 분석하여 정상 및 대장암 조직과 동맥경화혈관과 정상혈관의 차이점을 밝혀내는 연구를 수행하였다.