Research Of Colorectal And Esophageal Cancer Based On Multiphoton Microscopy Imaging And Raman Spectroscopy

Early nondestructive diagnosis of cancer is the most effective method to enhance the efficacy of cancer treatment, and is also the key problem of science. The multiphoton microscopic imaging technique (MPM) based on the two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) is very suitable for studying the microstructural morphology and internal composition spectrum characteristics of tissues and cells, thanks to its unique advantages such as "light-section" function, low cytotoxicity, deep imaging depth, high sensitivity and high spatial resolution, and so on,. Raman spectroscopy (RS) is another promising method to identify and distinguish the different material structure from the molecular vibration spectrum, which can provide the detailed information of certain intrinsic molecular components of biological tissue. The above two methods have their own advantages and can complement each other., in this dissertation, we try to establish the quantitative characteristic parameters of normal and cancerous samples at the molecular level and cell level by combining multiphoton microscopy with Raman spectroscopy. Two common cancers with high incidence in China, the colorectal and esophageal cancer, are mainly discussed in the study.. The morphological and spectral characteristics of both cancer tissue and blood samples are presented. The experimental results provide some evidences for cancer nondestructively early diagnosis by combing multiphoton microscopy imaging with Raman spectroscopy. The medical statistical data obtained in the study will also contribute to the cancer treatment evaluation., the research contents are presented detail as following:First of all, the morphology of colorectal and esophageal tissue are briefly introduceed, and the process of different TNM stages and pathological features are summarizedThe basic principle and experimental method of multiphoton microscopy imaging (MPM) and Raman spectroscopy (NRS/SERS) are displayed respectively. Second, taking isolated human colorectal and esophageal tissue as the experimental object, SHG and TPEF images of normal and cancerous tissue are obtained using multiphoton microscopy imaging technology. The optimization of laser parameters to improve imaging quality is explored, and the microstructure morphology changes between malignant and normal mucosa, submucosa and muscularis are systematic compared. the characteristic parameters of colorectal cancer and esophageal cancer are compared and featured to provide the evidences for the early diagnosis of colorectal cancer and esophageal cancer.Then, based on normal Raman spectroscopy and SERS technology with silver and gold colloid, The NRS and SERS spectrum of esophageal cancer and normal tissue were detected and compared, The feature Raman spectrum were extracted, based on which,, the feasibility, validity and reliability of NRS and SERS technology for cancer diagnosis are discussed with PCA-LDA multivariate statistical algorithm.Finally, Based on the membrane electrophoresis of plasma combined with SERS spectroscopy and taking gold nanoparticle as reinforced base, the plasma albumin and globulin samples are firstly studied. PCA-LDA multivariate statistics display that diagnosis effect of membrane electrophoresis united with SERS is with high accuracy, and the diagnostic sensitivity and specificity (97.1%and100%) of plasma albumin SERS spectroscopic is better than that of plasma globulin (90.9%and93.3%). Raman spectra indicates the changes of cancerous plasma protein structure, that is, the relative content of nucleic acid and phenylalanine is increasing, while the relative content of phospholipid and carbohydrates is decreasing in cancerous plasma protein.