Investgation Of The Modification Method Of Functionalized Slides And Application In The Cell Analysis

The process of cell adhesion is an important physiological process of organisms, and most kinds of cells have to depend on the effect of adhesion, stretching, reproductive behavior on the substrate surface to conduct normal physiological activities. In the liquid-based cytology technology for screening of cervical cancer, the slides are used as the substrate of cell adhesion substrate. The modified slides could enhance the adhesion of cells on the matrix to improve the diagnostic accuracy in the diagnosis of the disease. In this article, the glass slides are modified by the biocompatible materials modification, and applied into the liquid-based cytology system. The slides surface coated with poly-L-lysine, chitosan and collagen will enhance the adhesion of cervical exfoliated cells. And these slides would be applied into the ThinPrep system to increase the accuracy rate of diagnosis. Herein, the research papers are divided into the following three parts:Part Ⅰ The modification method of GOPS and poly-L-lysine modified glass slides and application in liquid-based cytology system 3-Glycidyloxypropyl trimethoxysilane (GOPS) was coated on the slides to form the epoxy-terminated slides, and then poly-L-lysine (PLL) was covalently modified onto the above slides to form GOPS-PLL double decorated slides. These different slides were characterized by X-ray photoelectron spectroscopy (XPS) and atomic force microscope (AFM). The cell adhesion ability was tested and compared with traditional PLL coated slides and the unmodified the slides by fixing the cervical exfoliative cells on the different slides to form the Pap smears. The cell morphology of adhered cervical exfoliative cells was observed under the microscope after the complex Papanicolaou staining. The number of cervical exfoliative cells on the unmodified slides, PLL coated slides and GOPS-PLL coated slides was1030±300,3283±226and4119±280(n=12), respectively. And the data among the three different modification methods showed significant differences after the statistical analysis (one-way analysis of variance, ANOVA test, P<0.05). The GOPS-PLL coated slide could be widely applied to the ThinPrep system for cervical carcinoma screening to increase the accuracy rate of diagnosis.Part Ⅱ Capture of Hela cells on chitosan functionalized glass slide and application in liquid-based cytology systemThe chitosan modified glass slides are conducted by the3-aminopropyl triethoxysilane (APTES) and cross-linked by the reaction of glutaraldehyde with chitosan. The surface topography of the slides is characterized by atomic force microscopy (AFM). The adhesion and growth of Hela cells on the chitosan-modified glass slide are studied. The slide under the favorable conditions was used for the production of cervical smears. The smears are placed under the microscope after HE staining. Control experiment was tested by the blank slide. The results showed that the cell morphology of the cervical exfoliative cells was easily to be observed and the cells was colored, which was benefit to the diagnosis of the cancer by the pathologists. The chitosan modified slides could be used in liquid-based cytology in the diagnosis of cervical cancer to improve the accuracy.Part Ⅲ Modification method of collagen functionalized slides and application in liquid-based cytology systemIn this part, two collagen coating methods (collagen grafted directly on the substrate and on the1%chitosan-modified substrate) were carried out, on which the adhesive behaviors of cervical exfoliative cells were studied. An atomic force microscope (AFM) was used to characterize morphologies of the slides surface. The cervical exfoliative cell adhesion ability was tested and compared with unmodified the slides by making cell smears. The cell morphology of adhered cervical exfoliative cells was observed under the microscope after the HE staining. The results showed that the adhesion ability of the cervical exfoliative cells was strengthened on the modified slides, and the morphology of colored cells was easily to be observed. This work might be helpful to the practical application of cell smears in the cervical cancer diagnosis to improve the diagnostic accuracy.