【摘要】：細(xì)胞粘附是一種對(duì)所有多細(xì)胞和單細(xì)胞生物都極其重要的基本現(xiàn)象,在組織生長(zhǎng)、細(xì)胞間交流、細(xì)胞遷移、細(xì)胞新陳代謝、發(fā)炎和感染中起著重要的作用。對(duì)細(xì)胞粘附力的研究有助于揭示生命的奧秘,促使新的疾病診斷和治療手段誕生,推動(dòng)納米生物力學(xué)的進(jìn)一步發(fā)展,甚至為生物的基因改造提供信息。在眾多的方法與技術(shù)中,原子力顯微鏡(Atomic Force Microscope,AFM)以其測(cè)力精度高、范圍廣且能在生理環(huán)境中檢測(cè)等優(yōu)勢(shì)成為納米生物力學(xué)中應(yīng)用最廣泛的研究手段之一�；谧灾餮邪l(fā)的納米機(jī)器人,本文應(yīng)用改進(jìn)的AFM單細(xì)胞力譜法對(duì)哺乳動(dòng)物細(xì)胞的粘附力進(jìn)行定量研究。在保持AFM單細(xì)胞力譜法高精度的同時(shí),采用中空探針吸附細(xì)胞,代替了傳統(tǒng)探針的化學(xué)修飾,提高了細(xì)胞粘附力的測(cè)試效率,為細(xì)胞力學(xué)的研究提供了新的測(cè)試技術(shù)與方法。主要研究?jī)?nèi)容包括:首先,本文改進(jìn)了傳統(tǒng)的AFM單細(xì)胞力譜法,采用中空探針代替?zhèn)鹘y(tǒng)AFM探針。設(shè)計(jì)了中空探針氣動(dòng)裝置,使中空探針內(nèi)產(chǎn)生負(fù)壓而吸附細(xì)胞,減少了固定細(xì)胞與標(biāo)定探針?biāo)ǖ臅r(shí)間,提高了測(cè)試的效率。同時(shí),該方法允許細(xì)胞和基底長(zhǎng)時(shí)間的接觸,擴(kuò)大了粘附力的測(cè)量范圍。接著,本文對(duì)中空探針負(fù)壓提拉細(xì)胞的過(guò)程進(jìn)行了模擬�；诒∧�-液體連續(xù)介質(zhì)粘附模型提出并建立了貼壁細(xì)胞的力學(xué)模型,利用有限元法對(duì)不同內(nèi)徑的中空探針負(fù)壓提拉細(xì)胞的過(guò)程進(jìn)行了仿真,得到了探針內(nèi)徑大小對(duì)細(xì)胞形變與應(yīng)力的影響。其次,本文分析了外力作用下粘附分子連接(Adhesion Molecular Bonds)的解離反應(yīng)�；谝后w中分子連接受力解離的布朗運(yùn)動(dòng)動(dòng)理學(xué)理論,分析中空探針負(fù)壓提拉細(xì)胞過(guò)程中粘附分子連接特性,得到了外力及其加載速率對(duì)粘附分子連接解離反應(yīng)速率、壽命、強(qiáng)度的影響規(guī)律。最后,本文采用改進(jìn)的AFM單細(xì)胞力譜法對(duì)細(xì)胞與基底的粘附力進(jìn)行測(cè)量。測(cè)量結(jié)果精確、效率高、范圍廣。應(yīng)用該方法可以得到細(xì)胞粘附力與探針上升速度關(guān)系以及細(xì)胞粘附力與時(shí)間的關(guān)系�；诩{米機(jī)器人系統(tǒng),改進(jìn)的AFM單細(xì)胞力譜法能夠在液相中對(duì)細(xì)胞進(jìn)行三維操作和粘附力測(cè)試,對(duì)納米生物力學(xué)的研究具有推動(dòng)效應(yīng),并為納米科學(xué)的研究提供了新的途徑。
[Abstract]:Cell adhesion is a basic phenomenon that is extremely important to all multicellular and single-celled organisms. It plays an important role in tissue growth, intercellular communication, cell migration, cell metabolism, inflammation and infection. The study of cell adhesion will help to reveal the mystery of life, promote the birth of new methods of disease diagnosis and treatment, promote the further development of nano-biomechanics, and even provide information for the genetic modification of organisms. Among the many methods and techniques, atomic force microscope (Atomic Force Microscope,AFM) has become one of the most widely used research methods in nano-biomechanics because of its advantages of high precision, wide range and can be detected in physiological environment. Based on the self-developed nanorobot, the adhesion of mammalian cells was quantitatively studied by modified AFM single cell force spectroscopy. While maintaining the high accuracy of AFM single cell force spectrum method, the hollow probe was used to adsorb the cells, instead of the chemical modification of the traditional probe, which improved the efficiency of the cell adhesion test, and provided a new testing technique and method for the study of cell mechanics. The main research contents are as follows: firstly, the traditional AFM single cell force spectrum method is improved and the hollow probe is used instead of the traditional AFM probe. A hollow probe pneumatic device was designed to produce negative pressure and adsorb the cells in the hollow probe, which reduced the time between the fixed cell and the calibrated probe, and improved the efficiency of the measurement. At the same time, the method allows long contact between the cell and the substrate, expanding the range of adhesion. Then, the process of hollow probe negative pressure pulling cells was simulated. Based on the membrane liquid continuum adhesion model, a mechanical model of adherent cells was proposed and established. The process of negative pressure Czochralski cells with different inner diameters was simulated by finite element method. The effects of probe diameter on cell deformation and stress were obtained. Secondly, the dissociation reaction of adhesion molecule attached to (Adhesion Molecular Bonds) under external force was analyzed. Based on the Brownian kinematics theory of molecular connection dissociation in liquid, the bonding characteristics of adhesion molecules in the process of negative pressure Czochralski cell with hollow probe are analyzed. The effect of external force and loading rate on the dissociation rate and lifetime of adhesion molecule is obtained. The influence law of strength. Finally, the adhesion between the cell and the substrate was measured by modified AFM single cell force spectroscopy. The measurement results are accurate, high efficiency and wide range. By using this method, the relationship between the cell adhesion force and the rising speed of the probe and the relationship between the cell adhesion force and the time can be obtained. Based on the nano-robot system, the improved AFM single-cell force spectrum method can perform three-dimensional operation and adhesion test of cells in liquid phase, which can promote the research of nano-biomechanics and provide a new way for the research of nanoscience.
【學(xué)位授予單位】：哈爾濱工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：Q27;TP242

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本文編號(hào)：2234824