本文關(guān)鍵詞：光譜與原子力顯微鏡方法研究金納米粒子細(xì)胞內(nèi)吞　出處：《東北師范大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 金納米粒子 細(xì)胞 吸收光譜 原子力顯微鏡 單分子示蹤

【摘要】：與傳統(tǒng)的光學(xué)探針相比,金納米粒子作為光學(xué)探針有著更好的穩(wěn)定性和易制備、修飾的特性,并且在光學(xué)信號(hào)和生物兼容性方面都有著很大的優(yōu)勢(shì),從而使金納米粒子在生物傳感器和生物成像以及在生物醫(yī)學(xué)領(lǐng)域和分析化學(xué)領(lǐng)域等方面有著廣泛的應(yīng)用。在單粒子光譜技術(shù)與單粒子成像的優(yōu)勢(shì)中,金納米粒子由于本身具有一定的光學(xué)特性,進(jìn)而在單細(xì)胞層面上可以與生物功能性分子相結(jié)合成為了研究學(xué)者們研究細(xì)胞的一種有力工具。本論文將紫外-可見吸收光譜技術(shù)、原子力顯微鏡單分子示蹤技術(shù)結(jié)合在一起,研究金納米粒子與細(xì)胞之間存在的細(xì)致相互作用,以及金納米粒子進(jìn)入細(xì)胞的動(dòng)態(tài)過程。具體結(jié)果摘要內(nèi)容如下:1.我們根據(jù)金納米粒子具有的吸收特性,通過采集金納米粒子吸收光譜來研究,金納米粒子與細(xì)胞之間的相互作用。通過實(shí)驗(yàn)發(fā)現(xiàn)納米粒子與細(xì)胞孵育以后也會(huì)存在一個(gè)明顯的吸收峰,此吸收峰和不與細(xì)胞作用的納米粒子吸收峰相比有微小的紅移。利用離散偶極子近似(discrete dipole approximation-DDA)理論,進(jìn)行理論模擬,得出了納米粒子與細(xì)胞孵育和納米粒子單獨(dú)存在時(shí)懸浮溶的液吸收峰。利用高斯擬合,得到吸收峰的峰值位置,理論模擬與我們?cè)趯?shí)驗(yàn)中所得的吸收光譜值存在5nm的誤差,這說明我們的實(shí)驗(yàn)是可行的。同時(shí)實(shí)驗(yàn)結(jié)果表明:金納米粒子與細(xì)胞孵育后,其吸收峰會(huì)發(fā)生紅移現(xiàn)象。吸收峰紅移說明:納米粒子周圍的介電環(huán)境發(fā)生了變化,即納米粒子只有與細(xì)胞發(fā)生了相互作用,周圍的介電環(huán)境才會(huì)發(fā)生變化。同時(shí)在納米粒子濃度降低時(shí),降低的金納米粒子與細(xì)胞孵育后的吸收峰在三個(gè)濃度之間沒有發(fā)生變化。而與單獨(dú)金納米粒子的吸收峰相比而言,它發(fā)生了吸收峰紅移的現(xiàn)象。這充分說明金納米粒子與細(xì)胞孵育以后會(huì)使吸收峰發(fā)生紅移,吸收峰紅移的原因是納米粒子周圍的介電環(huán)境變化所引起,金納米粒子周圍介電環(huán)境變化就充分說明納米粒子被內(nèi)吞到細(xì)胞內(nèi)部。2.用原子力顯微鏡單分子示蹤技術(shù)記錄了不同尺寸的單個(gè)金納米粒子進(jìn)入到細(xì)胞中的動(dòng)態(tài)過程。通過實(shí)驗(yàn)我們得出:分別為5nm、10nm、20nm的納米粒子與細(xì)胞作用的內(nèi)吞力的范圍大約在20p N至140p N之間。同時(shí),在納米粒子的粒徑逐漸增大時(shí),納米粒子與細(xì)胞之間的作用力與作用時(shí)間會(huì)隨著納米粒子與細(xì)胞接觸面積的增加而增加,同時(shí)進(jìn)入的平均速率會(huì)隨著進(jìn)入距離的增大而有所減小。做了對(duì)照和阻礙實(shí)驗(yàn),說明納米粒子與細(xì)胞之間存在一種特殊的作用力,而且納米粒子的內(nèi)吞是通過細(xì)胞質(zhì)膜上膽固醇的作用,依賴于網(wǎng)格蛋白或小窩蛋白的介導(dǎo)進(jìn)入細(xì)胞內(nèi)部。以上工作闡明:金納米粒子與細(xì)胞孵育以后,納米粒子會(huì)進(jìn)入到細(xì)胞內(nèi)部,并且在金納米粒子進(jìn)入細(xì)胞的過程中,將會(huì)與細(xì)胞膜上的物質(zhì)存在一定的關(guān)系。實(shí)驗(yàn)中根據(jù)紫外-可見吸收光譜的峰值變化,表明金納米粒子被內(nèi)吞到細(xì)胞中。同時(shí)我們利用原子力顯微鏡單分子示蹤技術(shù)研究不同尺寸單個(gè)納米粒子與細(xì)胞作用的動(dòng)態(tài)過程,這一工作為以后研究藥物轉(zhuǎn)運(yùn)具有重要意義。
[Abstract]:Compared with the traditional optical probe and gold nanoparticles as optical probe has better stability and easy preparation, modification and properties on the optical signal and bio compatibility have great advantages, so that the gold nanoparticles have been widely applied in biological sensors and biological imaging and in biomedical field and analysis in the field of chemistry. Single particle spectroscopy and single particle imaging advantage, gold nanoparticles due to its optical characteristics, and biological function and molecular combination of a powerful tool to study cell researchers at the single cell level. The UV Vis absorption spectroscopy technology atomic force microscopy, single molecule tracing technology together between gold nanoparticles and cell detailed interactions, and gold nanoparticles into The dynamic process of cells. The specific results are as follows: 1. according to the absorption properties of gold nanoparticles have, through the acquisition of gold nanoparticles to study the absorption spectra, the interaction between gold nanoparticles and cells. Experiments found that nanoparticles and cells were incubated with after there will be an obvious absorption peak, the absorption peak and not cells with nanoparticles, the absorption peaks are small redshift compared. By using the discrete dipole approximation (discrete dipole approximation-DDA) theory, theoretical simulation, obtained nanoparticles and cells were incubated with nanoparticles alone when suspension solution of liquid absorption. Using Gauss fitting, get the peak position of absorption peak, theoretical simulation of 5nm error our income and absorption spectra in the experiment, it shows that our experiment is feasible. At the same time, the experimental results show that the gold nanoparticles With the cells after incubation, the absorption peak red shift phenomenon. The absorption peak red shift that changes the dielectric environment around the nanoparticles, the nanoparticles only and cell interaction occurs, the dielectric surrounding environment will change at the same time. The concentration of nanoparticles decreased, gold nanoparticles and cells with reduced absorption peak after incubation between three concentration did not change. The absorption peak of gold nanoparticles with single contrast, it happened redshift of absorption peak phenomenon. This fully shows that the gold nanoparticles and cells were incubated after the absorption peak red shifted absorption peak red shift is a dielectric nanoparticles caused by environmental changes gold nanoparticles, around the dielectric environment can fully explain nanoparticle endocytosis to the interior of the cell.2. microscope single molecule tracing technique using atomic force records of different scale A single inch of gold nanoparticles into dynamic processes in the cell. Through experiments, we conclude that: respectively 5nm, 10nm, and 20nm nanoparticles range cell endocytosis capacity between about 20p N to 140P N. At the same time, the particle size increased in force, and action time between nanoparticles and the cell will increase with the increase of nanoparticles and cell contact area, the average rate at the same time will enter into with the increasing distance decreased. The control and block experiment shows that there is a special interaction between nanoparticles and cells, and endocytosis of nanoparticles is through the cytoplasmic membrane cholesterol, depend on clathrin or caveolin mediated entry into cells. The above work of gold nanoparticles and cells following incubation, nanoparticles will enter into cells, and In the process of gold nanoparticles into cells, there is a certain relationship and will be on the cell membrane material. The experiment according to the UV Vis absorption spectra showed that the peak value change, gold nanoparticles is internalized into the cell. The dynamic process at the same time we use the atomic force microscope study of single molecular tracer technique in different sizes of single nanoparticles and cells. The significance of this work for the future research on drug transport.

【學(xué)位授予單位】：東北師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：Q25;TB383.1

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本文編號(hào)：1420838