發(fā)布時間：2018-05-22 18:35

  本文選題：原位結晶 + 結晶調控　； 參考：《山東師范大學》2017年碩士論文

【摘要】：為了提高石英晶體微天平對生物活性分子的靈敏度,建立了基于石英晶體微天平的原位選擇性結晶方法。在之前的很多研究中,都對自組裝膜進行了科學的論述以及研究,自組裝膜的調控作用具有成熟的理論依據,并在實際應用中展現(xiàn)了良好的性能。在我們的試驗中所建立的自組裝膜主要是通過具有不同調控作用的官能團的暴露情況實現(xiàn)對碳酸鈣原位結晶的調控作用。在我們建立的信號放大機制中,選用的兩種功能基團分別為-N(CH3)3和-COOH,其中-N(CH3)3作為碳酸鈣在表面的結晶抑制劑,-COOH作為碳酸鈣在表面的結晶誘導劑,在這兩種功能基團的共同調配下,實現(xiàn)了對碳酸鈣在石英晶體微天平晶片表面原位結晶的調控,為我們的方法在信號放大方面提供了可能性。我們對其他常用的檢測腫瘤標志物的方法以及應用石英晶體微天平的信號放大技術做了一定的學習與比較。相比之下,我們建立的基于石英晶體微天平的原位選擇性結晶方法具有原理簡單、不依賴生物碼、靈敏度高和選擇性高的優(yōu)點,在癌癥的早期診斷方面展現(xiàn)了了良好的發(fā)展前景。本文的主要內容如下:1.碳酸鈣晶體在金片上的表面結晶過程可以通過石英晶體微天平檢測,通過調節(jié)兩種不同功能基團的用量比例可以實現(xiàn)晶體生長的定量控制。本實驗中碳酸鈣原位選擇性結晶過程用到的兩種功能基團分別為-N(CH3)3和-COOH,獲得的晶體具有均勻的尺寸大小,相同的形態(tài)及晶型。實驗發(fā)現(xiàn)隨著-COOH組分比例的增加,晶體的量呈現(xiàn)一定規(guī)律的增加趨勢。實驗證明對金表面晶體生長的定量控制可以作為一個有效的質量擴增機制,該擴增機制可以用于石英晶體微天平的信號放大,以此可以提高石英晶體微天平的分析靈敏度。2.將上述基于碳酸鈣原位選擇性結晶技術應用到QCM對DNA分子的檢測中,結果發(fā)現(xiàn),該方法將檢測DNA信號擴大了1000多倍,提高了DNA檢測的靈敏度。在檢測DNA時,最低檢測限LOD達到了2aM,并且線性檢測范圍寬,為10aM到1nM。該方法同時具有很高的選擇性,可以區(qū)分識別完全配對的DNA與1個堿基錯配的DNA,可應用于單核苷酸的多態(tài)性(SNP)檢測。3.基于碳酸鈣原位結晶的石英晶體微天平信號放大技術,在檢測DNA時展現(xiàn)了良好的性能。為了擴大其檢測領域,我們將其用于細胞的檢測,本實驗選用的是Ramos細胞。在引入適配體后,結合碳酸鈣原位結晶的石英晶體微天平信號放大技術檢測DNA的方法,實現(xiàn)Ramos細胞的檢測,其最低檢測限LOD為5個/ml,檢測范圍為5個/ml到6000個/ml。最后,我們將該方法與其他基于質量信號放大的石英晶體微天平檢測方法進行了對比,通過對比,證明了我們的實驗具有很好的靈敏度。4.我們對碳酸鈣原位結晶的石英晶體微天平信號放大技術進行了總結。該方法為石英晶體微天平技術的發(fā)展提供了新視角、新思路,基于該方法,可建立各種生物活性分子的通用檢測平臺,相信在研究者們的進一步研究下,該方法可以真正的應用于臨床,為癌癥的診療提供新技術。
[Abstract]:In order to improve the sensitivity of quartz crystal microbalance to bioactive molecules, an in-situ selective crystallization method based on quartz crystal microbalance was established. In many previous studies, the self assembled monolayers were scientifically discussed and studied. The regulation of self assembled films has a mature theoretical basis and is shown in practical applications. In our experiment, the self assembled monolayers are mainly regulated by the exposure of functional groups with different regulatory functions. In the signal amplification mechanism we have established, two functional groups are selected as -N (CH3) 3 and -COOH, respectively, of which -N (CH3) 3 is used as calcium carbonate. As a crystalline inhibitor on the surface, -COOH is a crystal inducer on the surface of calcium carbonate. Under the joint deployment of these two functional groups, the regulation of the in-situ crystallization of calcium carbonate on the surface of the quartz crystal microbalance wafer is realized. It provides a possibility for our method to amplify the signal. We have detected other commonly used tumor markers. In contrast, the in situ selective crystallization method based on quartz crystal microbalance has the advantages of simple principle, no dependence on biological code, high sensitivity and high selectivity, and is shown in the early diagnosis of cancer. The main contents of this paper are as follows: 1. the surface crystallization process of calcium carbonate crystal on the gold plate can be detected by quartz crystal microbalance, and the quantitative control of crystal growth can be realized by adjusting the proportion of two different functional groups. In this experiment, two kinds of work used in the process of selective crystallization of calcium carbonate are used in this experiment. The energy groups are -N (CH3) 3 and -COOH respectively. The obtained crystals have uniform size, the same morphology and crystal shape. The experimental results show that the amount of crystal increases with the increase of the proportion of -COOH components. The experimental results show that the quantitative control of the crystal growth of the gold surface can be used as an effective mass amplification mechanism. The increase mechanism can be used to amplify the signal of quartz crystal microbalance, so that the sensitivity.2. of quartz crystal microbalance can be improved by applying the CaCO3 in situ selective crystallization technology to the detection of DNA molecules by QCM. The results show that the method will expand the detection of DNA signals by more than 1000 times and improve the sensitivity of DNA detection. When detecting DNA, the minimum detection limit of LOD reaches 2aM and has a wide range of linear detection range, which has high selectivity for 10aM to 1nM.. It can distinguish completely paired DNA from 1 base mismatch DNA, and can be applied to single nucleotide polymorphisms (SNP) to detect quartz crystal microbalance signal based on calcium carbonate in situ crystallization. In order to expand its detection field, we use it to detect the area of DNA. In order to expand its detection field, we use it for the detection of cells. This experiment selects Ramos cells. The method of detecting DNA with the quartz crystal microbalance signal amplification technique combined with the in situ crystallization of calcium carbonate is used to detect the Ramos cells, and the lowest detection is made. The detection limit of LOD is 5 /ml, and the detection range is 5 /ml to 6000 /ml.. We compare the method with other quartz crystal microbalance detection methods based on the mass signal amplification. By contrast, we have proved that our experiment has a good sensitivity.4., we have the quartz crystal microbalance signal amplification by the calcium carbonate in situ crystallization. This method provides a new perspective and new idea for the development of quartz crystal microbalance. Based on this method, a general detection platform for various bioactive molecules can be established. It is believed that under the further research of the researchers, this method can be truly applied to the clinic and provide new techniques for the diagnosis and treatment of cancer.
【學位授予單位】：山東師范大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：O799

【參考文獻】

相關期刊論文 前10條

1 高嘉敏;張卓e，

本文編號：1923184