【摘要】：表面等離子體子共振(Surface Plasmon Resonance, SPR)成像技術(shù)被廣泛應(yīng)用于生物分子相互作用的研究。本技術(shù)具有實(shí)時(shí)、無(wú)標(biāo)記、靈敏度高的特點(diǎn),使其在提供分子相互作用的動(dòng)態(tài)信息上有巨大的優(yōu)勢(shì)。相應(yīng)地,研制高通量、高精度、精簡(jiǎn)實(shí)用的SPR檢測(cè)系統(tǒng),是實(shí)現(xiàn)SPR技術(shù)優(yōu)勢(shì)的關(guān)鍵。本論文從多個(gè)方面改進(jìn)了SPR檢測(cè)系統(tǒng)的性能,具體包括： 1、采用新穎的角度掃描結(jié)構(gòu),研制了一臺(tái)高分辨率的角度掃描SPR成像系統(tǒng)。本系統(tǒng)的掃描檢測(cè)平臺(tái)采用壓電陶瓷直線電機(jī)與四邊形的傳動(dòng)裝置的結(jié)構(gòu),其掃描角度的分辨率達(dá)10-4度。同時(shí)開(kāi)發(fā)操作控制界面,用于操控電機(jī)、相機(jī)等電子元件,并進(jìn)行數(shù)據(jù)處理分析。整個(gè)系統(tǒng)最終的重復(fù)精度為1×10-3度。 2、使用本SPR成像系統(tǒng)高通量的檢測(cè)caspase-3基因的堿基錯(cuò)配：使用自制的點(diǎn)樣儀制作DNA探針陣列,并與目標(biāo)DNA(caspase-3)雜交；結(jié)果顯示使用本系統(tǒng)可同時(shí)檢測(cè)多個(gè)不同堿基錯(cuò)配的目標(biāo)DNA,其檢出限達(dá)25pmol/L。使用該系統(tǒng)觀測(cè)癌細(xì)胞：在原儀器裝置的基礎(chǔ)上,組裝了一臺(tái)雙視場(chǎng)的SPR細(xì)胞成像裝置,在正常視場(chǎng)和SPR視場(chǎng)下對(duì)HepG2癌細(xì)胞同時(shí)成像；兩個(gè)視場(chǎng)的綜合比較可以排除許多外界干擾,并獲得癌細(xì)胞形態(tài)的信息。 3、開(kāi)發(fā)了一種適用于SPR檢測(cè)的無(wú)動(dòng)力流體注射系統(tǒng)。本進(jìn)樣系統(tǒng)利用液體自身的重力實(shí)現(xiàn)連續(xù)的進(jìn)樣。本重力引導(dǎo)的流體注射系統(tǒng)可提供相對(duì)穩(wěn)定的進(jìn)樣速度,僅需要簡(jiǎn)單的手動(dòng)操作。并在研究互補(bǔ)DNA單鏈雜交的過(guò)程中,成功地提取出雜交過(guò)程的動(dòng)態(tài)信息。最后,組裝了一臺(tái)手持的小型SPR成像裝置以用于現(xiàn)場(chǎng)檢測(cè)。
[Abstract]:Surface plasmon resonance (Surface Plasmon Resonance, SPR) imaging is widely used in the study of biomolecular interactions. This technique has the characteristics of real-time, unlabeled and high sensitivity, which makes it have a great advantage in providing dynamic information of molecular interaction. Accordingly, the development of high-throughput, high-precision, simplified and practical SPR detection system is the key to realize the advantages of SPR technology. In this paper, the performance of SPR detection system is improved from many aspects, including: 1. A high resolution angle scanning SPR imaging system is developed using a novel angle scanning structure. The scanning and detecting platform of this system adopts the structure of piezoelectric ceramic linear motor and quadrilateral transmission, and its scanning angle resolution is 10 ~ (-4) degrees. At the same time, the operating control interface is developed, used to control the motor, camera and other electronic components, and data processing and analysis. The final repetition accuracy of the whole system is 1 脳 10 ~ (-3) degrees. The SPR imaging system is used to detect the base mismatch of caspase-3 gene in high throughput. The DNA probe array is made by using a self-made spotting instrument and hybridized with the target DNA (caspase-3). The results show that the system can simultaneously detect multiple target DNAs with different base mismatches, and the detection limit is up to 25pmol / L. Using this system to observe cancer cells: on the basis of the original instrument, a dual-field SPR cell imaging device was assembled, and the HepG2 cancer cells were simultaneously imaged under the normal field of view and the SPR field of view. The comprehensive comparison of the two fields of view can eliminate many external interference and obtain morphological information of cancer cells. 3. A non-dynamic fluid injection system for SPR detection is developed. The system uses the gravity of the liquid itself to realize continuous injection. This gravity-guided fluid injection system provides a relatively stable injection rate, requiring only simple manual operation. The dynamic information of the hybridization process was successfully extracted during the study of complementary DNA single strand hybridization. Finally, a small hand-held SPR imaging device is assembled for field detection.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2013
【分類號(hào)】：R310

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