【摘要】：熒光顯微鏡是生命科學(xué)研究中的重要工具。在普通的寬場熒光顯微成像中,焦外的信號也會被探測器收集,嚴(yán)重降低成像對比度和分辨率。能夠克服焦外干擾的顯微鏡(如共聚焦、雙光子顯微鏡)往往成像速度較慢,設(shè)備昂貴�；谏吆途鶆蚬饣旌险彰鞯墓鈱W(xué)層析方法成本低,層析效果好,在小型化的設(shè)備上很有應(yīng)用前景。傳統(tǒng)的散斑和均勻光混合照明光學(xué)層析方法使用自由空間中的散射片散射激光來生成散斑,光路固定且所占體積較大,不利于與商業(yè)顯微鏡集成。本文利用多模光纖導(dǎo)光及振動光纖消散斑方法,改進(jìn)了傳統(tǒng)的混合照明層析方法,實(shí)現(xiàn)了一種基于光纖的混合照明光學(xué)層析熒光顯微成像系統(tǒng)。該系統(tǒng)的層析能力能夠接近共聚焦顯微成像;且具有光路封閉、裝置空間柔性、系統(tǒng)簡單、成本低(兩萬元左右),適合與商用顯微鏡結(jié)合等優(yōu)點(diǎn)。本文主要工作如下:首先,給出了光纖導(dǎo)光的混合照明光學(xué)層析熒光顯微成像系統(tǒng)原理圖;對散斑和均勻光混合照明光學(xué)層析方法進(jìn)行了理論分析,給出了具體算法;分析了振動光纖消散斑實(shí)現(xiàn)均勻照明的方法。其次,通過實(shí)驗(yàn)測試了不同參數(shù)對振動光纖消散斑效果的影響:振動電機(jī)驅(qū)動電壓越高,消散斑效果越好,并且隨著電壓升高提升效果越弱;光纖模式數(shù)越大,消散斑效果越好;光纖纏繞不提升消散斑效果,甚至有微小的惡化。根據(jù)實(shí)驗(yàn)結(jié)果,設(shè)計了振動光纖消散斑模塊及其工程封裝,得到一個即插即用、調(diào)節(jié)方便、成本低、可用于各類光學(xué)系統(tǒng)的消散斑模塊。最后,搭建了基于光纖的散斑和均勻光混合照明成像系統(tǒng);編寫了Matlab圖像處理程序;利用花粉粒和100μm厚小鼠腦片樣品對本方法進(jìn)行了測試,得到了顯著的層析效果。
[Abstract]:Fluorescence microscope is an important tool in life science research. In conventional wide-field fluorescence microscopic imaging, out-of-focal signals are also collected by detectors, which seriously reduce imaging contrast and resolution. Microscope, such as confocal microscope and two-photon microscope, which can overcome the external focal interference, is often slow and expensive. The optical tomography method based on speckle and uniform illumination has low cost and good performance. It has a good prospect in miniaturization equipment. The traditional optical tomography method of mixed illumination with speckle and uniform light uses scattering laser in free space to generate speckle. The light path is fixed and the volume is large, which is not conducive to the integration with commercial microscope. In this paper, the traditional hybrid illumination tomography method is improved by using multimode optical fiber light guide and vibratory optical fiber dissipation method, and a hybrid illumination optical tomography fluorescence microscopic imaging system based on optical fiber is implemented. The system has the advantages of close optical path, flexible device space, simple system, low cost (about 20,000 yuan), and suitable for combining with commercial microscope. The main work of this paper is as follows: firstly, the principle diagram of hybrid illumination optical tomography fluorescence microscopic imaging system is given, and the theoretical analysis and algorithm of speckle and uniform illumination optical tomography are given. The method of realizing uniform illumination of vibratory fiber dissipation spot is analyzed. Secondly, the effect of different parameters on the effect of vibration fiber dissipation is tested: the higher the driving voltage of vibration motor, the better the effect of dissipating spot, and the weaker the lifting effect is with the increase of voltage; The larger the number of fiber modes, the better the effect of dissipating speckle, and the less the effect of fiber winding is, the worse the effect is. Based on the experimental results, the vibratory fiber dissipation module and its engineering package are designed. A plug and play module with convenient adjustment and low cost is obtained, which can be used in various optical systems. Finally, a hybrid illumination imaging system based on fiber optic speckle and uniform light was set up, Matlab image processing program was compiled, and the method was tested by pollen grains and 100 渭 m thick mouse brain.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：TP391.41;TH742

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本文編號：2419776