本文選題：圖像流式　切入點(diǎn)：紅細(xì)胞含量　出處：《大連海事大學(xué)》2016年碩士論文

【摘要】：圖像流式細(xì)胞儀是一種用來檢測生物細(xì)胞的,尤其是用在統(tǒng)計(jì)細(xì)胞數(shù)量、檢測細(xì)胞狀態(tài)等方面的當(dāng)前較為流行的生物細(xì)胞檢測儀器。但是這種儀器的造價(jià)昂貴、體積龐大、結(jié)構(gòu)復(fù)雜,并不適用于普通生物實(shí)驗(yàn)室所要完成的細(xì)胞觀察及分類計(jì)數(shù)等簡單任務(wù)。所以急需設(shè)計(jì)出一種能夠?qū)ζ胀ㄉ飳?shí)驗(yàn)室顯微鏡進(jìn)行功能升級(jí)的圖像流式細(xì)胞儀。本論文中設(shè)計(jì)了一種新的基于微流控芯片及顯微鏡的圖像流式細(xì)胞儀,該圖像流式細(xì)胞儀以微流控芯片為樣品平臺(tái),通過單片機(jī)控制樣品平臺(tái)移動(dòng),實(shí)現(xiàn)樣品視野的轉(zhuǎn)換,結(jié)合普通顯微鏡實(shí)現(xiàn)微流控芯片內(nèi)細(xì)胞圖像的自動(dòng)獲取,通過PC機(jī)端的圖像處理算法及軟件實(shí)現(xiàn)細(xì)胞的輪廓跟蹤、識(shí)別和計(jì)數(shù)。本系統(tǒng)具有樣品需求量小、快速、準(zhǔn)確、成本低等優(yōu)點(diǎn),可以將實(shí)驗(yàn)室的普通顯微鏡升級(jí)成為圖像流式細(xì)胞儀,克服現(xiàn)有商用化圖像流式細(xì)胞儀價(jià)格昂貴、不便普及等缺點(diǎn)。本文的主要內(nèi)容包括系統(tǒng)的設(shè)計(jì),軟硬件測試和應(yīng)用:(1)按照系統(tǒng)的檢測原理對(duì)檢測系統(tǒng)進(jìn)行設(shè)計(jì)和搭建,本系統(tǒng)具有硬件和軟件兩個(gè)部分。其中,硬件部分主要由顯微鏡及攝像頭、微流控芯片、步進(jìn)電機(jī)控制模塊、系統(tǒng)機(jī)械結(jié)構(gòu)等構(gòu)成。軟件部分主要包括拍攝軟件和圖像處理軟件,圖像處理軟件用C++語言在VC++6.0開發(fā)平臺(tái)下編寫,包括圖形處理、文件管理、軟件界面等功能模塊。(2)完成了對(duì)本系統(tǒng)硬件部分的測試,主要是對(duì)由單片機(jī)控制的移動(dòng)平臺(tái)模塊進(jìn)行測試。在測試中,顯微鏡視野下的樣品隨芯片移動(dòng)的距離主要由目鏡及物鏡的放大倍數(shù)、步進(jìn)電機(jī)轉(zhuǎn)動(dòng)圈數(shù)及螺桿的螺紋間距所決定。經(jīng)過對(duì)這些參數(shù)的計(jì)算和進(jìn)行實(shí)驗(yàn),得到了靈活準(zhǔn)確的樣品移動(dòng)效果。(3)完成了對(duì)本系統(tǒng)軟件部分的準(zhǔn)確性的測試,主要包括對(duì)不同大小的顆粒的分類計(jì)數(shù)結(jié)果的測試,對(duì)人體血液樣品中的紅細(xì)胞的計(jì)數(shù)結(jié)果的準(zhǔn)確程度測試和對(duì)海藻細(xì)胞濃度檢測功能的測試。對(duì)不同大小顆粒測試的主要作用是評(píng)價(jià)系統(tǒng)對(duì)體積不同的細(xì)胞進(jìn)行檢測時(shí)的分類識(shí)別能力。對(duì)海藻細(xì)胞和血液細(xì)胞樣品的測試體現(xiàn)系統(tǒng)對(duì)樣品液濃度檢測和計(jì)算的準(zhǔn)確程度且體現(xiàn)了本系統(tǒng)的生物學(xué)意義。檢測結(jié)果表明:本系統(tǒng)能夠快速準(zhǔn)確地完成分類、計(jì)數(shù)和濃度計(jì)算等任務(wù)。
[Abstract]:Image flow cytometry (FCM) is a popular biological cell detection instrument which is used to detect biological cells, especially in the fields of cell count and cell status.However, this instrument is expensive, large and complex, and is not suitable for simple tasks such as cell observation and classification and counting.Therefore, it is urgent to design an image flow cytometry that can upgrade the function of ordinary biological laboratory microscopes.In this paper, a new image flow cytometry based on microfluidic chip and microscope is designed.The image of cells in microfluidic chip can be acquired automatically by using common microscope, and the contour tracking, recognition and counting of cells can be realized by the image processing algorithm and software of PC.The system has the advantages of small sample demand, fast, accurate and low cost. It can upgrade the ordinary microscope of laboratory to image flow cytometry, and overcome the disadvantages of high price and inconvenient popularization of commercial image flow cytometry.The main contents of this paper include the design of the system, software and hardware testing and application: 1) according to the detection principle of the system to design and build the detection system, the system has two parts: hardware and software.The hardware is mainly composed of microscope and camera, microfluidic chip, stepper motor control module, system mechanical structure and so on.The software part mainly includes shooting software and image processing software. The image processing software is written under VC 6.0 development platform with C language, including graphic processing, file management, etc.The hardware part of the system is tested, and the mobile platform module controlled by single chip microcomputer is mainly tested.In the test, the distance of the sample moving with the chip under the microscope is mainly determined by the magnification of the eyepiece and objective lens, the number of rotation coils of the stepping motor and the screw thread spacing.Through the calculation and experiment of these parameters, the flexible and accurate sample moving effect is obtained. The accuracy of the software part of the system is tested, mainly including the test of the classified counting results of different sizes of particles.The accuracy of erythrocyte count in human blood samples and the function of algal cell concentration were tested.The main function of particle testing is to evaluate the ability of classification and recognition of cells with different size.The measurement of algal cells and blood cell samples shows the accuracy of the system in detecting and calculating the concentration of the sample liquid and the biological significance of the system.The results show that the system can quickly and accurately complete the tasks of classification, counting and concentration calculation.
【學(xué)位授予單位】：大連海事大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：TH79

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