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荧光检测生物芯片在生命科学研究及诸多相关领域已经得到了广泛应用。利用荧光修饰核酸探针可以在液相态中检测到c-fos mRNA致癌基因信息或病毒性核糖核酸(RNA)。通过微细加工技术分别制造了变深度微沟道和变宽度微沟道,目的是找到微生物荧光检测芯片中的最佳检测宽度和深度,并对检测装置中光学滤波片进行了优化。芯片采用最优尺寸和装置采用优化过的荧光滤波片,不仅大幅缩短了检测时间,节省了荧光探针试剂,而且还提高了芯片荧光检测灵敏度。实验结果表明:当荧光检测系统的微沟道深度为500μm、宽度为200μm,并使用(480±15)nm的带通滤波片时,荧光修饰核酸探针的探测灵敏度从通常的20 fmol(即2×10-14 mol)减低到50 amol(即5×10-17 mol),提高了约400倍,大大提高了系统的检测精度。
Fluorescent detection of biochips in the life sciences research and many related fields has been widely used. Using fluorescence-modified nucleic acid probes, c-fos mRNA oncogene information or viral ribonucleic acid (RNA) can be detected in the liquid phase. The micro-channeling and the variable-width channeling were respectively fabricated by microfabrication technology. The purpose was to find out the best detection width and depth in the microbe fluorescence detection chip and to optimize the optical filter in the detection device. Optimized chip size and optimized fluorescence filter, not only significantly shorten the detection time, save fluorescent probe reagents, but also improve the chip fluorescence detection sensitivity. The experimental results show that the detection sensitivity of the fluorescence-modified nucleic acid probe increases from the usual 20 fmol when the microchannel depth of the fluorescence detection system is 500 μm and the width is 200 μm and the bandpass filter of (480 ± 15) nm is used 2 × 10-14 mol) to 50 amol (ie 5 × 10-17 mol), an increase of about 400-fold, greatly improving the system’s detection accuracy.