中国激光, 2004, 31 (s1): 307, 网络出版: 2013-01-29
生物芯片的激光共聚焦扫描检测
Fluorescence Detection for Biochips by Laser Confocal Scanning
激光技术 生物芯片 激光共聚焦扫描 荧光检测 分辨率 信噪比 灵敏度 laser technigue biochip laser confocal scanning fluorescence detection resolution signal-to-noise ratio detection limit
摘要
介绍了基于荧光标记的生物芯片扫描检测方法,主要分为两大类,以光电倍增管(PMT)为荧光探测器的共聚焦扫描检测方法和以CCD为荧光探测器的全视场扫描检测方法.重点介绍一种采用双波长(532 m及635 m)激光器作为激发光源,以激光共聚焦原理所设计的生物芯片荧光信息检测技术,由一个光电倍增管分时实现cy3与cy5两种荧光信号的检测.生物芯片的横向扫描由远心f-θ扫描物镜与振镜实现,纵向扫描由步进电机驱动精密导轨实现.实验结果表明,检测技术的分辨率可达到5μm,信噪比高达103,检测灵敏度最高为1 fluor/μm2,并且扫描速度快,cy3与cy5之间无串扰.
Abstract
Scanning detection of biochip based on fluorescence labeling is introduced in the paper, the detection method can be divided into two category: one is confocal scanning detection method in which a photomultiplier tube is used as fluorescence sensor, the other is flood illumination of the entire biochip and uses CCD as fluorescence sensor. A novel method of fluorescence detection for biochips is presented, which uses a 532 nm laser and 635nm laser to excite the fluorescent dyes according the principle of laser confocal scanning. The fluorescence emitted from cy3 and cyo is detected using a photomultiplier tube (PMT) sequentially. One dimension scanning of the biochips is performed by a telecentric f-θ objective with a moving coil optical scanner, the other dimension is scanned by a stepping motor driving the precision guidance. The experiment results state that the resolution of the presented technology is 5 μm, the signal-to-noise ratio can reach 10s and the detection limit is 1 fluor/μm2, and the cross-talk phenomenon is eliminated completely.
王立强, 倪旭翔, 陆祖康, 李映笙, 郑旭峰. 生物芯片的激光共聚焦扫描检测[J]. 中国激光, 2004, 31(s1): 307. WANG Li-qiang, NI Xu-xiang, LU Zu-kang, LI Ying-sheng, ZHNEG Xu-feng. Fluorescence Detection for Biochips by Laser Confocal Scanning[J]. Chinese Journal of Lasers, 2004, 31(s1): 307.