光学学报, 2011, 31 (6): 0618002, 网络出版: 2011-05-18   

近紫外波段频分复用荧光显微探测技术研究

Near UV-Band Frequency Division Multiplexing Detecting Technique with Fluorescence Microscopy
作者单位
上海理工大学上海市现代光学重点实验室, 上海 200093
摘要
分析了多路频分复用并行显微荧光探测的基本原理,即把激发光分成多束,对每一束进行不同频率调制,聚焦到生物样品的不同位置,激发产生相应频率的荧光信号,再对光电倍增管(PMT)接收的荧光信号进行分频解调,实现实时、并行和高分辨率的探测技术。搭建了激发光源为405 nm近紫外激光的双路方波调制荧光显微探测系统,实验探测了老鼠神经细胞显微形态,分析解调了双点荧光能量随时间变化的曲线。并研究了荧光显微成像系统放大率、时间分辨率等技术参数,还通过数值分析给出了避免各通道间信号串扰的条件。
Abstract
The principles of the microscopic fluorescence detection with frequency division multiplexing paralleling detection technology (FDMPD) is analysed. In the FDMPD system, the exciting laser beam is firstly divided into the multi-beams, and each beam is modulated with the individual carrying frequencies. The multi-beams exciting laser is focused on the surface of target cells to generate the multichannel fluorescent signals with corresponding carrying frequencies. Photomultiplier tube collected the fluorescent signals and transmitted the signal into the computer. Then individual channel fluorescent signal which varies with the time can be demodulated in a parallel and a high-resolution way. Employing 405 nm exciting laser sources, the two channel FDMPD system is constructed. The experiments explored micro-morphology of mouse nerve cells sample and demodulated the two channel fluorescence curves varying with the time. Furthermore, the basic parameters including the magnification, time resolution ability etc are analyzed, and the basic conditions to avoid the cross talk among multiply channels are also put forward.

张运波, 郑继红, 蒋妍梦, 侯文玫, 张仁杰, 庄松林. 近紫外波段频分复用荧光显微探测技术研究[J]. 光学学报, 2011, 31(6): 0618002. Zhang Yunbo, Zheng Jihong, Jiang Yanmeng, Hou Wenmei, Zhang Renjie, Zhuang Songlin. Near UV-Band Frequency Division Multiplexing Detecting Technique with Fluorescence Microscopy[J]. Acta Optica Sinica, 2011, 31(6): 0618002.

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