Multi-color optical microscopy using modulated excitation lasers and single-frequency demux
Multi-color optical microscopy using modulated excitation lasers and single-frequency demux
复制标题
使用调制激发激光器和单频解复用器的多色光学显微镜
DOI：
10.35848/1347-4065/ab67df
复制
发表时间：
2020-02
影响因子：
1.5
通讯作者：
Ruan Shuangchen
中科院分区：
文献类型：
--
作者：
Wang Nan;Kobayashi Takayoshi;Ruan Shuangchen
多色显微镜已经成为工业和生物医学应用的有力工具。传统上，通过不同颜色通道的图像采集必须分别使用单个检测器进行顺序检测或使用多个检测器进行同时检测。这种昂贵而复杂的过程可能会由于对齐而引入共定位错误。本文介绍了一种紧凑的多色显微镜，使用调制激发激光器和单电子频率检测器进行信号检测。为了分别标记样品中不同标记的荧光，激发激光器以不同的频率进行强度调制。检测到荧光后，用单个频谱分析仪对电子信号进行解复用。该方法允许以最少的检测元素实现多色成像。荧光球，金纳米颗粒和生物组织的多色成像进行。采用光纤耦合系统简化了光学对准，并对其焦色差进行了表征。
Multi-color microscopy has been a powerful tool for industrial and biomedical applications. Traditionally, image acquisition through different color channels has to be carried out separately using a single detector for sequential detection or multi-detectors for simultaneous detection. This costly and complex process may introduce colocalization errors due to the alignment. This paper describes a compact multi-color microscope using modulated excitation lasers and a single electronic frequency demux for signal detection. To mark fluorescence from different labels in the sample separately, the excitation lasers are intensity modulated with different frequencies. After fluorescence is detected, the electronic signals are demultiplexed with a single spectrum analyzer. This method allows the realization of multi-color imaging with minimal amount of detection elements. Multi-color imaging of fluorescent spheres, gold nanoparticles, and biological tissues are carried out. A fiber-coupling system has also been used to simplify the optical alignments, while its focal chromatic aberration has been characterized.