National University of Singapore, Faculty of Engineering, Department of Bioengineering, Optical Bioimaging Laboratory, 9 Engineering Drive 1, Singapore 117576.
J Biomed Opt. 2010 Mar-Apr;15(2):027010. doi: 10.1117/1.3374051.
We report on a novel acousto-optic tunable filter (AOTF)-based near-infrared (NIR) fluorescence excitation-emission matrix (EEM) spectroscopy technique for rapid multifluorophore characterization. We implement a unique light filtering module design by using cascaded AOTFs coupled with three orthogonally oriented polarizers to effectively remove the side-ripple artifacts of AOTFs as well as by using a pair of AOTFs coupled with two orthogonally oriented polarizers to improve detection efficiency for high-quality fluorescence EEM acquisitions. NIR fluorescence EEM spectroscopy (41 excitation wavelengths ranging from 550 to 950 nm in 10-nm increments; fluorescence emission from 570 to 1000 nm at 10-nm intervals) can be acquired from fluorescence dyes [e.g., diethylthiatricarbocyanine (DTTC) iodide, oxazine 750, and IR 140] within 10 s or even less, illustrating the potential of the AOTF-based NIR EEM technique developed for rapid multifluorophore analysis and characterization in biochemical and biomedical systems.
我们报告了一种基于声光可调滤波器（AOTF）的近红外（NIR）荧光激发-发射矩阵（EEM）光谱技术，用于快速多荧光团特征分析。我们通过使用级联 AOTF 与三个正交偏振器相结合的独特光滤波模块设计，有效地去除了 AOTF 的侧纹波伪影，并且通过使用一对 AOTF 与两个正交偏振器相结合，提高了检测效率，从而实现了高质量的荧光 EEM 采集。NIR 荧光 EEM 光谱（41 个激发波长，范围为 550nm 至 950nm，间隔为 10nm；荧光发射波长范围为 570nm 至 1000nm，间隔为 10nm）可以在 10s 内甚至更短的时间内从荧光染料（例如二乙基噻嗪羰花青碘化物、噁嗪 750 和 IR 140）中获取，这说明了所开发的基于 AOTF 的 NIR EEM 技术在生物化学和生物医学系统中用于快速多荧光团分析和特征分析的潜力。
