2021
DOI: 10.7554/elife.63129
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Simultaneous recording of multiple cellular signaling events by frequency- and spectrally-tuned multiplexing of fluorescent probes

Abstract: Fluorescent probes that change their spectral properties upon binding to small biomolecules, ions, or changes in the membrane potential (Vm) are invaluable tools to study cellular signaling pathways. Here, we introduce a novel technique for simultaneous recording of multiple probes at millisecond time resolution: frequency- and spectrally-tuned multiplexing (FASTM). Different from present multiplexing approaches, FASTM uses phase-sensitive signal detection, which renders various combinations of common probes f… Show more

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Cited by 3 publications
(2 citation statements)
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References 65 publications
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“…Gundogdu研究组 [63] 针对吸收光谱仪的应用场景, 结合峰值整形模拟前端电路设计, 提出了一种瞬态 锁相检测系统, 实验证明可将吸收光谱的灵敏度带 来数量级的提升; 德国明斯特大学的Brenker研究 组 [64] 介绍了一种将多通道锁相放大器用于膜电位 和离子荧光探针的各种组合, 可用于多路检测, 结 合光电倍增管进行弱荧光的锁相并行检测多细胞 信号, 以毫秒时间分辨率同时记录多个光电探针信 号, 实现细胞信号通路的原位动态监测; 借助锁相 放大器对噪声干扰的抑制, 靶标含量极低生物样品 的微弱发光信号也能被准确提取, 从而有望用于更 快更灵敏的核酸检测部件 [65] . 在环境监测应用中, 来自意大利罗马大学的Sarra等 [66] 报道将双通道 锁相放大器用于激光透射光谱检测, 来实现单分散 和多分散纳米颗粒的尺寸和浓度的快速测量, 结合 可变增益光谱校准方法能将颗粒度分析不确定度 降低1个数量级, 这给环境中生物团聚体检测提供 了先进的手段.…”
Section: 典型应用unclassified
“…Gundogdu研究组 [63] 针对吸收光谱仪的应用场景, 结合峰值整形模拟前端电路设计, 提出了一种瞬态 锁相检测系统, 实验证明可将吸收光谱的灵敏度带 来数量级的提升; 德国明斯特大学的Brenker研究 组 [64] 介绍了一种将多通道锁相放大器用于膜电位 和离子荧光探针的各种组合, 可用于多路检测, 结 合光电倍增管进行弱荧光的锁相并行检测多细胞 信号, 以毫秒时间分辨率同时记录多个光电探针信 号, 实现细胞信号通路的原位动态监测; 借助锁相 放大器对噪声干扰的抑制, 靶标含量极低生物样品 的微弱发光信号也能被准确提取, 从而有望用于更 快更灵敏的核酸检测部件 [65] . 在环境监测应用中, 来自意大利罗马大学的Sarra等 [66] 报道将双通道 锁相放大器用于激光透射光谱检测, 来实现单分散 和多分散纳米颗粒的尺寸和浓度的快速测量, 结合 可变增益光谱校准方法能将颗粒度分析不确定度 降低1个数量级, 这给环境中生物团聚体检测提供 了先进的手段.…”
Section: 典型应用unclassified
“…Owing to their ultrafast photochemistry (≤ 10 ns) (Eckhardt et al, 2002), physiological reactions can be triggered with sub-millisecond time resolution. Moreover, our caged NMPs offer excellent spectral properties: they do not require UV light for release and allow parallel monitoring of multiple cellular reactions using fluorescent dyes or proteins without release, i.e., stimulating and measuring the cellular response can be achieved independently (Kierzek et al, 2021). This cannot be achieved with optogenetic tools, such as channel rhodopsin, as their action or response spectra overlap with the excitation spectrum of most fluorescent reporters (Lin, 2011).…”
Section: Resultsmentioning
confidence: 99%