Phasor S-FLIM: a new paradigm for fast and robust spectral fluorescence lifetime imaging.
Phasor S-FLIM: a new paradigm for fast and robust spectral fluorescence lifetime imaging.
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DOI：
10.1038/s41592-021-01108-4
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发表时间：
2021-05
期刊：
影响因子：
48
通讯作者：
Gratton, Enrico
中科院分区：
文献类型：
--
作者：
Scipioni, Lorenzo;Rossetta, Alessandro;Tedeschi, Giulia;Gratton, Enrico
荧光寿命成像显微镜和光谱成像是显微镜中两种广泛应用的增维方法。然而，就获取和处理而言，它们的组合通常效率低且速度慢。通过结合技术和计算的进步，我们开发了一个稳健和无偏的光谱薄膜(S-薄膜)系统。我们的方法，相量S-Flim，结合了真正的并行多通道数字频域电子学和多维相量方法，以光学分辨率提取关于荧光样品光物理的详细和精确的信息。为了展示相量S薄膜技术的灵活性及其在生物和生物医学领域的应用，我们解决了四个常见但具有挑战性的问题：来自多个未知物种的光谱和寿命信号的盲混、生物传感器的无偏出血和无背景Förster共振能量转移分析、活细胞中环境敏感探针的光物理表征以及肿瘤球体中的并行四色薄膜成像。
Fluorescence lifetime imaging microscopy (FLIM) and spectral imaging are two broadly applied methods for increasing dimensionality in microscopy. However, their combination is typically inefficient and slow in terms of acquisition and processing. By integrating technological and computational advances, we developed a robust and unbiased spectral FLIM (S-FLIM) system. Our method, Phasor S-FLIM, combines true parallel multichannel digital frequency domain electronics with a multidimensional phasor approach to extract detailed and precise information about the photophysics of fluorescent specimens at optical resolution. To show the flexibility of the Phasor S-FLIM technology and its applications to the biological and biomedical field, we address four common, yet challenging, problems: the blind unmixing of spectral and lifetime signatures from multiple unknown species, the unbiased bleedthrough- and background-free Förster resonance energy transfer analysis of biosensors, the photophysical characterization of environment-sensitive probes in living cells and parallel four-color FLIM imaging in tumor spheroids.
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影响因子：
3.8
作者：
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影响因子：
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作者：
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DOI：
10.1073/pnas.1814965116
发表时间：
2019-04-09
影响因子：
11.1
作者：
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通讯作者：
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