Fluorescence-encoded vibrational spectroscopy has attracted increasing attention by virtue of its high sensitivity and high chemical specificity. We recently demonstrated fluorescence-encoded time-domain coherent Raman spectroscopy (FLETCHERS), which enables low-frequency vibrational spectroscopy of low-concentration fluorophores using near-infrared (800–900 nm) light excitation. However, the feasibility of this study was constrained by the scarcity of excitable molecules in the near-infrared range. Consequently, the broader applicability of FLETCHERS has not been investigated. Here we extend the capabilities of FLETCHERS into the visible range by employing a noncollinear optical parametric amplifier as a light source, significantly enhancing its versatility. Specifically, we use the method, which we refer to as visible FLETCHERS (vFLETCHERS), to individually acquire Raman spectra from five visible fluorophores that have absorption peaks in the 600–700 nm region. These results not only confirm the versatility of vFLETCHERS for a wide range of molecules but also allude to its widespread applicability in biological research through highly sensitive supermultiplexed imaging.

中文翻译：

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可见光范围内的荧光编码时域相干拉曼光谱

荧光编码振动光谱凭借其高灵敏度和高化学特异性而引起越来越多的关注。我们最近展示了荧光编码时域相干拉曼光谱（FLETCHERS），它可以使用近红外（800-900 nm）光激发实现低浓度荧光团的低频振动光谱。然而，这项研究的可行性受到近红外范围内可兴奋分子稀缺的限制。因此，FLETCHERS 的更广泛适用性尚未得到研究。在这里，我们通过采用非共线光学参量放大器作为光源，将 FLETCHERS 的功能扩展到可见光范围，显着增强了其多功能性。具体来说，我们使用称为可见 FLETCHERS (vFLETCHERS) 的方法，从五个可见荧光团中单独获取拉曼光谱，这些荧光团在 600-700 nm 区域具有吸收峰。这些结果不仅证实了 vFLETCHERS 对于多种分子的多功能性，而且还暗示了它通过高灵敏度超多重成像在生物研究中的广泛适用性。