Frequency Encoding of Upconversion Nanoparticle Emission for Multiplexed Imaging of Spectrally and Spatially Overlapping Lanthanide Ions,Journal of the American Chemical Society

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Frequency Encoding of Upconversion Nanoparticle Emission for Multiplexed Imaging of Spectrally and Spatially Overlapping Lanthanide Ions
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2021-11-15 , DOI: 10.1021/jacs.1c07691
Mikkel Baldtzer Liisberg ₁ , Satu Lahtinen ₂ , Ane Beth Sloth _{3,

4} , Tero Soukka ₂ , Tom Vosch ₁

Affiliation

We present frequency encoded upconversion (FE-UPCON) widefield microscopy, an imaging approach that allows for multiplexed signal recovery based on frequency encoding of selected upconverted lanthanide ion emission rather than separation based on energy or time. FE-UPCON allows for the separation of luminescence from spectrally and spatially overlapping trivalent lanthanide ions (Ln³⁺) in upconversion nanoparticles (UCNPs). Utilizing the numerous electronic energy levels of Ln³⁺, one can generate a frequency encoded signal by periodic coexcitation with a secondary light source (modulated at a chosen frequency) that, for a particular wavelength, enhances the luminescence of the Ln³⁺ of interest. We demonstrate that it is possible to selectively image spectrally overlapping UCNPs co-doped with Yb³⁺/Ho³⁺ or Yb³⁺/Er³⁺ by FE-UPCON in cells up to 10 frames per second on a conventional widefield microscope with the simple extension of an additional secondary light source and a chopper wheel for modulation. Additionally, we show that FE-UPCON does not compromise sensitivity and that single UCNP detection is obtainable. FE-UPCON adds a new dimension (frequency space) for multiplexed imaging with UCNPs.

中文翻译：

上转换纳米粒子发射的频率编码用于光谱和空间重叠镧系离子的多路成像

我们提出了频率编码上转换 (FE-UPCON) 宽场显微镜，这是一种成像方法，它允许基于所选上转换镧系元素离子发射的频率编码而不是基于能量或时间的分离来进行多路复用信号恢复。FE-UPCON 允许从上转换纳米粒子 (UCNPs) 中光谱和空间重叠的三价镧系元素离子 (Ln ^{3+ ) 中分离发光。}^{利用 Ln 3+}的众多电子能级，可以通过与辅助光源（以选定频率调制）的周期性共激发来生成频率编码信号，该辅助光源对于特定波长可增强 Ln ^3+的发光出于兴趣。我们证明，通过 FE-UPCON 在具有高达 10 帧/秒的传统宽场显微镜的细胞中，可以选择性地对与 Yb ³⁺ /Ho ³⁺或 Yb ³⁺ /Er ³⁺共掺杂的 UCNPs 进行选择性成像。一个额外的辅助光源和一个用于调制的斩波轮的简单扩展。此外，我们表明 FE-UPCON 不会影响灵敏度，并且可以获得单个 UCNP 检测。FE-UPCON 为使用 UCNP 的多路成像增加了一个新维度（频率空间）。

更新日期：2021-11-24

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