Varied Length Stokes Shift BODIPY-Based Fluorophores for Multicolor Microscopy.
Varied Length Stokes Shift BODIPY-Based Fluorophores for Multicolor Microscopy.
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DOI：
10.1038/s41598-018-22892-8
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发表时间：
2018-03-15
影响因子：
4.6
通讯作者：
Gibbs SL
中科院分区：
文献类型：
--
作者：
Bittel AM;Davis AM;Wang L;Nederlof MA;Escobedo JO;Strongin RM;Gibbs SL
定位和观察亚细胞系统的复杂性所需的多色显微镜工具受到当前荧光团技术的限制。虽然商用荧光团覆盖了从紫外线到近红外区的光谱空间，并针对传统的基于带通的荧光显微镜进行了优化，但它们不是高度复合荧光显微镜的理想选择，因为它们往往具有较短的斯托克斯位移，从而将单个样品中可以检测到的荧光团数量限制在四到五个。在这里，我们合成了95个新的基于BODIPY(BODIPY)的荧光团的文库，并对它们的光物理、光学和光谱性质进行了筛选，以期在多色显微镜中应用。我们基于BODIPY的荧光团的一个子集产生了不同长度的斯托克斯位移探针，首次使用共聚焦激光扫描显微镜的单次激发创建了五色图像。将这些新的荧光团与传统的荧光团相结合，可以使用基于线性分解的显微镜方法来促进多达九到十种颜色的成像。
Multicolor microscopy tools necessary to localize and visualize the complexity of subcellular systems are limited by current fluorophore technology. While commercial fluorophores cover spectral space from the ultraviolet to the near infrared region and are optimized for conventional bandpass based fluorescence microscopy, they are not ideal for highly multiplexed fluorescence microscopy as they tend to have short Stokes shifts, restricting the number of fluorophores that can be detected in a single sample to four to five. Herein, we synthesized a library of 95 novel boron-dipyrromethene (BODIPY)-based fluorophores and screened their photophysical, optical and spectral properties for their utility in multicolor microscopy. A subset of our BODIPY-based fluorophores yielded varied length Stokes shifts probes, which were used to create a five-color image using a single excitation with confocal laser scanning microscopy for the first time. Combining these novel fluorophores with conventional fluorophores could facilitate imaging in up to nine to ten colors using linear unmixing based microscopy approaches.
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