Department of Pharmacological Sciences , Icahn School of Medicine at Mount Sinai , New York , New York 10029 , United States.
Medical School for International Health, Faculty of Health Sciences , Ben-Gurion University of the Negev , Be'er Sheva , 84105 , Israel.
ACS Comb Sci. 2018 Nov 12;20(11):653-659. doi: 10.1021/acscombsci.8b00101. Epub 2018 Oct 30.
Ultraviolet-to-infrared fluorescence is a versatile and accessible assay modality but is notoriously hard to multiplex due to overlap of wide emission spectra. We present an approach for fluorescence called multiplexing using spectral imaging and combinatorics (MuSIC). MuSIC consists of creating new independent probes from covalently linked combinations of individual fluorophores, leveraging the wide palette of currently available probes with the mathematical power of combinatorics. Probe levels in a mixture can be inferred from spectral emission scanning data. Theory and simulations suggest MuSIC can increase fluorescence multiplexing ∼4-5 fold using currently available dyes and measurement tools. Experimental proof-of-principle demonstrates robust demultiplexing of nine solution-based probes using ∼25% of the available excitation wavelength window (380-480 nm), consistent with theory. The increasing prevalence of white lasers, angle filter-based wavelength scanning, and large, sensitive multianode photomultiplier tubes make acquisition of such MuSIC-compatible data sets increasingly attainable.
紫外至红外荧光是一种多功能且易于使用的检测方法，但由于发射光谱的广泛重叠，因此很难进行多重检测。我们提出了一种称为基于光谱成像和组合学的荧光多重检测（Multiplexing using Spectral Imaging and Combinatorics，MuSIC）的方法。MuSIC 由通过将单个荧光团共价连接组合来创建新的独立探针组成，利用目前可用的广泛探针库和组合学的数学能力。可以从光谱发射扫描数据中推断混合物中的探针水平。理论和模拟表明，MuSIC 可以使用当前可用的染料和测量工具将荧光多重检测提高约 4-5 倍。实验原理验证证明，使用约 25%的可用激发波长窗口（380-480nm），可以稳健地对 9 种基于溶液的探针进行解复用，与理论一致。白光激光器、基于角度滤波器的波长扫描以及大型、灵敏的多阳极光电倍增管的日益普及，使得获取这种 MuSIC 兼容的数据集越来越可行。
