Alkaline Phosphatase-Catalyzed Amplification of a Fluorescence Signal for Flow Cytometry.
Alkaline Phosphatase-Catalyzed Amplification of a Fluorescence Signal for Flow Cytometry.
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DOI：
10.1021/acs.analchem.7b03893
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发表时间：
2018-01
影响因子：
7.4
通讯作者：
T. Nobori;Kenta Tosaka;A. Kawamura;Taisei Joichi;Kenta Kamino;A. Kishimura;E. Baba;Takeshi Mori;Y. Katayama
中科院分区：
文献类型：
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作者：
T. Nobori;Kenta Tosaka;A. Kawamura;Taisei Joichi;Kenta Kamino;A. Kishimura;E. Baba;Takeshi Mori;Y. Katayama
尽管流式细胞术的使用不断扩大，但其检测限对于许多具有低拷贝数的抗原蛋白并不令人满意。在此，我们描述了一种基于碱性磷酸酶（AP）的技术，以放大细胞染色应用的荧光信号。我们设计了一种荧光底物，其在AP脱磷酸化后获得膜通透性。通过使用该底物，可以成功地放大流式细胞术中细胞的荧光信号，以给出比使用常规荧光团修饰的抗体标记的细胞强得多的信号。
Despite the expanding use of flow cytometry, its detection limit is not satisfactory for many antigen proteins with low copy numbers. Herein, we describe an alkaline phosphatase (AP)-based technique to amplify the fluorescence signal for cell staining applications. We designed a fluorescent substrate that acquires membrane permeability upon dephosphorylation by AP. By using the substrate, the fluorescence signal of cells in flow cytometry could be successfully amplified to give a much stronger signal than the cells labeled using a conventional fluorophore-modified antibody.