Open AccessAlkaline Phosphatase-Catalyzed Amplification of a Fluorescence Signal for Flow Cytometry
Author(s) -
Takanobu Nobori,
Kenta Tosaka,
Akira Kawamura,
Taisei Joichi,
Kenta Kamino,
Akihiro Kishimura,
Eishi Baba,
Takeshi Mori,
Yoshiki Katayama
Publication year - 2017
Publication title -
analytical chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.117
H-Index - 332
eISSN - 1520-6882
pISSN - 0003-2700
DOI - 10.1021/acs.analchem.7b03893
Subject(s) - chemistry , flow cytometry , fluorescence , fluorophore , alkaline phosphatase , detection limit , dephosphorylation , staining , substrate (aquarium) , microbiology and biotechnology , biophysics , phosphatase , biochemistry , chromatography , enzyme , medicine , physics , oceanography , quantum mechanics , pathology , biology , geology
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.
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