Development of a label-free ATP image sensor for analyzing spatiotemporal patterns of ATP release from biological tissues

SENSORS AND ACTUATORS B-CHEMICAL

卷:335

Article

Doi, Hideo¹  Parajuli, Bijay²  Horio, Tomoko¹  Shigetomi, Eiji²  Shinozaki, Youichi²  Noda, Toshihiko¹  Takahashi, Kazuhiro¹  Hattori, Toshiaki¹  Koizumi, Schuichi²  Sawada, Kazuaki¹ 

[1] Toyohashi Univ Technol, Dept Elect & Elect Informat Engn, Toyohashi, Aichi 4418580, Japan

[2] Univ Yamanashi, Interdisciplinary Grad Sch Med, Dept Neuropharmacol, Kofu, Yamanashi 4093898, Japan

关键词: Adenosine 5'-triphosphate;    Enzyme membrane;    Label-free detection;    Hydrogen ion image sensor;    Hippocampus;   

DOI  :  10.1016/j.snb.2021.129686

来源: Elsevier

【 摘 要 】

We investigated the output characteristics of a label-free, 37.3 mu m pitch, 128 x 128 pixel adenosine-5'-triphosphate (ATP) image sensor, in which different thicknesses of ATP-degrading enzyme apyrase were deposited using the poly-ion complex (PIC) method. ATP was detected based on the changes in the concentration of hydrogen ions (H+) generated by enzymatic reactions. Enzyme films of different thicknesses were formed on the surface of the sensor by controlling the volumes of the PIC solution. A reduction in the enzyme film thickness (0.2 mu m thickness), improved the sensitivity of the ATP sensor, and a clear image than that obtained with a sensor with a thicker film (2.3 mu m thickness) was obtained. Additionally, the detection limit for the 0.2-mu m-thick sensor was as low as 10 mu M, whereas that of the 2.3-mu m thick sensor was 100 mu M. To examine ATP release from biological tissues, we placed a mouse hippocampal slice on the sensor with the 0.2-mu m-thick film and electrically stimulated the nerve fiber (i.e., the Schaffer collateral). The neuronal stimulation evoked significant changes in the output signals of the hippocampal slice but not in the sensor area which was not in contact with tissue. An elevation in the output signal was initiated at the stimulated site with a maximum increment of 5 mV, and the signal traveled along the neuronal fibers. By reducing the film thickness, we successfully developed a high-sensitivity label-free ATPo image sensor with a high spatiotemporal resolution, which allows imaging of endogenous ATPo released from the hippocampus.

【 授权许可】

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