ビデオ顕微分析法の開発と好中球刺激応答系解析への応用

概要

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The real time sequence of phagocytosis-exocytosis in a single neutrophil was directly visualized by video-enhanced contrast differential interference contrast (VEC-DIC) microscopy. Exocytotic responses were investigated with special reference to their dynamic and spatio-temporal properties. By using a dual imaging microscope that allowed one to observe DIC and fluorescence images simultaneously at high magnification, exocytotic responses and cellular calcium concentration ( [Ca^<2+>]_i) were directly investigated. [Ca^<2+>]_i transients were more closely related to the extension of pseudopodia for engulfing a foreign particle and not directly to exocytosis. A chemiluminescence microscope equipped with a photon-counting camera was further applied for the detection of active oxidants. The production of active oxidants during phagocytosis was detected at the sub-cellular level. The chemiluminescence was observed exclusively in the region of the phagosome, suggesting that active oxidants involved in killing of foreign particles were released directly into the phagosome. Superoxide anions (O_2^-) generated by stimulated neutrophils were specifically detected and quantified. One count obtained by this system was equivalent to 59 amol of O_2^-. Maximum O_2^- production was observed at 6〜8 min after stimulation and was estimated as 1.9 fmol/min/cell on average.
社団法人日本分析化学会の論文
1995-02-05