Fukuda J, Fujita Y, Ohsawa K
J Neurosci Methods. 1983 Jul;8(3):295-302. doi: 10.1016/0165-0270(83)90042-0.
ATP content in a nerve cell isolated from dorsal root ganglia of adult guinea-pigs by collagenase was measured by a newly developed technique modified from the conventional luciferin-luciferase methods. A small volume (4 microliters) of the nerve cell suspension, which contained 10-300 nerve cells (3-100 X 10(-4) microliters of cellular volume) under view of an inverted, phase-contrast microscope, was heat-treated for about 1 s by flame of an alcohol lamp. This heat-treated cell suspension was then reacted with a luciferin-luciferase solution. Light flux from the bioluminescence thus elicited gave an ATP content in single nerve cell, 27 pg (mean) +/- 10 pg (S.D.). ATP concentration in a nerve cell was calculated as 1.7 mM (mean) +/- 0.6 mM. The ATP content in a nerve cell was reduced when the nerve cells were exposed to KCN (5 microM) or dinitrophenol (20 microM), respectively.
采用一种新开发的、从传统荧光素-荧光素酶方法改良而来的技术，测定了用胶原酶从成年豚鼠背根神经节分离出的神经细胞中的ATP含量。在倒置相差显微镜下，取一小体积（4微升）含有10 - 300个神经细胞（细胞体积为3 - 100×10⁻⁴微升）的神经细胞悬液，用酒精灯火焰进行约1秒的热处理。然后将经此热处理的细胞悬液与荧光素-荧光素酶溶液反应。由此产生的生物发光的光通量给出了单个神经细胞中的ATP含量，为27皮克（平均值）±10皮克（标准差）。神经细胞中的ATP浓度经计算为1.7毫摩尔（平均值）±0.6毫摩尔。当神经细胞分别暴露于5微摩尔的氰化钾或20微摩尔的二硝基苯酚时，神经细胞中的ATP含量降低。
