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Cerebellar granule cells equilibrated with &gt;1 \u03bcM hydroethidine (dihydroethidium) which is oxidised to the fluorescent ethidium cation by O<jats:sup>\u22c5\u2212<\/jats:sup>\n<jats:sub>2<\/jats:sub> showed a large increase in fluorescence on protonophore addition. However, controls showed the fluorescent enhancement to be a consequence of release of unbound preformed ethidium from the mitochondrial matrix within the cell with resultant fluorescent enhancement. This ambiguity was removed by the use of low (1 \u03bcM) concentrations of dye in which case generated ethidium remained bound within the mitochondria. Under these conditions antimycin A, but not protonophore addition, produced an increase in fluorescence. It is concluded that excess ethidium acts as an indicator of mitochondrial membrane potential obscuring the monitoring of O<jats:sup>\u22c5\u2212<\/jats:sup>\n<jats:sub>2<\/jats:sub> and that certain experiments employing this indicator in cells may require re\u2010evaluation.<\/jats:p>","DOI":"10.1016\/s0014-5793(97)01088-0","type":"journal-article","created":{"date-parts":[[2002,7,25]],"date-time":"2002-07-25T19:16:52Z","timestamp":1027624612000},"page":"21-24","source":"Crossref","is-referenced-by-count":192,"title":["Mitochondrial membrane potential and hydroethidine\u2010monitored superoxide generation in cultured cerebellar granule cells"],"prefix":"10.1002","volume":"415","author":[{"given":"Samantha L.","family":"Budd","sequence":"first","affiliation":[]},{"given":"Roger F.","family":"Castilho","sequence":"additional","affiliation":[]},{"given":"David G.","family":"Nicholls","sequence":"additional","affiliation":[]}],"member":"311","published-online":{"date-parts":[[1997,11,7]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1017\/S0033583500005795"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-9861(85)90293-0"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi9612982"},{"key":"e_1_2_6_5_1","unstructured":"Liu S.S. and Huang J.P. 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